ACTIVATION OF IRON IN PLANTS BY MANGANESE AND
OTHER CHEMICALS IN A LIME-INDUCED CHLOROSIS

A THESIS

presented to the Faculty of Michigan State College
of Agriculture and Applied Science in partial
fulfillment of the requirements for the
Degree of Doctor of Philosophy

George Donald Sherman

East Lansing
1940

ProQuest Number: 10008426

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ACKNOWLEDGMENT
The writer is grateful to Dr, Paul M. Harmer
and to Dr* C. E, Millar for guidance in the work
reported in this paper and for their constructive
criticisms in the preparation of this manuscript*
He wishes also to express appreciation to the mem­
bers of the Chemistry Experiment Station staff for
their cooperation and worthy suggestions and to
Dr. R. A, Gortner of the University of Minnesota for
his kind suggestions in the more technical phases of
this problem*

127461

TABLE OF CONTENTS
Introduction

Page
1

Historical

3

Soil Factors

5

Total Analysis

11

Successive Extraction

13

Exchangeable Bases

19

Extractable Manganese

21

Water-soluble Manganese

21

Exchangeable

22

Easily Reducible MnO^

23

Aqua Regia Extraction of Soil Ash

23

Discussion of Experimental Results

24

Seasonal Trend of Replaceable, Total
Manganese and Reducible MnO^

33

Effect of Manganese Fertilizer on the Mineral
Composition of the Crop

43

Analysis of Mature Crop

44

Discussion of Data

54

Mineral Composition of Normal and Chlorotic
Plants at Different Stages of Growth

55

Normal and Chlorotic Peas

59

Seasonal Trend of Mineral Content
of Barley Plants

61

Seasonal Trend of Mineral Composition
of Legumes

69

Page
Fallacy of Present Theories

94

Hypothesis

102

The Effect of Minor Element Fertilization
on Oxidation-ReductionSystems Within Plant

113

General Discussion

130

Summary

137

Bibliography

141

Introduction
A great deal of attention has recently been given to
the role of minor elements in soil fertility and plant
nutrition.

At present, investigation is being directed

not only to the determination of the quantities of these
elements present or required for crop production, and to
the factors affecting their availability to the plant, but
also to their utilization by animals which will ultimately
feed upon these plants.

Manganese is one of the more

prominent of the elements belonging to this group, to which
much consideration is being paid, both in nutritional and
fertility studies.

A lack of sufficient manganese is likely

to cause a chlorotic condition in the plant.

Manganese-

deficiency symptoms vary greatly among different species of
plants, due to the inherent characteristics of the plant*
It appears very doubtful if there are any specific symptoms
which will definitely characterize manganese deficiency,
unless it be some of the malnutritional diseases such as
"grey speck disease" in oats*
In Michigan, manganese deficiency occurs on certain
neutral to alkaline organic soils* which alkalinity has
*

Since the definition of organic soils varies from region
to region, it becomes necessary to define certain terms as
they are used in Michigan. Organic soils of Michigan are
divided into two groups, mucks and peats. They are differeniated on the basis of decomposition and not upon their
mineral and organic matter composition. Muck is a well
decomposed organic soil in which the original material from
which it was derived cannot be easily recognized. Peat
refers to the organic soil in which the original material
iB readily recognizeable*

often been produced by burning.

These alkaline organic

soils are decidedly less productive than are the acid
organic soils.

The productiveness of these soils can be

restored, either by the application of manganese salts or
by restoration of the soil acidity by tne application of
sulphur.

The distribution of such alkaline organic soils

in Michigan is considerable and they may occupy the whole
or only portions of the organic soil areas in practically
any part of the state.

These manganese—deficlent soils

present a real problem to the farmer, vegetable grower and
the fertility specialist as to what is their best manage­
ment.

This study was made with the purpose of securing a

better understanding of these problems, which information
might lead to a more profitable and economical management
of these areas.
Historical.
Manganese deficiency in organic soils of Michigan has
produced a lime-induced chlorosis in many crops similar to
that of the soybean leaves in Plate 1.

It has also pro­

duced the characteristic Mgrey speck diseaseH in oats which
is shown in Plate 2.

This chlorosis has varied from crop

to crop, so no definite description is possible which would
cover all cases*
Chlorosis was first recognized by Gris (25) in 1844,
when he caused yellow plants to become normal by the appli­
cation of soluble iron to the soil.

In 1895, Degrully (16)

2a.

Plate 1.

A*
Bi.

Soybeans leave exhibiting the lime-induced chlorosis.
A normal soybean leaves from a plant grown on a soil
receiving manganese sulfate.
Plate 2.

A
B.'
C.
D„

A leaf showing early stages of Grey Speck disease.
A normal leaf from a plant growing on soil receiving
mang ane se sulf at e .
Intermediate stage of Grey Speck*
Advanced stage of Grey Speck.

3.
discovered that sulphuric acid, applied to the soil, would
also cure this chlorotic condition.
The actual use of sulphur for this purpose was not
made until 1911, and then by Bertrand (8 ).

The first

applications of manganese sulfate were made on rice
plantations in Japan-, in 1903, by Aso (4) and in 1903, by
Hagoaka (56).

The first application of manganese to organic

soil probably was made in Sweden by von Feilitzen (75) in
1907.

In 1909, Sjollema and Hudvig (68) found that manganese

salts would cure "grey speck disease" in oats.
Many theories had been advanced by 1916, regarding the
function of manganese, both as to its use as a fertility
element and to the part it plays in plant growth.

Bertrand

was perhaps the first to claim that manganese is indispen­
sable for plant growth which theory of its essentiality as a
plant nutrient has been generally accepted.

Gilbert (23)

stated that it was essential since iron did not cure the
chlorotic condition.

Culture solutions have presented ample

proof of its necessity as a plant nutrient.

In recent years,

some workers (13) have suggested that the benefit of manga­
nese to the plant lies in its control of the intake of other
ions or in its making possible the utilization of some of
the other essential ions by the plant.

Kelley (41) reported

that the addition of manganese fertilizers resulted in a
marked increase in the availability of calcium and magnesium.
Sullivan and He id (71) found that manganese additions

4•
increased the oxidative power of the soil and that it could
he associated witn the productivity of the soil.
Many workers held that the function of manganese was
one of regulating the biological oxidation and reduction
reactions within the plant.

Bertrand (7) considered that

manganese increased the oxygen-carrying power of oxidase
enzymes.

Hopkins (34) believed that the function of manga­

nese was one of activation of iron, in that iron was in the
reduced form in the organism and manganese served to reoxidize it.

Willis (79) considers that there is a relation­

ship between manganese and iron which theory is also
supported by Scholz (65) who considers that a relationship
existing between manganese, iron and calcium in plant nutri­
tion.
wGrey speck11 disease has been recognized in Europe,
Australia and Canada for some time and is known in certain
localities as dry speck, dry spot or halo blight.

It attacks

principally oats and barley but there are reports of similar
disturbance in wheat and rye.

Although known for some time,

there is still doubt as to whether it is a bacterial disease
or is due to an abnormal soil condition.

For 30 years or

more, the application of manganese sulfate has been recog­
nized as a corrective of this condition.

It occurs on soils

that have a pH of 6.3 to 7.8, usually organic soils or sands
high in organic matter.

The majority of workers, including

Davies and Jones (15) of Wales; Maschhaupt (51) of the
Netherlands and Samuel and Piper (62) and Wild of Australia(77)

5*

are quite positive that grey speck is due to manganese
deficiency in soils*
Gerretsen (19) is of the opinion that grey speck is
caused by bacteria.

He further states that the precipi­

tation of insoluble manganic oxides in the soils seems to
be caused by specific organisms which are active between
the pH 6.5 and 7.8, which corresponds to the limits of grey
speck in oats.

This has been substantiated by Leeper and

Swaby (44).
Because of the rather widespread distribution of manga­
nese-deficient organic soils and the large acreage of organic
soils in Michigan, it was considered that this problem war­
ranted this detailed study regarding (1 ) the chemical nature
of the soils before and after application of manganese salts
and elemental sulphur; (2 ) the changes in chemical composit­
ion in the plants due to these treatments; and (3) the role
played by manganese in plant nutrition*

Soil Factors.
The literature relating to soils which have exhibited
manganese deficiency reveals many contradictory opinions
as to the kinds of soil in which this problem is manifested.
Greater still are the various opinions as to the nature and
properties of manganese compounds in the soil.

Manganese

deficiency has been reported from many parts of Europe,
Australia, Japan and the United States.

Leeper (43), in his

recent paper, states that this deficiency occurs on certain

Australian soils which have naturally a neutral or alkaline
reaction or soils which originally had a strongly acid
reaction hut which have been made neutral or alkaline by the
application of lime.
The question of availability of soil manganese is a very
live one.

Leeper (43) concludes that, in general, the total

quantity of manganese in the soil has little bearing on the
occurrence of the deficiency.

This opinion naturally brings

up the question of availability.

Piper (59) believes that

manganese is absorbed by the plants as the manganous ion and
that, in the form of MnOg, it is unavailable to plants.

He

holds further that, under decidedly alkaline conditions,
manganese exists as the bivalent exchangeable form or in the
form of MnOg.

The equilibrium which exists between these

forms swings towards the bivalent form under conditions of
reduction and towards MnOg if oxidation prevails.

Steenbjerg

(70) has a similar opinion and states that the available manga
nese is controlled by reaction and oxidation-reduction con­
ditions.

Mann (48) holds that lime represses the solubility

of manganese to such an extent that it becomes unavailable.
He worked with soils similar to those which Gilbert et al,
(23) investigated, in that they were strongly acid and leach­
ed soils which had been limed.

According to Leeper, mangan­

ese occurs as the bivalent form in strongly acid soils and
in this form may be leached from the soil.

He holds that

liming a soil until it has an alkaline reaction might reason­
ably cause a manganese deficiency.

Not all of the work reported, however, has dealt with
manganese deficiency.

Considerable attention has been given

to soils that have a highly soluble manganese content, which
situation quite often causes a chlorotic condition or other
metabolic disturbance in the plant.

Johnson (38) and others

(41) attribute the chlorosis in Hawaiian pineapple to a very
high pyrolusite (MnOg) content in the soil which, he holds,
results in the oxidation of the iron in the soil to the
ferric condition, a form very unavailable to plants.
Bortner (10) cites a similar condition with tobacco in
Kentucky; as did also Jacobson and Swanbeck (37) in
Connecticut.

The brown spotting disease of potatoes found

in Russia is reported by Arkhangelskaya (3) not to be due to
soil acidity but rather to a high content of soluble manga­
nese in the soil*
In Michigan, according to Harmer (31) manganese defi­
ciency appears on high-lime organic soils which have been
burned; on those which are fed by alkaline spring water; on
those which have a marl deposit near the surface and on those
which originally were acid but have been made alkaline by the
application of lime.

It should not be construed that every

alkaline or burned organic soil in Michigan is manganese
deficient.

Occasionally, decidedly alkaline organic soils

have never given any crop response to the application of
manganese salts.

Results with these Michigan soils agree

quite favorably with those from soils in other countries in
that the reported deficiency generally occur at a pH above

8.

6*7, although in Michigan, benefits have sometimes been noted
down to 6.2.

A strongly acid organic soil deficient in manga­

nese has not been encountered, even though some acid soils
show very little manganese upon total analysis.
Harmer, in his efforts to study this very interesting
problem, laid out an experimental area in 1934, on the
alkaline portion of the Michigan State College Muck Experi­
mental Plots at East Lansing.

On these plots a series of

treatments were made with the following purposes in view;
(1 ) to determine the effect of the application of manganese
salts on crop growth on an alkaline organic soil.

(2 ) to

determine the comparative results secured by acidifying the
soil with an application of sulphur flour, both with and
without the added application of manganese.

Twelve plots

were laid out and the annual treatments applied as shown in
Table 1.
During the period of this experiment, Harmer has used
many different crops.

The crops can be divided into two

general groups, namely, those that are definitely responsive
to manganese fertilizers and those that show little or no
response.

In the first group, such crops as barley, rye,

oats, potatoes, alfalfa, onions, peas, radishes, spinach,
leaf lettuce and many other crops gave very marked response
to the application of manganese salts.

If these salts were

absent, a chlorotic condition developed in the plant.

There

were some plants, however, which did not develop this
chlorosis in the absence of manganese, namely, sugar beets,

peppermint, sweet clover, Swiss chard, strawberries and
rhubarb.

Either these plants have a greater power to

extract manganese from the soil, or the nature of their
physiological processes are such that the need of manganese
is not acute.

There were instances, however, when even

these crops showed some improvement during growth due to
these treatments.

From Harmer8s unpublished data (31), the

yields of the several years indicate that the highest yields
generally were obtained on the plots receiving manganese
salts in combination with sulphur.

The highest application

of sulphur has at times given the highest yield, which
usually was in the neighborhood of the yield secured with a
combination of sulphur and manganese.

Manganese salts alone

did not give as high yields as did sulphur or the two in
combination.

In general, it may be stated that manganese

applications gave yields which are from 50 to 85 per cent of
those of the sulphur or the combination applications.

The

nature of the prevailing weather during the growing season
of the several years of this experiment has influenced the
response to these treatments.

A cold wet season has resulted

in the greatest response to manganese, with the control and
copper sulfate treated plots producing a near crop failure.
Under high temperature and near drouth conditions, the yields
from the controls and copper sulfate treated plots may nearly
equal those of the treated plots with certain crops and in the
case of spinach may sometimes be even better.

10.

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1, showing arrangement of plots of the M^ugaasse- Sulphur Series of the
College Muck Experimental Fields on which the bulk of the studies
presented in this thesis were made* All applications were made
broadcast in the spring of each year and prior to seeding of crops*

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11.
In view of the numerous conceptions as to the part
played by manganese in the soil, it appeared advisable to
study the various soil factors affecting the yields.

This

study began with the total analysis of the soil followed by
investigations of the various soil equilibriums, such as the
salts extractable by successive leachings with water, and a
determination of exchangeable bases.

This in turn was

followed by a study of the soil manganese in the various
horizons of the soil, with special attention to the amounts
extractable by various reagents.

The results of these ex­

periments and a consideration of work of Leeper (43) and
Piper (59) led to a seasonal trend study to follow the
transformation of the soil manganese from its manganous to
manganic form.
Total Analysis.
The 13 plots of the manganese-sulphur series of the
College Muck Experimental Plots were sampled in November of
1937.

Each plot was sampled to a depth of 6 inches and at

8 sites.

The samples from the 8 sites were thoroughly

mixed to form a composite sample.

Portions of these samples

were dried at 100°C and a 10-gram sample taken from each for
analysis.

These were ashed in an electric furnace and the

following constituents determined; insoluble ash, soluble
ash, organic matter, RgOg group, and ferric, calcium,
magnesium and manganese oxides.

Soluble ash included the

portion of total ash soluble in aqua regia.

The sesquioxides

(R2O3 ) were determined as the ammonium precipitate.

Ferric

iron wag determined by the titration of the ferric ion in a
thiocynate solution with a standard titanous chloride;
calcium volumetrically as the oxalate by the standard tit­
ration with permanganate solution.

Magnesium was determined

as the pyrophosphate and manganese, colorimetrically as the
permanganate by a method described by Williard and Greathouse
(78).
The data (Table 2) show very little variation in deter­
mined constituents as a result of soil treatment, except in
the case of MngC^ which was markedly increased when manganese
sulfate was applied.

The plots receiving manganese sulfate

are higher in most constituents, which may be due to the
higher mineral composition of the soil on these plots.

There

is a suggestion in the data that the application of sulphur
has caused a loss of MgO from the soil.
Successive Extractions.
The total analysis was followed by studies for the
purpose of securing an insight as to the various soil equi­
libriums which might exist.

A series of successive extract­

ions were made to determine the ability of the soil to give
up various ions.

The procedure was as follows;

a 25-gram

sample of soil was weighed out and transferred to a 500 ml.
Erlenmeyer flask. 250 ml. of carbon dioxide-free distilled
water was added and flasks were tightly stoppered.

These

were shaken at frequent intervals for 72 hours at which time
the soil—water mixture was filtered through a Buchner funnel.

13 •

Table 2,

Treat­
ments
Plot to 1937

showing total per cent of various constituents in soils
from Manganese-Sulphur Series on the College Experimental
Muck plots at East Lansing*
Insol­ Sol­
uble
uble
ash
ash

Org­
anic
matter CaO

MfiO

1 MnSO.400 4

19.21

15.63

65.16

6.63

2.26

4.82

1.04

.023

2 Control

17.03

14.68

68.29

5.94

1.96

4.37

.94

.016

3 s-iooo
MnS04400

14.35

12.11

73.04

5.32

1.32

3.50

.79

.035

4 MnS04800

14.57

15.22

70.21

6.70

1.82

4.14

.87

.056

5 Control

15.22

14.75

70.03

6.58

1.82

3.99

.88

.010

S-1000
MnSO.800

13.72

14.58

71.90

6.16

1.58

3.90

.83

.059

7

S-1000

13.56

13.78

72.66

6.21

1.55

3.76

.80

.011

8

Control

11.90

13.54

74.56

6.18

1.42

3.53

.74

.009

9

S—3000

13.72

13.36

72.92

6.03

1.28

4.07

.97

.010

10

S—2000

15.26

15.41

69.33

6.67

1.28

4.59

1.00

.013

11

Control

14.95

14.59

70.46

6.37

1.50

4.03

.90

.010

12

CuS04-200 16.70

17.26

66.04

7.54

1.78

4.81

1.10

.013

6

Treat­
ments
Controls
MnS04
S + Mn
S

Insol­ Sol­
uble
uble
ash
ash
14.77
16.89
14.28
14.18

14.38
15.42
13.24
14.18

FepOz

Averages in Per Cent
Org­
anic
matter CaO
MgO
RpO^ FepO^
70.34
67.68
72.46
71.64

6.27
6.66
5.74
6.30

1.68
2.04
1.45
1.37

3.98
4.48
3.70
4.14

.37
.96
.81
.92

Mn^04 ......

Mn?;04
.0112
.0395
.0470
.0115

The soil was washed with four 50 ml. portions of carbon
dioxide-free distilled water and allowed to filter quite
dry between each wash.

The filtrate was transferred to

a 500 ml. volumetric flask and made up to volume.

The

soil was returned to the original flask and allowed to
stand 72 hours, at which time the whole procedure was
repeated.

In aliquots of the filtrade, CaO, MgO and S04

were determined by methods described previously.

Attempts

were made to determine manganese and iron, but in such a
water extract they were in such small concentrations that
accurate analysis was impossible with the technique employ­
ed.

KgO was determined gravimetrically as the chloroplati-

nate salt.

The same soil samples were used for this study

as were used in the total analysis.
The most striking feature of these data is the behavior
of calcium.

Figure 1 shows that in the first three ex­

tractions, there appears to be a repression of the solu­
bility of the calcium ion, followed by a tremendous release
on the fourth extraction.

A possible explanation lies in

the common ion effect, as the fourth extraction shows the
last measureable quantity of the sulfate ion.

Since a soil

is a very heterogensis substance, a definite explanation is
quite difficult and such a repression of solubility might be
the result of several factors.

The calcium data show that

the treatments have affected its solubility in that, although
the total analysis shows practically no difference in CaO
content of these samples, the sulphur as shown in the graph,

15.
has markedly decreased the solubility of calcium, even
after the disappearance of water-soluble sulfate.

This

point becomes difficult to explain, since there is an
apparent conversion of calcium to gypsum and the data
show no difference between control plots and those treated
with sulphur in the amount of sulfate ions.

Leaching would

explain it if there were some indication in the total CaO
content of these plots that such leaching occurred.

The

data show that the water-soluble calcium content was lowered
almost two-fifths by the application of sulphur.

On the

other hand, the manganese applications apparently increased
the solubility of calcium.
Magnesium gave curves which were in general more
uniform but which have some points of interest.

A small

jump in the totals extracted came in most cases on the fifth
extraction.

The application of sulphur tended to lower the

soluble magnesium, with very little difference existing
between control plots and those plots which received manga­
nese sulfate.
Figure 3, presenting the quantities of water-soluble
KgO shows that a large quantity was obtained in the first
extraction and then it dropped to a low constant value.
The plots receiving manganese gave the highest values and
the sulphur applications the lowest value of KgO.
The value of successive extractions can be shown better
later in this work.

If the effect of the various treatments

has had a bearing on the solubility of any one element, that

16.

Graph 1, showing accumulated totals of CaO extracted
by successive (1-10) soil-water leachings.

1300

CaO
p *p .m.

9000

1100

* Control

MnS04 **
Sulphur

1000

3000

5000

7000

Sulphur

Numbers of extractions

17.

Graph 2, showing accumulated totals of MgO extracted
hy successive leachings.
MgO
p .p.m.

MnS04

1300

y+

^
t

MnS04
sulphur
Sulphur

^

300

500

700

900

1100

tr

100

— ^

1

_

3

_

3

L

4

_

5

i

^

6

^

7 8

m m

9

10

11

Control

12

G rbayp hsuccessive
3, showingleaehings#
accuinuXated totals of

**° e x ^ o tetX
ao
•■P*222.

1

^80^

8

Mn304 *

3

c°ntiv,i

4

Su^Phur

3ulPhUr

19.
element is calcium.

This would strongly indicate that the

soil equilibriums are dominated by calcium.

Ligon (46) in

his work on the effects of overliming has shown that, in
periods when soils were recently drenched with rains, the
quantity of calcium in solution was greatly increased.

With

a high content of calcium in solution the solubilities of the
other ions are greatly influenced.

The bases capable of

taking part in base exchange reactions are repressed by
calcium under such conditions.

Lastly, it is not unreason­

able to suspect that the nutrition of plants is greatly
affected even to the extent that a physiological disturbance
may be the result.
Exchangeable Bases.
Any study of soil equilibrium should include a study of
exchangeable bases.

It is to be realized that, in alkaline

organic soils, the method of analysis employed will influence
the values obtained.

Because of the low calcium carbonate

content of these soils, it was considered unnecessary to use
an alcoholic solution of the replacing reagent, avoidance of
which is desirable whenever possible.

Although it was

realized that the method has some deficiencies, the neutral
normal ammonium acetate was chosen as the replacing agent.
The method used was essentially the one proposed by
Schollenberger and Dreibelbis (63).

In aliquots of the

extract, the following exchangeable bases, calcium, manganese,
magnesium, iron and potassium, were determined by methods
previously described.

