ZINC LEVELS IN SOILS AS RELATED TO
ZINC UPTAKE AND YIELD OF
PHASEOLUS VULGARIS

Thesis for the Degree of Ph. D.
MICHIGAN STATE UNIVERSITY
JAMES RAY MELTON
1968

 

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Zinc Levels in Soils as Related to Zinc Uptake
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ABSTRACT

ZINC LEVELS IN SOILS AS RELATED TO ZINC UPTAKE
AND YIELD OF PHASEOLUS VULGARIS

by James Ray Melton

Experiments were conducted to survey the zinc status
of various Michigan soils, to study the effect of zinc
fertilization on yield and zinc uptake by pea beans, to
correlate various zinc soil tests with yield by pea beans,
and to study the effect of lime applications on yield and
zinc uptake by pea beans. Field, greenhouse, and laboratory
studies were initiated in order to fulfill these objectives.

Measurements of total (12.0 g hydrochloric acid) and
extractable (0.1 §_hydrochloric acid) zinc did not appear
to separate those soils on which yield reSponses to zinc had
been obtained from those on which no responses had been
obtained.

On all soils, highest yields were obtained when the zinc
concentration in plants was between 25 and 54 parts per
million. Yields were depressed by zinc toxicity when plant
zinc concentration was greater than 50 parts per million.

On the basis of both greenhouse and field studies, a

0.1 §_hydrochloric acid extraction procedure was found to

James Ray Melton

be a good soil test for plant available zinc in Michigan.
It should be desirable, however, to initiate further field
studies to further evaluate the 0.01 M disodium ethylene—
diaminetetraacetate + 1.0 M ammonium carbonate extractant as
a zinc soil test.

Generally,yields were increased when zinc fertilizers
were applied after liming soils which tested pH 6.3 or below.
It is quite probable that zinc deficiency could be induced

by liming certain Michigan soils.

ZINC LEVELS IN SOILS AS RELATED TO ZINC UPTAKE

AND YIELD OF PHASEOLUS VULGARIS

 

BY

James Ray Melton

A THESIS

Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of

DOCTOR OF PHILOSOPHY

Department of Soil Science

1968

To My

Wife

ii

ACKNOWLEDGMENTS

Sincere appreciation is expressed to Drs. B. G. Ellis
and E. C. Doll for their helpful guidance throughout the
course of this investigation, and to Dr. E. C. Doll in the
preparation of the thesis.

The writer is also grateful to Dr. B. D. Knezek for
his suggestions during the investigation and preparation
of the thesis.

Grateful acknowledgment is given to Dr. L. S. Robertson
for his helpfulness in determining sites for collection of
soils and to Professor I. F. Schneider for classification
of these soils.

A note of appreciation is extended to Dr. Don F. Wagner
and Mr. Fred Brinkerhoff for taking photographs of various
portions of the greenhouse experimentation.

Appreciation is extended to Mrs. Nellie Galuzzi and to
Mrs. B. H. Quinlan for their efforts in statistical analysis
of data.

To the other faculty members and fellow graduate stu-
dents in the Soil Science Department who assisted during this

study go my profound thanks.

 

iii

TABLE OF CONTENTS

INTRODUCTION . . . . . . . . . . . . . . . . . .
REVIEW OF LITERATURE . . . . . . . . . . . . .

Role of Zinc in Plants. . . . . . . . . . .
Zinc in Soil. . . . . . . . . . . . . . . .
Factors Affecting Zinc Availability . . .
PhOSphorus . . . . . . . . . . . . . .
Nitrogen . . . . . . . . . . . . . . .
Other Nutrients. . . . . . . . . . . .
Soil Reaction. . . . . . . . . . . . .
Carbonates . . . . . . . . . . . . . .
Type of Nutrient on Clay . . . . . . .
Temperature. . . . . . . . . . . . . .
Procedures for Extracting Zinc From Soils .
Extraction of Total Zinc . . . . . . .
Extraction of Available Zinc . .

EXPERIMENTAL METHODS . . . . . . . . . . . . . .

Soil Analytical Methods . . . . . . . . . .
Soil pH. . . . . . . . . . . . . . . .
Extractable PhOSphorus . . . . . . . .
Extractable Potassium, Calcium, and

Magnesium . . . . . . . . . . . .
Zinc Determinations. . . . . . . . . .

Greenhouse Studies. . . . . . . . . . . . .

Plant Analytical Methods. . . . . . . . . .

Field Studies . . . . . . . . . . . . . . .

Statistical Procedures. . . . . . . . . . .

RESULTS AND DISCUSSION . . . . . . . . . . . . .
Total and Extractable Zinc in Sixty-Five
Michigan Soils . . . . . . . . . . . .

Yields and Zinc Content of Pea Beans in the
Greenhouse . . . . . . . . . . . . . .

iv

Page

TABLE OF CONTENTS

Soil

- Continued

First CrOpping. . . . . . . . . . . . . .
Yield. . . . . . . . . . . .
Zinc Concentration . . . . . . . . .
Zinc Uptake. . . . . . . . . . . . .
Second CrOpping . . . . . . . . . . . . .
Yield. . . . . . . . . . . . . . . .
Zinc Concentration . . . . . . . . .
Zinc Uptake. . . . . . . . . . . . .
Third CrOpping. . . . . . . . . . . . . .
Yield. . . . . . . . . . . . . . . .
Zinc Concentration . . . . . . . . .
Zinc Uptake. . . . . . . . . . . . .
Between CrOps . . . . . . . . . . . . . .
Yield. . . . . . . . . . . . . . . .
Zinc Concentration . . . . . . . . .
Zinc Uptake. . . . . . . . . . . . .
Zinc Extraction Procedures. . . . . . . .
Zinc Extracted From Soils Before CrOpping
Zinc Extracted from Soils After Final
Cropping . . . . . . . . . .'. . . .

Relationship Between Extractable Soil Zinc and

Yield and Zinc Uptake by Pea Beans. . . .

Field Results. . . . . . . . . . . . . . . . .

SUMMARY AND CONCLUSIONS . . . . . . . . . . . . .

LITERATURE CITED. . . . . . . . . . . . . . . . .

APPENDIX.

Page

52
52
57
38

38
40

4O
43
43
45
44
44
45
45
46
46
47
47

52

55
61

65
72

82

LIST OF TABLES

Table

l.

10.

1A.

Extractable nutrients, pH, and lime requirement
of 20 Michigan soils used in greenhouse and
laboratory evaluations. . . . . . . . . . . . .

Methods of extracting soil Zn to correlate with
yields of pea beans . . . . . . . . . . . . .

Total and extractable Zn and pH at various
depths in 65 Michigan soils which include 35
soil series from 8 soil suborders . . . . . . .

. Yield and Zn content of successive crops of pea

beans grown on 20 different soils at 5 levels
of Zn application . . . . . . . . . . . . . . .

Effects of greenhouse cropping and of lime ap-
plication on pH values of soils . . . . . . . .

Amount of Zn extracted from Michigan soils by
the various procedures listed in Table 2 prior
to crOpping . . . . . . . . . . . . . . . . . .

. Amount of Zn extracted from Michigan soils by 4

extractants before and after growing 3 crops of
pea beans. Rates of lime application are given
for those soils that were limed . . . . . . . .

Linear and multiple correlation coefficients
between relative yield of the first crop of pea
beans and extracted soil Zn (Zn) and extract-
able soil Zn plus soil pH (Zn 4 pH) . . . . . .

Linear correlations between extractable Zn, pH,
and Zn Uptake by pea beans on 20 Michigan soils
used in greenhouse and laboratory evaluations .

Yield of pea beans, soil pH, and concentration
of Zn extracted from 15 Michigan soils utilized
for field studies . . . . . . . . . . . . . . .
Suborders and series, texture, depth, pH, ex-
tractable Zn, and county of 65 Michigan soils .

vi

Page

21

24

30

33

41

48

53

57

62

63

82

LIST OF FIGURES

Figure Page

1.

Relation between Zn concentration and relative
yield of the first crop of pea beans grown in
the greenhouse. . . . . . . . . . . . . . . . . 39

Relation between relative yield and 0.01 M
EDTA + 1.0 M_(NH4)2C03 extractable Zn at pH 7.0
and pH 7.5. . . . . . . . . . . . . . . . . . . 59

Relation between relative yield and 0.1 N HCl
extractable Zn at pH 7.0 and pH 7.5 . . . . . . 60

Relation between relative yield in the field
and Zn extracted from soils by 0.1 N HCl. . . . 64

vii

INTRODUCTION

The essentiality of zinc (Zn) for growth of the fungus

Aspergillis niger was first shown by Raulin (1869), and later

 

confirmed by Bertrand and Javillier (1911). Brechley (1914)
described Zn deficiency in higher plants, but Zn toxicity
was considered more important at that time.

In the higher plants, Mazé (1915) reported that Zn was
essential for the growth of corn. The need for Zn in higher
plants was not universally accepted, however, until Sommer
and Lipmann (1926) demonstrated the essentiality of Zn for
sunflower, broad beans, kidney beans, and barley. Bonner
and Varner (1965) recently reported that Zn is also essential
for many Species of algae and for many nonphotosynthetic
organisms.

Zinc deficiency in Phaseolus vulgaris var. Sanilac (pea

 

beans) has been identified in Michigan (Ellis et al., 1964a).
Many pea beans (approximately 1/2 million acres) are grown
on soils which test above pH 7.0 in the lake-plain area of
Eastern Central Michigan. Heavy rates of phOSphorus (P)
applied to preceding sugar beet crops are believed to accentu-
ate Zn deficiency symptoms.

Zinc fertilizers that have been effective for pea beans

include the sulfate, chloride, carbonate, nitrate, oxide,

oxysulfate, and phOSphate salts of Zn, and Zn-containing
materials such as blast furnace slag, stripping acid resi-
dues, and frits. Polyaminocarboxylic acid chelates and
organic extracts such as polyflavanoids have been success-
fully used as organic carriers of Zn. Zinc sulfate was
found to be one of the most effective and economical sources
of Zn in Michigan (Brinkerhoff et al., 1966 and 1967; Judy
et al., 1965).

Michigan experiments concerned with the effectiveness
of various kinds of Zn fertilizers for plant growth, the
depressing effect of high P levels on plant uptake of Zn,
and the increase in uptake of Zn by plants as temperature was
increased have previously been reported (Judy, 1965: Brinker-
hoff et al., 1966 and 1967; and Ellis et al., 1964). However,
the relation between the amounts of Zn extracted from Michigan
soils by various methods and the yield and Zn Uptake by plants
have not been adequately established. Consequently, the in-
vestigations reported herein were conducted to:

1. Survey the Zn status of various Michigan soils.

2. Study the effect of Zn fertilization on yield and

Zn uptake by pea beans.
5. Correlate various Zn soil tests with yield of pea
beans.
4. Study the effect of lime applications on yield and

Zn uptake by pea beans.

REV IEW OF LITERATURE

Most soils contain micronutrients in sufficient quanti-
ties to sustain normal plant growth. Deficiencies occur on
some soils, however, because of crop removal, leaching,
chemical fixation, erosion, or an initial lack of primary

minerals which contain these nutrients.
Role of Zinc in Plants

A concentration of approximately 20 ppm Zn in plant tOps
appears to be optimal for normal plant growth and metabolism
(Bonner and Varner, 1965). Zinc deficiency symptoms usually
develop when Zn content is below 15 to 20 ppm (Hiatt and
Massey, 1958; Nelson, 1956; Viets et al., 1954). Rosetting
of fruit trees, mottle leaf of citrus, little leaf of beans
(Stiles, 1946), white bud of corn (Barnette et al., 1936,
Stiles, 1946), and fern leaf of potato (Boawn and Leggett,
1965) are typical of the types of Zn deficiency symptoms
that have been reported.

The primary role of Zn in the plant is as a catalyst
(Schutte, 1964). Zinc is located in the prosthetic group of
carbonic anhydrase (Day and Franklin, 1946) and is associated
with aldolase activity in subterranean clover (Quinlan-Watson,

1953). The activity of NADase decreases in fungi as Zn

concentration is increased (Nason and Erans, 1951). Hagi
and Vallee (1960) suggested that Zn might act as a bridge
between protein and pyridine nucleotide in yeast alcohol

dehydrogenase.

Zinc may be involved in chlorophyll synthesis and rate
of transpiration (Schutte, 1964). Chester and Robinson
(1951) postulated that Zn may be involved in the activity
of the auxin 6-indole acetic acid (IAA). Tsui (1948) and
Nason et al. (1951) also believed that Zn was related to
auxin synthesis as deficient plants were low in the IAA pre-
cursors tryptophan and tryptophan synthetase.

Zinc is essential for seed production in many plants,
and is a component of glycylglycine dipeptidase and dihydro-
peptidase, enzymes involved in protein metabolism (Seatz and
Jurinak, 1957). Smaller quantities of free amino N, amide
N, and inorganic P compounds were found in plants which con-
tained adequate Zn (Possingham, 1954; Reed, 1946).

Several morphological and physiological changes were
found in plants when the level of metabolically active Zn
was inadequate (Seatz and Jurinak, 1957). Among these
changes were: 1) palisade cells of leaves were abnormally
large and were transversely rather than columnarly divided,
2) number of chloroplasts were reduced, 3) starch grains
were absent, 4) oil droplets formed in chloroplasts, and
5) phenolic materials and calcium oxalate crystals accumu-

lated in leaves. Visual deficiency symptoms in plant t0ps

are interveinal chlorosis, necrosis of lower leaves, and
shortening of internodes. Deficiency of zinc in roots was
indicated by abnormal shape and large amounts of fats and
tannins but no starch (Seatz and Jurinak, 1957).

Some examples of plant response to Zn were: 1) the
top:root ratio was greater in plants adequately supplied
with Zn (Millikan, 1963), 2) zinc was not translocated in or
from older tissue (Shaw et al., 1954), 3) upper leaves of
corn contained more Zn in both normal and deficient plants
than in lower leaves (Viets et al., 1953), 4)zinc was located
mostly in and around the primary veins in the corn leaf
blade (Sayre, 1952), 5) meristamatic tissue of pineapple
contained the largest concentrations of Zn (Lyman and Dean,
1942), 6) zinc requirement of vegetative parts of peas and
beans was much lower than during seed production (Reed,
1942), and 7) an equivalency of Zn and other bases was found
throughout snap bean plants near early bloom stage (Seatz

et al., 1956).

Zinc In Soil

In most soils, more Zn is found in the A1 horizon than
in any of the lower horizons. Total content of Zn varies
greatly between soil groups (Mitchell, 1964). The total Zn
content of soils varies from 10 to 300 ppm, and only part
of this Zn is available for plant growth (Swaine, 1955).

In the U. 8., Zn deficiencies have been reported in 31 states

(Berger, 1962). Soils with adequate available Zn contained
more than 10 ppm available Zn, and deficient soils contained
less than 2 ppm Zn available for plant growth as determined
by the Aspergillus niger extraction (Bould et al., 1963).

Zinc is commonly found in the primary minerals biotite,
hornblende, magnetite, and the ferromagnesian group (Bould,
1963). Smithsonite, willemite, Zn blende, and calamine,
which are primary Zn-containing minerals, occur in the soil
in minute quantities. More available Zn is found in acid
soils derived from granite than in calcareous soils derived
from limestone (Thorne et al., 1942).

According to Bould (1963), Zn is adsorbed as a divalent
cation on clays or complexed by organic matter after release
from the minerals. Elgabaly (1950) stated that Zn may enter
the inner layer of the electrical double layer of a micelle
where it cannot be exchanged for neutral inorganic salts
such as ammonium acetate. Elgabaly and Jenny (1943) postu-
lated that Zn clay has a mosaic surface capable of independent
cation and anion exchange, and that non-replaceable Zn is
inside empty oxygen and hydroxyl octahedra of the brucite
layer of the montmorillonite clay crystal. However, Nelson
and Melsted (1955) stated that Zn was not fixed on cation
exchange positions and was not adsorbed as a complex ion in
the electrical double layer in Illinois soils.

Bingham et al. (1964) postulated that Zn is precipitated

as Zn(0H)2 in the clay systems, and Bernheim and Quintin

(1950) reported that the bi-zincate ion, Zn02=, became more
prevalent as the base concentration increased. DeMumbrum

and Jackson (1956) found that Zn reacted with the octahedral
hydroxide in layer silicates but did not react with kaolinite.
Jurinak and Thorne (1955) prOposed that both chemical and
strong clay absorption complexes and Zn hydroxide were

formed in soils.

Carboxyl and phenolic groups in soil organic matter have
been shown to be involved in chelation (Broadbent et al.,
1952; Himes et al., 1963). According to Himes and Barber
(1957), removal of organic matter from soil by oxidation with
hydrogen peroxide destroyed Zn-chelating ability of these
carboxyl and phenolic groups. Zinc saturation of peat frac-
tions resulted in numerous shifts in the double bond region
of the infrared spectrum, indicating a chelation with N=0
and C=0 groups (DeMumbrum and Jackson, 1956). Randhawa and
Broadbent (1965) found that humic acid complexed very little
Zn at pH values less than 3.6, but complexing ability in-
creased rapidly with an increasing hydroxyl ion concentration
up to pH 8.5. Miller and Ohlrogge (1958) discovered that
water-soluble chelating agents in organic materials complexed
more Zn at higher pH values. Ark (1936) reported that
steam sterilization of Zn deficient soils released sufficient
Zn to correct the deficiency, thus implying fixation by
microbial action. Mortensen (1963) concluded that surface
adsorption, chelation, ion exchange, and peptidization were

involved in complexing of Zn by soil organic matter.

According to Hibbard (1940a), removal of organic

matter from soils by ashing or H202 treatment did not affect
the solubility of soil Zn. Hodgson et al. (1966) found that
organic matter complexed 28 to 99 percent of the Zn in the
soil solution; however, they found only a small proportion
of total Zn in the soil complexed by soil organic matter.
Geering and Hodgson (1966) showed that addition of citrate
to a carbonate-saturated water system drastically increased

Zn movement.
Factors Affecting Zinc Availability

Soil Zn may be classified as water-soluble, replaceable,
and nonreplaceable (Jones et al. 1936). In addition to
organic matter and clay, such factors as replaceable bases,
carbonates, phOSphates, type of nutrient on clay, and soil
temperature influence Zn distribution within these three

fractions.

PhOSphorus

Bingham et al. (1958 and 1960) related soil type to
P-induced Zn deficiency; high levels of residual or ferti-
lizerl’were required before yields were reduced on some soils.
A reduction in both P and Zn uptake was found in kidney
beans, corn, and tomatoes when P was applied to limed soils
(Burleson et al., 1961; Ward et al., 1963). This reduction
in uptake seldom occurred on unlimed soils. Millikan (1963)

proposed that depression in plant growth was due to a high P

concentration rather than a low Zn concentration. Boawn and
Leggett (1963 and 1964) suggested that Zn deficiencies were
more closely associated with high P:Zn ratios than with low
concentrations of soil Zn. The critical P:Zn ratio was 400:1
for Russet Burbank potatoes (Boawn et al., 1964) and 300:1
for corn (Watanabe et al., 1965). Some researchers (Ward
et al., 1963: Burleson et al., 1961; Stukenholtz et al.,
1966) suggested a P-Zn antagonism within the root, but
Bingham (et al., 1963) concluded that a reaction outside the
root contributed to this antagonism.

Boawn et al. (1954) stated that the amount of extract-
able P in the soil was not related to appearance of
Zn deficiency symptoms in field beans. Seatz et al. (1959)
showed that up to 436 pounds of applied P per acre had no
effect on Zn reSponse. Zinc uptake by citrus increased as
more P was applied (Bingham, 1963) and soluble Zn was in-
creased by additions of 900 pounds of K, H, NH4, and Ca
phOSphates (Bingham et al., 1960). Nelson et al. (1965)
found that wheat showed no Zn deficiency symptoms or yield
depression at several levels of applied P when grown on a

soil testing pH 7.4.

Nitrogen

Many crOps have shown an increased Uptake of soil Zn
after nitrogen application (Ellis et al., 1964: Nelson et al.,
1962). This enhancement of uptake was attributed to the

acidification effect of the nitrogen fertilizer (Boawn et al.,

10

1960). Zinc uptake by subterranean clover, however, was
reduced by nitrogen application (Ozanne, 1955), and this
affect was attributed to formation of Zn-protein complexes

in the root.

Other Nutrients

 

Magnesium has been shown to increase Zn Uptake by bean
plants (Seatz, 1960), and a mutual substitution effect be-
tween Zn and Mg has been prOposed (Barrows et al., 1960;
Merrill et al., 1953). Other nutrient-relationship studies
showed that a high K:Ca ratio in a calcareous soil resulted
in Zn deficiency symptoms while a low K:Ca ratio resulted
in Fe deficiency symptoms (Greenwood et al., 1951). Also,
Jurinak and Thorne (1955) showed that Na and K increased Zn
solubility, but Ca decreased Zn solubility in soils. Ward
et al. (1963) reported that K level in the soil was a very

important factor governing P-induced Zn deficiency.

Soil Reaction

 

Camp (1945) found the best utilization of Zn from soils
at pH values of 6.0 to 6.5. Shaw and Dean (1951) observed
that non-chelated Zn was practically unextractable at pH
7.0 to 8.5 (dithizone extraction). Jurinak and Thorne (1955)
showed Zn solubility in Na and K bentonite systems to be
lowest between pH 5.5 and 6.7, but increased at higher pH
values. However, Zn solubility in a Ca bentonite system

did not increase at higher pH values; in fact, minimal

11

solubility was found at pH 7.6. The authors suggested that
these effects were due to differential solubility of alkali

zincates and of calcium zincate.