Exchangeable sodium was determined by

Table 3, showing exchange capacity and milliequivalents of exchange­
able bases* Values are expressed as milliequivalents per 100 grams
of soil.
Treatments
applied in 1938
in lbs. per acre

Exchangecapac­
Ca
ity
PH

Bite

K

Na

1

200 MnS04

7.5

118

100

15.6

.94

.15

1.18

.13

2

Control

7.8

123

105

15.5

to
CD
*

.31

1.29

.07

3

200 MnS04, 1500
Sulphur

7.4

128

110

14.0

.53

.18

3.07

.22

4

400 MnS04

7.8

138

121

15.1

.69

.17

.81

.23

5

Control

7.2

130

109

15.6

.56

.39

4.35

.10

6

400 MnS04, 1500
Sulphur

6.8

128

108

14.0

.41

.25

4.40

.96

7

1500 Sulphur

6.4

126

105

14.1

.34

.23

6.26

.07

8

Control

7.2

125

107

15.5

.47

.22

1.71

.10

9

Sulphur-3500

5.1

123

99

10.3

.92

.29

12.37

.12

10

2500 Sulphur

6.7

113

95

9.3

.41

.29

9.88

.12

11

Control

7.6

122

105

15.1

.56

.25

1.00

.09

1Z

50 Cu S04

7.6

123

105

15.1

.73

.32

1.77

.08

H

Mn

Averages
Controls

125

106

15.4

.60

.29

2.09

.09

MnS04

128

110

15.3

H
CO
«

.16

1.00

•
H
00

Plot

MnS04 ! Sulphur

128

109

14.0

.47

.20

5.73

•59

121

99

11.2

.56

.27

9.50

.10

Sulphur

20.
the zinc uranyl acetate method proposed by Barber and
Kolthoff (5) and adapted by Bray (11).
The data show very little difference in the base
exchange capacity of soils from the various plots, with
the exception of that from plot 10, which appears slightly
lower.

The average for all the samples shows that they

have a base exchange capacity of slightly over 120 milli­
equivalents.

It may be said that no essential difference

between the capacity of the soil from various plots exists.
The outstanding feature of the exchangeable-base data
is the high saturation of calcium and magnesium in the soil
complex, which in most cases make up more than 100 milli­
equivalents of the exchangeable bases-

The calcium ion

accounts for more than 90 milliequivalents, which is not
surprising when one considers the high quantity of soluble
calcium obtained in the successive leaching data.

The

change of reaction, due to the application of sulphur,
appears to have reduced both calcium and magnesium, the
latter being reduced as much as a third.

Their reduction

is represented in an approximately equal gain in exchange­
able hydrogen.
The exchangeable ions, sodium, potassium, manganese
and iron, together, make up less than a milliequivalent
of the exchange capacity.

The application of manganese

salts causes an increase in exchangeable manganese.
Exchangeable iron occurred in such small quantities that it
was impractical to measure it.

The evidence points to the

21.
low affinity for exchangeable potassium or sodium ions in
organic soils.

This contradicts some of the work of

Enfield and Conner (17) who attempted to show a fixation of
potassium in organic soils.

In these soils, the highly

soluble calcium content may be a prime factor in the low
values for these ions.

The organic soils demand great

quantities of these monovalent ions and, due to the great
annual need for crop production, it is quite apparent that
these ions are rapidly leached from the soil.
Extraction of Manganese from Soil.
To study the relationship of manganese to soil fertility,
it is necessary to investigate its solubility in soils under
various conditions.

To accomplish this end, a series of ex­

tractions were designed to determine the quantities of manga­
nese soluble in water, weak sulphuric acid, strong nitric
acid, a saturated solution of carbon dioxide, and the replace­
able and the easily reducible manganese in the soils of the
various plots of the manganese sulphur series of the College
Muck Experimental Field.

The samples were collected December

6 , 1938, from layers representing 0 - 5
and 10 - 16 inches.

inches, 5 - 1 0 inches

The samples were air dried and passed

through a 4 mns. sieve.
Methods of Extraction.
Water soluble manganese.

A 25-gram sample of air dry

soil is taken and placed in a 500 ml. Erlenmeyer flask;
250 ml. of carbon dioxide-free water is added and the flask

stoppered tightly to prevent entrance of air.

The flask

is shaken at intervals and after 48 hours the contents
filtered through a Buchner funnel.

The filtrate is

evaporated to a small volume and it is then transferred to
crucibles and evaporated to dryness.

The soil is returned

to the Erlenmeyer flask to use in further extractions.
After the filtrate has reached dryness, the crucibles are
placed in an electric furnace and are ignited over night,
after which time they are removed from furnace, allowed to
cool; aqua regia is added and is evaporated to dryness on a
steam bath.

The residue is then taken up with sulphuric

acid (1-1 ) and evaporated to dryness to volatilize the
chlorides.

The residue is taken up with a nitric acid (1-4),

filtered and 20 ml. of concentrated nitric acid is added with
approximately 0.3 gram of potassium periodate and the solution
boiled until color of the permanganate developes.

The solution

is then cooled and is compared in a colorimeter with previous­
ly prepared standards.

The manganese is reported in parts

per million.
Exchangeable Manganese.

Exchangeable manganese is the

manganese which can be replaced in the soil complex by
cation exchange.

The exchangeable medium used is a normal

neutral ammonium acetate solution adjusted to pH 7.0.
To the soil sample used in determining water-soluble
manganese, which was returned to the original flask, is
added 250 ml. of neutral normal ammonium acetate.

The flask

is tightly stoppered and then shaken at frequent intervals.

23.
At the end of 24 hours, it is assumed that equilibrium has
been attained.

The mixture is filtered through a Buchner

funnel, the soil washed with portions of ammonium acetate
solution and the soil again returned to the original flask.
The filtrate is treated exactly as in the determination of
wat er-soluble mangane se.
Easily Redueible Mangane se Dioxide.

Easily reducible

manganese is the quantity of manganese dioxide that can be
reduced by a 0*2 per cent solution of hydroquinone in a
buffered solution of neutral, normal ammonium acetate after
the water-soluble and replaceable manganese have been ex­
tracted.

The theory of this extraction was proposed by

Leeper (43), in 1935, when, in his work on calcareous soils
of Australia, he found he could pick out the manganese de­
ficient soils by this method.

His contention is that this

represents the quantity of manganese dioxide that the plant
roots can reduce and which becomes available for their nutri­
tional processes.
To the soil in the flask from which both the watersoluble and replaceable manganese has been extracted, is
added 250 ml. of normal ammonium acetate solution containing
0.2 per cent of hydroquinone and buffered to the pH 7.0.

The

flask is tightly stoppered and shaken at frequent intervals.
At the end of 24 hours, the content is filtered through a
Buchner funnel, and the filtrate is treated in the same manner
as the filtrate in the water-soluble manganese determination.
Total Mangane se in Organic Soils.

The method here is

applicable only to organic soils and really expresses the

24 •
quantity of manganese that is soluble in aqua regia*
5-gram sample is weighed into a crucible*
in an electric furnace at red heat.

A

It is then ignited

After complete ignition,

the crucible is cooled and then placed on a steam bath and
digested with aqua regia*

The residue is taken up with

sulphuric acid (1-1 ) and chlorides are expelled by heating
on a hot plate.

If much calcium is present, it is better

to filter before expelling the chlorides*

After the removal

of the chlorides, 20 ml* of nitric acid (1-4) is added and
the solution digested a short time*

If a residue or any

cloudiness is present, cool and filter.

To the filtrate

is added 30 ml* concentrated nitric acid and 0.3 gram of
potassium periodate and the solution boiled until color of
the permanganate appears.

After cooling, the color compari­

son is made with previously prepared standards*
Discussion of Experimental He suit s
Very few tests have been proposed for the determination
of the manganese available for plant growth in organic soils.
Muckenhirn (55) proposed the use of .005 N sulphuric acid
for such a determination while Steenbjerg (69) used a normal
solution of magnesium nitrate as a means of measuring the
exchangeable manganese, since he considered the magnesium ion
a better replacing agent.

The so-called quick-test methods

are of little value in determining available manganese in
organic soils, as the organic matter in the extracted solution
prevents the development of the required permanganate color*

35.
Leeper (43) proposed the use of 0.2 quinol solution buffered
with neutral normal ammonium acetate, which, according to
his theory would extract the easily reducible manganese
dioxide by reducing it to the bivalent form.

All of these

methods measure three forms of soil manganese, namely:
(1) The portion soluble in weak acids.
12) The portion taking part in the cation exchange.
(3) The portion in the form of easily reducible
mangane se dioxide.
The various extractions listed above were made on the
soils being studied and it was found, as shown in Table 4,
that the amounts of water-soluble manganese, of the manganese
soluble in .005 H. sulphuric acid and of that soluble in a
saturated carbon dioxide solution, were so small as to be of
no value.

The data obtained, when five per cent nitric acid

was used, are interesting in that they show a marked increase
in soluble manganese with the application of manganese sul­
fate.

The plots receiving sulphur show, in general, slightly

less nitric acid-soluble manganese than do soils from the
controls.

This difference becomes more marked in the 5-10

inch layer, while in the 10-16 inch layer the manganese is
almost half that of the control.

The manganese soluble in

.05 H. sulphuric acid is about one-half that of the nitric
acid-soluble and bears approximately the same relationships
as the nitric-acid soluble does in respect to treatments.
Many investigators ascribe a low exchangeable manganese
supply as the cause of insufficient manganese for plant growth..
Steenbjerg (70) states that it is not only the amount of
exchangeable manganese that is important but also the tenacity

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26.
of retention of the exchangeable manganese by the soil
colloids.

He states further that the exchangeable manganese

is decreased by liming to a high pH; by excessive aeration
and by deficiency of moisture and is increased by chemical
reducing agents as readily decomposable organic matter;
starch and waterlogging.

He holds that heavy soils have a

high exchangeable manganese content and a low retention of
the manganese by the soil colloids, which is perhaps the
reason why ngrey speckw disease in cereals never occurs on
this type of soil.

In a later paper, he states that, on

soils having a total exchangeable manganese content of
0.5 milliequivalents per 27.5 kg. of air dry soil, **grey
speck11 disease would occur but that, if there were 2.0 milli­
equivalents, there would be no signs of the disease.
A recent paper by Ruether and Dickey (61), working in
Florida on mineral soils, states that two pounds per acre of
exchangeable manganese was found insufficient for normal
growth of tung trees, that 2.3 to 3.4 pounds per acre gave
normal trees and that an excess would be toxic.

This does

not hold true for the manganese—deficient soil on the College
muck plots, since a very conservative figure shows that the
Boil of the control plots contain at least three to four
pounds of exchangeable manganese per acre in the surface five
inches.
The determination of exchangeable manganese shows that,
where manganese sulfate has been added, the exchangeable
manganese is increased, while a sulphur application tends to

27.
decrease it in respect to the total manganese of the soil
and, in actual quantities, it is only slightly higher than
the exchangeable manganese in the controls*

The data of

the 10-16 inch layer are not entirely reliable, inasmuch as
the underlying clay enters this layer section in plots 1 , 2 ,
3, 11 and 12.
Leeper (43) in his work in Australia, found that a low
content of easily reducible manganese dioxide gave rise to
the so-called manganese deficient soils.

The soils that he

worked on were very calcareous and he considers that the
water-soluble and exchangeable manganese in these soils are
so low that they would be of little value to the plant.

His

theory is that the plants can reduce colloidal manganese
dioxide, a reasonable assumption, since other workers (36)
have reported benefits from the application of manganese
dioxide.

He suggested the use of a 0.2 per cent solution of

quinol in a buffered solution of neutral normal ammonium
acetate for the extraction of reducible manganese.

This

extraction is made after the exchangeable manganese has been
removed.

He found that soils having 100 parts per million or

more of manganese grew healthy plants; those having 15 or less
had a very severe chlorosis.

Leeper apparently holds that

the range between these figures is the range of recovery.

It

is also interesting to note that he states the best yields
were not obtained on those plots which received manganese but
on those which received sulphur which correlates with condi­
tions that occur on the plots under consideration in this work.

The results of the extractions using 0*2 per cent hydroquinone
in neutral ammonium acetate are very interesting.

Those plots

receiving 200 pounds of manganese sulfate per acre showed 27
and 61 parts per million of manganese hy this test, while
those receiving the 400-pound application gave 111 and 99
parts per million.

The controls yielded only 9 and the

sulphur treated plots 12 parts per million.

In the 5 - 1 0

inch layer, the plots receiving manganese sulfate still
showed more than 15 parts per million while the controls and
sulphur contained 7 and 6 , respectively*

The two mineral

soils from Minnesota, high in manganese, gave 98 and 492
parts per million, the latter being the soil high in pyrolusite.

It is very likely from these results that this

reagent can be used to measure the active manganese dioxide
content of the soil*
Considering Steenbjerg*s (70) work again, the difference
in retention of manganese by the soil colloid may be the
very factor as given here.

In Table 5, it will be noticed

that a Minnesota soil high in pyrolusite, when extracted
with the 0.2 per cent hydroquinone reagent, yielded a
quantity of manganese approximating that soluble in the five
per cent nitric acid.

It is apparent, then, that the pyro­

lusite content of a soil might be a factor in the quantity
of manganese available to plants.

29.
Table 5, showing the analysis of two mineral soils from the
Red River Valley in Minnesota.
Depth
Ex­
Easily
of
.005N .05N change­ reduci­ Nitric
sample K z B O a H2.SQA. able
ble
acid
"

Soil Type
<i>
1. Semi-hydro­
morphic assoc­
iate of Barnes

0-6

2

148

50

98

625

2. Dolomite Sand
(Averill Area,
22-38
492
Clay Co.)
32
<£)1. Serai-hydromorphic associate of Barnes Series.
calcareous containing 44$ OaCOg, also some CaSO^.
of this soil is 7.89.

522
Very
The pH

Plants growing on this soil do not

respond to applications of manganese salts up to 1250
pounds per acre and above this figure yields are depressed.
Copper Sulfate and Gypsum also depress yields.

The appli­

cation of ferrous sulfate has shown beneficial results
with such crops as red clover and spinach.
2.

A glacial lake sand in which the carbonates have been

altered to dolomite by the action of ground water rich in
magnesium.

This sample is taken from the C horizon of the

Ulen Series 3 miles southeast of Averill, Clay County,
Minnesota.

This subsoil has from 25 to 30 per cent carbon­

ates of which 90 to 95 per cent is in the form of dolomite.
The pH of this soil ranges from 7.4 to 7.6.

This subsoil

has many pyrolusite concretions and upon heavy mineral
separation, some carbonates in the form of ankerite are
found with the heavy minerals.

Ankerite is the double

carbonate dolomite in which either the calcium or magnesium
ion has been replaced by either iron or manganese.

Ho minor

30.
element studies have been conducted on this soil but, from
the analysis, it undoubtedly has sufficient manganese to
meet the requirement of plants under these conditions.
Table 6 , showing the Mn extracted from various alkaline
mucks by the different reagents.
Location
of
muck

To'fcal
aqua
Reaction regia

Alkaline 1070
Boysen
n
Oalhoun
tf
Richmond
n
3234
Hubbard
w
150
Wyn
ft
Born
Very acid
Rich

w

■■

nitric Water
soluble
acid
715
—

—

500
1667
111
217
6

0.6
trace
trace
0.6
trace
0.4
trace

Exchange­
able
33
34
69
80
S
35
none

Easily
reduci­
ble
53
40
22
302
trace
16
none

Table 6 , showing the analysis of various mucks from
different parts of Michigan, is very interesting in respect
to these various extractants.

In field and greenhouse ex­

periments in past years, Harmer found that all of these
alkaline mucks, except the Hubbard, responded to the appli­
cation of manganese salts.

The Hubbard and Boysen mucks

are both very high in iron content, the chief difference
between them other than the above analyses being that the
Boysen muck was very high and the Hubbard very low in carbon­
ates •
The Hubbard muck has three times the manganese content
of the Boysen, yet they show little differences in watersoluble manganese.

The Hubbard has 80 parts per million of

exchangeable manganese against 33 for the Boysen and the
difference in the reducible is still greater, 302 against 53.
The data support both theories that the exchangeable or

reducible manganese is the important factor but, when one
takes into account the results of Richmond's muck which
proved very responsive in Harmer's trials to both manganese
or sulphur applications; this muck has almost as high an
exchangeable manganese content as the Hubbard muck, but has
a lower reducible manganese content than Boysen,

This fact

gives a great deal of weight to the reducible manganese
dioxide theory but it appears to be only applicable to
alkaline conditions.

Rich's muck, which is very acid shows

no exchangeable or reducible manganese.

Coupling this with

Harmer's field experiments, it appears definite that the
plants demand for manganese is much less on acid organic
soils than on alkaline soils.

This is in agreement with

trials on mineral soils and with Martin's work (50) on sugar
cane with nutrient solutions which showed that manganese was
beneficial at pH of 6.0 or higher.
Table 4, which presented the various quantities extract­
ed by various reagents, showed a much higher concentration of
manganese near the surface.

This may be indicative of a

migration of manganese in the soil, or it may be due to a
negative enrichment.

The work of McCool (53) will lend some

support to the former theory that, during reducing conditions,
the bivalent manganese is set free and during drouthy condi­
tions may migrate to the surface, where it is oxidized into
colloidal pyrolusite.

If such a process takes place the sur­

face soil will show a higher manganese content in the colloid­
al fraction.

This theory is supported by the following table

from the work of Alexander, Byers and Edgington (1).

33 •

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O
cd
d
-P

m
T3

O toH

£
r
H
•H

a
o
d
Cm

d
o
p p
d i—1
<D CD
f**J
^
pH Cd
p d O
I&4 ho rH

I
Q
o o

t
!

O
CM

cm
00

•

O
i—1
«

CM
. O
•

<d -p

8

1

'O
CD
d
pq

M

O
rH

O P
P rH
d P
CD CO

rH
CQ

CM

pq

to
PQ O

I H1
I
O•

|

rj

Na

o cd
■P o
CO (H
d
CD p
tlO rH
cd P
W co
&
g: cd
O rH
p O
CO
d
<d p

a

fciOrH 3
.f i P °
tQ w rH

cm

•

p

I uo

I

o

•

to

I

03
I
O
1

r

d
o
W

d
o
N

rH

CM

CM

A

cd

to
O
•

A

05

tCM

tO
O
•

PQ

CQ

to

m

o

* Data taken from U,S«D.A, Tech, Bui, 678, by Alexander, Byers and Edgington

g

33
Soils.
1.
2.
3.
4.
5*
6.
7.
8.
9.
10.

Hagerstown silty clay loam, State College, Pennsylvania*
Hagerstown silt loam, Hagerstown, Maryland.
Frederick silt loam, Fairfield, Virginia.
Maury silt loam, Ashwood, Tennessee.
Dewey silt loam, Russellville, Alabama.
Decatur clay loam, Russellville, Alabama.
Greenville sandy loam, Pretoria, Georgia.
Fullerton gravelly loam, Lawrence, Alabama.
Hagerstown silt loam, Ha^el Run, Missouri.
Lebanon silt loam, St. James, Missouri.
Seasonal Trend of Replaceable,Total Manganese
and of Reducible Manganese Dioxide
The apparent enrichment in manganese of the upper part

of the soil profile leads one to suspect that a change in
the chemical state of manganese is taking place in the soil.
The question naturally arises as to what becomes of the
manganese sulfate after its application.

What climatic

factors appear to play a part in the chemical changes of the
manganese?

Of special interest would be the effect upon

the equilibrium between the exchangeable manganese and the
active manganic oxides.

This is of considerable importance,

since there are two schools of thought as to which of these
two forms of manganese are most important to plant growth.
Beginning with May 9, 1939, samples of muck were collect­
ed every fifteen days until September 22.

The area sampled

was that portion of the plots seeded to alfalfa in the spring
of 1938.

The samples were rapidly air dried so as to keep

chemical changes after sampling to a minimum.
The application of manganese sulfate to plots 4 and 6 ,
was made May 1, 1939.

The treatments of these plots including

the 1939 application are as follows;

34.
Plot

1&38 Treatment

1§39 Treatment

1934-193^ Treatment

4

MnS04-400

Mh304-400

MnS04—1600

6

MnS0A-400
Sulphur-500

MnSO^-400

MnS04-1600
Sulpnur-1500

8

Control

Control

Control

9

Sulphur-500

No treatment

Sulphur-3500

The samples were analyzed for total manganese, exchange­
able manganese and reducible MnOg by methods described
previously in this paper.
Figure 4 shows the results obtained from such a study
in respect to total manganese.

The uniformity of the total

manganese curve suggests that manganese is fixed rather firm­
ly in the soil.

This supports the results shown in the

horizon study in which there was very little evidence of
enrichment of the lower horizons due to manganese sulfate
applications to the surface.

The decided uniformity of the

curve of the total manganese on the control plot and on the
plot receiving sulphur is in general what one would expect.
Any enrichment due to migration from a lower layer would
have very little effect upon total content, due to the small
amount present.
The curves representing the seasonal changes in exchange­
able manganese are of an entirely different nature.

In gen­

eral, as the season progresses, the exchangeable manganese
becomes less and less.

It would appear that the tendency

during the season is for the manganese to be oxidized apparent­
ly to the manganic forms.

The treatments indicate that the

35

CM
CM
j

x

Table 8,

showing the seasonal trend of the manganese existing as reducible MnQg and >
changeable cation and the total manganese in the soil at each sampling date
during the season of 1939* Manganese sulfate applied approximately a week
before first sampling.

Q>

CD

CO

P

P
U
Q>

CD
P

CM
CD

0

OO
c>
rH

CD to
rH to
CM
rH

to
P

IP
CD

p

O
rH

to CM
to to
rH
rH

vl<

05

to

03

CM
to
rH

CD

to
p
p

00

O

to

00

tO

O D- CM
LO 00 CM
P

05

to

CO

04
5
^

P to
CO
rH P
rH

CD
CO
P

CD
CO

CM LO
rH CM
CM
rH

to
to

p

'j}*

p

O- to
C-~
rH P
rH

CM

0

CO
CO

CO

ip

'■Ct* C*O co
to p

O
CM

00

10

IP

O
rH LO
rH p
1
—1

O
05

p

CD

to

CD

to
to
to

to
to

o
o
to

rH
CM

CD

CO rH
c
CM

I

»

O LO
CO P
rH

to
to
1—1

CD
lO
CM
T—

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IP

CM

co

O

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to

CQ

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04
©

CQ
to
cm

I

rH

oo
so

rH

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CO
OS

CM

£
P
05

to
to

6

o-3
t

rH

rH
to

to
to

rH

to
rH

00

t>-

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CM
H

*
05
to
05
rH
|

1

to
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CD

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to

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to
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to

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to

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pH

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0
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C£

Figure 4, showing seasonal
of Mn-S Series,

trend of total manganese

in 1939 of certain plots

36.

o

o
o

-Kl

H>

o>

o
o

o
o
02
rH

O
O
O
H

o

O
o
«o

o
o

June 9

July

of exchangeable manganese

June 24 July 9

trend

rH

rH

o
10

May 9

May 25

Figure 5, showing seasonal
p.pjuplots of Mn-S Series.

O

CM

o

rH

to

O
24 Aug. 8 Aug. 23

in 1939 of certain

H

a

Sept. 7 Sept. 22

37.

O
o

O

o
<£>

o

Figure 6 , showing seasonal
plots of Mn-S Series*

trend of reducible HnOg during 1939

of certain

38.

&

0

O

O

t—I

»H

to

o

O

CM

rH

CM

CM

O
O

tO

•p
rH

to

o
o

rH

HD

CM

CM

to

CM

CM

O

o

o

29,
application of sulphur tends slightly to increase the ex­
changeable manganese or rather it tends to hold it at a
slightly higher level*

During the season of increasing soil

temperatures and lowering soil moisture content, the ex­
changeable manganese falls from 6 parts per million to 0*6
part per million.

It is reasonable to assume that, during

the winter season, the reverse action will take place and
that the level of exchangeable manganese on the controls from
one spring to the spring of any following year will be
reasonably constant.
There is a gain during the summer In the amount of active
MnOg, as measured by Leeper*s method.

In other words, the con­

clusion can be drawn that, as the exchangeable manganese
decreases, the active MnOg increases.

It seems probable, then,

that during the winter part of the season the reverse of this
reaction is possible.

It appears that the manganese sulfate

application gives the highest level in these treatments and
that sulphur treatments give values which are practically
comparable with the control.

This contradicts in a measure

Leeper*s theory since, in this case, the crop yields do not
correlate with his test.

At this point it might be well to

consider Leeper*s theory more seriously.

Fundamentally, he

has divided the soil manganese into several groups and, in a
broad sense, they are exchangeable and manganic compounds.
He states (page 24): (43)
"It appears that manganic oxides may form a continuous
series from the most active to the most inert; for convenience
one may separate these oxides into four classes -

40.
(a) extremely active, capable of oxidizing quinol at
pH 7 and of oxidizing the organic matter of the soil rapidly
in sulphuric acid suspension (pH 1.5 to 2.0);
(b) also very active, capable of oxidizing quinol at
pH 7, but only slowly attacking organic matter at pH 1.5 to
2 .0 ;
(c) moderately active, capable of oxidizing quinol at
about pH 2 or Na2 S2 04 at pH 7;
(d) inert (including all the remaining manganic com­
pounds).*1......."analogous to the most resistant residues
of pyrolusite, which dissolve in oxalic and sulphuric acids
only after prolonged boiling."
He believes that the various categories of manganese
are in a dynamic equilibrium as he expresses its
^

(
...

"The dynamic equilibrium, .............. Exchangeable
unavailable Mn02 of former workers, however, may
■> now be replaced by some such series as
Exchangeable Mn
■
MnOo (a) <•--- . Mn02
(b) *---- Mn02 (c) 4— •■■y; inert Mn02 (d)"
This equilibrium will be shifted from right to left or

vice versa by the conditions existing in the soil.

Piper

(59) has shown that water logging will shift it from right
to left and, in the seasonal trend of soil manganese present­
ed here, we have shown that, under increasingly drouthy condi­
tions, the equilibrium is shifted in the reverse direction.
The relationship which exists between exchangeable and active
MnOg certainly substantiates this theory of dynamic equili­
brium in the soil.

The rapidity of the oxidation and the

lack of leaching shows that the direction of this equilibrium
is strongly in the direction of the inert MnOg.

This perhaps

explains the low values received when this soil was extracted
with water or weak acids.

41.
It is interesting to note that, if one were to compare
the total manganese in acid organic soils and alkaline
organic soils in Michigan, one would at once he struck by
the great difference in the two types of soils.

Harmer’s

unpublished data (31) show that a majority of the acid mucks
of Michigan have a total manganese content which is very
small, in most cases, much lower than Leeper*s critical
values of active Mn02 and even lower than some of the critical
values given for exchangeable manganese.

A great deal has

been attributed to the soil reaction, since, at a low pH, the
organic matter will reduce MnOg easily, with the result that
an increase occurs in bivalent Mn which is subject to leach­
ing.

Leeper (43) states in mineral soils,
"since reserves of "active Mn02 " fall off rapidly
with a fall in pH, one might forecast that, other
things being equal, it is more dangerous to lime a
highly acid than a moderately acid soil. It may
be remarked that many of the soils that have become
"deficient" after liming have naturally a very acid
reaction".
This perhaps explains some of the observations in the

past in which it is possible to have a medium acid soil
under conditions which subject it to long periods in which
reduction is predominating.

Under such a condition the

manganese can be leached from the soil.

However, as long as

this soil is acid in reaction, no deficiency of manganese
will appear, but, whenever it is limed, this deficiency will
appear as soon as the active MnOg becomes too low.

It is also

possible to have a medium acid soil which does not have periods
of reduction or, if it does, the manganese is not leached from

42
the soil and, under such a condition, over liming will not he
possible.

43.
THE EFFECT OF MANGANESE FERTILIZER
OH THE MINERAL COMPOSITION OF THE CROP

A large amount of work has been done on the effect of
the application of certain fertilizers upon the mineral
composition of the crops grown on mineral soils.

Very little

work has been done on the composition of the crops produced
on organic soils.

Recent studies of Thomas and Mack (73)

working with mineral soils, have shown that considerable
more attention should be paid to the time of season that
samples are collected for analysis.

Further, the work of

McCool (53) has demonstrated that light intensity and kinds
of light determines to a great extent the quantity of ions
present.

Thus, it is evident that there are many factors

playing a part in the utilization of soil nutrients other
than availability of the nutrients themselves.
The difference between the composition of normal and
chlorotic tissue will be considered very thoroughly in this
work.

As has been pointed out, plants growing on the area

under investigation are chlorotic unless they either receive
manganese or the soil is made acid by the application of
sulphur.

The assumption has prevailed in the latter case

that a change in soil acidity will cause the manganese to
go into solution and be available to plants.

In order to

secure definite information regarding this point, it was con­
sidered desirable to make mineral analysis of plants receiv­
ing these respective treatments.

It was deemed of equal

interest to see if there were any effects upon the other

44.
mineral constituents other than the contents of manganese.
Scholz (65) states in his work with nutrient solutions on
lupines, that manganese appears to he essential and that
there exists some relationship between calcium, manganese
and iron.

Hoffer*s work (33) indicates a possible associ­

ation between iron and potash and Willis (79) intimates
that there may be a relationship between manganese and iron.
It was believed that chemical analysis may give valu­
able information as to the validity of these points.

For

this study samples of plant material were taken for analysis
during the seasons of 1937 and 1938.

The samples taken at

the date of harvesting of the crops, should shed some light
on amounts of the various materials removed in the crop.
Early in the summer of 1938, a crop of canning peas offered
excellent material for a comparative study of chlorotic
and normal tissue.

Since there are seasonal changes in

composition due to various seasonal factors, a seasonal trend
study was considered advisable.

During the season of 1938,

samples of barley plants likewise were taken, at ten-day
intervals during their growing season.

Samples also were

gathered at fifteen-day intervals on a legume experiment
which included alfalfa, alsike clover and sweet clover.

De­

termination of CaO, MgO, Fe^Og, Mn^O^, S, ^3^5 > ^2^’ and
!Tao0 were made upon the crops sample in 1937.

The seasonal

trend study included only Mn^O^, Fe^Og, CaO and MgO.

In

some cases a few analyses were made to determine its possible
influence on still other constituents.

The plants were

45.
sampled, weighed immediately, and then dried rapidly at
60°C.

The dried plant tissue was ground in a Willey Mill*

A portion of the mixed ground material was then placed in
a drying oven and dried approximately six hours at 105°C.
At this point, samples were taken for analysis.
Calcium was determined volumetrically by titrating the
oxalate with a standard permanganate solution, and magnesium
gravimetrically as the pyrophosphate.

The iron determination

was made volumetrically by dissolving the ash by hydrochloric
acid and then converting the iron to the ferric state by
cautiously adding a weak permanganate solution.

Ten milli-

leters of a ten per cent solution of ammonium thiocyanate
solution was added and the ferric iron reduced by titrating
with a freshly standardized solution of titanous chloride*
Manganese was determined by the method described for the
determination of total manganese in the organic soils.
Sulphur and phosphorous were determined in a magnesium nitrate
ignition, sulphur gravimetrically as barium sulfate and phos­
phorous volumetrically by dissolving the yellow ammonium
phoepho-molybdate precipitate with an excess of a standard
alkali and titrating the excess base with a standard acid.
Sodium and potassium determinations were made by methods
similar to those employed in exchangeable ions.
The following crops were analyzed by these methods:
onion bulbs, potato tubers, potato vines, sugar beets,
sugar beet tops (including crowns), strawberry leaves, and

46.
raspberry leaves.

Strawberries and sugar beets are crops

non-responsive to manganese and sulphur applications.

The

following tables will show the data which were obtained in
these analyses.

Table

9,

showing the mineral composition of mature onions produced
in 1937 under varying applications of manganese and sulphur
on various plots of the Mn-S Series at the College Mck plots.

Total
applications HgO
1954 - 1957
(a)
MnS04 -400
91.34

6.59

.083

.022

.00117

.00017

.176

.0025 .046 .287

MnS04 -400
S - 1000

91.32

6.67

.083

.024

.00111

♦00012

.212

.0054 .041 .289

4

MnS04 -800

91.15

6.39

.086

.023

.00112

.00020

.206

.0039 .038 .284

5

Control

90.01

6.62

.073

.024

.00140

.00008

---

6

MnS04 -800
S-1000

90.29

6.03

.079

.024

.00096

.00024

.194

.0022 .039 .239

7

S - 1000

90.51

5.74

.078

.022 ..00109

.00013

.212

.0024 .037 .245

8

Control

90.59

6.43

.079

.029

.00236

.00007

.245

.0023 .050 .285

9

S - 3000

90.58

6.19

.091

.026

.00102

.00011

.223

.0034 .044 .276

11

Control

90.00

6.27

.062

.030

.00137

---

--

Plot
1
3

Ash

Percentage calculated on field weight basis*
CaO
MgO Fej?03
Mn504
KgO
Nag0
S p2°5

--

---

\
Average composition in per cent.
NagO
MgO FegO 3 Mn304
KgO

.058 .320

-- .335

H&0

Ash

CaO

Control

90.20

6.44

.071

♦028

.00171

.00007

.245

.0023 .054 .313

MnSO^

91.24

6.49

.084

.022

.00114

.00018

.191

.0031 .042 .285

MnS04 + S

90.80

6.35

.081

.024

.00103

.00018

.205

.0028 .040 .264

S

90.54

5.96

.084

.024

.00105

.00012

.217

.0029 .040 .260

Treatment

*

Ash calculated on water-free basis

(a) Pounds per acre.

S p2°5

1

Total
applications
.
1954 - 1957
(a)
MnS04 -400
9.63

3.97

.98

.045

.00266

1.12

.246

.156

.99

2

Control

9.85

4.37

.73

.058

.00146

.96

.232

.144

1.07

5

BlnS04-400
S - 1000

9.07

3.27

.85

.038

.00294

1.27

.364

.205

1.11

4

MnS04-8Q0

8.81

3.38

.97

.035

.00319

1.02

.246

.176

1.02

5

Control

9.91

4.30

00

showing the mineral composition of potato vines produced
under varying applications of manganese and sulphur on
the Mn-S Series of the College Muck plots*

*

Table 10,

.072

.00242

.91

.244

.128

1.12

6

MnS04-800
S-1000

9*47

3.61

.93

.039

.00374

1.09

.252

.150

1.17

7

S-1000

9.28

5.88

.74

.043

.00261

.74

.256

.151

1.14

8

Control

8.98

3.81

.87

.064

.00242

.93

.243

.118

1.16

9

S—3000

7.66

3.42

.72

.043

.00277

(a)

.74

.235

.120

1.11

Average composition in per cent
MgO FegOg B&15O4 KgO
IJsgO
S

P2O5

Treatment

Ash

CaO

Control

9.59

4.16

.73

.065

.00215

.95

.240

.130

1.12

MnS04

9.22

3.67

.97

.039

.00292

1.07

.246

.166

1.00

MnS04 + S

9.27

3.44

•
00
CO

Plot

Percentage calculated on oven dry weight basis
CaO MgO FegOg Mh304
KgO
Na20
S
PgOg

.038

.00334

1.18

.308

.178

1.14

S

8.47

3.65

.73

.043

.00269

.74

.245

.135

1.12

pounds per acre*

Table 11,

showing the mineral composition of potato tubers produced
in 1937 under varying applications of manganese and sulphur
on Mn-S Series of College Muck plots.

•Total
applications
Plot 1954 - 1937
(a)
1
MnS04-400
84.32

Percentage calculated on field weight basis*
Ash Ca0
MgO FegOg Mn304
KgO
NagO

S P2O5

7.04

.018

.047

.0016

.00012

.515

.022

.045 .392

2

Control

84.48

7.18

.018

.049

.0018

.00008

.504

.024

.058 .371

3

MnS04-400
s-1000

84,68

6.99

.017

.044

.0016

.00011

.539

.037

.055 .446

4

MnS04-800

84.09

6.96

.022

.050

.0016

.00015

.584

.044

.057 .391

5

Control

86.02

7.36

.018

.046

.0018

.00008

.471

.021

.053 .382

6

B&1SO4-8OO
s-1000

85.17

6.66

.020

.048

.0016

.00013

.593

.036

.057 .442

7

S-1000

84.16

6.89

.019

.046

.0019

.00008

.541

.041

.058 .411

8

Control

86.69

7.20

.017

.046

.0018

.00006

.443

.015

.051 .355

9

S—3000

85.30

6.66

.018

.045

.0016

.00007

.555

.029

.057 .389

Average composition in per cent
Treatment

h 2o

Ash

CaO

MgO

Feg03

Mngp4

KgO

NagO

S Pg05

Control

85.75

7.25

.018

.047

.0018

.00007

.473

.019

.054 .369

MnS04

84.20

7.00

.020

.048

.0016

.00013

.549

.033

.051 .391

MnS04 + S

85.92

6.82

.018

.046

.0016

.00012

.566

.036

.056 .444

S

84.75

6.77

.018

.045

.0017

.00007

.548

.035

.057 .400

* Ash calculated on water-free basis
(a) pounds per acre.

Table 129

Total
applications
ttppxicatiions"
1934- 1937

Percentage calculated on field weight basis*
Ash
CaO
MgO Fe2°3
Mhg04
KgO Na^O
S F2°5

I
1

i
o
o

■
’lot

showing the mineral composition of sugar beet roots produced
in 1937 under varying applications of manganese and sulphur
on Mn-S Series of College Muck plots.

1

77.8

3.79

.035

.099

.00027

.00041

.366 .043 .014 .229

2

Control

77.4

5.43

.033

.092

.00019

.00031

.372 .039 .013 .238

3

MnS04-400
S-1000

78.5

3.59

.037

.099

.00028

.00059

.397 .035 .014 .221

4

MnS04-800

77.2

3.51

.033

.094

.00027

.00068

.370

5

Control

76.9

3.19

.034

.109

.00019

.00070

.369 .030 .014 .252

6

MnSO4-8OO
S-1000

76.7

3.19

.043

.095

.00022

.00084

.392 .036 .013 .227

7

S - 1000

76.4

3.16

.030

.104

.00023

.00057

.362 .038 .016 .235

8

Control

77.2

4.01

.036

.102

.00019

.00036

.406 .040 .014 .232

9

S - 5000

76.5

3.16

.035

.099

.00026

.00046

.402 .036 .016 .276

10

S - 2000

77.2

3.72

.038

.103

.00022

.00037

.406 .041 .013 .244

11

Control

75.5

3.40

.038

.114

.00019

.00037

.394 .038 .015 .230

12

CuS04-200

77.9

4.28

.037

.101

.00016

.00031

.405 .046 .012 .250

.057 .015 .235

Average composition
MgO

Fe2°3

.035

.104

.00019

.00043

.385 .037 .014 .238

3.65

.033

.096

.00027

.00054

.368 .050 .014 .232

77.6

3.59

.040

.097

.00025

.00071

.394 .036 .013 .224

76.7

3.52

.034

.102

.00024

.00047

.390 .038 .015 .252

%0

Ash

Cao

Control

76.7

3.51

MnS04

77.5

Treatment

MnS04
S

S

* Ash calculated on water-free basis
(a) pounds per acre.

Mng04

KgO NagO

S PgOs

Table IS, showing the mineral composition of sugar beet tops and crown
produced in 1937 under varying applications of manganese and
on Mn-S Series of College Muck Plots.

1

Total
applications HgO
1954 - 1937
(a)
MnSO4-400
70.5

16.88

.541

.620

.0200 .00105 .944 .242 .062 .512

2

Control

70.3

14.09

.480

.622

.0164 .00119 .715 .225 .056 .499

3

MnS04-400
S - 1000

72.8

14.29

.475

.630

.0168 .00240 .761 .242 .046 .494

4

MnS04-800

72.1

14.69

.403

.436

.0125 .00235 .915 .236 .059 .497

5

Control

73.2

13.03

.338

.439

.0107 .00140 .777 .234 .049 .464

6

MnS04-800
S - 1000

70.8

13.77

.523

.492

.0132 .00283 .716 .251 .050 .472

7

S - 1000

72.0

14.49

.429

.485

.0137 .00094 .728 .222 .057 .486

8

Control

75.2

14.96

.411

.451

.0131 .00125 .721 .230 .054 .483

9

S - 3000

75.0

14.54

.440

.441

.0151 .00149 .733 .227 .060 .455

10

S - 2000

73.1

14.56

.422

.507

.0150 .00131 .712 .234 .057 .472

11

Control

75.3

13.51

.284

.348

.0112 .00082 .711 .218 .049 .461

IS

CuS04-200

74.9

14.25

.335

.432

.0135 .00096 .712 .210 .052 .454

Plot

Percentage calculated on field weight basis*
Ash
CaO
MgO FegOg Mh304 KgO Nag0 S p2°5

Average composition
Treatment

HgO

Ash

73.5 13.90
Control
71.3 15.78
MhS04
71.8 14.03
MnS04 + S
73.4 14.55
S
* On water—free basis.
(a)pounds per acre.

CaO

MgO

.378
.472
.499
.430

.465
.528
.561
.478'

FegOg Mng04 KgO NagO
.0128
.0162
.0150
.0146

.00166
.00170
.00261
.00091

.731
.929
.738
.724

.227
.239
.246
.228

S PgOg
.052 .477
.060..504
.048 .483
.058 .471

Table 14,

showing the mineral composition of strawberry leaves
collected in late fall of 1937 on Mn—S Series of
College link Plots.

2

Total
applications
1954 - 1937
(a)
Control

7.68

2.33

.63

.072

.0034

.115

1.27

4

MnS04 - 800

7.81

2.41

.63

.076

.0079

.125

1.20

5

Control

7.30

2.43

.63

.057

.0049

.121

1.09

6

MnS04 - 800
S - 1000

7.18

2.46

.70

.053

.0097

.118

1.33

8

Control

8.52'

3.05

.60

.068

.0027

.141

1.14

9

S - 3000

8.29

2.26

.67

.073

.0035

.139

1.27

Average of Controls
7.83

2.60

.62

.066

.0037

.126

1.13

Plot

Percentage calculated on water--free basis
Ash
Mn5°4
CaO
MgO
S
P2°5
Fe2°3

(a) Pounds per acre*

Table 15,

showing mineral composition of raspberry leaves
collected in October, 1957, from College Plots,

Total
applications
1954 - 1957
(a)
Control

7,09

1.78

.95

.089

.0057

.069

1.14

MnS04-400
S - 1000

7.14

1.92

1.00

.053

.0066

.069

1.09

8.91

1.78

1.00

.057

.0074

.080

1.09

6,98

1.77

.99

.075

.0079

.044

.99

MnSO--800
S - 1000

7.92

1.97

.91

.057

.0105

.059

.84

7

S - 1000

7.55

1.71

.90

.079

.0125

.063

1.10

8

Control

7.84

1.79

.88

.085

.0063

.070

1.21

9

S - 5000

7.72

1.82

.88

.082

.0086

.070

1.20

Control

7.95

1.75

.86

.091

.0063

.102

1.04

S

P2°5

Plot
2
5
4
5
6

11

MnS0.-800
4
Control

Percentage calculated on water—free basis
Ash
CaO
MgO FegOg
M113O4
S
P2O5

Average composition in per cent
Treatment

Ash

CaO

MgO

Control

7.47

1.77

.95

.085

.0066

.071

1.09

MnS04

8.91

1.78

1.00

.057

.0074

.080

1.09

MnS04 + S

7.55

1.94

.95

.055

.0085

.064

.96

S

7.52

1.76

.89

.080

.0104

.066

1.15

(a)

pounds per acre.

Fe2°3 ^n3^4

54.
Discussion of Data.
The data showed very few differences due to treatments;
however, there were some interesting points which were con­
sidered important.

As already mentioned, the plant material

which was analyzed in this study was taken at the time when
the crop was harvested and consequently may not show as
great differences as might have been found during the grow­
ing season.

The data indicated that the treatments had

little or no effect upon the per cent of water, ash, calcium,
magnesium, sodium, and sulphur contents of the plants.

In

the majority of the cases, the application of manganese sul­
fate increased the manganese content of the plant, especially
in the case of the combination of manganese sulfate and
sulphur treatment.
The most outstanding result of the analysis is that
given by iron.

The crops which developed the chlorotic

condition had a high iron content unless they were treated
with either manganese sulfate or sulphur.

The non-

responsive crops, sugar beets and strawberries, failed to
show similar results and, in the case of sugar beets, the
lowest iron content was in the controls.

These data would

be indicative of some influence of the manganese upon the
iron in the plant.
Although these tables showed instances of apparent
effect upon the phosphorous and potassium content of the
crop by the soil treatment, later work failed to substanti­
ate these results.

Table 10 showed a substantial increase

55.
in potassium content with the application of manganese
sulfate, hut the contrary appeared to he the result with
the application of sulphur.

Later work has never produced

similar results; however, this table reflects to some ex­
tent the data obtained by successive water extractions in
graph 3.

Mineral Composition of Hormal and Chlorotic Plants at Differ­
ent Stages of Growth.
Due to the interesting data on iron and manganese con­
tent of the mature crops, an investigation into the mineral
composition of normal and chlorotic plants at different
stages of growth was made during 1938.

Hormal plants were

produced on those plots receiving either manganese sulfate,
sulphur or the combination of the two.

During the spring

of 1938, additional treatments were applied to the treated
plots, as follows:

plots 1 and 3 received a 200-pound-per-

acre application of manganese sulfate, and plots 4 and 6,
400 pounds of the same salt; plots 3, 6, 7, 9, and 10 each
received an additional treatment of a 500-pound-per-acre
application of sulphur.