Carbonates

 

The success by Nelson et al. (1959) in evaluating Zn
status of soils suggests that CaCOa content, as well as soil
pH, may help to determine Zn availability in soils. Rogers
et a1. (1948) postulated that liming reduced Zn uptake in
plants by increasing the pH.

Jurinak and Bauer (1956) found that 10 percent of the
adsorption sites on calcite were occupied by Zn when the
aqueous Zn++ equilibrium concentration was 0.90 x 10'6 M_at
25.10 c. The relative degree of affinity for Zn was found
to be: magnesite > dolomite > calcite. Endothermic heats
of adsorption and very large positive entrOpies of adsorp-
tion indicate that aqueous Zn++ is dehydrated when adsorbed
by dolomite and magnesite. Compatability of Zn++ with the
MgCOs crystal lattice was thought to be a possible reason
for the stronger Zn interaction with dolomite and magnesite

than with calcite.

Type of Nutrient on Clay

Pretreatment of clays with Ca, K, and Cu solutions re—
sulted in Zn adsorption in the order: K-clay > Ca-clay >
Cu-clay (Mangaroo et al., 1965). Ammonium acetate could

replace Zn added to a H-saturated soil system but could only

12

partially replace Zn added to a Ca—saturated soil system.
When Zn was applied to a Ca—saturated soil system, the
amount of Zn extracted with NH4OAc decreased with time

(Nelson et al., 1955).

Temperature

 

Ellis et al. (1964) showed that yield, Zn concentration
in plant tissue, and Zn Uptake by corn decreased when soil
temperature decreased from 750 F to 550 F. Also, Martin
et al. (1965) found that low soil temperature accentuated

P-induced Zn deficiency.
Procedures for Extracting Zinc From Soils

The average Zn concentration in the lithOSphere is
estimated to be about 80 ppm. Viets et al. (1953) found
that total Zn in a loam soil averaged 66 ppm to a depth of
3 feet. They calculated that this soil contained 400 pounds
per acre total Zn which is approximately 2,000 times the
annual needs of an agronomic crOp. Since much of this total
Zn is not available to the plant, methods of determining

both total Zn and plant available Zn were needed.

Extraction of Total Zinc

Wahhab and Bbatti (1959) digested several West Pakistan
soils in 72 percent perchloric acid, and then extracted with
1.0 N_HCl. Amounts of total Zn extracted were between a
trace and 15.0 ppm with sands, and from 17.5 to 87.5 ppm

with the sandy loams and clays.

13

Nair and Mehta (1958) who analyzed 58 soils from
Western India, reported total Zn values ranging from 49.5
to 80.6 ppm, as determined by the HNOa-HF method of Holmes
(1943).

Holmes (1943) extracted from 11 to 140 ppm total Zn
from soils throughout the United States. Southwestern soils
contained 18 to 135 ppm, Central soils contained 40 to 130
ppm, Southeastern soils contained 11 to 147 ppm (only 2
New Hampshire soils studied) and Northwestern soils contained
66 to 105 ppm total Zn. Total Zn was fairly uniform between
most profiles studied, eSpecially those from the Central
United States.

Alben and Bogg (1936) reported that soils utilized for
pecan orchards in Texas and Louisiana had from 18 to 252 ppm
total Zn, with an average of about 60 ppm total Zn in the
tOpsoil (determined by fusion of soil with potassium pryo-
sulfate and disintegrated in hot HCl). Only about one-half
as much total Zn was extracted from soils at 0 to 6 inches
as was extracted from soils at a depth of 12 to 72 inches
in the soil profile.

Sherman and McHargue (1942) extracted from 80 to 513
ppm total Zn from Kentucky soils with a mixture of hydro-
fluoric and perchloric acids. Total Zn content of these same
soils was 78 to 502 ppm as determined by Na2C03-K2C03
fusion. Takazaua and Sherman (1947) found no correlation
between total Zn determined by the mixed Na2C03-K2C03 fusion

and plant available Zn.

14

According to Woltz et al. (1953), the total Zn content
of twenty agricultural soils of New Jersey varied from 10
to 225 ppm. Zinc content of Appalachian soils averaged 100
ppm while that of Coastal Plain soils averaged 41 ppm.
Heavy-textured soils generally contained more total Zn than
light-textured soils.

Total Zn in 42 surface soils from Southern and Central
Wisconsin varied from 4 ppm in a peat soil to 109 ppm in a
silt loam. The average total Zn content of approximately
60 ppm agrees closely with reported data from other soils

throughout the United States (Stewart and Berger, 1965).

Extraction of Available Zinc

Total Zn determinations have not adequately evaluated
the level of plant available Zn in soils. Therefore,
scientists have investigated soil extractions with micro-

organisms such as ASpergillus niger, weak extracting agents

 

like water, ammonium acetate, and magnesium sulfate, or
stronger extracting agents such as HCl, dithizone, and EDTA
to devise a suitable procedure for measuring plant available
Zn in soils. Many soil testing laboratories have based
their Zn—application recommendations on quantity of Zn ex-
tracted by some chemical reagent and other considerations
such as pH, carbonate content, kind of crOp, humus content,
or soluble P.

Burd and Martin (1923) percolated water through soils

(previously moistened approximately to field capacity) in

15

filtration tubes and determined the quantity of certain
macronutrients in the leachate. Hibbard (1940), utilizing
a similar experiment, concluded that pure water has almost
no solvent power for soil Zn. However, he extracted less
Zn with water from soils previously giving evidence of Zn
deficiency, the range being from 0 to 21 ppb (parts per
billion).

A dithizone (diphenylthiocarbazone) method for measure-
ment of small quantities of Zn was deveIOped by Hibbard
(1937). Epstein and Stout (1951) used dithizone to extract
Zn from dilute clay su5pensions. Shaw and Dean (1951) modi-
fied the extraction procedure by directly extracting Zn from
soil samples with a two-phase system of aqueous NH4OAc and
carbon tetrachloride (CC14) containing dithizone. Of the 52
soils studied by Shaw and Dean, less Zn was generally ex-
tracted by dithizone from soils deficient in Zn for plant
growth than from soils sufficient in Zn for plant growth.
There was from 0.5 to 17.0 ppm Zn extracted, with most soils
releasing from 0.3 to 3.0 ppm Zn. These values are similar
to those that other workers (Stewart et al., 1965: Massey,
1957) reported when using the same dithizone extraction pro-
cedure (Wisconsin and Kentucky soils). Massey (1957) found
a good correlation between Zn Uptake by corn plants and
dithizone extraction.

Wear and Sommer (1948) found a good correlation between

the occurrence of Zn deficiency symptoms and quantity of Zn

16

extracted with 0.1 N_HC1 or 0.04 N acetic acid (HOAc) from
acid soils of Alabama. On Zn deficient soils, 0.05 to 0.09
ppm Zn was extracted with 0.1 N_HC1 and 0.00 to 0.50 ppm Zn
was extracted with 0.04 N_NH4OAC. However, on soils with
sufficient Zn, 1.20 to 4.70 ppm Zn was extracted with 0.1 N
HCl and 0.05 to 3.50 ppm Zn was removed by 0.04 N NH4OAc.

Hibbard (1940) found that either HCl or H2804, in con-
centrations as dilute as 0.01 N, extracted much more Zn than
that present in the soil solutions. Other investigators
(Nelson et al., 1959; Hoover, 1966) found that quantities of
Zn extracted from soils with 0.1 N_HC1 did not correlate well
with crop reSponse, eSpecially on highly calcareous soil.
However, Nelson et al. (1959) found a high correlation be-
tween plant deficiency symptoms and 0.1 N HCl extractable Zn
in calcareous soils if the "titratable alkalinity" was taken
into consideration. Stewart and Berger's (1965) data from
0.07 to 13.70 ppm Zn extracted from Wisconsin soils with 0.1
N HCl agreed closely with the 0.9 to 12.0 ppm extracted by
Nelson et al. (1959).

The additional Zn extracted by 0.1 N_HC1, as compared to
that extracted with dithizone, was suggested by Martens et al.
(1966) to be held by organo-clay complexes. They concluded
that the Zn held in these complexes was less available to
plants than that extracted by dithizone.

Zinc extracted from Texas soils with 0.1 M_copper sul-

fate (CUSO4) and 0.1 N_HC1 was significantly correlated with

17

Zn content of sorghum leaves (Hoover, 1966). Zinc levels
in soils used in this study varied from 0.3 to 17.8 ppm Zn.
Trierweiler and Lindsay (1966), using an ammonium
carbonate (NH4)C03-LEDTA extraction, found a good correla-
tion between extractable Zn and plant reSponse on Colorado
soils. Hoover (1966) found that extraction of Zn with 1
percent EDTA was significantly correlated with Zn content

of sorghum leaves from plants grown on Texas soils.

Of the numerous solvents used by Hibbard (1940) for
extracting Zn from soils, a saturated water solution of
carbon dioxide (C02) and 0.5 N_KC1 plus 0.04 N_HOAc was
selected. He later used 0.05 N KCl adjusted to pH 3.2 with
HOAc. In general, only 1 to 5 ppm Zn was extracted from
California soils by this procedure: however, Up to 32.7
ppm Zn was removed from one soil at the 0—2 inch depth
(Hibbard 1940a). Lyman and Dean (1942) found Zn extracted
by C02 saturated water was highly correlated with soil pH
but not with Zn Uptake by pineapple plants. Koter et al.
(1965) found that the quantity of Zn extracted with 1.0 N
KCl correlated well with the quantity of Zn taken up by
wheat, oats, and rye plants.

A good correlation was found between Zn deficiency ex-
hibited by pineapple plants and soil Zn soluble in NH4OAc
(HOAc brought to a pH of 4.6 with NH4OAc). The Hawaiian

soils contained from 0.5 to 3.5 ppm extractable Zn.

18

Stewart and Berger (1965) found a higher correlation
between Zn Uptake by plants and Zn extracted from the soil
by 2.0 N_MgC12 than with 0.1 N HCl or NH40Ac-dithizone combi-
nations. Surface samples from 42 locations in South and
Central Wisconsin were used in this study: extractable Zn
in these soils varied from 0.60 to 3.98 ppm.

Martens et al. (1966) reported highly significant rela-
tionships between Zn uptake by corn plants and Zn extracted
from the soil by each of the following methods: 0.2 M_MgSO4,
dithizone, Aspergillis niger, and total Zn. Organic carbon
content was also highly significantly correlated with Zn Up-
take by corn plants. These workers felt that 0.2 M_MgSO4
was the most satisfactory extractant.

Vlasyak and Zimina (1954) reported that the lowest
amount of available Zn in Russian soils was 18.5 to 24.1 ppm
in the cultivated podzols, and that the highest amount was
87 to 140 ppm in the Solonetz and Solonchak soils. This ob-
servation suggests that either available Zn levels in Russian
soils are much higher than those in the United States soils
or that they are actually determining total Zn.