In addition to these treatments,

all plots received an 800-pound-per-acre application of a
0-8-24 fertilizer and 350 pounds per acre of common salt
(NaCl).

Analytical samples were collected at various times

during the season from the responsive crops, barley, rasp­
berries, peas, alsike clover and the non-responsive crops,
sweet clover and peppermint.

Samples of peas were taken on June 23, 1938, for a
study of the mineral composition of the chlorotic tissue,
as compared to that of the normal tissue.

The plants from

plots 2, 5, and 8 were chlorotic, inasmuch as the lower
leaves and stems were yellow and only a few upper leaves
and stems had the normal green color.

The plants on the

plots 1 and 4 receiving manganese sulfate; 7 and 9, sulphur
and 3 and 6, the combination of sulphur and manganese sul­
fate had a normal green color throughout the vegetative
growth.
The plants which were chlorotic were divided into
chlorotic tissue and normal tissue.

The plants from the

plots producing the normal plants were divided in a manner
so as to give comparable parts to those of the chlorotic
plants.

This material was analyzed for ash, calcium,

manganese, potassium, iron and nitrogen, and the results
are shown in the following table;

57.

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Table 17,

showing averages of preceding data

Dry
Treatment________ Matter

Ash

RaO

Fe203

BfasQ4

KgO

N

.651
.610

.815
.480

Control

upper
lower

11*59
14*20

12.34
15.75

.184 .00384 .00011
.553 .00883 .00013

MnSO„4

upper
lower

15*95
14.61

10.95
12.90

.285
.579

.00497
.00441

.00029
.00029

.601
.563

.688
.479

BifnSO^ "f S upper
lower

15*61
14.55

12*12
15*05

.272
.664

.00343
.00471

.00036
.00054

.596
.518

.721
.474

upper
lower

15*66
15*95

11.16
15.53

.238
.598

.00330
.00496

.00015
.00013

.598
.551

.770
.467

s

59.

Peas are a crop in which the chlorotic condition
develops

in the lower leaves first and progressively

approaches the growing tip.

There is always a clear-cut

boundary between the growing parts which are normal and
the older tissue which is chlorotic.

For this reason, the

chemical data should give very clear-cut evidence as to
the chemical factors that may be involved.
standing point is that of iron.

Again the out­

Hormal tissue has marked­

ly less iron than the chlorotic tissue which has two and a
half times as much as does the normal tissue.

These data

clearly and conclusively indicate that the effects of either
the sulphur or the manganese sulfate treatments are one of
activation of the iron.
The calcium content of the plant appears to increase
with age of the plant tissue.

The active growing portions

of the plant have about half as much calcium as is found in
the lower portion of the plant.

The treatments do not

appear to have any influence upon the calcium content.
Potassium has an opposite relationship in respect to the age
of tissue, with the newer tissue having the higher content.
Treatments do not have any influence upon its content in the
plants.

Wo important difference appears in either the con­

tent of dry matter or nitrogen, while the ash content is
higher in the case of the chlorotic tissue.
Treatments have a marked effect upon the manganese
content of the plants.

The lower portion of those on the

sulphured plots show an increase of 2 0 per cent over the

60.
controls; those receiving manganese sulfate, 342 per cent
increase; while those receiving the combination, show a
374 per cent increase in comparable tissue.
These data suggest a similarity to the work of
Oserkowsky (58) in which he showed a higher total iron
content in lime—induced chlorotic tissue as compared to
normal tissue.

Data of Marsh and Shive (49) revealed a

similar condition existing in comparable tissue of soybeans
in which chlorosis is caused by lowering the hydrogen ion
concentration in nutrient solutions.

A study was made as

to factors which might have caused such observations.

The

pH of the cell sap of the chlorotic and normal tissue of
canning peas reported in Table 16 was determined by a
Beckman potentiometer.

The chLorotic tissue in every case

had a higher pH, varying from 6.40 to 6.65, while the normal
tissue varied from 6.15 to 6.30.

Hoffer (33) associated

the higher pH with deposition of iron in nodes of corn.
Allyn (2) is of the opinion that a high soluble calcium con­
tent in the soil may disturb the metabolism of iron within
the plant.

Since these reports indicated some kind of a

break down in the metabolism of iron within the plant, it
was considered advisable to establish where the iron was
concentrated in the plant tissue.

Using the technique of

Kliman (43) for staining ferric and ferrous iron in plant
tissue, some very interesting results were obtained in the
tissue of canning peas and soybeans*

The ferric iron present

was found deposited heavily in the veins of the chlorotic

61.
leaves.

The normal leaves showed no ferric iron in the

veinal tissue and a mere trace in the rest of the leaf.
The ferrous iron in the normal leaves was uniformly dis­
tributed throughout the leaf, while, in the chlorotic leaf,
the stains indicated considerable ferrous iron but an
extremely uneven distribution with a tendency to become
concentrated near the veinal tissue and adjacent to the
ferric iron.

These results would indicate that the benefi­

cial results of manganese and of sulphur are due to their
effect on the state of oxidation of the iron in the plant.

Seasonal Trend of Mineral Content of Barley plants
Taken at Ten-day Intervals.
Barley was seeded on a portion of the manganese-sulphur
series for the purpose of studying the mineral composition
of the crop at different stages of its growth.

It was seed­

ed on May 5, 1938, and the first samples for analysis were
taken on June 13.

Thereafter it was sampled at ten-day

intervals throughout the season.

The final sampling was

made on August 2, at which time the barley was ripe.

It

should be noted here that at the mature stage most of the
plots were heavily infested with stem rust which was very
severe in the cases of plots 2, 5, 8, 11 and 12, so much
so that they failed to mature properly.

Plots 3, 6, 7 and

9 matured normally.
Early in the growth of the barley, the plants on plots
2

5, 8, 11 and 12 became chlorotic, the chlorosis closely

62.
resembling that produced by nitrogen starvation.

As growth

progressed, the yellowing gradually became more pronounced
up to the hot weather of July, at which time a few plants
turned dark green on the control plots and especially on
plot 5 on which dark green plants became very numerous.

At

heading time, July 20, all plants on all of these plots, with
the exception of 12, were dark green and on this plot a week
later they also became green.
In the first five samplings, the area taken for a
sample was determined by the size of the sample needed for
analysis.

The August second sampling was taken with the

idea of obtaining a comparative yield on each plot.

An

area six feet long and two feet in width was taken.

Calcium,

manganese, and iron were determined in these samples by
methods previously described.
The highest grain yield, (Table 18), was obtained on
plots receiving only sulphur.

The three plots showed a

600 per cent increase in yield over the controls.

The

manganese sulfate and sulphur combinations showed a 568
per cent increase and the plots receiving only manganese
sulfate showed a 386 per cent increase.

The yields of

straw have approximately the same relationship.
The date given in Table 19 on the dry matter content
indicate that on all dates before maturity, the content
of the several constituents is higher on all treated plots.
At maturity none of the treatments varied from the controls
to any significant degree in percentage of dry matter.

Table 18*

Plot

showing yields of barley in 1937 on
Mn-S Series of College Muck Plots taken from
from one row comprising an area of 12
square feet.

Treatment-1938

(a)

1

MnS04 - 200

2

Control

3

Number Wt. of
of
heads —
heads
grams

Yield of straw
per area —
grams

127

172

174

26

19

68

MnS04 - 200
S - 1500

196

259

246

4

MnS04 - 400

151

211

225

5

Control

73

65

91

6

MnS04 - 400
S - 1500

207

307

276

7

S -

249

356

317

8

Control

57

61

96

9

S -

3500

229

301

293

10

S -

2500

208

229

238

11

Control

74

58

108

12

CuS04 - 50

2

1

13

1500

(a) pounds per acre.

Table 19,

showing the effects of manganese and sulphur, applied
on an alkaline muck on the seasonal trend of dry matter
content of barley plants grown thereon in 1938.

_____ Percentage calculated on oven dry basis_____
Plot Treatment
June 15 June 25 July 3 July 15 July 25 Aug. 2
(a)
1 MnS04 - 200
11.99
12.35
16.83
23.89
26.60
89.84
2

Control

11.04

10.75

11.95

17.01

21.71

90.48

3 MnS04 - 200
S - 1500

12.46

13.52

14.36

25.06

27.78

90.12

4 MnS04 - 400

12.29

12.12

14.16

23.60

25.22

89.70

5

Control

10.86

9.95

10.89

19.55

23.15

89.55

6

MnS04 - 400
S - 1500

11.67

11.72

12.52

25.63

26.73

90.20

S -

11.89

10.72

13.60

23.55

25.96

89.75

8 Control

10.57

9.80

10.36

17.39

19.89

89.80

9

11.24

12.12

14.31

25.44

24.45

89.88

7

1500

S - 3500

Averages
June 13 June 25 July 3 July 13 July 23

Aug 2

Control

10.82

10.17

11.07

17.98

21.58

89.94

MnS04

12.14

12.23

15.49

23.74

25.91

89.77

12.06

12.62

13.44

25.54

27.25

90.16

11.56

11.42

13.95

24.49

25.20

89.81

MhS04
S

S

(a) pounds per acre.

The chemical data given in Tables 30, 31 and 33, show
several interesting points.

There appears to be no real

differences between the calcium contents of the barley re­
sulting from the various treatments.

In the case of the

manganese, all the treatments produced higher^Jp: cent ages
than did the controls.

One of the highest values for manga­

nese in crops grown on the sulphur plots was obtained in the
barley.

These results gave the strongest evidence offered

by all crops analyzed in support of the theory that the soil
acidity increased the solubility of the manganese.

The un­

usual data that appeared in the August 3 analysis is hard to
explain.

The control plots gave the highest manganese con­

tent of the crop at that date for which no explanation can
be given.

The iron data are in keeping with those that had

been obtained in other crops.

From June 13 to July 13 the

iron content is in inverse order to the yields, with the
chlorotic plants from control plots having the highest values.
On July 30, when the plots had become green, no difference
existed between the iron content of these plants and of those
on the treated plots.

This relationship definitely estab­

lishes the iron factor as the determining one in chlorosis
on alkaline soil.

66.
Table 20,

Plot

showing the effects of manganese and sulphur, applied on an
alkaline muck, on the seasonal trend of the CaO content of
barley plants

Treatment
Lbs. per
acre

June
13
*

June
23
1

July
3

.

..

July
13
%

Straw
July
Aug.
23
2
%
%

Heads
July
Aug.
23
2
&

Calculated on over-dry basis
1

Mc S04_200

1.112

1.295 1.118

.945

1.162

1.186

.371

.189

2

Control

1.041

1.309 1.116

1.414

1.302

1.086

.295

.315

5

MnSO -200
S - 1500

1.119

1.176 1.044

.903

1.029

.997

.252

.210

4

MnS04-400

1.099

1.155 1.160

.938 1.190

1.044

.329

.189

5

Control

1.067

1.491 1.303

1.085

1.141

1.052

.315

.283

6

MnS04-400
S - 1500

1.177

1.253 1.022

.707

1.064

1.013

.224

.194

7

S - 1500

1.132

1.281 1.141

.833

1.022

1.034

.273

.200

8

Control

1.161

1.491 1.406

1.309

1.330

.934

.301

.283

9

S - 3500

1.293

1.260 1.047

.840 1.029

1.002

.238

.194

Averages on over-dry basis
2,5,8 Control

1.090

1.430 1.275

1.260

1.258

1.024

.304

.294

1,4

Mn

1.105

1.225 1.159

.941

1.176

1.115

.349

.189

3,6

Mn + S

1.148

1.214 1.033

.805

1.046

1.005

.238

.202

7,9

Sulphur

1.212

1.270 1.094

.836

1.025

1.018

.255

.197

Averages calculated on field basis
2,5,8 Control

.118

.145

.140

.223

.269

.925

.088

.270

1,4

Mn

.154

.163

.175

.219

.305

1.004

.123

.172

3,6

Mn + S

.138

.153

.158

.199

.285

.905

.091

.184

7,9

Sulphur

.140

.144

.152

.201

.258

.917

.091

.179

67.
Table 21, showing the effects of manganese and sulphur on the seasonal,
trend of the Mn^O^ content of barley plants.

Pint Trfifl+.monf Tnwo
15
%

June
25

July
5

July
15
%

July
25
%

Straw
Aug. July
2
25
%
%

Heads
Aug.
%

Calculated on oven-dry basis
1

MnS04_200 .00217 .00508 .00277 .00198 .00298 .00246 ►00187 .00505

2

Control

3

MnS04_200
S - 1500 .00556 .00585 .00462 .005^7 ^fi409 .00502 .00505 .00454

4

MnS04_4Q0 *00584 .00454 .00421 .00502 TO0454 .00502 .00259 .00554

5

Control

6

HnS04-400
S - 1500 .00615 .00661 .00661 .00555 •00554 .00555

.00159 .00251 .00186 .00150 .00206 .01085 .00065 .01532

.00126 .00171 .00257 .00168 .00255 .00267 .00125 .01194
00288 .00579

7

S - 1500

.00251 .00257 .00289 .00212 .00264 .00246 ,00264 .00385

8

Control

.00126 .00128 .00169 .00121 .00271 .00296 ,00065 .01332

9

S - 5500

.00264 .00519 .00584 .00222 .00547 *00215

00259 .00454

Averages on oven-dry basis
Control

.00150 .00177 .00204 .00146 .00244 .00549 .00084 .01286

Mn

.00500 .00371 .00349 .00250 .00366 .00274 .00213 .00329

Mn + S

.00485 .00523 .00561 .00451 .00471 .00428 .00296 .00506

Sulphur

.00247 .00288 .00556 .00217 .00505 .00230 .00251 .00409
Averages on field basis

Control

.00014 .00018 .00023 .00026 .00052 .00497 .00025

Mn

.00037 .00048 .00053 .00058 .00094 .00247 .00075

Mn

S

Sulphur

.00058 .00065 .00085 .00112 .00128 .00587 .00116
.00028 .00053 .00047 .00052 .00077 .00208 .00091

68.
Table 22,

showing the effects of manganese and sulphur, applied on an
alkaline muck, on the seasonal trend of the FepOK content of
barley plants grown thereon in 1958.
°

Plot Treatment

June
15
________

June
23
%

July
3
%

July
13
%

Straw
July
Aug.
23
2
$
Jo
%

Heads
July
Aug.
23
2
%
%

Calculated on oven- dry basis
1

MnS04-200

.0252

.0204 .0177

.0148 .0155

.0194

.0101

.0117

2

Control

.0574

.0383 .0268

.0242 .0148

.0226

.0114

.0058

5

MnSO _£00
S - 1500

.0236

.0204 .0191

.0124 .0128

.0141

.0117

.0094

4

MnS04_4QQ

.0225

.0255 .0210

.0114 .0160

*0166

.0088

.0091

5

Control

.0288

.0396 .0288

.0177 .0167

.0217

.0068

.0094

6

MnS04-400

.0227

.0204 .0191

.0106 .0153

.0148

.0127

.0117

7

S - 1500

.0232

.0223 .0171

.0106 .0114

.0180

.0117

.0127

8

Control

.0337

.0355 .0294

.0191 .0160

.0130

►0076

.0127

9

S - 2500

.0236

.0210 .0160

.0114 .0153

.0173

.0104

.0105

Averages on oven-dry basis
Control

.0333

.0571 .0285

.0203 .0158

.0208

.0086

.0093

Mn

.0228

.0229 .0193

.0131 .0147

.0180

.0094

.0104

Mn + S

.0231

.0204 .0191

.0115 .0140

.0144

.0122

.0105

Sulphur

.0234

.0216 .0165

.0110 .0133

.0176

.0110

.0116

Averages on field basis
Control

.00560 .00357 .00314 .00358 .00341 .00187 .00251

Mn

.00277 .00301 .00297 .00305 .00381 .00162

Mn t S

.00279 .00257 .00278 .00285 .00581 .00151 .00471

Sulphur

.00270 .00248 .00252 .00265 .00534 .00158 ,00590

00530

69*
Seasonal Trend of Mineral Composition of Legumes Sampled at
T5-day "Intervals,
The following legumes, alfalfa, alsike clover, and
sweet clover, were seeded in July, 1938, on the ManganeseSulphur Series of the College Muck Plots.

These three

legumes represent three degrees of responsiveness to
Manganese fertilizers.

Alfalfa is a very responsive crop;

alsike clover, mildly responsive; and sweet clover, nonresponsive.

The first samples were taken August 27, 1938,

and samples at approximately fifteen-day intervals there­
after.

In the case of alfalfa, very marked differences due

to treatment could he observed.

CaO, MgO, MngO^ and Fe^O^

were determined in the samples collected.
In Tables 23, 24 and 25, is shown the dry matter
content of these three legumes at each date of sampling.
For the most part, no differences could be recognized as
resulting from treatments.

In most cases, the plants from

the controls were slightly higher in dry matter than were
those from the treated plots.

Table 25,

Plot

Mn - S series legume experiment showing per cent of dry
matter content of alfalfa sampled at approximately 15—day
periods during 1958.

Treatment
lbs. per acre

Aug. 27

£ept. 15

Oct. 1

Oct. 15

h'ov. 1

1

MnS04_200

25.99

22.66

50.01

59.56

58.98

2

Control

24.40

25.67

50.25

42.05

58.59

5

MnS04_200
£ - 1500

25.55

22.68

28.50

51.72

56.29

4

MnS04_4Q0

22.19

21.57

50.95

29.52

57.74

5

Control

24.12

22.52

28.59

52 *18

55.58

6

MnSO -400
8 - 1500

22.98

22.12

27.78

55.64

38.15

7

£ - 1500

25.24

22.41

27.78

55.87

58.50

8

Control

24.68

22.85

28.99

28.04

54.51

9

£ - 5500

25.55

22.91

28.86

52.65

54.79

10

£ - 2500

24.95

21.55

27.91

52.86

55.94

11

Control

25.92

22.42

50.14

54.81

56.88

12

Cu£04-50

24.59

21.09

27.15

29.88

57.19

Averages
Control

24.78

22.86

29.44

37.15

56.29

Mn

23.09

22.12

50.47

54.54

58.36

Mn + £

24.15

22.40

28.04

55.68

57.21

Sulphur

25.24

22.22

28.18

53.12

36.54

Ratiosi to control
Mn
Control
Mn - S
Control
Sulphur
Control

.95

.97

1.05

.92

1.06

.97

.98

.95

.91

1.03

1.02

.97

.96

.90

1.00

71.
Table 24,

Mn—S series legume experiment shovd_ng per cent of* dry matter
content of* alsike clover sampled at approximately 15—day
periods during 1958.

Plot Treatment
lbs. oer acre

. ..Aug. 27

Sent. 15

Oct. 1

Oct. 15

Nov. 1

1

MnS04-200

20.55

19.05

22.28

26.91

35.80

2

Control

20.67

18.02

22.55

27.56

56.25

5

MnS04_200
S - 1500

22.11

18.52

25.88

25.65

36.14

4

MnS04-400

22.67

17.57

23.25

26.98

39.52

5

Control

24.42

18.55

24.26

28.83

38.68

6

MnS04_400
S - 1500

21.02

19.42

24.95

27.56

40.05

7

S - 1500

24.76

19.25

25.68

26.26

38.07

8

Control

25.90

17.76

23.94

25.82

52.97

9

S - 2500

21.82

18.12

25.59

28.95

58.00

10

S - 2500

22.08

18.76

21.12

26.79

34.71

11

Control

25.58

18.28

22.54

27.28

34.94

12

CuS04-50

25.52

17.89

20.86

24.08

36.59

Averages
Control

25.14

18.10

25.22

27.37

55.70

Mn

21.51

18.51

22.76

26.94

37.66

Sulphur

22.89

18.70

22.80

27.55

56.93

Mn 4- S

21.58

18.87

24.40

26.49

38.08

Ratios to control
Mn
Control

.95

1.01

.98

.98

1.05

Mn + S
Control

.95

1.04

1.05

.97

1.07

Sulohur
Control

.99

1.05

00
CD*

1.00

1.05

72

aac

25,

Mn—S series legume experiment showing per cent- of dry matter
content of sweet clover taken at about 15-day intervals*

Treatment
Pl(

W V

V t

-JL-

X'iKJ V

•

X

1

MnS04-200

17.55

16.25

21.91

24.69

27.65

2

Control

17.05

17.24

25.05

25.26

28.98

3

S - 1500
MnS04»2Q0

18.79

15.91

21.05

24.05

29.52

4

MnS04~400

18.82

16.54

19.94

24.41

27.86

5

Control

17.45

16.70

21.85

25.04

29.66

6

Mn£04-400
S - 1500

18.21

16.20

22.54

24.68

27.55

7

5 - 1500

18.55

16.50

22.78

25.86

28.71

8

Control

17.11

16.09

22.69

24.87

27.42

9

S - 5500

18.65

16.62

21.79

25.22

29.68

10

S - 2500

18.56

16.81

21.86

25.80

29.09

11

Gontrol

17.19-

16.05

21.96

25.28

28.55

12

CuSO -500
4

16.69

16.54

19.04

24.87

30.70

Averages
Control

17.19

16.52

22.58

25.11

28.60

Mn

18.18

16.29

20.92

24.55

27.74

Mn + £

18.50

16.05

21.68

24.35

28.33

Sulphur

18.57

16.64

22.14

24.96

29.16

Ratios to control
Mn
Control
Mn -r S
Control

1.06

.99

.95

.98

.97

1.08

.97

.97

.97

.99

1.08

1.01

.99

.99

1.02

C

Control

.

Perhaps one of the "best indications of the part played
by calcium in this problem will be seen in Tables 26 and 27,
and graphs 7 and 8, which show the effect of soil moisture
and temperature upon the CaO content of the responsive crop,
alfalfa, as compared to the non—responsive crop, sweet clover.
It is to be remembered that the greatest response of manga­
nese fertilizers occurred when soils were cold and wet.

On

September 13, 1938, these plots received a heavy rain and it
is interesting to note that the calcium content in the samples
gathered on September 15, made a very great increase in the
alfalfa; samples of sweet clover from the same date gave no
increase in calcium content*

In the samples taken October 1,

the calcium content of the alfalfa was lower than in those
taken September 15, while the sweet clover showed practically
no fluctuation.

Many investigators have reported the effect

of lime as a cause of chlorosis, yet, as far as one has been
able to observe in the data of this investigation, this was
the only instance in which calcium played a part.

It is

possible that its intake may be associated with certain cli­
matic factors.

Jones (40) reported that when the soil tempera­

ture fell below a certain minimum, 18°C., gardenias became
chlorotic.

At temperatures above 30°0. they were normal.

A

higher calcium content of the chlorotic gardenias is shown
in his analyses.

A low soil temperature would be conducive

to a higher soluble calcium level in the soil, with attendant
general lowering in concentration of all other elements.
Under conditions of this nature, a plant, in order to maintain

74.
its cell sap concentration, may take in abnormal quantities
of calcium, which would result in a higher pH of the sap.
This in turn would decrease the availability of the iron
in the plant.

A higher pH of the cell sap has been associ­

ated with a lower available iron content.

A high assimi­

lation of calcium will result in the precipitation of iron
in the cells of plants according to McGeorge (54).

If a

high temperature should prevail again, the calcium content
of the soil solution would decrease, as would also the cal­
cium content of the plant in respect to the other elements.
The result would be the disappearance of the chlorotic condi­
tion.
The manganese contents of these crops were similar to
those obtained in other crops#

The manganese sulfate and

sulphur treatments gave the highest manganese level for all
sampling dates as shown in Tables 29 and 31 and graphs 9
and 10.

Manganese sulfate treatment resulted in considerable

increase in manganese content of crops over that of plants
from control in the case of alfalfa and alsike clover and
slightly higher in the case of sweet clover.

The controls

and sulphur treatments were very similar and in some cases
the plants on the control plot were higher in manganese than
those from the sulphured plots#
The difference in manganese content in plants harvested
at different dates reveals that it dropped to some extent
after rains, with those on the plants receiving manganese
sulfate showing the greatest drop.

However, plants on the

75.
sulphur—treated plots and controls showed very little
difference "between dates, irrespective of rainfall.
Unquestionably the information given by the data in
Tables 33 and 34 and graphs 11 and IS, on the seasonal
trend of iron content is in line with the previous data*
The uniformity of the iron content in sweet clover, a nonre sponsive crop indicates that no disturbance has taken
place in its metabolism of iron.

In the case of alfalfa,

the high iron content occurred in those plants taken from
plots having the chlorotic plants, the control plots.
The seasonal trend of Fe^Og was not very variable in
the case of sweet clover; mildly variable in the case of
alsike; and in alfalfa, it appeared to be affected by pre­
vailing climatic conditions.

On September 15, after the rain,

the alfalfa plants showed very small differences in percent­
age of iron and also contained the lowest quantity in the
seasonal trend at this point.
to Fe was the greatest.

At this date, the ratio of Oa

The abruptness in the rise of the

iron in the control plants after that date establishes clear
evidence as to the accumulation of iron, due to its desposition in an unavailable form.
The most unusual and unexpected data of this study came
in the magnesium content of these plants.

It was unexpected

inasmuch as the data so far had given no indication that
magnesium might be a factor, and unusual in that there
appeared to be almost a complete opposite relationship be­
tween the non—responsive plants and the responsive plants.

The magnesium content correlates very well with the iron
content in that the chlorotic plants contain the larger
quantities*

There was very little difference between the

effects of the sulphur, manganese sulfate, and the combin­
ation treatments in the case of the alfalfa*

A comparison

of the data for alfalfa and sweet clover shows that the
plants from the control plots of the sweet clover had the
least magnesium and the order of the magnesium contents
of the sweet clover from the various treatments were in
reverse order to that of alfalfa.

Haas (28), working with

walnut yellows, found that the diseased leaves were higher
in ash, magnesium, and iron which would be very comparable
to the results obtained in these analyses*

Analysis of

peppermint plants gave data very similar in all respects
to that for the non-responsive crop, sweet clover, while
data obtained from raspberry leaves were comparable to
those for barley and alfalfa*

77 .
Table 26,

Plot

Showing content of CaO in alfalfa plants sampled at 15-day
intervals on Mn-S series

Treatment
lbs * per acre

Percentage calculated on oven-dry basis
Aug. 27

Sept. 15

Oct. 1

Oct. 15

Nov. 1

1

MnS04-200

1.985

2.829

1.856

2.165

2.184

2

Control

2.184

5.089

1.979

2.496

2.557

5

MnS0,-200
S - 1500

2.170

3.195

2.079

2.571

2.4-20

4

MnS04_400

2.184

£.195

2.065

2.444

2.505

5

Control

2.856

5.619

2.244

2.288

2.551

6

MnSO -400
S - 1500

2.482

5.528

1.959

2.496

2.482

7

S - 1500

2.252

5.205

1.875

2.509

2.580

8

Control

2.455

5.580

2.155

2.685

2.441

9

S - 5500

2.219

5.546

1.921

2.256

2.599

10

S - 2500

2.087

5.401

1.908

2.288

11

Control

2.177

5.078

1.862

2.496

2.505

12

CuS04-50

2.198

5.057

2.024

2.517

2.571

—

Averages
Control

2.407

5.291

2.059

2.491

2.458

Mn

2.084

5.011

1.959

2.505

2 •545

Mn + S

2.526

5.260

2.019

2.455

2.451

Sulphur

2.186

5.585

1.901

2.278

2.630

Mn
Control
Mn
S
Control
S
Control

.87

.91

.95

.92

.95

.97

.99

•
to
00

Ratio to <control

.98

1.00

.91

1.05

.92

.91

1.07

78 *
Table 27,

Plot

showing t-he content of CaO in sweet clover plants sampled
at 15-day intervals on Mn-S series

Treatment
lbs. oer acre

Aug. 27

Percentage calculated on oven-drv basis
Sent. 15
Oct. 1
Oct. 15
Nov. 1

1

MnS04-2 0 0

2 .6 1 4

2 .9 6 7

2 .5 6 1

5 .5 4 9

5 .1 9 6

2

Control

2 .9 0 5

2 .5 8 0

2.0 2 4

5 .0 8 5

2.877

5

MnSO.-sOO
S - 1500

2 .8 0 1

2 .9 5 2

2.5 4 8

5.446

3 .2 0 5

4

MnSO^-400

2 .7 0 4

2 .9 8 1

2 .5 4 8

2 .9 4 7

5 .0 6 5

5

Control

2 .7 5 9

2 .7 1 1

2 .5 9 5

5 .1 8 9

5 .5 9 7

6

MnS04_400
S - 1500

5 .0 5 0

2 .9 1 2

2 .5 0 2

5 .1 5 5

3 .1 6 1

7

S - 1500

2 .9 4 7

2 .7 7 5

2.2 2 4

5 .1 8 9

3 .2 3 1

8

Control

2 .9 1 2

2 .5 5 5

2 .1 0 2

5 .2 4 5

2 .9 9 5

9

S - 5500

2 .9 4 7

5 .0 0 9

2 .5 8 7

5 .1 9 6

5 .3 2 8

10

S - 2500

2 .7 5 9

2 .7 7 5

2 .5 6 0

5 .0 5 2

2 .9 7 5

11

Control

2 .9 6 7

2 .5 5 7

2 .2 9 6

5 .1 2 0

5.2 6 5

12

CUSO4-5O

2 .8 5 7

2 .5 5 7

2 .2 7 6

2 .9 4 7

2 .7 9 4

Averages
Control

2.886

2.595

2.204

3.160

3.133

Mn

2.659

2.974

2.554

5.148

3.130

Mn t S

2.915

2.922

2.525

3.500

3.182

Sulphur

2.878

2.852

2.590

5.146

5.178

Ratio to control
Mn
Control
Mn .+ S
Control
S
Control

.92

1.15

1.16

1.00

1.00

1.01

1.15

1.15

1.04

1.02

1.00

1.10

1.08

1.00

1.01

79.
table 28,

Plot

showing the content of CaO in alsike clover plants sampled
at 15-day intervals*

Treatment
lbs* oer acre

Aue. 27

Percentage calculated oh oven-dry basis
Sent. 15
Oct. 1
Oct. 15
Nov. 1
2.400
1.820
2.288
5.120

1

MnS04-200

2.592

2

Control

2.427

2.400

1.755

2.519

2.790

5

MnSO -200
S - 1500

2.794

2.554

2.655

2.850

5.654

4

MnS04-400

2*415

2.290

2.273

2.798

2.919

5

Control

2.974

2.684

2.118

2.725

5.231

6

M11SO4-4OO
S - 1500

2.557

2.560

1.869

2.496

2.865

7

S - 1500

2.808

2.787

1.791

2.267

2.628

a

Control

2.884

2.231

1.953

2.278

2.628

9

S—5500

2.670

2.341

1.753

2.586

2.836

10

S

2*556

2.561

1.850

2.150

2.551

11

Control

2.850

2.057

2.031

2.184

2.461

12

CuS04-50

2.926

2.141

1.891

2.592

2.961

- £500

Averages
Control

2.784

2.293

1.959

2.376

2.777

Mn

2.402

2.545

2.041

2.545

5.019

Mn ^ S

2.575

2.457

2.052

2.673

5.258

Sulphur

2.605

2.563

1.785

2.354

2.665

Ratio to control
Mn
Control
Mn + S
Control
S
Control

.86

1 .0 2

1.04

1.07

1.09

.92

1.07

1.05

1.13

1.17

.94

1.12

.91

.98

.96

Figure 7, showing CaO content in sweet clover sampled
at 15-day intervals.

Mn
Ok
Q

Oct. 1

Aug. 27

Oct. 15

Hov. 1

Figure 8, showing CaO content in alfalfa sampled
at 15-day intervals.
Mn
Ok
— = Mn + S
/^

A u g . 27

Sept. 15

Oct. 1

Oct. 15

81.
Table 29,

Plot

showing the content of Mn„0. in alfalfa plants sameled at
15-day intervals on Mn-S series

Treatment
lbs* ner acre

Percentage calculated on oven-drvbasis
Aug. 27
Sent. 15
Oct. 1
Oct. 15
Nov ■ 1

1

MnS04-2 0 0

*00410

.00507

.00433

.00334

.00381

2

Control

*00222

.00242

.00173

.00247

.00216

3

MnS04_200
S - 1500

.00632

.00455

.00547

.00412

.00569

4

MnS04-4 0 0

.00693

.00541

.00364

.00511

.00541

5

Control

.00288

.00307

.00277

.00394

.00465

6

MnSO -4 0 0
S - 1500

.01304

.00724

.00992

.00140

.01082

7

S - 1500

.00258

.00277

.00364

.00284

.00374

8

Control

.00199

.00273

.00277

.00299

.00547

9

S - 3500

.00305

.00242

.00301

.00288

.00260

10

S - 2500

.00277

.00260

.00204

.00256

.00260

11

Control

.00199

.00260

.00173

.00216

.00260

12

CuS °4_50

.00231

.00216

.00192

.00225

.00211

.00225

.00289

.00522

.00423

.00461

.00726

.00826

.00276

.00296

Averages
Control

.00228

.00271

Mn

.00502

.00425

Mn + S

.00968

.00590

Sulphur

.00280

.00260

.00599
.00670
.00290

Ratio to control
1 .7 7

1 .4 6

1 .4 3

00
iH•

Mn
Control
■ Mn t 5
Control
S
Control

2 .9 7

2 .5 1

2 .5 6

.9 6

1 .2 9

.95

.93

2 .4 3

1 .5 7

4 .2 5

2

1 .2 3

82

SO,

showing Mrig04 content of alsike clover plants sampled at 15—day
intervals on Mn-S series

Treatment

Percentage calculated on oven-dry basis
v

m

1

MnS04_200

.00485

.00585

.00420

.00478

.00513

2

Control

.00185

.00182

.00251

.00277

.00279

5

MnSG4_200
S - 1500

.00528

.00496

.00577

.00659

.00728

4

MnS04_400

.00624

.00461

.00527

.00572

.00680

5

Control

.00225

.00211

.00547

.00554

.00347

6

MnS04_4oo
S - 1500

.00747

.00862

.00759

.00818

.00902

7

S - 1500

.00255

.00298

.00251

.00329

.00388

8

Control

.00218

.00201

.00547

.00395

.00515

9

S - 5500

.00277

.00241

.00255

.00529

.00312

10

S - 2500

.00260

.00274

.00208

.00347

.00578

11

Control

.00194

.00248

.00208

.00364

.00378

12

Cu S04_5q

.00218

.002.16

.00251

.00277

.00347

Averages
Control

.00205

.00210

. 00285

.00347

.00379

Mn

.00555

.00425

.00475

.00525

.00596

Mn f S

.00657

.00679

.00658

.00739

.00815

Sulphur

.00265

.00271

.00225

.00535

.00559

Ratio to control
Mn
Control

2.70

2.01

1.67

1.51

1.57

Mnt S
Control

2.10

5.22

2.52

2.12

2.15

S
Control

1.28

1.29

.79

.96

.95

83.

Tal

Pl<

y

showing Mng04 content of sweet clover plants taken at 15-day
intervals on Mn—S series

Treatment
lbs. oer acre

Percentage calculated on oven—dry basis_______
Aug* 27
Sept. 15
Oct. 1
Oct. 15
Nov. 1

1

MnS04_200

.00216

.00250

.00182

.00242

.00501

2

Control

.00199

.00177

.00175

.00190

.00232

5

MnSO -200
S - 1500

.00427

.00225

.00256

.00277

.00515

4

MnSQ4_400

.00555

.00248

.00260

.00270

.00547

5

Control

.00249

.00251

.00216

.00356

.00322

6

MnSO -400
S - 4 1500

.00604

.00461

.00485

.00554

.00694

7

S - 1500

.00260

.00208

.00164

.00246

.00256

8

Control

.00160

.00187

.00156

.00211

.00255

9

S - 5500

.00199

.00177

.00175

.00208

.00232

10

S - 2500

.00205

.00209

.00173

.00252

.00232

11

Control

.00177

.00209

.00157

.00190

.00260

12

CuS04_50

.00194

.00210

.00189

.00197

.00246

Average
Control

.00197

.00201

.00176

.00232

.00267

Mn

.00274

.00239

.00221

.00256

.00524

Mn + S

.00516

.00342

.00361

.00406

.00504

Sulphur

.00222

.00198

.00170

.00229

.00253

Ratio to control
Mn
Control

1.39

1.19

1.26

1.10

1.21

Mn 4- S
Control

2.62

1.70

2.05

1.75

1.89

Control

1.13

.99

.97

.98

.87

Figure 9, showing M*04 content in alfalfa sampled
at 15-day intervals.

.008$

006$

Mn
♦

*002$

Aug. 27

Sept. 15

Oct. 1

Oct. 15

ft & i x e ,
at 7® iOj
aies<% Q°^t
e^t

■*2j

siree*
ov,e?

86.
■Table 52,

Plot

showing the content of Fe205 in alfalfa plants sampled at
15 day intervals on Mn—S series

Treatment
lbs* oer acre

Percentage calculated on oven—drv basis
Aug. 27
Sent. 15 Oct. 1
Oct. 15

Nov. 1

1

MnS04_g00

.04730

.03362

.03788

.04812

.04738

2

Control

.8188

.04037

.04018

.06935

.07844

5

MnS04_goO
S - 1500

.06443

.03622

.03458

.05985

.04537

4

MnS04_400

.05321

.05725

.02899

.04450

.05628

5

Control

.08633

.04242

.05066 v

.04865

.05924

6

MnS04_400
S - 1500

.06503

.03002

.02282

.04140

.05352

7

S - 1500

.07390

.04037

.05071

.04140

.05430

8

Control

.09164

.05520

.04018

.06210

. 04936

9

S - 5500

.07330

.03622

.05114

.04760

.04787

10

S - 2500

.06739

.05105

.03229

.04450

.04887

11

Control

.08558

.04057

.04018

.05590

.05574

12

CuS04 _ 50

.08070

.05777

.04548

.05072

.05054

Averages
Control

.08636

.03959

.04280

.05900

.06159

Mn

.05025

.03543

.03343

.04631

.05138

Mn T S

.06473

.03512

.02870

.04062

.04934

Sulphur

.07153

.03588

.03138

.04450

.05035

.58

.89

.78

.78

.75

.84

.67

.69

•

S
Control

.83

.91

.73

.75

.82

CD

Mn
Control
Mn r S
Control

•
00
03

Ratio to control

87

33,

showing the content of Fe^Og in alsike clover plants sampled
at 15-day intervals on Mn—S series

Treatment

Percentage calculated on oven—d r y basis
V/v

-4.

»*' W *

4.V

V •

J

1

MnS0^-200

.03547

.01837

.02023

.02608

.03603

2

Control

.03793

.01952

.02511

.02980

.03751

3

MnS04-200
S
1500

.03448

.01569

.01937

.02442

.03702

.03253

.02009

.02025

.02608

.03751

.03942

.02009

.02224

.03146

.04147

MnSO. -400
S - 1500

.03448

.01837

.01937

.02608

.03949

7

S

.03448

.01837

.02023

.03064

.03949

8

Control

.04779

.02124

.02152

.03230

.03553

9

S - 3500

.03547

.01952

.01995

.03064

.03949

10

S - 2500

.03841

.01908

.01995

.02732

.03751

11

Control

.03547

.02009

.02267

.02816

.03849

12

CuS0A - 50

.04926

.02124

.02267

.02650

.05034

-

4
5
6

MnSO. -400
4
Control

-

1500

Averages
Control

.04015

.02023

.02288

.03057

.03825

Mn

.03400

•01923

.02023

.02608

.03677

In t 8

,03448

•01700

.01937

.02525

.03825

Sulphur

,03621

.01899

.02004

.02953

.03883

Ratio to control
Mn
Control
Mn -t- S
Control
S
Control

.85

.95

.83

.85

.96

.86

.84

.85

.83

1.00

.90

.94

.88

.97

1.02

88.
«-4,

showing the content of Fe^Og in sweet clover plants sampled
at 15-day intervals on Mn-S series*

Treatment
lbs. Der acre

Au £. 27

Sept. 15

Oct. 1

Oct. 15

Nov. 1

1

MnS04_200

.02267

.01758

.01822

.02070

.02175

2

Control

*01970

.01656

.01822

.01864

.02270

5

MnSO.—200
S - 1500

.02167

.01758

.01607

.02070

.02571

4

MnS04-400

.02464

.01822

.01822

.01946

.02571

5

Control

.02464

.01698

.01756

.02070

.02571

6

Mn804-400
S - 1500

.02267

.01988

.01550

.01946

.02571

7

S - 1500

.02167

.01822

.01478

.02028

.02614

8

Control

.02167

.01576

.01894

.02070

.02571

9

S - 5500

.02167

.01780

.01756

.01864

.02571

10

S - 2500

.02267

.01656

.01650

.01904

.02571

11

Control

.02267

.01490

.01578

.02070

.02571

12

CuS04-50

.02565

.01614

.01794

.01988

.02468

Pl(

Averages
Control

02217

.01605

.01757

.02018

.02546

Mn

02565

.01780

.01822

.02008

.02272

Mn -t* s

02217

.01865

.01579

.02008

.02571

Sulphur

02201

.01755

.01621

.01952

.02452

Ratio to control
Mn
Control
Mn + S
Control
S
Control

1.07

1.11

1.04

1.00

.97

1.00

1.16

.90

1.00

1.01

.99

1.09

.92

.96

1.05

Figure 11, showing Fe30g content in sweet clover

02#

Aug. 27

Sept. 15

Oct. 1

Oct. 15

Fe ,0 content in alfalfa sampled
Figure 12,
12 showing Fe^Og
at 15-day intervals

06#-

02#

—

—

i

Aug. 27

,

1

n '
j

Sept• 15

Oct. 1

Oct. 15

Hov. 1

90.
.Table 35, showing the content of MgO in alfalfa plants sampled at
__________ 15-day intervals on Mn-S series

Plot

Treatment

—

1

MnS04-200

.465

.402

.457

.598

.442

2

Control

.485

.478

.480

.472

.466

5

MnSO^-200
S - 1500

.461

.407

.405

.599

.591

4

MnS04_400

.485

.411

.591

.418

.442

5

Control

.579

.452

.517

.441

.418

6

MnSO^-400
S - 1500

.519

.407

.449

.394

.452

7

S - 1500

.485

.412

.449

.390

.377

8

Control

.502

.468

.497

.498

.452

9

S - 5500

.551

.420

.455

.420

.432

10

S - 2500

.485

.458

.406

.420

.458

11

Control

.565

.458

.471

.462

.437

12

CuS0^-50

.485

.478

.485

.478

.403

Percentage calculated on oven—dry basis
<V »
L / V M U * 4»t;
ut. U • -iiWV • J.
Uw V • <1*0

Averages
Control

.552

.459

.491

.468

.445

Mn

.475

.406

.414

.408

.442

Mn + S

.490

.407

.427

.396

.411

Sulphur

.499

.425

.450

.410

.422

.87

1.00

.89

00
00
•

.84

Mn +- S
Control

.92

.89

.87

•
CD
ta

.93

S
Control

.94

.92

.87

.95

\

Mn
Control

*
00
CO

Ratio to control

91 •
'Tal

56,

Pli

Treatment
.lbs, per acre

showing the content of MgO in alsike clover plants sampled
at 16-day intervals on Mn-S series
_Percentage calculated on oven-dry basis
Ausr. 27
Sf>n+._ 1a
rm+ l
nn+ k
W V v t

1

MnS04_goo

.480

.414

.454

.481

.545

2

Control

.572

.452

.414

.482

.558

5

MnS04—200
S - 1600

.567

.414

.449

.512

.605

4

MnS04-400

.514

.414

.444

.485

.551

5

Control

.657

.452

.456

.482

.551

6

MnS04_4Q0
S - 1500

.611

.401

.470

.474

.551

7

S - 1500

.555

.421

.450

.484

.551

8

Control

.565

.452

.422

.444

.468

9

S - 5500

.570

.406

.574

.444

.495

10

S - 2500

.565

.406

.405

.466

.502

11

Control

.640

.414

.442

.460

.502

12

CuS04_ 50

.554

.428

.410

.478

.551

Averages
Control

.605

.427

.428

.467

.510

Mn

.497

.414

.448

.485

.557

Mn + S

.589

.407

.459

.495

.567

Sulphur

.565

.411

.409

.465

.509

Ratio to control
Mn
Control
Mn t S
Control
S
Control

.82

.97

1.05

1.04

1.05

.98

.95

1.07

1.06

1.11

*95

.96

.96

1.00

1.00

93.
37,

showing the MgO content of sweet clover plants sampled at
15-day periods on Mn-S series

Treatment
lbs, ner acre

_ Percentage calculated on oven-drv basis_______
Aug.. 27
Sept. 15
Oct. 1
Oct. 15 Nov. 1

1

MnS04_200

.707

.536

.521

.471

.471

2

Control

.635

.501

.478

.495

.471

3

MnS04-200
S - 1500

.736

.540

.534

.483

.483

4

MnS04-400

.693

.548

.553

.507

.471

5

Control

.659

.499

.505

.495

.471

6

MnS04_400

.719

.555

.507

.507

.538

S -

1500

7

S - 1500

.712

.541

.461

.471

.483

8

Control

.536

.476

.447

.464

.434

9

S - 3500

.679

.522

.492

.464

.492

10

S - 2500

.621

.518

.453

.519

.524

11

Control

.579

.462

.437

.422

.444

12

CuSO -50
4

*591

.476

.488

.434

.471

Averages
Control

.602

.484

.467

.469

.455

Mn

.700

.542

.537

.489

.471

Mn r S

.727

.547

.520

.495

.510

Sulphur

.671

.527

.469

.485

.500

Ratio to control
Mn
Control

1.16

1.12

1.15

1.04

1.03

Mn
S
Control

1.21

1.13

1.11

1.05

1.12

1.11

1.09

1.00

1.04

1.10

S
Control

Figure 13, showing MgO content in alfalfa sampled
at 15-day intervals♦

5#

Aug * 27

Oct. 1

Oct. 15

Nov . 1

Figure 14, showing MgO content in sweet clover sampled
at 15-day intervals

Mn
Ck

5<%-

♦

Aug. 27

Sept• 15

Oct. 1

Oct, 15

Nov ♦ 1

94.
FALLACY OF PRESENT THEORIES

The data thus far presented do not appear to support
the theory that manganese is an essential nutrient.