Nair and Mehta (1958) found 1.79 to 4.57 ppm dithizone-
extractable Zn in 58 soils of Western India. These values
agree closely with those found in United States soils using
the same extractant.

Ravikovitch et al. (1968) used 7 different methods to

extract Zn from 15 calcareous soils of Israel. Ammonium

19

nitrate (1.0 N), 1.0 N KCl, and 0.01 N_disodium ethylene-
diamine di (O-hydroxyphenyl acetic acid) in 1.0 N_NH4OAc
gave the most significant multiple correlation coefficients

with Zn Uptake by six crOps.

EXP ER IM ENTAL METHODS

The levels of extractable Zn in the soils of Michigan
were evaluated by obtaining samples from various depths at
65 locations which varied in pH and texture. These soils
(Table 1A in the Appendix) represent 34 soil series from the
Udalf, orthod, aquept, baralf, aqualf, aquod, aquoll, and
Udoll soil suborders (see Soil Survey Staff, 1960 if defini-
tions are desired).

Twenty of the original 65 soils which varied in extract-
able Zn level, soil pH, and soil texture were selected for
further study in both the laboratory and in the greenhouse
(Table 1). Approximately 150 pounds of soil was obtained

from the Ap horizon at each of the 20 locations.
Soil Analytical Methods

The 65 soils were sampled by horizons at depths given
in Table 1A and placed in paper bags, air dried, passed
through a 20 mesh plastic sieve, and stored in cardboard
containers. Soil pH, lime requirement, and available P, K,
Ca, and Mg were determined by the Michigan State University

Soil Testing Laboratory.

Soil pH

Ten grams of soil were mixed with 10 ml of water (1:1

ratio). After 15 minutes, the mixture was stirred again,

20

21

Table 1. Extractable nutrients, pH, and lime requirement of
20 Michigan soils used in greenhouse and laboratory
evaluations.*

—A__._.

Levels of Nutrients Extracted pH Lime

 

 

Soil Suborder P K Ca Mg Requirement
and Series pp2m pp2m pp2m pp2m pp2m
Aqualf:
Conover 63 189 4766 493 7.1 0.0
Metamora I 44 248 1604 101 6.2 4,000
Aquept:
Breckenridge 84 118 5472 644 7.4 0.0
Charity I 79 312 8135 375 7.7 0.0
Charity II 47 432 8848 405 7.9 0.0
Hettinger 91 248 6621 966 7.5 0.0
Sims I 21 264 6336 950 7.3 0.0
Sims II 96 112 3910 375 7.6 0.0
Wisner I 47 204 6480 405 7.8 0.0
Wisner II 167 264 5760 934 7.8 0.0
Aquod:
Brimley I 43 126 4057 433 0.0
Brimley II 135 118 3763 331 7.4 0.0
Aquoll:
Colwood I 71 232 6336 1016 7.5 0.0
Lenawee I 63 132 5616 757 6.9 0.0
Orthod:
Kalkaska 54 43 800 29 5.9 3,000
Karlin III 234 70 400 43 5.7 6.000
Udalf:
Hillsdale 20 84 1752 130 6.6 0.0
Hodunk 84 183 1456 259 6.3 3,000
Locke 43 197 800 101 5.8 3,000
Miami 58 162 4490 302 7.2 0.0

 

*
Determined by the Michigan State University Soil Testing
Laboratory.

22

and the pH of the SUSpension determined using a Beckman
Zeromatic glass electrode pH meter. The lime requirement
of samples testing below pH 7.0 was determined by the method

of Shoemaker, McClean, and Pratt (1961).

Extractable PhOSphorus

PhOSphorus was extracted for 1 minute from samples
with Bray P-l reagent (0.025 N HCl and 0.05 N NH4F), using
a 1:8 soil-solution ratio. Activated charcoal was used to
remove organic matter from the filtrate. PhOSphorus in the
extract was determined by the molybdophOSphoric blue method,
using 1,2,4-aminonaphtholsulfonic acid as the reductant

(Jackson, Method IV, p. 148, 1958).

Extractable Potassium, Calcium and Magnesium

Cations were extracted for 1 minute with 1.0 N NH40Ac
(pH 7.0) using a 1:8 soil-solution ratio. Potassium in the
extract was determined by means of a Coleman Model 21 flame
photometer, Ca by means of a Beckman Model DU flame Spectro-
photometer, and Mg by means of a Perkin Elmer Model 290
atomic absorption Spectrophotometer as described by Doll

and Christenson (1966).

Zinc Determinations

Total soil Zn was determined on all 65 soils by boiling
5 grams of? soil in 50 ml of 12 N HCl. The su5pension was
boiled until approximately 5 ml of solution remained, then

filtered (Whatman No. 3 filter paper) into 200 ml volumetric

23

flasks. Zinc was determined in the resulting solution using
a Perkin Elmer Model 303 atomic absorption SpectrOphotometer.
Zinc removed by this method was nearly equal to that removed
by hydrofluoric and perchloric acids (Sherman and McHargue,
1962) and to that removed by sodium carbonate fusion (Jackson,
1958).

The different procedures listed in Table 2 were compared
to determine the most effective method of extracting available
soil Zn. Zinc in the extracts was determined as described

above.
Greenhouse Studies

Greenhouse studies, using the 20 soils listed in Table 1,
were conducted to determine the effect of Zn applications and
pH levels on yield and Zn Uptake, and to determine the rela-
tionship between Zn extracted and plant yield and Zn uptake
.by pea beans. Three successive crOps were grown.

The greenhouse eXperiment was laid out in a completely
randomized design with each treatment on each soil being con-
ducted in quadruplicate. One-gallon galvanized cans, lined
with plastic bags, were used as containers. Soils were air-
dried and passed through a 4-mesh stainless steel sieve; 3.5
kilograms were placed in each container.

Three levels of Zn (0, 7.5, and 15 pp2m) were established
on each soil by applying ZnSO4 in a circular band 2.5 inches

deep and one and one-half inches from the outside of the can.

24

Table 2. Methods of extracting soil Zn to correlate with
yields of pea beans.

 

 

Extracting Soil-Solution Method of Extraction
Solution Ratio Extraction Time (minutes)
0.1 N HCl 5:50 Shaking 30

0.2 N_Mg(N03)2 5:50 Shaking 60

0.2 N M9804 5:50 Shaking 60

H20 40:50 Percolation Variable*
0.2 N Ca(N03)2 5:50 Shaking 60
0.01 N EDTA +

1.0 N (NH4)2C03 10:20 Shaking :50H
1.0 N_NH4OAC 2.5:20 Shaking 60

0.4 N MgSO‘ 27.5:150 Shaking 1440+

 

*Two 25-ml increments of double-distilled H20 (total of
50 ml.) were added to 40 g. of soil, and Zn concentration
in leachate determined.

**Method of Trierweiler and Lindsay (1966).

+Method of Martens et al. (1966).

25

On all soils testing above pH 6.5, 20 pp2m Mn were applied
as MnSO4.

Prior to planting, 300 pp2m N as NH4N03 were applied in
the fertilizer band. Fifty pp2m N from the same sources were
added weekly (8 times) during the eXperiment. A total of 700
pp2mN were applied during the eXperiment.

PhOSphorus was applied as monobasic calcium phOSphate on
each soil in an amount so that soil test P plus applied P
equalled 100 pp2m, K as KCl so that the total K equalled 200
pp2m, and Mg as M9804 so that the total Mg equalled 100 pp2m.
All fertilizers applied prior to planting were mixed and
banded as described above (together with the Zn and Mn
fertilizers).

On September 18, 1966, 10 pea bean seeds were evenly
Spaced one-half inch below the soil surface at three inches
from the outside of each can. The fertilizer band was lo-
cated 2 inches below and 1.5 inches to the side of each seed.
Four hundred milliliters of deionized water were added to each
can of soil after covering the fertilizer with 2 inches of
soil (the water was applied before planting the seed to pre-
vent crusting of the soil surface). No more water was applied
until after germination. No supplemental lighting was used.

After seed germination, deionized water was applied daily
as needed, and the pots were brought to a constant volume of

water weekly.

26

Plants were thinned to four per pot two weeks after
emergence. When necessary, 3 foot plastic stakes were insert-
ed into the pots to support the plants.

On November 13, 1966, stakes were removed, plant tOps
were cut with stainless steel razor blades, and dried at 600C
in paper bags. Plant samples were ground in a stainless steel
Wiley mill.

For the second crop of pea bans, P was applied as mono-
basic calcium phOSphate on each soil in an amount so that
original soil test P plus applied P equalled 500 pp2m; all
other fertilizers were reapplied to all soils at the same
rates for the first pea bean crop. Supplemental artificial
fluorescent lighting was used. Lime (CaC03) was uniformly
mixed in certain soils with pH values less than 6.5 at
approximately twice the rate recommended for field crOps by
the Michigan State University Soil Testing Laboratory.

Beans were planted December 10, 1966 and harvested February
5, 1967.

For the third crOp of pea beans, no fertilizers were
added except for the weekly applications of 50 pp2m of N,
and no artificial lighting was used. Any soils below pH 7.0
were heavily limed (20,000 to 28,000 pp2m) prior to the
third crOpping (Table 5). Beans were planted April 1, 1967
and harvested May 27, 1967.

Plants were thinned to 3 per pot in both second and

third plantings.

27
Plant Analytical Methods

One gram of dry ground plant sample was ashed in a muffle
furnace at 5000 C for 4 hours. Five ml of 2 N HCl was slowly
added to the plant ash until effervescence ceased, and the
resulting SUSpenSion was filtered (Whatman No. 3 filter paper)
into 50 ml. volumetric flasks for Zn determination as described

above.
Field Studies

Field experiments to determine the reSponse of pea beans
to applied Zn were conducted at 20 locations from 1964 to
1967 inclusive. Varieties used, soil types, and fertilizer
treatments have previously been reported (Judy et al., 1965:
Brinkerhoff et al., 1966 and 1967: Vinande et al., 1968).
Yields from these experiments were correlated with levels of
extractable soil Zn (0.1 N_HC1), and the results are reported

herein.
Statistical Procedures

Statistical analyses were conducted using a Controlled
Data Corporation (CDC) 3600 computer. Yield and Zn concen-
tration and Uptake data were analyzed by means of the analysis
of variance: significant differences between treatments were
determined by using Tukey's HSD values (Federer, 1955).

The HSD values give a more rigorous test of significance than

28

do the more commonly used LSD values. Multiple regression
coefficients were determined by means of standard computer

programs utilizing the method of least Squares.

RESULTS AND DISCUSSION

Total and Extractable Zinc in Sixty-five
Michigan Soils

Total and extractable (0.1 N HCl) Zn were determined in
65 soils in order to evaluate Zn distribution in Michigan
soils. These soils were from 35 series, and represented 8
soil suborders. Adjusted average values, together with the
range in values, for total and extractable Zn and soil pH at
various depths are given in Table 3 for each suborder: data
for each soil are given in Table 1A in the Appendix. Adjusted
averages were computed because the depth of sampling was not
constant at all locations.