Per­

haps one would he safer to say that, if it is essential,
it also plays an accessory function in plants, which
function may he of far more importance than the part it
plays as a nutrient.

Analysis of plants show that there

is a great variability in the manganese content in plants.
The variation between the quantities of manganese present
in two samples of a plant species obtained from different
sources may easily be tenfold.

This situation may occur

under one set of conditions with no effect upon the meta­
bolic processes of the plant yet, under another, serious
nutritional disturbance may result.
It is the contention here that the data do not support
the nutrient theory of manganese and that it functions more
in an accessory capacity than as a required nutrient.
There are several factors which form the basis for this con­
tention, one of which is the relationship between crop yield
and manganese content of the crop.

The data concerning the

manganese content of crops presented in this thesis show
that the treatments combining manganese sulfate and sulphur
give the highest values.

In the case of manganese sulfate

alone, it is high as compared to the controls.

Little

difference exists between the controls and the sulphur treat­
ments; in some crops the sulphur treatment is slightly higher

95.

and in others, it is slightly lower.

Crop yields do not

follow the manganese content of the crop since the sulphur
and the combination of sulphur and manganese sulfate give
consistently the highest yields and also superior quality.
Even the 400—pound application of manganese sulfate appears
to be on the average only two—thirds as efficient in its
effect on yields as are the above treatments.

If manganese

were the only limiting factor, there would be no reason for
this inconsistency and, for this reason, the function of
either manganese or sulphur must be an influence upon the
other factors of plant nutrition.
Further support of this theory is given in some of
Harmer*s unpublished data (31) which showed increasing yields
with increasing amounts of manganese sulfate applied up to
3,000 pounds per acre.

His applications were 100, 250, 500,

1,000, 2,000 and 3,000 pounds per acre and onions were grown
as a test crop.

The crops were grown during the spring and

early summer season.

From the other analytical work it is

known that, in all onions receiving more than a 250-poundper-acre application, the manganese content would be high
and therefore the increase in yield resulting from applicat­
ion above this quantity would be suggestive of the possibility
of an intensity factor.
Peculiar responses of several crops were observed in
the plant growth during the growing season, such as chlorotic
plants becoming normal during period of hot weather and dry
soil conditions.

When the jars of soil, used in the greenhouse

experiment reported above were sown to spinach in August,
they produced a crop in which the controls outyielded the
treated jars by three—fold.

Soybeans grown in the green­

house during August on the same manganese—deficient muck
were chlorotic on those jars receiving manganese salts,
while such treatments as copper sulfate and ferric sulfate
produced normal plants.

Later in the fall very remarkable

growth was produced by spinach treated with copper and zinc
separatively.

These occurred on a soil which gave very

marked response to manganese in the greenhouse during the
spring.

This situation is substantiated by results secured

by Harmer who found in past years that the results he
secured with manganese and sulphur applied in greenhouse
experiments on other alkaline mucks in early winter was no
indication as to what results would be secured on the same
soils in the spring.

Strongly acid organic soils have been

found which have only a mere trace of manganese upon total
analysis; in many cases, as mentioned before, they were often
lower than the critical amounts given by some investigators.
It may be concluded from this that the essentiality of manga­
nese under these conditions would not be as great as under
neutral or alkaline conditions.

Sometimes, indeed, one might

expect that there is no need for it under these conditions, a
thought which is suggested by Martin’s work with sugar cane
shoots (50).
It has been shown by several workers, that there is a
disturbance in the distribution of iron in the chlorotic

97.
plants.

Normal plants appeared always to have a lower iron

content than did the chlorotic plants, regardless of whether
the material is reported on oven-dry basis or on its field
condition.

Kliman (42) found that iron in plant tissue ex­

isted as the ferrous and anionic forms.

In the work report­

ed in this paper it has been shown that ferric iron is con­
centrated in veinal tissue of chlorotic leaves, a disturbance
in metabolism of iron is evident.

Oserkowsky (58) has

divided iron in the plant into two classes, available and
unavailable.

The iron which he considered available was

that portion which was soluble in half-normal hydrochloric
acid.

Using this test, he found that chlorotic tissue con­

tained less available iron than normal tissue.

Ingalls and

Shive (35) made a study of the relationship between the dis­
tribution of iron as related to hydrogen ion concentrations
of tissue fluids.

They reported that (1) Hydrogen ions con­

centration of tissue fluid corresponded to light intensity;
(2) all plants showed differences in hydrogen ion concentrat­
ion between leaves and stem; and (3) soluble and total iron
content of tissue sap was greatly influenced by its hydrogen
ion concentration.

In the metabolism of iron an equilibrium

between ferrous and ferric iron is set up, the direction of
which in normal plants is strongly towards the ferrous form.
The intensity of the directional trend can be shifted by
internal and external factors which factors, if in dominance,
may cause physiological disturbances within the plant.

If

sufficiently strong, these will cause a complete break down
of the plant tissue.

Much of* the recent work has shown support of the
theory that the function of manganese is that of an acti­
vation of iron,

Hopkins (34) has stated that the effect

of manganese lies in the activation of iron and he believes
that the iron is reduced in photosynthesis and that manga­
nese reoxidizes it*

The data presented in this paper strong­

ly supports this theory.

The iron apparently has entered

the plant and has failed to function properly in the meta­
bolic processes of plant growth due possibly to (1) an un­
favorable mineral balance; (3) an unfavorable hydrogen ion
concentration or (3) an unfavorable oxidation-reduction
system of such intensity as to disrupt the functioning of
iron, or perhaps a combination of any two or of all three.
This paper is in full agreement with Willis and Piland*s idea
that copper and manganese will produce opposite effects and
therefore both could function as activators of iron.
Since it is firmly believed that at least some of the
minor elements play a part in the functioning of iron, and
since some workers have suggested that climate may be a factor
(30), it would be interesting to study the geographical locat­
ion of the reported deficiencies of manganese, copper, iron,
boron, zinc and cobalt.

After mapping the distributions of

these locations, one immediately notes that (1) the concen­
tration of these deficiencies are in rather definite areas
such as the humid climate regions which have a considerable
precipitation during the cool part of the season, as western
Europe, England, Australia, Hawaii, South Africa, Southern

99.

France, and northern Italy, the Baltic countries, Japan,
Eastern Coast of United States and Canada, country adjacent
to the Great Lakes, Florida, Gulf Coast of Texas, California,
Oregon, Washington and several other western states; (2) in
most regions, two or more minor element deficiencies occur;
(3) many of these regions are regions in which nutritional
troubles occur in animals, such as bush sickness and anemia.
Since the indications are that, in a lime—induced
chlorosis, the metabolism of iron in the physiological pro­
cesses breaks down, and the plants actually suffer from the
lack of iron at the points where it is vitally necessary, it
is quite apparent that such a break down necessarily involves
a state of oxidation of the iron.

For that reason, any factor

which would affect this oxidation-reduction system or direct­
ion of the intensity of this system, would probably upset
this equilibrium to the extent that a physiological disturb­
ance would result.

Several factors that may affect these

intensities may act as follows^

(1) Abnormal mineral composi­

tion may upset this equilibrium and produce a chlorosis.
Several of these are known; manganese-induced chlorosis;
lime-induced chlorosis and recently Curini-Galletti (14)
describes a copper-induced chlorosis.

Camp and Euether

report a boron-induced chlorosis in Florida (12); Nemec and
Babecka (57) describe a chlorosis induced by excessive co­
balt ; chromium—induced chlorosis occurs in South
according to Van der Merve and Anderson (74).

Africa

Greenhouse

work shows that excessive amounts of zinc and titanium causes

the chlorotic condition to become more severe, a result to
be expected since they are capable of taking part in oxi­
dation—reduction reactions.

(2)

External factors such

as climate and perhaps quality of light may play a very
important part in the direction of the oxidation—reduction
intensities in metabolic processes of plants.

Little is

known of the nature of these factors and their effects.
Haas and Quayle (30) suggest the climatic factor; the work
of Jones (40) tends to show that soil temperature is
extremely important in that, when soil temperature is below
18°0., gardenias become chlorotic.

Recent work shows that

light intensity may be a very important factor, since it
produces changes in hydrogen ion concentration of the plant
sap.

Hydrogen ion concentration influences the oxidation-

reduction intensities.

These points are very important

since, first, it influences the physiological processes of
the plant which is important since the break down is within
the plant.

Second, the chemical elements under considerat­

ion are capable of resisting changes in oxidation-reduction
intensities due to changes in hydrogen ion concentration.
The recent work of Cooper, Paden and Smith (13) shows that
the quality of radiant energy and some of the oxidationreduction systems materially affect the intake of ions by
plants.

They hold further that those reactions, which occur

at approximately the same energy level and which may be
effective in bringing about certain oxidation—reduct ion

101.
reactions necessary in physiological processes, can he
arranged into groups.

They state,

11It has been observed that, on certain soils rela­
tively high in nitrates, there has been a marked
response to applications of copper and in some in­
stances to manganese. These responses are probably
related to their effect upon oxidation-reduction
reactions. The greatest growth response observed
from additions of boron to nutrient media seems to
be in complexes where iron may not be readily
available. There is a relation between magnesium
chloride and the reduction of the carbonate ion,
which is a very important reaction in photosynthesis.
Some of the important reactions such as the reduct­
ion of nitrates to nitrites, formation of hydrogen
peroxide, the free energy decrease in the formation
of cupric chloride and the change of valence of
manganese from Mn * * to Mn * * + are on approxi­
mately the same energy level ................... w,
A third theory is the one Willis proposes (81) in
which microbial activity in the soil Influences the oxidat­
ion-reduction potential of the soil by depletion of the
available oxygen.

The work of Turk (72) gives very little

support to this theory since he shows no effect by minor
elements on the carbon dioxide or ammonia production in
well decomposed organic soils.

Bacterial activity should

accelerate either one or both of these processes.

Fourth,

the effects of water logging may bring about chlorotic
plants in that it causes highly reducing conditions which
release bivalent ions, and also that it depletes the oxygen
supply in the soil.

A lack of available oxygen can be a

possible fundamental factor, however, it is not in harmony
with the fact that the deposition of iron is in the ferric
form in the plant tissue.

102.
After careful consideration, it was considered that,
fundamentally, only two of the above factors were signifi­
cant in this problem, namely, abnormal balance of the multivalent group of minor elements and second, the climatic
factors.

The combined effect of these two under natural

soil conditions, is not only possible but highly probable.
The problem now resolves itself into the following questions.
1.

Do minor elements. either alone or under specific

climatic factors» affect the oxidation-reduction reactions
of plants in such a manner as to disturb the state of oxi­
dation of iron in the plant?
2.

Do the minor elements alone or In combination affect

some of the oxidation-reduction systems in the plant?
To establish the answer to these questions, various
minor elements in systems which would produce various oxi­
dation intensities were used and a study was made as to
their effect upon the plants, paying special attention to
features of climate or light factors.
of the iron was carefully observed.

The state of oxidation
To evaluate the effect

upon oxidation-reduction systems in the plant, reduced and
oxidized ascorbic acid, and reduced and oxidized glutathione
were determined in chlorotic and normal tissue.
Experimental
This experiment was designed to determine the possi­
bility of manganese acting in the oxidation-reduction processes

of plant nutrition*

It is assumed that, if it does, another

ion capable of oxidation and reduction reactions would be
capable of functioning in its place.

Furthermore, if a

reducing agent were added to the so-called manganese-defi­
cient organic soils the chlorotic condition of the plants
would become more pronounced.

The application of manganese

sulfate was taken as the basis of the oxidizing condition.
It was assumed that the system Mn +' + ______ Mn *

* was

set up by this application and that it would give a potenti­
al of -1.5.

The system of Fe + + ______ Fe + ^ + has a

potential of -.74, therefore it should take twice as much
ferrous sulfate to produce the same oxidation-reduction
system as is produced by manganese sulfate.

Titanous chlor­

ide, potassium chromate, chromic sulfate and ferrous sulfate
were applied in this manner.

The reducing agents were not

added in quantities that would produce identical reducing
conditions.

The systems used are as follows:

Oxidation-Re duct ion Systems.
Mn * +
0r + + +
Cr04
pe+ +
g + .*
Cu +

Fe +
Zn +

+

..

.Mn
— ■-— > or
’ Cr04Fe+
. Ti

* * **
* * *+
**■ +
* + * '*’

* Ou +

Fe
! Zn *
*

An alkaline muck was selected for the experiment,
having a pH of about 7.8.

All pots were given 1,000 pounds

104.
per acre of an 0—8—34 fertilizer made up of chemically pure
chemicals.

Each treatment was replicated four times so as

to majce a statistical analysis of yield data possible.
Eight spinach plants were planted in each jar.

Titanous

chloride was found to be mildly toxic to the plants in the
early stages of their growth and it also affected the
physical condition of the soil by causing it to become
granular.

The spinach was harvested April 7, 1939, and the

following data was obtained and is shown in Table 38.
In considering these data, one is immediately impressed
by the high yield produced by manganese sulfate which is not
only significantly greater than that of the control treat­
ment but also than of all other treatments, including manga­
nese acetate, on the five per cent basis.

Significant

increases over the control were obtained in the two treat­
ments with manganese, and two with chromium while the one
with ferrous sulfate was on the border line of five per cent
significance.

The first four were also significant on the

one per cent basis.

The zinc acetate treatment produced

yields that were significantly lower than these produced on
the control and, since this treatment has the highest reduc­
ing possibilities, this is to be expected if the oxidationreduction theory holds good.
Since color of the plants indicates the cure of the
chlorotic condition, it must be considered as a part of tne
data.

For this reason the ferrous sulfate treatment should

be considered as giving positive results, especially since

Table 38*

yield? of spinach on Oxidation-Reduction Experiment
Green Weight in grams.
Block
A

Block
B

70.2

85.8

74.7

376# MnS04

142.2

350# Mn Acetate

Treatment
Check

Block Block
C
D

Ave.

Color of
plants

Signif­
icance

62.7

122.2

130.7

73.3 Greenish yellow
125.lx ]30.1 Dark green

**

126.8

102.7

107.6X 106.3 no.9 Dark green

**

450# Zn Acetate

49.2

69.7

49.6

32.4

50.2 Yellow

-H*

1200# Crg(S04)s

102.7

110.4

86.0

90.6

97.4 Green

**

1125# FeS04

82.2

97.4

91.2

87.7

89.6 Green

750# TiClg

62.7

55.2

77.8

65.2

65.2 Green

320# CuS04

88.4

88.0

56.0

52.8

250# Feg(S04)3

66.8

58.4

70.0

73.0

71.3 Greenish yellow
67.0 Yellow

111.3

103.4

81.0

630# KgCr04

**
112.0 101.9 3 green
1 greenishyellow

x Data supplied by missing value formula by W. D. Baten.
Analysis of variance
Source

D.F.

S S

Var.

39

25824.9

662.2

Replication

3

685.1

228.4

Treatment

9

21603.9

Total

Exp, Error

2400.4**

3535.9
130.9
27
F - 18.3
Difference for significance between means
Error

1% - 22.4
5% = 16.6

106.
those plants treated with ferric sulfate were yellow.

The

titanous chloride treatments gave plants which are healthy
in all respects except that the injury in their early
growth stunted their final development.

the

In Table 39

is shown yields of Mandarin soybeans on

same jars

asreported

in Table 38.

The treatments were

altered and, in some cases, the additions brought the total
application of the two to double the original.

Application

of 500 pounds per acre of a chemically pure 0-8—24 ferti­
lizer was applied before planting the seed.

The data ob­

tained in the experiment gave very conclusive evidence for
the manganese salts which were the only treatment which re­
sulted in normal plants.
In Table 40
the

is shown the yields of Cayuga soybeans on

same jarsused in the preceding table.

The jars again

received 500 pound-per-acre applications of a chemically
pure 0-8-24 fertilizer before seeding.

Very unusual

responses were obtained in these jars.

The jars receiving

manganese salts were chlorotic during most of the growing
season of this crop.

Potassium chromate grew the largest

plants of all treatments with copper sulfate, chromic sul­
fate and the iron salts following in the order named.

How­

ever, It must be mentioned that the plants receiving many
of these treatments were chlorotic at certain stages of
growth; those receiving the chromium salts became partially
chlorotic in the later stages of their growth.

Those

Table 59,

yields or Mandarin soybeans on Oxidation—
Reduction Experiment. Seed sown April 10,
1959. Harvested June 5, 1959* Green weight
in grams.

Treatment

Block A

Block B

Block C

Block D

SignifiAve• cance

Control

41.2

38.1

35.7

38.8

38.4

750# MnS04

46.2

51.5

50.1

78.0

56.4

700# Mn Acetate

50.5

65.0

81.3

56.2

63.2

450# Zn Acetate

35.6

34.0

42.5

40.7

38.2

£380# Crg(S04)5

40.2

38.5

36.8

35.8

37.8

2250# FeSO.

36.5

33.4

37.2

33.8

34.7

1500# TiClg

37.9

40.8

31.1

40.2

37.5

640# CuS04

55.9

35.8

37.3

40.2

57.3

340# Feg(S04)3

31.8

43.1

34.0

36.9

36.4

1380# KgCr04

33.4

30.0

37.5

41.8

33.2

•#*

Analysis of Variance
Source_____ D.F.

S S_____ Var.

59

5114.6

Replications

3

143.6

Treatment

9

3591.7

399.08

27

1379.3

51.09

Total

Error

S.D.

7.15

F = 7.81
Difference for significance between means 1% » 14.00
5% - 10.57

Table 40,

yields of Cayuga soybeans (grain) on OxidationReduction Experiment. Sown June 10, 1939,
harvested Aug. 20, 1939. Weight in grams.

Treatment

Block A Block B

Block C Block D

Average

Control

12.0

12.7

14.0

13.3

13.00

750# Mn S04

12.4

12.2

13.6

10.0

12.04

700# Mn Acetate

10.6

11.8

13.6

10.9

11.72

450# Zn Acetate

10.8

11.0

11.8

10.4

11.00

2380# Crg(S04)g

16.8

13.0

15 .4

13.6

14.70

2250# F©S04

16.4

12.0

14.3

15.5

14.55

1500# TiClg

4.8

4.9

6.3

4.2

4.40

640# CuS04

17.2

15.5

14.2

14.7

15.40

340# Fe2(S05)4

14.4

14.4

15.1

14.8

14.67

1380# K2Cr04

17.7

17.7

20.5

19.5

18.85

Analysis of Variance
Source_____ D. F*

S. S._____ Var.____ S. _D.

39

519.9

Replication

3

11.6

Treatment

9

470.8

52.31

27
F ~ 37.26

37.5

1.39

Total

Error

Difference for significance between means

1.18

1% = 2.31

m - i.7i

Signif­
icance

*

**

109.
receiving copper sulfate and ferric sulfate were very
chlorotic during their early growth but became normal in
appearance during late growth.
sulfate were very irregular.

Those receiving ferrous
The titanium treatment pro­

duced very injurious effects.
In Table 41 is shown data received on these jars when
sown to spinach in the fall.

On two blocks of this experi­

ment, red spider destroyed the spinach to the extent that
they were of little value as data.

The remaining blocks

gave very interesting results but some of them were entirely
observational and are not expressed in the final data.

Zinc

acetate treatments stimulated the growth of the spinach in
the early growth.

This stimulation did not last long, as

these plants became very chlorotic later.

Marked response

was obtained on those jars receiving ferrous sulfate and
copper sulfate.

The manganese salts were beginning to show

beneficial effects late in their growth.

Ascorbic acid and

glutathione determinations were made on these plants and
they are shown later in Tables 49 and 50, respectively.
Wolverine oats were seeded in these jars January 5,
1940, and the following data secured as given in Table 42.
A very marked response was obtained from the manganese salts
which was significant over that from all other treatments.
The results from the zinc and titanium salts were signifi­
cantly lower.

The oats receiving the manganese sulfate had

a healthy green color, while with all other treatments, the
leaves showed infestation of grey speck disease.

Table 41, showing yields of spinach on OxidationReduction Experiment. Sown Sept. 1, 1939, har­
vested Nov. 27 to Dec. 1, 1939. Green weight in
grams.
Block A

Block B

Average

59.7

50.4

55.0

750# MnS04

57.1

45.6

51.3

700# Mn Acetate

38.0

72.7

50.3

450# Zn Acetate

54.0

44.0

3380# Or2 (S04 )3

51.6

46.8

49.2

3350# FeS04

60.8

76.7

68.7

1500# TiOlg

55.0

31.4

43.2

640# 0uS04

63.5

74.2

68.3

340# Fe (S04 )3

40.6

45.5

43.0

1380# K2Cr04

43*2

51.0

47.1

o

Check

CD
•

Treatment

Table 42,

showing yields of Wolverine oats on OxidationReduction Experiments in Greenhouse. Sown
Jan. 5, and harvested Febr. 26, 1940.

Treatment_____ Block A Block B

Block C Block D

Average

Significance

Control

17.5

24.7

24.5

20.4

21.7

750# MnS04

54.6

58.9

50.1

56.4

55.0

■H-K-

700# Mn Acetate

59.6

61.6

54.0

61.1

54.1

4f*

450# Zn Acetate

15.2

15.7

15.5

18.5

15.7

2550# Crg(S04)g

23*7

22.0

26.5

16.8

22.3

225(# FeS04

16.0

25.5

23.8

18.8

21.0

1500# TiClg

11.7

15.6

16.7

12.0

14.0

640# CuS04

18.1

18.2

22.2

22.5

20.3

500# Feg(S04)5

26.5

27.6

27.0

27.4

27.1

1380 K2Cr04

19.6

20.7

27.9

21.0

22.3

Analysis of Variance
Source_________D. F.___ S. S.____ Var.____S. D »
39

8460.1

Replications

3

160.9

Treatment

9

7862.9

27

456.2

Total

Error

875.7

4.02

16.16

Difference for significance between means

1% = 7.87
5% = 5.85

*

112.
Grey speck disease made its first appearance on the
cultures receiving zinc and titanium salts.

Those re­

ceiving manganese sulfate treatment showed the greatest
resistance to the disease, with manganese acetate nearly
as efficient.