Total Zn varied greatly between the different soil sub-
orders; average total Zn in the aquoll (humic gley) suborder
was more than twice that in the orthod (podzol)--possibly
because of the greater content of organic matter and clay of
soils in the aquoll suborder. However, differences in total
Zn within a given suborder tended to be greater than average
differences between suborders, as indicated by the ranges
given in Table 3.

Total Zn was as high or higher in the surface layer than
at lower depths in all suborders, and these findings were

similar to those of Thorne (1942). No consistent differences

29

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31

between suborders were noted with reSpect to the distribution
of total Zn throughout the profile.

Extractable Zn was highly variable between and within
soil series, both within the same suborder and between sub-
orders. Most of the Zn reSponseS in Michigan have been noted
on soils within the aquept suborder; however, average levels
of extractable Zn in this suborder did not consistently differ
from that in the other suborders.

Extractable Zn always decreased with depth. About one—
tenth of the total Zn was extracted with 0.1 N_HCl in the
surface layer, while much lower proportions of the total Zn
were extracted at the lower depths. This decrease in the
prOportion of extractable Zn with depth may in part be due
to increases in soil pH with depth, but it may also in part
be due to the lower organic matter content of the lower
horizons.

Soil pH increased with depth on most of the soils, with
considerable variations both within and between suborders.
The level of extractable Zn in the different suborders was
not related to soil pH, as some of the lowest levels of
extractable Zn were noted in the aquod and orthod suborders,
in which the average pH values differed by about 2.0 pH units.

Soils varied greatly in total and extractable Zn, both
within and between suborders and series. Measurements of
total and extractable Zn, together with soil pH, did not ap-

pear to separate those soils on which yield reSponses had

32

been obtained with Zn fertilizers from those on which no
reSponses had been noted.
Yields and Zinc Content of Pea Beans
in the Greenhouse

Twenty of 65 soils referred to previously were selected
for more intensive greenhouse and laboratory studies because
measurements of total and 0.1 N HCl extractable Zn in Michigan
soils did not adequately evaluate their Zn status. Three
Zn levels were established on each soil and 3 successive
crOpS of pea beans were grown. The effectiveness of various
methods of extracting soil Zn was evaluated by correlating
extracted soil Zn with total dry weight of the plant tOps

(yields) and Zn uptake by the pea beans.

First CrOpping

zgglg; In general, yields of the first crOp were in-
creased by Zn application in soils testing above pH 7.1, and
were either not affected or decreased on soils testing 7.1
or lower (Table 4). Marked yield decreases were noted when
Zn was applied to soils testing below pH 6.3. Yields were
decreased slightly when Zn was applied to the Hodunk soil
(pH 6.3), but the growth of plants on this soil was stunted
by residual effects of atrazine which had been applied to
the soil prior to sampling.

Yields of beans were increased by Zn application on all
soils in the aquept and aquod suborders except for Hettinger

silty clay loam, the Colwood loam of the aquoll suborder, and

33

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u0S\m1 ESSI u0S\m u0S\m1, ESSIIu0S\m DOSVmiI, ESS DOSVm ENSS, moflumm USS
SN m0 SN m0 SN mo SN mo SN m0 SN mo SN nwpuonsm HHom
armed: .oeoo Sawfly armed: .ucoo nemem armed: .ocoo page» emaHSSS
SOHO SMASH SOHO OSoomm SOHO umuflm wo mumm

 

emeeaacoo II a means

35

UOSSHHSOO

 

 

 

 

m.mO n.0m N.m ¢.OO> m.>va N.m N.emm 0.00 ¢.m O.ma
d.ma m.aa ¢.a m.m>m m.mm m.m m.amm 0.0m m.OH m.>
0.0H O.HN m.O ¢.mOa O.Hm O.m 0.00N m.wm N.HH 0.0 *OHSOmHHHm
"HHSOD
O.m m.ON m.O m.wm m.mm 5.0 m.mmm m.mem ¢.H O.ma
m.m >.On w.O N.Ha m.mm m.O ¢.mmm m.mwm m.m m.>
m.> >.HH N.O m.ma O.ma m.a 0.0mm 0.00 #.O 0.0 OHHH SHHme
m.ma >.em >.O m.ma m.ON ¢.O N.HOO m.amm e.N O.ma
d.ma H.ON m.O 0.5 O.ma 0.0 m.mwm O.mea m.m m.>
m.m ¢.ma 0.0 m.O >.NH m.O m.w¢m m.m¢ N.> 0.0 omxmmxamm
"OOSHHO
m.Oom m.wm ¢.NH O.>Nm m.mm >.wa N.N¢m 0.0m >.m O.mfi
n.mam 0.5a a.ma N.mmm H.NN >.ma m.mmm >.mm N.m m.>
>.Oma m.md 0.0 m.OHN >.¢H >.¢H m.ama m.ma O.m 0.0 *H OO3SSOA
m.~me m.mm m.¢a m.mmm O.mm N.ma m.mmm «.mm O.Ha O.ma
>.maw m.am N.ma m.Oma a.¢a m.ma O.>mfi 0.0N m.m m.>
>.mmm «.mm m.Oa >.mma ¢.Oa m.ma N.Oma ¢.ma O.m 0.0 *H O003HOO
”HHOSO<
HOS\m1 ESS HOS\O HOS\m1 ESmI HOS\O HOS\O1 ESS HOS\O ENSS mOHHOm USO
SN HO SN m0 SN mo SN mo SN mo SN mo SN HmOHOQOm HHOm
mxmuSD .OSOO OHOHw mxmuSD .OSOO UHOHS mmeSD .OSOO OHOHS OOHHSSd
SOHO OHHSB SOHO OSOOOm SOHO umHHm mo Oumm

 

OOSSHHSOO II w OHQSB

36

.OSHHSSHS OSOOOm OH HOHHS CUEHA

+

.OSHHSMHS OHHSH OSm OSOOOm OH HOHHS SUEHAO

.OSHHSMHS OHHSH OH HOHHS OmfiHq
.x.

 

 

 

 

 

N.¢HH a.ma >.m N.OO m.aa ¢.m O.Hma m.Ov m.a HHOm 0Emm OSH
SO mHSOEHmmHH
SN SOO3HOm
m.mma O.mfi a.¢ N.OMH H.>a a.m m.mma 0.00 N.N .SN HO
HO>OH 05mm Hm
mHHOm SOO3HOm
“Am0.0V 9mm
m.Oma N.ma O.h >.¢O¢ m.mm >.ma H.mmm m.a¢ O.m O.ma
0.5m m.ma a.m N.>Nm O.mm m.ma O.NMN m.¢m O.m m.>
O.m >.da 0.0 N.O¢d N.OH m.ma O.mm m.ma m.n 0.0 *HEOHZ
H.¢Oa m.mm >.m O.mmm ¢.Ne O.m N.amm >.mmm m.N O.ma
m.mna >.HN ¢.O O.m>H m.a¢ >.¢ O.mm> O.>Oa ¢.¢ m.>
0.0H ¢.ma m.O O.¢N 0.0a a.m m.mw¢ m.mm O.m 0.0 OOxOOA
¢.Nm¢ H.Ow m.aa d.mmm O.am O.m a.m>m m.m>a N.N O.ma
m.mON m.HN O.m >.mem >.mm 0.0a O.mwm 0.0m O.N m.>
w.noa O.¢a h.> 0.0> w.ma a.¢ m.mm O.Hm O.N 0.0 +xSSUOm
"HHMOD
HOS\m1 ESSI HOS\O HSm\m1I ESSI HOS\m HOS\mm ESS HOS\m ENSS mOHHOm USS
SN HO SN HO SN m0 SN mo SN mo SN HO SN HOUHOnsm HHOm
mmeSD .OSOO OHOHM ONMHSD .OSOO OHOHM mmeSD .OSOO OHOHN OOHHSmd
SOHO OHHSB SOHO OSOOOm SOHO HmHHS mo OHmm

 

OOSSHHSOO II S manna

37

Miami loam in the udalf suborder. Yields obtained when 7.5
pp2m Zn were applied to soils were nearly as high as those
when 15.0 pp2m Zn were applied, except for Sims II clay loam
in the aquept suborder and Colwood loam in the aquoll sub-
order.

Yields were decreased by Zn application on all soils in
the aqualf, orthod, and udalf suborders except for the Miami
and Conover loams. Large yield depressions were obtained
when 7.5 pp2m Zn were applied to soils in the orthod suborder
and to Locke loam in the udalf suborder. Further yield de-
pressions were noted on all other soils in these 3 suborders
when 15.0 pp2m Zn were applied.

Zinc concentration: Zinc concentration in bean tissue

 

was increased by Zn application on all soils, except Wisner II
clay loam in the aquoll suborder. The Zn concentration was
possibly lower after addition of Zn because of the large
yield increase (41 percent) on this Wisner soil.

Since actual yields varied Significantly on the different
soils, relative rather than actual yields were used in all
correlations between yields and Zn concentration in pea beans.

Relative yields were calculated using the following equation:

. . _ Actual Yield without Zn
Relative yield _ Maximum Yield with Zn X 100

 

Yields were increased when Zn was applied to 12 of the soils.
Highest yields were obtained when Zn concentration in the

tissue was between 11 and 32 ppm; the average concentration

38

of Zn in plants from the highest yielding treatments was 25
ppm (Figure 1).

Yields were decreased when Zn was applied to 8 of the
soils. Highest yields were obtained when Zn concentration
in the tissue was between 26 and 60 ppm; the average concen-
tration of Zn in plants from the highest yielding treatments
was 34 ppm. The yield depressions usually resulted from a
Zn concentration greater than 50 ppm.

Zinc Uptake: Zinc Uptake by beans was increased by Zn

 

application of all soils; however, no definite relationship
was observed between increases in Zn uptake by the plants

and increments of Zn fertilizer added to these soils.
Approximately 0.2 to 3.0 percent of the Zn application of

7.5 pp2m and approximately 0.1 to 2.0 percent of the Zn appli—
cation Of 15.0 pp2m was recovered from the different soils by
pea bean plants.

Total uptake of Zn by beans was significantly greater
when grown on light-textured soils (sandy loams, loamy sands,
and sands) with low pH values (below 6.7) than when grown on
cmher soils. Soils in the former category were Metamora I,
Kalkaska, Karlin III, Hillsdale, and Locke. Total uptake of

Zn was not strictly related to yield.

Second Cropping
Zinc was again applied to all soils at a rate equivalent
to the Zn added for the first crOp. Lime was applied to the

Metamora I, Kalkaska, Karlin III, Hodunk, and Locke soils

39

.OmSOSSOOHm OSH
Se S30Hm mSMOn SOS mo SOHO HmHHH OSH Ho UHOH>

O>HHmHOH USS SOHHSHHSOOSOO SN SOO3HOQ SOHHOHOm .H OHSOHS

2.33 5 SN and

 

com omv omv com ovm com cow ONN om. cc. co. co ON
I q A q 1 q 4 d I d d d d d d d d d d u d d d d d d
4
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Q 0
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OSHN HOHHM OaOHh SH OmmmHOSH n O c o:am d

 

 

O.

ON

On

9.

On

Om

Om

OO.

013M 3A|1V138

40

after yield depressions were obtained with Zn application
for the previous pean bean crop (Table 5).

Yields: ReSponses to Zn addition with the second crOp
were similar to those of the first crop (Table 4). Greatest
yield increases were obtained for the first increment (7.5
pp2m) of Zn fertilizer while increasing the rate of Zn appli-
cation from 7.5 to 15.0 pp2m did not result in significant
increases except on the Charity II soil.

After liming the Locke and Hodunk soils in the udalf
suborder and the Metamora soil in the aqualf suborder (Table
6), yields were increased when Zn was applied, while in the
first crop, yields on these soils were decreased when Zn was
added. Increasing the rate of Zn application from 7.5 to
15.0 pp2m Zn increased yields in the Locke soil, but not on
the Hodunk or Metamora soils. On the Kalkaska and Karlin
soils, liming appeared to depress yields, and Zn application
was not effective in increasing yields.

Zinc concentration: Zinc concentration in plant tissue

 

was again increased by Zn addition on all soils in which
yield increases were obtained for the first crOp. Consider-
ing all these soils, highest yields were obtained when Zn
concentration in the tissue was between 14 and 36 ppm: the
average concentration of Zn in plants from the highest yield-
ing treatments was 25 ppm.

Zinc concentration in plant tissue was also increased

by Zn application on all limed soils. On these soils, highest

41

UOOSHHSOO

 

e.a m.m ooo.mm m.m III 0 e.a HH amHaHnm
O.N 0.0 OO0.0N 0.0 III O m.n H mmaeflHm
"Uosvd
n.> III O m.> III O m.m HH HOSmHZ
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HOHHS OHOwOm HOHHS HOHH< OHOme HOHHS mS
mm mm emcee ween mm mm peeve mqu

 

 

 

.mHHOm mo mODHm> mS SO SOHHOOHHSSO OEHH mo USS OSHSSOHO OmSOSSOOHO HO mHOOmmm .m OHQMB

 

42

m.h 0.5 OOO.dN n.m III O N.> HEMHZ

 

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"HHODO<
ENSSI, ENSS NSHSSOHO mOHHOm USm
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HOHHN OHOHOm HOHH< HOHHH OHOHOm HOHHS mS
mm me one? meg SS me. Senna msHS

 

 

UOOSHHSOO II m OHQMB

43

yields were obtained when Zn concentration in the tissue was
between 20 and 42 ppm: the average concentration of Zn in
plants from the highest yielding treatment was 27 ppm,
approximately the same concentration as was obtained from
plants on unlimed soils.

Zinc Uptake: Zinc uptake was increased by Zn fertiliza-

 

tion of all soils; however, no definite relationship was ob-
served between increases in Zn Uptake by the plants and
increments of Zn fertilizer added. From 0.03 to 1.0 percent
of the fertilizer Zn applied at 7.5 pp2m and 15.0 pp2m rates,
reSpectively, on unlimed soils was recovered from the dif-
ferent soils by pea bean plants.

Zero to 1 percent of the Zn applied at 7.5 pp2m and 0.03
to 1 percent of that applied at 15.0 pp2m was recovered from
the limed soils by the plants. Uptake of Zn by beans from
the limed soils was much lower than that absorbed from the

same soils by the first crop.

Third CrOpping

 

Zinc fertilizer was not applied again for the third crOp.
Lime was applied to all soils on which pH values were below
7.0 after the second pea bean crOpping; these included all
soils in the aqualf, aquod, aquoll, orthod, and udalf sub-
orders and Breckenridge and Sims I in the aquept suborder
(Table 5).

Yields: Yields were increased by Zn fertilization on

all unlimed soils on which similar increases were obtained

44

for previous crOps (Table 4). Greatest yield increases

were again obtained for the first increment of Zn fertilizer
while increasing the rate of Zn application from 7.5 to 15.0
pp2m did not result in significant yield increases.

Yields were increased by Zn addition on 11 of the 13
limed soils, but on the Kalkaska and Karlin soils, Zn ferti-
lizers were not effective in increasing yields. Increasing
the rate of Zn application from 7.5 to 15.0 pp2m did not
result in significant yield increases except on the Sims I
and Hillsdale soils.

Zinc concentration: Zinc concentration in bean tissue
was again increased by Zn application on all unlimed soils.
Highest yields were obtained from crOps grown on the unlimed
soils when Zn concentration in the tissue was between 19 and
28 ppm, and the average concentration of Zn in plants from
the highest yielding treatments was 25 ppm.

Zinc concentration in plant tissue was also increased
by Zn fertilization on all limed soils except the Hillsdale.
Highest yields were obtained from crops on soils where Zn
concentration in the bean tissue was between 22 and 49 ppm;
the average concentration of Zn in plants from the highest
yielding treatments was 25 ppm.

Zinc Uptake: Zinc Uptake by pea beans was increased

 

by Zn fertilization of all soils; however, no definite
relationship was observed between increases in Zn uptake by

the plants and increments of Zn added to these soils.

45

Approximately 0.2 to 1.4 percent of residual Zn at the first
increment and 0.4 to 0.7 percent of residual Zn at the
second increment was recovered from the unlimed soils by
the plants.

Between 0.007 and 0.8 percent of residual Zn at the
first increment and 0.004 to 0.7 percent of residual Zn at
the second increment was recovered from the limed soils by

bean plants.

Between Crops

Yields: Yields of the second crOp increased more than
yields of the first crOp after applying Zn to the unlimed
soils (Table 4). The differences in yields may have been
due to higher P rates for the second crOp. Higher yields
On soils with both 7.5 and 15.0 pp2m Zn applications may have
also been due to higher P rates (Doll et al., 1967).

Yields were increased by Zn addition on all limed soils
except those in the orthod suborder. Liming progressively
increased relative yield differences obtained from soils at
the 0 and at the 15.0 pp2m Zn applications for both the
second and third crops. For example, yields of the first
crOp were only one-third as great after Zn fertilization on
the Locke soils, while yields of the second and third crops
were 4 times and 7 times greater, reSpectively, after Zn
fertilization of this soil. A smaller increase in yield was
Obtained after applying Zn to the Hodunk soil for the third

crOp than after applying Zn to the second crop (lime applied

46

before the second crOp but not before the third crop).

Yields at each Zn level usually decreased when large
quantities of lime were applied to soils. The actual yields
between crOps often decreased while relative yields increased
with Zn fertilization.

Zinc concentration: Highest yields for all crops of
pea beans were obtained on both limed and unlimed soils at
similar levels of plant Zn concentration (approximately 25
ppm). No other definite trends in Zn concentration were
found between crOps on these soils.

Plant Zn concentration was decreased by applying lime
to soils. Liming soils for both the second and third crOp
resulted in a progressive decrease Of Zn concentration in
the plant. For example, on the Locke soil, Zn concentration
in plants decreased from 324 to 42 ppm from the first to
second crOps, and to 33 ppm in the third crOp. A larger Zn
concentration was obtained in the third crOp than in the
second crop of pea beans from the Hodunk soil (lime only
applied for second crOp). Highest yields were obtained on
limed soils at similar ranges and averages of Zn concentra-
tion for both the second and third crOps.

Zinc Uptake: A gradual increase in Zn Uptake by beans

 

was generally obtained at each Zn treatment on unlimed soils
from the first to the third crOpS. Similar quantities of

Zn were extracted from these soils by pea bean crOps (approxi-
mately 0.1 to 3.0 percent at both 7.5 and 15.0 pp2m Zn appli-

cation).

47

Zinc Uptake by beans was decreased when lime was applied.
Liming soilstr'both the second and third crOpS resulted in
a progressive decrease in Zn uptake by plants. For example,
at 15.0 pp2m Zn application on the Locke soil, Zn uptake by
beans decreased from 921 to 336 ug/pot from the first to
second crOps to 184 ug/pot for the third crOp. A larger Zn
Uptake was obtained in the third crOp than in the second crOp
from the Hodunk soil (lime only applied for second crop).
Similar quantities Of Zn fertilizer were extracted from these
soils by bean crOps (approximately a trace to 1.0 percent at

both 7.5 and 15.0 pp2m Zn application).
Soil Zinc Extraction Procedures

Zinc was extracted from soils using 9 different extrac—
tion procedures (Table 6). Five of these procedures had
been previously reported as suitable for determining avail-
able soil Zn. All of these procedures were adaptable to
routine analyses as are used in soil testing laboratories.
The amount Of Zn removed from the different soils by the
various extractants ranged from 11 ppb (parts per billion)

with H20 to 63 ppm with 12 N_HCl.

Zinc Extracted from Soils Before Croppgng

No definite relationship was noted between Zn extracted
from soils by 0.1 N HCl (extractable) and that extracted by
12 N HCl (total), but much more Zn was extracted with the

12 N HCl than with 0.1 N HCl (54 times greater in the

48

UOOSHHSOO

 

OH0.0 NN.O ON.H O>.O mm.O mH.O m¢.O N.N 0.0¢ ¢.h HH HOHEHHm
MH0.0 Om.O HN.H O0.0 HN.O HN.O Hw.O N.N O.wm m.> H mmHEHHm

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HH0.0 No.0 Nm.H h¢.O Hm.O wH.O >¢.O m.O N.Nw N.N HH HmSmHS
«H0.0 NN.O >¢.H No.0 om.O OH.O O¢.O >.m 0.0N N.N H HOSmHZ
OH0.0 mm.O OO.H HN.O NN.O HH.O H¢.O N.¢ H.Hm O.n HH mEHm
OH0.0 NN.O ON.H NN.O Om.O OH.O om.O m.¢ 0.0¢ m.> H mEHm
HN0.0 OO.H ON.H NN.O NN.O OH.O Ow.O 0.0 ¢.¢¢ m.> HOOSHHHOE
SH0.0 m0.0 Om.N O¢.O NN.O mH.O >¢.O m.O 0.0¢ N.N HH mHHHmnO
mH0.0 O0.0 ON.N m¢.O NN.O OH.O Ne.o N.N ¢.N¢ N.N H >HHHMSO
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mH0.0 >H.H Hm.m Om.O mm.O OH.O m¢.O m.m N.mO H.> HO>OSOO

"MHODOH
ESS IESS ESQ, .I EmS.l ESQ .I ESS, .I ESS, ESS ESS mOHHOm USO
omm .z o.H z «.0 z N.o z N.o 2 m6 m H.o m 0.3 mm Heeuonsm HHom

oSOemz Seam «ommz m Anozv m: m Anozvmo HOS

 

mSOHHSHOm mSHHOMHme

 

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SH UOHmHH mOHOUOOOHS msoHHm> OSH >9 mHHOm SSOHSOHE EOHH UOHOMHHxO SN HO HSDOE<

.O OHQMB

49

 

OH0.0 mm.o OH.H 05.0 Om.o mH.O om.o H.N m.>N N.N HEMHZ

 

meo.o 0H.H oo.m ea.H om.o an.o Hm.H m.m m.mH m.m exuoq
omo.o mm.o me.H em.o mm.o mm.o mm.o H.H m.om m.m xcsoom
nmo.o mm.o mm.H em.o He.o Hm.o mm.o m.m m.em w.m mHnenHHHm
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SSW, ESS 8mm, .I ESS.I SSS, .l EmS ESS ESS ESS mOHHOm USm
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ueoemz Seam aommz mAnonmz «Anozvmo HOS

 

mSOHHSHom OSHHOSHme

 

 

UODSHHSOO II O OHQMB

50

Wisner II silty clay loam). As previously mentioned, total
and extractable Zn were highly variable within and between
suborders.