Table 43 gives some idea as to effect of

treatment upon rate of infection.
Table 43, showing per cent of plants exhibiting
grey speck disease at different stages of
growth. Sown Jan. 5, 1940.

Treatment

Date of Counts
Feb. it
Feb. 24
34

98

Mangane se sulf at e

1

1

Mangane se acet at e

3

3

Zinc acetate

99

100

Chromic sulfate

22

91

Ferrous sulfate

14

79

Titanous chloride

91

100

Copper sulfate

24

98

Ferric sulfate

25

100

Potassium chrornate

15

76

Control

Ascorbic acid and glutathione were determined in
samples of this oat and the data is reported in Tables
51 and 52•

Tire Effect of Minor Element Fertilization on OxidationReduction Systems Within Plant.
The recent findings of various workers indicates that
the application of various minor elements to nutrient
solutions may affect the oxidation-reduction systems of
ascorbic acid and glutathione.

Euler, Myrbock and

Larrson (18) have shown that the application of copper,
manganese and nickel ions increased the oxidation of
ascorbic acid; also that cobalt inhibited this oxidation.
These results are substantiated by Green, McCarthy and
King (24) who found that the aerobic oxidation of ascorbic
acid is very sensitive to catalysis by copper protein com­
plexes in a great variety of expressed plant press juices;
also they found that both copper and ascorbic acid are
found in relatively high concentrations in green leaves or
other tissue capable of photosynthesis.

Further, they

believe that although the evidence is far from complete,
it is not unreasonable to believe that copper and ascorbic
acid may be concerned directly or indirectly with both
respiration and photosynthesis in green plants.

In another

report (52) the same authors describe various systems in
which a rapid oxidation of ascorbic acid is due to the
catalytic effect of copper combined with proteins.

They

enumerated other plant and animal oxidative catalysts as
those that lead to the quinone formation, the cytochrome
systems and the hemochromogens, all of which can obviously

114
act as catalysts for the oxidation of ascorbic acid.

They

conclude that the dominant active agent in many plants
appears to be copper.

Guzman Barron, Demeio and Klemperer

(26) , from their studies of the oxidation of ascorbic acid
by copper and hemochromogens, propose the following
equations:

(1 )

H H H
I l i/>
HOOC-C-C
C=0 + 4 Cu+ +
1 I \ /

H CH 0-0
/

/

CH OH

20.

(2)

4 Ou + + 4H

(3)

2 H203 _____ ^ 2 H20 - 03

H H H
1

1

HOOC-C-C
0=0
I I \
i
H CH 0 - 0
it
it
0
0

4 Ou

4 Cu+

+>
H2°2

Euler indicated that he obtained similar results with
the use of manganese in his experiments.

Rudra (60)

reports that nutrient solutions containing manganese pro­
duced more ascorbic acid in the plants.

In reviewing the

work of these investigators, it will be found that those
reporting benefit from manganese worked at a pH above 6*0.
The significance of this will be brought out in later disc ussion.
Glutathione exists in plants and animals in both the
reduced and oxidized form and, for this reason, should re­
flect the oxidation-reduction intensities that exist.

This

115.
system might be expressed by the following equations: —
2 HOOO.CHNHg.OHgSH JDx^ HOOC.CHNHg.GHgS-SHgC.CHHHg.COOH
*Ked
or G-S-S-G the oxidized form and G-Sh the reduced form.
This system has been studied considerably in the blood of
animals.

Lyman and Guzman Barron (47) studied the oxi­

dation-reduction systems in living cells.

They state,

MGlut athi one and ascorbic acid seem to possess closely re­
lated properties in the chemical activities of biological
systems, maintaining graded levels of reduction intensity
necessary for the performance of certain biochemical pro­
cesses. tt If glutathione is similar to ascorbic acid in
properties, it will not be surprising to find that it enters
into oxidation-reduction reactions with copper and manganese.
They also bring out this reaction of glutathione and iron: 3G3H ____ ^ 3 Fe * * + G— S-SH + 2H +
<---3H + + i 02 ____ * 2 Fe + + + t H20

(1) 3 Fe + + * +
(3) Fe + + +

< -----

These reactions are intensely interesting, since it
shows the possibility of an oxidation-reduction tie-up
between iron and glutathione.

These findings may have much

to do with either chlorophyll formation or its function in
plants.
This study has for its purpose the exploration of the
possibility of the minor elements, manganese, copper, iron,
chromium, zinc influencing the oxidation-reduction

116.
equilibriums within the plant.

From the greenhouse studies

and various field observations, it has been noted that
these various elements give marked responses to plant
growth, the response depending upon the environmental condi­
tions.

These conditions appear to manifest themselves either

directly or indirectly as soil temperature.

The application

of a manganese salt to an alkaline muck in the spring, when
the soil temperature is low, will cause chlorotic plants to
become normal.

If the temperature of the soil were raised

to a higher level, these same plants would become chlorotic.
At this point, an application of copper salt would cause the
plants to become normal again.

It follows therefore that

two types of chlorosis exist; namely, a minimum temperature
chlorosis, characterized by a mottled leaf with prominent
dark green veins, and the other, maximum temperature chlor­
osis, characterized by a yellowish-green leaf with almost
transparent veins.

In

both cases the iron content appears

to be affected and the results of staining many leaves in­
dicate that the difference appears to be in the state of
oxidation of iron.

Since iron is a catalyst to the format­

ion of chlorophyll, it must serve in this function, setting
up an unbalanced oxidation-reduction equilibrium which is
essential to the living plant.

It therefore follows that

any factors affecting the oxidation-reduction intensity of
the plant will shift the direction of this equilibrium and,
if sufficient, might and should cause a physiological dis­
turbance of the living processes of the plant.

The extent

117.
of this change of direction of the equilibrium is determined
by the resistance to such a change.

This is analogous to

buffering capacity, as related to pH, and may be pictured
as follows;
Zn
Ou
Ti

vs
*♦*

Mn
Co
Fe**- +
Cr

Fe4, * ^Fe* + *
The results of the investigations carried on in green­
house (oxidation-reduction experiment) and also those made
under field conditions support this theory to some degree.
Determination of reduced and oxidized ascorbic acid
and reduced and oxidized glutathione

should give data

which would reflect the effect of minor elements in plant
nutrition when it is subjected to conditions which produce
a lime-induced chlorosis#

Methods of Analysis
Ascorbic Acid: - The method used was essentially the
one used by Bessey and King (9).

A two-gram sample of

plant material was extracted twice by a solution, which
was about eight per cent tri-chloro-acetic acid and two
per cent in respect to metaphosphoric acid.

The strength

of the acid was adjusted, so that the pH of the extract was
maintained close to 4.5 and not more than 5.0.

The plant

material was ground in a mortar under 10 to 13 ml. of the

118*

to
l
O
02

§

showing yields in order of their percentage of the
control♦ Control is taken as 100 per cent.

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119.
acid*

A small amount of quartz sand was added to facilitate

the grinding of the tissue.

After thorough grinding, the

mixture was transferred to centrifuge tuhes and centrifuged*
The clear liquid was poured off and the extraction repeated*
The filtrates from hoth extractions were combined and the
ascorbic acid was titrated with a standardised 2, 6, dichlorobenzenoneindophenol, to a pink end point which was
permanent*

For total ascorbic acid, hydrogen sulfide gas

was passed through the combined filtrates until all oxidized
ascorbic acid was reduced.

The excess hydrogen sulfide was

washed from the solution by passing carbon dioxide gas
through it.

The titration for total ascorbic acid is the

same as described for the reduced acid.

In colored plant

extracts the end point is determined in a chloroform layer
described by Lewis (45).
Glutathione:

The method used for determining total,

reduced and oxidized form of glutathione was a modification
of that proposed by Woodward and Fry (82).

Two grams of

green material were weighed out and to this 16 ml. of dis­
tilled water were added in a porcelain mortar.

The material

was crushed and allowed to stand a few minutes at which time
2 ml. of 22 per cent sulfosalicylic acid was added slowly
with grinding.

The mixture was transferred to a centrifuge

tube and centrifuged.
repeated.

The liquid was decanted and extraction

The combined filtrates contained 2 per cent sulfo-

salicylic acid and had a pH below 2.0.

^o the combined fil­

trates were added 2.5 ml. of 4 per cent sulfosalicylic acid

120.
and 2.5 ml. of 5 per cent potassium iodide solution.

Two

or three drops of a one per cent starch solution were added
and the filtrate titrated with a .001 IT potassium iodate
solution.

This gave the reduced form of glutathione.

Total glutathione was determined by taking the combined
filtrates of the extraction containing 2 per cent sulfosalicylic acid and adding to it 30-40 mg. of zinc dust.

It

was then allowed to stand at room temperature for 20 minutes.
Excess zinc was removed by centrifuging.
same as for the reduced form.

Titration was the

Total glutathione-reduced

glutathione - oxidized glutathione.
-=mg. of glutathione
ml of KIQg
x
;
3.26
weight of sample
per gram of plant tissue. If divided by 100 would give per
cent of glutathione.
Experimental
Spinach was sown on jars of soils taken from 3 five-inch
layers of plat 8 of the Manganese-Sulphur Series of the
College plots.

These layers corresponded to the horizon

layers which were analyzed for various forms of manganese in
the soils (Table 4).

Three treatments were made; control;

250 pounds per acre of Manganese sulfate and 100 pounds per
acre of copper sulfate.

This spinach was grown during the

fall of the year and showed a response to both copper and
manganese salts.

On the 10—16 inch layer, however, in the

spinach died on the jar receiving manganese sulfate.

This

121.
layer is acid which may account for such a reaction to the
treatment.

Ascorbic acid and glutathione were determined

on the spinach from these jars and the data are given in
Table 45.

Copper treatment resulted in an increase in

reduced ascorbic acid in every case.

There is an increase

in the ascorbic acid content of the spinach on the jar of
the 0-5 inch layer receiving manganese sulfate but it is not
as high as in those receiving copper sulfate.

Glutathione

was determined in the spinach from the 0-5 inch layer only.
The glutathione content of this spinach gives data very
similar to that received in the case of ascorbic acid.
Plates 3, 4 and 5 show this spinach as it appears at the
time of sampling.

121a.

Plate 3.
Spinach on the 0-5 inch layer of plot 8 of Mn-S
Series. Soil pH = 7.2.

A A A -

1 Control
2 250 pounds
3 100 pounds

of MnS04
of CuSO^

121b.

Plate 4.
Spinach on the 5-10 inch layer of plot 8 of the
Mn-S Series. Soil pH = 7.3.

D - 1 (ceutei
B - 2 (right
B - 3 (left

Control
250 pounds of MnSO.
100 pounds of CuSO^

121c .

Plate 5.
Spinach on the 10—15 inch layer of plot 8 of the
Mn-S Series. Soil pH = 6 .8 ,

C C C -

1 (center) Control
2 (right ) 250 pounds
3 (left 5 100 pounds

of MnS04
of CUSO4

Table 45,

Depth
Jar No, In,

showing the content of ascorbic acid (Vit. C) in
Long Standing Bloomsdale spinach. Sown Aug, 12,
harvested Oct, 16, 1939,

Treatment

A - 1

0-5

Control

A - 2

0-5

A - 3 0-5

Wt./j ar
gms.

Mg. Vit. C Units of
per
Vit. C
100 eras. per gm.

Units of
Vit. C
per iar.

74.3

39.35

7.87

585

250# MnS04

113.8

60.00

12.00

1366

100# cuso4

135.6

73.35

14.67

1989

B - 1

5-10

Control

107.3

35.00

7.00

751

B - 2

5-10

250# MnS04

103.6

30.00

6.00

622

B - 3

5 — 10

100# CuS04

88.0

60.55

12.10

1065

C - 1 10 - 16

Control

19.3

60.10

12.02

232

C - 2 10 - 16

250# MnS04

died

----

----

C - 3 10 - 16

100# CuS04

49.9

72.20

14.44

Table 46,

Depth
Jar No. in.

721

showing the content of glutathione (G-SH) in
spinach grown in greenhouse. Sown Aug. 12 , harvested Oct. 16, 1939.

Treatment

Wt. of spin­
ach / jar"
gms.

Mg. G-SH/
gm
spinach

Mg. G-SH/
jar

74.3

2.18

162

A -- 1

0-5

Control

A --2

0-5

250# MnS04

113.8

2.72

310

A --3

0-5

100# CuS04

135.6

2.48

336

123.
During the fall, samples of fall spinach were collect­
ed from various plots of the Manganese— Sulphur Series of
the College plots.

The plants collected do not represent

the treatment, those taken on the plots receiving manga­
nese sulfate, sulphur and the combination of the two were
healthy normal plants.

Those selected from the control

plots were the most chlorotic plants on the plot.
chlorosis would not "be considered severe.

This

This analysis

should be considered a comparison between normal plants
and those exhibiting a mild case of chlorosis.

The data

show that the chlorotic tissue contains one-half to twothirds as much ascorbic acid as the normal tissue.
Table 47, showing ascorbic acid (Vit. C) content
in units per gram of normal and mildly chlor­
otic spinach. Samples taken Oct. 22, 1939.
Condition

Treatment
Manganese sulfate
tf

fi

Manganese sulfate
& sulphur
Mang ane se sulf at e
& sulphur
Sulphur
»
Control
it

Uormal green

Ascorbic Acid
units/gram
29.52

ii

it

30.28

n

n

30.03

n

n

ii

it

31.37
22.72

tt

it

24.08

Mildly chlorotic
it

it

14.87
16.67

All determinations were made on leaves from the inner part
of the rosette.

124.
Cayuga soybean leaves were collected, which showed two
degrees of chlorosis, namely, chlorotic leaves which are
nearly yellow and leaves which show some mottling of yellow
in the interveinal tissue.

These were analyzed for reduced

glutathione and the data show that glutathione (G-SH) con­
tent was reduced with the increased severity of the chlorosis.
Table 48, showing glutathione (G-SH) content in normal and
chlorotic soybean leaves.
Condition
of
leaves

ilgm. G-SH/gm.
Sample 1

Mgm. G—SH/ gm. Average
Sample 3
Mgm. G-SH

Mgm. G-SH
per 100 gm.

Hormal

4.01

4.10

4.055

406

Chlorotic
(mottled)

3.60

3.52

3.560

356

Chlorotic
(severe)

3.11

2.88

2.995

300

The content of ascorbic acid in the spinach, reported
in Table 41, was determined and the data is given in Table
49.

The greatest yield per jar was secured on those jars

producing the most spinach, namely, those receiving ferrous
and copper sulfates.

The highest value per gram was pro­

duced by copper sulfate and manganese acetate treatments.
Zinc acetate, titanous chloride and ferric sulfate treat­
ment gave lowerdd yields on both production bases.

Table 49,

showing Ascorbic Acid (vit. C) content in spinach
plants grown in greenhouse. Sown Sept. 1, 1939,
harvested Dec. 1.

Av. of 2
Mg.
titration
Vit. C Units of
units of
Jar
units of
per
Vit. C
Vit. C
•______ Treatment___ Vit. C*
Ave. 100 gm. per gm._____ per .1ar

0-2
0-12

750$finS04

0-5
0-13

700#m(CgH50?.)2

??
H

H
H H
1 1
O O

Control

450#Zn(CgH50g)2

U5
W H
t I
O O

2350#Crg(S04)g

0-6
0-16

2250#FeS04

0-7
0-17

1500# TiClg

0-8
0-18

640#CuS04

0-9
0-19

500#Feg(S04)3

7.40
11.85

9.62

48.10

9.62

529

10.02
10.40

10.21

51.05

10.21

524

12.20
14.45

13.32

66 .60

13.32

670

4.76
6.10

5.43

27.15

5.43

266

14.39
6.67

10.53

52.65

10.55

518

9.90
11.67

10.79

53.95

10.79

741

7.55
8.12

7.83

39.15

7.33

338

9.22
14.90

12.06

60.30

12.06

824

4.90
7.17

6.03

30.15

6.03

259

136.
Oxidized and reduced glutathione was determined in
the spinach from two of the treatments.

The spinach

treated with ferrous sulfate was normal in appearance
while that receiving zinc acetate was very chlorotic.
Very little difference existed in total glutathione, while
the reduced form was higher and the oxidized form much
lower in those receiving ferrous sulfate.
Table 50, showing content of various forms of glutathione
in spinach.

Treatment

Reduced
Total
gluta­
gluta­
thione
thione
in mgm. mgm. per
/gm.
___ gm.._

Oxidized
gluta­
thione
mgm. per
_gaw
.

G-SH
Ratio -------G-S-S-G

...

450 Zn Acetate

1.244

.854

.390

2.19

2250 FeS04

1.390

1.341

.049

29.48

Reduced and oxidized ascorbic acid and reduced gluta­
thione determined on the Wolverine oats reported in Table
42, is given in Table 51.

The manganese treatments which

gave the highest yields showed the highest content of re­
duced ascorbic acid and reduced glutathione and the lowest
oxidized ascorbic acid.

The zinc acetate and titanous

chloride treatments which produced the lowest yields gave
the lowest content of reduced ascorbic acid and reduced
glutathione and the highest values for oxidized ascorbic
acid.
The ratio of reduced to oxidized ascorbic acid indi­
cates some effect of that treatment.

Very little difference

127.
existed between the other treatments and the controls.
The data appear to support the theory that manga­
nese salts when giving beneficial results also have a
favorable effect upon oxidation—reduction systems within
the plant.

It also appears that any minor element pro­

ducing a beneficial effect upon the plant growth will
affect the oxidation-reduction systems in a favorable
manner.

128,

Table 51,

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showing the reduced and oxidized ascorbic acid content in Wolverine oats on
Oxidation-Reduction Experiment, Sown Jan, 5, harvested Febr, 26, 1940, Ascorbic
acid reported as units per gram of green oats.

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Table 52,

showing average contents of reduced and oxidised forms of ascorbic
acid and milligrams per 100 grams of glutathionb in Wolverine oats
on Oxidation-Eeduction Experiment*

tic

130,
General Discussion
1^ is very difficult to give a very definite explan­
ation as to the role that manganese plays in soil fertility
and plant nutrition#

In the face of all the essential

nature of manganese in plant nutrition, it would he ex­
tremely foolish to say that it is not an essential element.
A very comprehensive study of these data and other data
strongly suggests the possibility of an accessory function,
of an intensity nature.

It has been previously pointed

out that certain observational facts, yield data, and
chemical analysis failed to support the essential element
theory.

It is therefore necessary to study the data given

here and the data of other investigators in order to ob­
tain a comprehensive insight into its possible functions.
In the first place, certain soil conditions have been
associated with manganese deficiency, such as alkalinity,
high lime content and organic matter supply.

It is the

contention of this paper that the pH of the soil is not a
causative but rather an attendant factor.

Such a relation­

ship is hardly tenable, since pH is a resultant of many
processes including such reactions as leaching and oxidation —
reduction which may alter the availability of, or the
essentiality of, the manganese.

These may be the resultant

of natural processes or subsequent processes initiated by
the nature of soil management or by change in environmental
conditions.

It generally has been accepted that if the soil is
made more acid or if it is water logged, the solubility
or availability of the soil manganese is increased.

Leeper

has pointed out that this is true only if the soils are
high in active manganic compounds.

His method of determin­

ing manganese—deficient soils appears to be very reliable.
On the other hand, this work presents a very contradictory
phase to Leeper*s active manganese theory.

In general,

the soils made acid by application of sulphur have given
either the highest or approximated the highest yields of
crops grown on these plots.

The plants have been just as

completely cured by this treatment as those receiving the
manganese salt.

The disturbing element is that this treat­

ment does not show an increase in active manganese nor does
it show an increase in bivalent manganese over that of the
control which has produced chlorotic plants.

The analysis

of plant material fails to show a consistently higher manga­
nese content in the plants from sulphur tested plots than
on those from controls and quite often it appears to be
definitely lower.

This eliminates the possibility of small

increases with an accelerated velocity of the chemical re­
action.

Such a contradiction does not necessarily refute

Leeper»s theory.

It is highly probable, however, that with

a higher hydrogen ion concentration, the need for manganese
becomes less.

This idea is supported in Harmer’s unpublish­

ed data (31) which shows strongly acid organic soils which
contain but mere traces of manganese upon total, analysis and

132.
are still capable of producing perfectly normal crops.

A

survey of the literature shows that there is a great vari­
ation in the quantity of manganese sulfate required to be
added to the soil for a desired benefit.

Further, the

amounts considered necessary for toxic effects also vary
greatly,

it is therefore contended that the soil»s need

for active Mn02 is quite variable and it can be concluded
that, in the apparent presence of exchangeable hydrogen, the
need for manganese may be very small and is not so acute.
The data show that in making these soils more acid,
the solubility of the calcium is decreased.

The water-

soluble and exchangeable calcium are lowered by the appli­
cation of sulphur.
The analysis of the chlorotic plant tissue shows that
the mobility of the iron has become impaired and that the
plant suffers from a lack of iron at points where it is
vitally necessary.

McGeorge (54) has pointed out that,

when plants assimilate luxury amounts of calcium iron will
be precipitated in the cells.

Wilcox and Kelley (76) state

that, under such conditions, there is an abundance of cal­
cium oxalate crystals in the plant cells.

The data of the

seasonal trend tend to support McGeorge •s theory in that,
during periods of low temperature in soils and with high
moisture conditions, the plant takes in excessive amounts
of calcium.

It is during these periods that severe chlor­

osis prevails which chlorotic condition is curable by the
application of manganese or sulphur.

Either of these

133.
treatments cause the iron to function properly in the
plants.

There is no reason why another chemical other

than manganese should not he capable of producing the
same effect upon iron.

The functioning of iron which is

affected by the hydrogen ion concentration and the calcium
content of the plant tissue is subject to the favorable
and unfavorable influence of the ions capable of taking
part in oxidation-reduction reactions.

It is therefore

plausible and very probable that, under different sets of
conditions, different members of this group of ions will
give favorable responses to plant growth.

Excesses of any

members of this group will in all probability be expressed
in a disturbed state of oxidation of the iron.

The results

obtained by staining the iron in plant tissue gives very
remarkable support of the effect upon the state of its oxi­
dation.

The non-responsive crops such as sweet clover,

sugar beets and peppermint do not show these effects.
Since the precipitation of iron appears to be the re­
sultant of disturbed oxidation-reduction equilibrium, it
would be expected therefore, that the other oxidationreduction systems within the plant would be affected.

In

a normal healthy plant, the ascorbic acid and glutathione
are predominantly in the reduced form.

The fact that they

occur in this form substantiates the theory of an effect
upon the oxidation-reduction system by the manganese and
other minor elements capable of taking part in such
reactions.