More than twice as much Zn was usually extracted from
soils with 0.2 N Ca(N03)2 as compared with 0.2 N_Mg(N03)2
indicating that Ca++ was more effective than Mg++ in replac-
ing exchangeable Zn++ or (ZnOH)+ at low salt concentrations.
The Ca++ ion was possibly more effective than Mg++ in reduc-
ing the zeta potential because of the smaller hydrated size
of Ca++. Also, about twice as much Zn was extracted from
soils by 0.4 N MgSO4 as was extracted by 0.2 N_MgSO4, pos-
sibly because of the higher concentration of Mg ions and—-
to a much lesser extent--a longer extraction time and larger
soil:solution ratio. More Zn was generally extracted by 0.2
N M9804, than by 0.2 N Mg(N03)2.

Higher amounts Of Zn were extracted by Ca and Mg salts
from the Metamora soil in the aqualf suborder, the soils in
the orthod suborder, and the Locke soil in the udalf sub-
order than from other soils. The pH values of these soils
ranged from 5.7 to 6.2 with an average of 5.9. Lesser
amounts of Zn were extracted from soils in the aquept, aquod,
and aquoll suborders where the pH values of these soils
ranged from 6.9 to 7.9, with an average of 7.5. The quantity
of Zn extracted from soils by Ca and Mg salts was fairly
consistent between soils within the various suborders, except
for the Locke soil in the udalf suborder which was higher in

extractable Zn than other soils in this suborder.

51

Greater quantities of Zn were extracted by 0.01 N
EDTA + 1.0 N (NH4)2C03[EDTA] from soils in the aqualf and
orthod suborders and the Locke soil in the udalf suborder
than from other soils; however, the 2 Charity soils in the
aquept suborder contained larger quantities of EDTA extract-
able Zn than the other soils in that suborder. Perhaps
some Zn was removed by the EDTA chelate from carbonate
minerals of these highly calcareous soils. The amount of Zn
removed from soils by this extractant was generally not
consistent between soils within various suborders.

Greater amounts Of Zn were extracted by 1.0 N_NH40AC
from the soils which contained large amounts of EDTA extract-
able Zn, except for the Charity soils. No consistency was
Observed for amount of 1.0 N NH4OAc extractable Zn from
soils within various suborders.

The Metamora soil in the aqualf suborder, soils in the
orthod suborder, and soils, except Miami, in the udalf
suborder contained the largest amount of H20 extractable Zn.
The pH values of these soils ranged from 5.7 to 6.6 with an
average of 6.0. Smallest amounts of Zn were extracted from
the Conover soil in the aqualf suborder, soils in the aquept
suborder, soils in the aquod suborder, soils in the aquoll
suborder, and the Miami soil in the udalf suborder. The pH
values of these soils ranged from 6.9 to 7.9 with an average
Of 7.5. The quantity of Zn removed from soils by H20 was

only consistent within the aquoll and orthod suborders.

52

The amount of Zn extracted by Ca and Mg salts, EDTA.
1.0 N_NH4OAC, and H20 generally increased as soil pH
decreased: however, total and 0.1 N_HCl extractable Zn were
not correlated with soil pH values. Therefore, the Ca and
Mg salts, EDTA, 1.0 N_NH4OAc, and H20 removed largest amounts
of Zn from soils in the orthod suborder, the Locke soil in
the udalf suborder, and the Metamora soil in the aqualf sub-
order than from other soils. In addition, H20 extracted
comparatively large amounts of Zn from the Hodunk and Hills-
dale soils in the udalf suborder, 1.0 N NH4OAc extracted
large amounts of Zn from Conover soil in the aqualf suborder,
and EDTA extracted large amounts of Zn from the Conover soil
and the 2 Charity soils in the aquept suborder. The amount
of Zn extracted by Ca and Mg salts was fairly consistent for
soils within the same suborder, but the amount of Zn removed
by other extractants (listed in Table 6) was generally not

consistent for soils within the same suborder.

Zinc Extracted From Soils After Final CrOpping

Liming soils after the first and/or second crOp of pea
beans generally resulted in a significant decrease in Zn ex-
tracted by H20, 1.0 N_NH4OAc, 0.1 N HCl, and EDTA after the
third cropping (Table 7). The amount of Zn extracted by
these 4 extractants was not appreciably altered during crop-
ping of unlimed soils.

Soils with pH values less than 6.4 (Metamora I soil in

the aqualf suborder, soils in the orthod suborder, and all

USDSHHSOO

 

 

 

 

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mSHSSOHO SHOHSQ USHOSHHNS SN SSS

 

 

 

UOSSHHSOO II 5 SHQSB

55

soils except Miami in the udalf suborder) contained only
about one-half as much H20 extractable Zn after liming as
they did prior to liming; other soils contained about three-
fourths as much water extractable Zn after liming. Only
one-half as much 1.0 N NH4OAc extractable Zn was removed
from soils in the aquept, aquod, and aquoll suborders after
liming as before liming, but other soils contained variable
amounts of 1.0 N_NH4OAc extractable Zn after liming. Soils
in the aqualf suborder contained only 50 percent as much EDTA
extractable Zn after they were limed while other limed soils
contained approximately 75 percent as much EDTA extractable
Zn as they did prior to liming.
Relationship Between Extractable Soil Zinc
and Yield and Zinc Uptake by Pea Beans

Of the 20 soils used in the greenhouse eXperiment (dis-
cussed in previous section), yields were increased or not
affected by Zn fertilization on 12 soils, but were decreased
by Zn fertilization on the remaining 8 soils. In order tO
relate Zn soil tests to pea bean yields, only the results
from the 12 soils where yields reSponded positively were
used. Since actual yields varied significantly on the dif-
ferent soils, relative rather than actual yields were used
in all correlations with soil tests. Relative yields again
were calculated using the equation previously given.

Highest linear correlations between relative yield and
extractable Zn were Obtained when soil Zn was extracted with

1.0 N NHgOAc, H20, and 0.1 N EDTA + 1.0 N (NH4)2C03 [EDTA]

56

while significant correlations were obtained when Zn was
extracted with 0.2 N Ca(N03)2 and 0.2 N Mgso4 (Table 8) .
While the correlation coefficient was statistically signifi-
cant, the correlation between relative yield and 0.2 N
Mg(N03)2 extractable Zn was not real, as it indicated that
yield decreased as extractable Zn increased. NO linear
correlation between yield and extractable Zn was noted when
soils were extracted with 0.1 N_HCl, 12 N_HCl, or 0.4 N MgSO4.
In order to determine if a non—linear relationship
existed between relative yield and extractable soil Zn,
multiple correlations between extractable Zn and relative
yield were calculated using the following equation (Zn =

extractable soil Zn):
Relative Yield = a + b(Zn)+ C(Zn2)

With the use of this quadratic equation, higher correlations
between relative yield and extractable Zn were obtained when
soil Zn was extracted with 0.2 N_Mg(N03)2, 12 N_HCl, and 0.4
N_MgSO4 than were Obtained by fitting linear regressions,
but correlations between Zn removed by other extractants
and relative yield were not greatly improved (Table 8). No
multiple correlation coefficients were given for 0.1 N_HCl,
since the curve fitted using the computer program was not
compatible with established yield reSponse data.

Since yield reSponse to Zn application in the greenhouse,

together with the factors discussed in the preceding

57

Table 8. Linear and multiple correlation coefficients between
relative yield of the first crop of pea beans and
extracted soil Zn (Zn) and extractable soil Zn plus
soil pH (Zn + pH).

 

 

Correlation Coefficients

 

 

 

Extracting Agents Linear Multgple
Zn Zn Zn + pH
r R R
Distilled water 0.52** 0.53** --—
0.2 N Mg(N03)2 -0.32* 0.52** ---
0.2 N Ca(N03)2 0.38** 0.45** -—-
0.01 N EDTA + 1.0 N (NH4)2C03 0.45** 0.50** 0.71”
0.1 N HCl 0.28 --- 0.61**
12 N HCl 0.09 0.34* --—
0-2 N M9804 0.29* 055* ---
1-0 N NH4OAC 0.55** 0.55** --—
0.4 N M9504 0.02 0.35* --—

 

*-
Significant at 0.05 level.

*

*
Significant at 0.01 level.

58

discussion section concerning extractable Zn on the different
soils, indicated that Zn availability was related to soil pH,
another series of multiple regression equations were fitted

using the following equation (pH = soil pH):

Relative Yield = a + b(Zn) + C(Zne) + d(pH) + e(pH2)

+ f(Zn - pH)

Higher correlations were obtained with EDTA and 0.1 N HCl
when soil pH was included in the correlations (Table 8).
Multiple correlation coefficients are not reported for H20
and 1.0 N NH4OAc extractable Zn, since the curves fitted
using the computer program were again not compatible with
yield reSponse data.

With the curve fitted using the computer program, yields
were increased on soils on which Zn extracted by EDTA and
0.1 N_HCl increased when soil pH values of 7.0 and 7.5 were
included in the correlations (Figures 2 and 3). Highest
yields could be expected from soils at pH 7.0 on which 1.5
and 3.5 ppm Zn were extracted with EDTA and 0.1 N_HCl
reSpectively. Highest yields could be eXpected from soils
at pH 7.0 on which 2.5 and 7.0 ppm Zn were extracted with
EDTA and 0.1 N HCl reSpectively. Only small increases in
yield of pea beans would be expected after addition of Zn
to similar Michigan soils with pH values equal to or less
than 7.0, but increases in yield after Zn application would
be expected to vary greatly with amounts of EDTA or 0.1 N_HCl

extractable Zn from soils with pH values of approximately 7.5.

IOO
90
80
70
60
50
40
30
20
IO

RELATIVE YIELD

 

59

 

I / 1 I

 

0.5 IIO Ifs 2:0 2.5 3.0
PPM Extractable Zn (EDTA)

Figure 2. Relation between relative yield and 0.01.N

EDTA + 1.0 N (NH4)2C03 extractable Zn at pH
7.0 and pH 7.5.

RELATIVE YIELD

IOO
90
80
7O
60 '-
50 -
4O -
30 -
20 -

 

 

60

l l L l l l l l l l 1 l

 

Figure 3.