134.
The part played by climatic factors appears to be
indirect in that it probably influences the intake of
calcium.

It is also plausible that temperature might be

capable of playing a part in the direction intensities
oxidation-reduction reactions which is partially sup­
ported by the lower ascorbic acid content under higher
prevailing temperature ranges.

The elements of climatic

environment need further investigations under more control­
led conditions than has been possible in this study.

The

observations made of these factors in this study suggest
that they are very important.
Oxidation-reduction has been stressed considerably
in recent work.

The data obtained in this study indicate

that oxidation-reduction potential of the soil is a minor
factor in the plant growth.

The prevailing oxidation-

reduction systems may influence the balance of nutrients
available to plants which in its natural condition may or
may not have an injurious influence upon plant growth.
This influence may become greatly aggravated when the condi­
tion is altered by treatments, such as liming or by other
means.

Oxidation-reduction systems within the plant are of

great importance to its physiological processes.

The kind

of ions taken in by the plant and the environmental condi­
tions have an influence upon these systems.

The state of

oxidation of iron can be altered by an excess of the multivalent ions or precipitated by excessive calcium intake.

135.
The oxidation-reduction experiment supports this hypothesis
inasmuch as when manganese gave beneficial results, zinc
gave very detrimental effects and on the other hand when
manganese salts were detrimental to plants, copper, chrom­
ium and other ions were beneficial.

It is not to be construed

that the ions other than those of the multivalent group can­
not produce similar disturbance.

A lack of potash in the

soil will cause iron to deposit in nodes of corn.

Recent

work of Schmitt (67) would substantiate this as he found
that a lack of potash in plant nutrition produced plants
which were low in reduced form of ascorbic acid and high
in the oxidized form.
The question naturally arises as to the importance of
the above relationships to plant and animal nutrition.

In

case of the former it gives added weight of the necessity
of harmony of nutrients.

Chlorosis can be considered the

resultant of an unbalanced condition in the nutrients
available to the plant.

This unbalanced condition may be

due to the lack of one or a group of other ions or to the
excess of one or more ions.

It is reasonable to theorize

that many of the toxic conditions described are probably
in all reality a failure of iron to function or to the
breakdown of other oxidation-reduction systems within the
plant.

These physiological breakdowns result in plants

which are extremely susceptible to disease.
The relationship of this problem to animal nutrition
is extremely important as many of the mineral malnutritions!

136.
diseases are associated with areas where minor element
deficiencies occur in plants.

Investigators in nutrition

research are confronted with the question of available
iron in foods.

Many foods are high in iron and yet fail

to produce hemoglobin when fed to anemic animals.

Chlor­

osis in plants produces a high total iron and a low avail­
able iron content.

Since mild chlorosis cannot be readily

ascertained it would be expected that plants having such a
condition would reach the channels of animal nutrition.
Spinach is a very good example of such a crop and is also
one of the most susceptible crops to lack of proper ferti­
lization.

Great emphasis should be given to proper ferti­

lization as it reflects directly upon nutritional quality
of the crop.
Finally it is reasonable to speculate from results
here that the role of minor elements is that of the acti­
vation of iron.

SUMMARY
As a result of a chemical study of the crops produced
on an alkaline organic soil, and of the soil itself, as
affected by the application of manganese sulfate, sulphur
and other chemical materials, the following conclusions
were reached.
A.

The Soil
1.

The application of sulphur decreases the
exchangeable calcium and magnesium, the
water-soluble calcium and to a lesser
degree, the water-soluble magnesium. It
also markedly increases the exchangeable
hydrogen.

2.

The application of manganese salts increases
the total, exchangeable and easily reduci­
ble manganese in the soil but does not
appreciably influence the exchangeable and
the water-soluble calcium and magnesium or
the exchangeable hydrogen.

3.

The soil complex of this alkaline muck is
highly saturated with calcium and magnesium
with the greater portion of the exchangeable
cations as the former.

4.

Manganese salts applied to the soil are
fixed firmly in the surface layers. There
is no evidence of enrichment of subsoil
layers by an application of a manganese salt
to the surface layer. The fixation appears
to be due to the oxidation of the manganese
to the manganic oxide.

5.

The system of Mn * +
■> Mn ^
+ + is
very important in soils. There is a cor­
relation between crop growth and the
quantity of the right-hand member of this
equilibrium that can take part in a reaction
which goes from right to left.

6.

The hydroquinone—ammonium acetate extraction
is a reliable means of measuring the manganic
manganese which takes part in the manganousmanganic equilibrium of alkaline and neutral
organic soils.

138.
7.

The water—soluble manganese is very low in
these soils, even in those receiving manganese
salts.

8.

The application of sulphur tends to increase
the exchangeable more than the reducible man­
ganese in the soil. The application of manga­
nese salts tends to increase the reducible more
than the exchangeable manganese.

9.

The seasonal trend of the exchangeable manganese
in this soil shows a decrease as soil tempera­
ture increases and at the same time the easily
reducible manganese dioxide increases.

10.

B.

The seasonal changes in soil temperature and soil
moisture affect the state of oxidation of the
manganese. Under low soil temperature and high
soil moisture content the manganese tends to­
wards the bivalent form. Under conditions of
higher soil temperature and generally lower soil
moisture content the equilibrium swings towards
the manganic form.

The Plant.
1.

Chlorotic plants show a higher iron content
than normal plants. The application of manga­
nese salts or sulphur produces normal plants
which have a lower total iron content.

2.

There is a difference in the state of oxi­
dation of the iron in the chlorotic and normal
plant tissue. In chlorotic plants, ferric iron
is abundant in the veins of the leaves and
vascular tissue of the stems. Ferric iron
could not be found in similar tissue of normal
plants.

3.

The hydrogen ion concentration is higher in the
normal than in the chlorotic tissue.

4.

Minor
element deficiency of the multivalenced
group has to do with the state of oxidation of
the iron in the plant. This being the condi­
tion, any factor affecting the oxidation-re­
duction capacity of the plant will affect the
oxidation-reduction equilibrium of the ferrousferric system. This system can be and is
affected by these elements and, if one is in
excess then another producing an opposite condi­
tion will counteract it. Further, the state of

139
oxidation existing in the plant is the re­
sultant of the actions of many physical and
chemical processes occurring in the plant;
or externally, or lastly indirectly through
its localized environment; the result of
these forces can be expressed in the oxidat­
ion level of the ferrous—ferric system in
the plant. It follows, while, under one
condition, one element may be much more
efficient than another, it does not elimin­
ate from causing the same result,

C.

5*

The content of magnesium is higher in
chlorotic than in normal plant tissue,

6,

The application of manganese salts to the
soil increases the manganese content of the
plant. A still greater increase is obtained
when both manganese salts and sulphur are
applied to the soil together,

7,

In general, the application of sulphur to the
soil did not increase the manganese content
of the plants over that of the plants from
the control,

8,

Climatic factors, precipitation, soil tempera­
ture and soil moisture affect the plants
intake of calcium. A high soil moisture con­
tent and a low soil temperature cause certain
plants to take in abnormal amounts of calcium.

The Oxidation-Re duct ion Systems.
1.

In a healthy plant ascorbic acid and gluta­
thione axe predominantly in the reduced form.

2.

In chlorotic plants a marked increase occurs
in the proportion of ascorbic acid and gluta­
thione in the oxidized form. The reduced
form of ascorbic acid is much lower in chlor­
otic plants,

3.

Total ascorbic acid content of the plant is
decreased by a chlorotic condition. The
limited data on the content of glutathione
does not show a decrease due to chlorosis.

4.

Minor elements influence the oxidation—reduct­
ion equilibrium within the plant. If the minor
element is beneficial to plant growth, the oxi­
dation—reduction equilibriums under considerat­
ion are strongly towards the reduced forms. If

140.
the minor element causes a chlorotic condi­
tion or causes an existing chlorotic condi­
tion to become more severe the direction of
these equilibriums is towards the oxidized
form.

141.
BIBLIOGRAPHY
1.

3.

A1fXm?de?* Lyle L*’ Horace
Byers and Glenn Edgington.
A Qjiemical Study of Some Soils Derived from Limestones
u.s.d.a. Tecfrrmr: stbt nrero.-------------— ----Allyn, W. P. The Relation of Lime to the Absorption of
Plants. Indiana Acad. Science Proceedings 43
pp. 405-409, 1927.
B

3.

Arkhangelskaya, N. New Method of Studying the Brown
Spotting Disease in Potatoes. Oompt. rend. Ac ad. Sci.
U.S.S.R. 19 pp. ITl-14, 1938.

4.

Aso, K. On the Physiological Influence of Manganese
Compounds in plants. Bui. Coir. Agr. Tokyo Imp. Univ.
5 pp. 1771X85, 1903. From minor element abstracts.

5.

Barber, H. H. and I. M. Kolthoff, Determination of sodium
as sodium zinc uranyl acetate. Jour. Amer. Chem. Soc.
50 pp. 1635, 1938.

6*

Baten, W. D. Formulas for Finding Estimates for Two and
Three Missing Plots in Randomized Block Layouts.
Mich. State College Sgr. Exp. Sta. Tech. Bui. 165, 1939.

7.

Bertrand, G. On the Oxidizing Action of Manganese Salts
and Chemical Composition of Oxidases. Compt. rend.
Acad. Sci. Paris 124
pp. 1355-58,1897.

8.

____________. The Role of Infinitely Small Amounts of
Chemicals in Agriculture. American Fertilizer 3Y ~
pp. 37, 38, 1912.

9.

Bessey, 0. A. and C. G. King. Vitamin C Distribution.
Journal of Biological Chemistry, Vol. 103,"pp“. 687,
1933.

10 . Bortner, C. E. Toxicity of Manganese to Turkish Tobacco
in Acid Kentucky Soils_. Soil Science 39
pp. 15-33,
1535.
11 . Bray, R. H.

The Quantitative and Qualitative Determination
of'Replaceable Sodium, in Alkali and Non-Alkali Soils.
Jour, of Amer. Soc. Agronomy 20 pp. 1160, 1928.

13.

Camp, A. F. and Walter Ruether. The Yellowing of Citrus
Leaves. Citrus Industry 17. No. 5, 1936.

142.
13.

Cooper, H. p., W. R. paden and R. L. Smith. Intensity
of^Removaljof Cations from Cotton, Corn, and Soybean
—
Fractional Electrodialysis. Plant PhvfiinU
ogy 12 p p ~ 9 % - 9 8 7 ; 1937.------ -----

14.

> A*
jggyr — n£

Yellowing of Wheat Seedlings
Rev. Applied My col". 16 pp. 800,

15.

Davies, D^ W. and E. T. Jones. Grey Speck Disease of
QQjj'l» Welsh Journal of Agriculture y , pp. 549-35^7

16.

Degrully, L. Treatment of Chlorosis by Sulphuric Acid.
Prog. Agr. et Vit. 12, 1895^
-----------

17.

Enfield, G. H. and S. D. Conner. The Fixation of Potash
by Muck Soils. Journal American Society ofAgronomy
ZB
pp. 146—155, 1936.

18.

Euler, H . , K. MyrbtJck and H. Larrson. Sauerstoffaufnahme
Durch Vitamin C Halige Organe and Durch Gluco-Hedukton.
Z. Physiol. Chem. 217, 1933.

19.

Gerretsen, F. C. The Effect of Manganese Deficiency on
Oats in Relation to Soil Bacteria. Trans. 3rd Intern.
Congr. Soil Science 1 pp. 189-91, 1935.

20.

.
______________. The Causes of the Grey Speck Disease
of Oats. Verslag. landb. Onderzoek Rijkslandbouwproefsta No. 42A pp. 1-67. C.A. 30-6118, 1936.

31.

. Manganese Deficiency pf Oats and Its
Relation to Soil Bacteria. Am. BotanyHTn. s.) 1
p p 2 0 7 - 3 U 7 1937.

2 2 . Gilbert, B. E. Normal Crops and the Supply of Available
Soil Manganese. Rhode Island St a. Bui. 24§““ p p . 15,
1934.
23.

, F. L. McLean and L. J. Hardin. The
Relation of Manganese and Iron to a Lime-induced
Chlorosis. Soil Science 22 pp. 437-446, 1926.

24.

Green, Lowell F., James F. McCarthy and C. G. King.
Inhibition of Respiration and photosynthesis in
^Chldrella Pyrenoidosa1'.by Organic Compounds that
Copper Catylizes. Journal of Biol. Chemistry 128
ppTT?77lt^

25.
Sci . 19

p ~ 1118-1119, 1844T

143 *
36.

.??an ®arro^, E. S., R. H. DeMeio and Frudrich
Klemperer. Studies on Biological Oxidations. Jour.
Biological Chemistry, VolY 113, pp. 625-640, 1936.

37.

Haas, A. R. 0. Innurious Effects of Manganese and Iron
Deficiencies on the Growth of Citrus. Hilerardis 7
pp. ±81-206, 19327

38.

_____________ « Walnut Yellows in Relation to Ash
Composition, Manganese, Iron and Other Ash Constituents.
Bot. Gaz. 94 pp. 495-511, 193F7

39.

_____________ . Deficiency Chlorosis in Citrus.
Science 43 pp. 435-443, 1936.

30.

______________ and H. J. Quayle. Copper Content of
Citrus Leaves and Fruit in Relation to Exanthema
and Fumigation Injury. Hilgardia 9 pp. 143-177, 1935.

31.

Harmer, Paul M.

33.

Hoffer, G. N. and R. H. Carr. Iron Accumulation and
Mobility in Diseased Cornstalks. Abst. in Phyto­
pathology 10, 1930.

33.

34.

Soil

Unpublished Manuscript.

________ and J. F. Trost. The Accumulation of Iron
and" Aluminum Compounds in Corn Plants and Its Probable
Relation to Root Rots. Journal Amer. Soc. Agron. 15
pp. 323-331, 1933.
Hopkins, E. F. The Necessity and Function of Manganese
in the Growth of ,,Chlorella,,. Science 73 pp. 609-610,
1 3 3 0 7 ------------------------------

35.

Ingalls, R. A. and J. W. Shive. Relation of H-ion Con­
centration of Tissue Fluids to the Distribution of Iron
in Plants. Plant physiology 6 pp. 103-125, 1931.

36.

Iyer, C. R. Harihara, R. Rajagopalan and V. Subrahmanyan.
Role of Organic Matter in Plant Nutrition II Oxidizing
Agents as Fertilizers. Proc. Indian Acad. Science
pp. 106l§2, 1934.

37.

Jacobson, H. G. M . , and T. R. Swanbeck.
in Tobacco. Science 70, 1939.

38.

Johnson, M. 0. Manganese as a Cause of the Depression of
the Assimilation of Iron b£ pineapple Plants^ Journal
Ind. and Engr. Chem. 9 pp. 47-49, 1917.

39.

______________ . Manganese Chlorosis of Pineapples, Its
Cause and Control. Hawaii Sta. Bui. 52, 1924.

Manganese Toxicity:

4

144.
40.

Jones, Linus H., Relation of Soil Temperature to
Chlorosis of Gardenias. Journal of Agr. Research
1938.

41.

Kelley, W. P., The Function of Manganese in Plants.
Bot. Gaz. 57 p p . 213-27 , l 9 1 4 ~ --------------

42.

Kliman, Stephan. The Importance of Ferrous Iron in
Plants and Soils. Soil Science Soc. of Am. p r o ~
ceedings 3 pp. 385-92, 1937.

43.

Leeper, G. W. Manganese Deficiency of Cereals, Plot
Experiments and a New Hypothesis. Proceedings
Royal Soc. Victoria 47 II pp. 225-61, 1935.

44.

___________ _ and R . J . Swaby . The Oxidation of Manganous Compounds by Microorganisms in the Soil. Soil
Science 49 pp. 163-170, 1940.

45.

Lewis, Ralph W., Ascorbic Acid in Fungous Extracts.
Proc. Mich. Acad, of Science,“7£rts and Letters
Vol. 24, 1939.

46.

Ligon, W. S. Solubility of Applied Nutrients in Muck
Soils and the Composition and Quality of Certain Muck
Crops as Influenced by Soil Reaction Changes and
Moisture Conditions. Mich. State College Agr. Exp.
Sta. Technical Bulletin 147, 1935.

47.

Lyman, Carl M. and E. S. Guzman Barron, The Oxidation
of Glutathione with Copper and Hemochrongens as
Catalysts. Jour, of Biol. ChemT lSl ppT 75, 1937.

48.

Mann, H. B. Availability of Manganese and of Iron as
Affected by Applications of Calcium and Magnesium
Carbonates to the Soil_. Soil Science 30 pp. 117-141,
1930.

49.

Marsh, R. P. and J. W. Shive. Adjustment of Iron Supply
to Requirements of Soybean in Solution Culture. Bot.
Gaz. 79 p p . 1-27~J 1925".

50.

Martin, J. P. Growth of Sugar Cane in Nutrient Solutions.
Hawaiian Planters Record 39 pp. 79-96, 1935.

51.

Maschhaupt, J. G. The Enigma of Leaf-Speck Disease.
Z. Pflanzenernahr. Dungung Bodenk 13B pp. 313-20, 1934.

52.

McCarthy, James F., Lowell F. Green and C. G. King.^ The
Substrate Specificity and Inhibition Characteristics^of
Two Copper Protein ^Sxidases11. Jour. Biol. Chem. 128
pp. 455, 1939.

145.
53.

McCool, M. M.

54.

McGeorge, W. T. The Chlorosis of Pineapple Plante Grown
og^Manganiferous Soil. Soil Science 16 'ppTSgg-gfrl,"

55.

Muckenhirn, R. J. Response of Plants to Boron, Copper
Manganese. Jour. Amer. Soc. Agronomy 28 pp. 824—
42, 1936•

56.

ITago aka, M.^ The Stimulating Action of Manganese Upon
Rice. Bui. Coll. Agr. Tokyo Imp. Univ. 5 pp. 467T
472, 1903*

57.

Neraec, B., and J. Babicka. Chlorosis in Plants Caused
hy Cobalt Salts. Zolastini Otisk Z. Vest Kial Ces
Spal. Nauk. Tran. II R.O.C., 1934.

58.

Oserkowsky, J. Quantitative Relation Between Chlorophyll
and Iron in Green and Chlorotic Pear Leaves. Plant
Physiology 8 pp. 449-468, 1933.

59.

Piper, C. S. The Availability of Manganese in the Soil.
Jour. Agr. Science (England) 2T "ppT 762-779, 193TT

60.

Rudra, M. N. The Role of Manganese in the Biological
Synthesis of Ascorbic Acid. Nature 141 pp.~20J3, 1938.

61.

Ruether, Walter and R. D. Dickey. Preliminary Report on
the Frenching of Tung Trees. Florida Agr. Exp. Sta.
Bui. 318, 1937.

63.

Samuel, G. and C. S. Piper. Grey Speck (Manganese
Deficiency) Disease of Oats. Jour, Dept. Agr. South
Australia. 31 p p . 696-705, 1928.

63.

Schollenberger, C. J., and F. R. Dreibelbis. Effect of
Cropping with Various Fertilizers, Manures and Lime
Treatments Upon Exchangeable Bases of Plot Soils.
Soil Sci. 29 p . 371, 1930.

64.

Scholz, Werner. Chlorosis of Flax in Relation to Iron
and Manganese. Z. Pf lanzenernahr. Dungung Bodenk
34A pp. 296-311, 1934.

65.

The Chlorosis of Blue Lupine and
Serradella in Relation to Iron and Manganese.
Z. Pflanzenernahr. Dungung Bodenk A 35 pp. 88-101,
1934.
______________ . The Chlorosis of Hydrangea in Relation
to Iron. Z. Pf lanzenernahr. Dungung Bodenk 41
pp.“12^-64, 1935.

66.

Effect_ o£ Light Intensity on the Manganese
- Oontrih. Boyce-Thompson Inst . 7
pp. 427-435, 1935.

146.
67.

68

.

Schmitt, L. The Importance of Supplying German Soils
With Potash. Die Ernahrung Der Pflanse~35' No. 12
1939.
SJollema, B. and J , Hudvig. Inquiry Into the Causes
of the Decrease in Fertility of Some Soils in the
Groningen and Drenthe Moor Colonies. Verslag.
Landouwk. Onderzoek. Riikslandbouwproefstat. 5
pp. 29-157, 1909.

69.

Steenbjerg, F. Investigations on the Manganese Content
of Danish Soil. Part III__^ The Relation Between Plant
Growth and the Amount of Exchangeable Manganese in
the Soil. Tids. Planteavl 40 pp. 797-824, 1935.

70.

_____________ • The Exchangeable Manganese in Dani sh
Soils and Its Relation to Plant Growth. Trans. 3rd
International Congr. of Soil Sc. Oxford pp. 198-201,
1935.

71.

Sullivan, M. X. and F. R. Reid. Oxidation in the Soil.
Jour. Indus, and Engr. Chem. 3 pp. 25-30, 1911.

72.

Turk, L. M. Effect of Certain Mineral Elements on Some
Microbiological Activities in Muck Soil. Soil Science
47 pp . 42511446 , 1939.

73.

Thomas, Walter, and Warren B. Mack. Foliar Diagnosis in
Relation to Development and Fertilizer Treatment of the
Potato. Jour, of Agr. Science, 1938.

74.

Van der Merve, A. J. and F. G. Anderson. Chromium and
Manganese Toxicity. Is it Important in Transvaal
Citrus Growing?
Farming South Africa 12 p p . "439-440,
1937

75.

von Feilitzen, H. The Stimulating Effect of Manganese
gaits on Crops. Jour. Landw. 55 pp. 289-292, 1907.

76.

Wilcox, E. V. and W. P. Kelley. The Effect of Manganese
on Pineapple Plants and Ripening of Pineapple Fruit.
Hawaii Agr. Exp. Sta. Bui. 28, 1912•

77.

Wild. A. S. Field Experiments with Manganese as a
firmtrol of Grev Speck Disease in Western Australia.
PP • 223-5, 1934.

78.

Williard, H. H. and L . H. Greathouse, Oolormetric
Determination of Mangane se Using Per i°di£^Acid^as^
0 x idi zing Agent. Jour. Amer. Chem. Soc. 39 pp. 23bb,
1917.
Willis, L. G. The Effect of Liming; Soils, on the Avail­
ability of Manganese and Iron. J. Amer. Soc. Agron.
24 ppT 716-726, 1932.

79.

147
80.

81.

82.

Willis, L. G. and. J. R. Pi land. The Function of Copper
in Soils and. Its Relation to the Availability of Iron
and Manganese. J~. Agr. Research 52 pp. 467-76, 1936.
and
. Some Recent Observations
on the Use of Minor Elements in North~Carofina Agri­
culture . Soil Science 44 pp. 251-63, 1937.
Woodward, Gladys E., and Edith G. Fry. The Determination
of Blood Glutathione. Jour. Biol. Chem. 97 pp. 465482, 1932.