I 2 3 4 5 6
PPM Extractable ZnIO.1NHCII

Relation between relative yield and 0.1 N HCl
extractable Zn at pH 7.0 and pH 7.5.

61

Highly significant linear correlations between plant
uptake of Zn and soil extractable Zn were obtained when the
extractants used were H20, EDTA, or 1.0 N_(NH4)20AC (Table
9). No correlation between Zn Uptake and extractable Zn
was noted when these 20 soils were extracted with 0.1 N_HCl.
Amount (If Zn removed from soils by beans was highly linear-
ly correlated with soil pH, but curves fitted using the
computer program for multiple correlations were not compat-

ible with established Zn Uptake data.

Field Results

Extractable Zn (0.1 N_HCl) was determined for 15 soils
where field eXperiments with pea beans were located from
1964 to 1967, inclusively (Table 10). Relative yields were
highly correlated with 0.1 N HCl extractable Zn (Figure 4,

r = 0.60). Multiple correlations which included soil pH
were not determined, Since pH only varied from 7.2 to 8.0.

Using the equation calculated for the field eXperiments,

maximum yields were obtained when the soil contained 8.2

ppm extractable Zn.

62

Table 9. Linear correlations between extractable Zn, pH,
and Zn Uptake by pea beans on 20 Michigan soils
used in greenhouse and laboratory evaluations.

 

 

 

Independent Variables Linear Correlation
Coefficients
r

H20 O.84**
0.01 N EDTA + 1.0 N (NH4)2C03 0.4.7;H

0.1 N_HCl -0.04

1.0 N_NH4OAc 0.52**

pH -0.79**

 

**Significant at 0.01 level.

63

 

 

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"HHO:O<
o.m H.5 m.Hm H.mm ecmHeHz HmmH HHHn HmHeHnm
m.m H.e m.mm m.mH naneHz mmmH HH eeoH HHHn HmHsHHm
m.m o.m o.mm m.e eaneHz man H anoH aHHn HmHeHnm
"UOSO<
m.o m.e m.em H.0H anchnm HmmH anoH HnHu HHHHn Hmanz
5.0 o.m IIII o.o sechmm mme EnoH HnHu NHHHn Hmanz
N.m e.e e.em 0.5 anchnm emmH enoH SnHo HmenHz
m.m 5.5 H.®H m.NH aneHmnm HemH anoH HnHo nmean
m.o m.e H.em o.o 3neHmnm mmmH snoH HnHu HHHHn nmanz
N.m 5.H H.om e.mH mHounsH emmH amoH sedans
m.e m.e 0.0m N.Nm Hem meaH HH anoH HnHu neHm
N.m N.N e.em m.mm aneHmnm mmmH H enoH HnHo nsHm
H.m a.» m.mm m.mH engage HmmH amoH HnHo eHoHHUHa
H.e e.5 H.0N m.mH unease mmmH SnoH HmHu ewowone
5.4 e.5 n.0N o.om uncmee mmmH anoH HnHu SHOHHUHS
«HSSSOS
ESS OS\SQ OS\SQ SS>B USS
HOm m H.O mS SN SHHz SN HSOSHHS >HSSOO HSSEHHOSxm HSUHOQSM HHom
HQ USHOSHme EDEHNSE SOSHO>¢ HO HSmw

SN

 

 

0H EOHH USHOSHme SN HO mSOHHSHHSSOSOO USS

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.OH mHSme

64

IOO?

RELATIVE YIELD

 

 

l l l

I 2 3 4 5
PPM Extractable Zn (0.1N HCII

o —._°°L 1

Figure 4. Relation between relative yield in the field
and Zn extracted from soils by 0.1 N HCl.

SUMMARY AND CONCLUSIONS

Sixty—five Michigan soils varied greatly in both total
(12 N_HCl) and acid extractable (0.1 N_HCl) Zn, both within
and between suborders and series. Soil pH increased with
profile depth on most of the soils, and considerable vari-
ation was observed within and between suborders. Measure-
ments of total and extractable Zn together with soil pH, did
not identify those soils where additional Zn might be needed
for beans.

Since measurements of total and 0.1 N_HCl extractable
Zn in Michigan soils did not adequately evaluate their Zn
status, 20 of the 65 soils originally investigated were
chosen for more intensive greenhouse and laboratory studies.
Three Zn levels were established on each soil and 3 succes-
sive bean crops were subsequently grown on each soil. The
effectiveness of various methods of extracting soil Zn was
evaluated by correlating extracted soil Zn with yields and
Zn Uptake by the plants.

Yields of the first pea bean crOp were increased by Zn
application in soils testing above pH 7.1, and were not
affected on soils testing 7.1 or lower. Marked yield de-
creases were noted when Zn was applied to soils testing
below 6.3. The amount of Zn extracted by Ca and Mg salts,

0.01 N EDTA + 1.0 N (NH4)2C03 [EDTA], 1.0 N NH40AC, and H20

65

66

generally increased as soil pH decreased, but total and 0.1
N_HCl extractable Zn were not correlated with soil pH values.
Total concentration and Uptake of Zn by pea beans was signifi-
cantly greater in plants grown on coarse textured soils with
pH values below 6.7 as compared to plants grown on the other
soils. On the basis of these findings, pea beans would not be
expected to reSpond to Zn application on soils with pH values
below 7.0.

On soils where yields of the first crOp were increased
or not changed when Zn was applied, large increases in yields
were obtained when Zn concentration in the tissue was in-
creased from approximately 10 to 30 ppm. On all soils, high-
est yields were obtained when the Zn concentration in the
plants was between 25 and 34 ppm.

Marked yield depressions resulted from a Zn concentration
greater than 50 ppm. These yield depressions were attributed
to Zn toxicity since yields were decreased on the same soil by
the addition of Zn. This suggests that over 50 ppm Zn concen-
tration may also be injurious to beans grown in the field.

The narrow range of 25 to 34 ppm Zn concentration for
maximum yields of pea beans grown in the greenhouse was quite
similar to Zn concentration found for maximum yield in the
field. However, insufficient quantities of Zn were applied
to soils to get significant yield depressions in the field.

It is very probable that large field applications (10 pp2m or

greater) of Zn on some Michigan soils, especially those with

67

coarse textures and pH values less than 7.0, would result in
injury due to toxicity.

Yields obtained in the greenhouse for the first crop
were generally as high when 7.5 pp2m Zn were applied to soils
as when 15.0 pp2m Zn were applied. Only one-half (4 pp2m) as
much Zn as ZnSO4 was needed to obtain highest yields in the
field. Zinc concentration and Uptake by plants increased
with increasing rates of Zn fertilization in both the green-
house and field.

Of the 20 soils used in the greenhouse experiment,
yields of the first crop were increased or not affected by Zn
fertilization on 12 soils, but were decreased by Zn fertili-
zation on the remaining 8 soils. In order to relate Zn soil
tests to yields, only the results from the first 12 soils
were used. Since actual yields varied significantly on the
different soils, relative rather than actual yields were used
in all correlations between yields and soil tests. Relative
yields were calculated using the following equation:

Actual Yield without Zn x 100
Maximum Yield with Zn

Relative Yield =
Highest linear correlations between relative yield and extract-
able Zn were Obtained when soil Zn was extracted with 1.0 N
NH4OAc, H20, and EDTA.

Since yield reSponse to Zn fertilizer in the greenhouse,
together with other factors concerning extractable Zn on the

different soils, indicated that Zn availability was related

to soil pH, another series of multiple regression equations

68

were fitted using the following equation (pH = soil pH):

Relative Yield = a + b(Zn) + C(Zna) + d(pH) + e(pH2)

+ f(Zn-pH)

Higher correlations were obtained with EDTA and 0.1 N_HCl
when soil pH was included in the correlations. With the
curve fitted using the computer program, yields were in-
creased on soils on which Zn extracted by EDTA and 0.1 N
HCl increased if soil pH values of 7.0 and 7.5 were included
in the correlations. Only small increases in yield would be
expected after Zn fertilization of similar Michigan soils
with pH values equal to 7.0, but increases in yield after Zn
application would be eXpected to vary greatly with amounts
Of EDTA or 0.1 N HCl extractable Zn from soils with pH
values of approximately 7.5. A similar relationship was ob-
served on 15 soils when 0.1 N HCl extractable Zn was linearly
correlated with relative yields in the field, but multiple
correlations which included soil pH were not determined since
pH only varied from 7.2 to 8.0. Using the equation calcu—
lated for the field experiments, maximum yields were obtained
when the soil contained 8.2 ppm extractable Zn. Assuming a
pH Of 7.6 in the greenhouse eXperiment, maximum yields would
rave been Obtained when the soil contained approximately 7 ppm
extractable Zn. On the basis of both greenhouse and field
studies, it was suggested that the 0.1 N HCl extraction pro-
cedure was the most desirable of all the Zn soil tests uti-

lized in these experiments. It should be desirable, however,

69

to initiate further field studies to better evaluate the EDTA
extractant as a Zn soil test for Michigan soils.

For the second crOp, lime was applied to all soils on
which yield depressions were obtained after Zn application
for the first crOp. After liming the Locke and Hodunk soils
in the udalf suborder and the Metamora soil in the aqualf
suborder, yields were increased when Zn fertilizer was applied.
0n the Kalkaska and Karlin soils, liming appeared to depress
yields, and Zn fertilizers were not effective in increasing
yields. It is quite probable that liming some of these low
pH soils of Michigan to pH values of 6.5 (the pH value recom-
mended for most agronomic crops) would result in a Zn de-
ficiency for Zn reSponsive crOps such as pea beans.

For the third crOp, lime was applied to all soils on
which pH values were below 7.0 after the second crOp. Yields
were increased by Zn fertilization on 11 of the 13 limed
soils. 0n the Kalkaska and Karlin soils, Zn fertilizers
were again ineffective in increasing yields.

Liming for both the second and third crOps progressively
increased relative yield differences obtained from soils at
the 0 and at the 15.0 pp2m Zn applications. The difference
may be due to an increase of an insoluble compound such as
calcium zincate with liming. Yields at each Zn level gen-
erally decreased when large quantities of lime were applied
to soils. Increasing the rate of Zn application from 7.5
to 15.0 pp2m generally did not result in significant yield

increases.

70

Zinc concentration and uptake by pea beans were gener-
ally increased by Zn fertilization on limed soils; however,
liming soils for both the second and third crOps resulted
in a progressive decrease of Zn concentration and uptake
in the plant. Similar quantities of Zn fertilizer were ex-
tracted from these soils by pea bean crOps (a trace to 1.0
percent at both 7.5 and 15.0 pp2m Zn application). Liming
soils after the first and/or second crOp resulted in a sig-
nificant decrease in Zn extracted by H20, 1.0 N_NH4OAC, 0.1
N_HCl, and EDTA.

Yields of the second crOp of beans were increased more
than yields of the first crOp after applying Zn to the un-
limed soils. The greater reSponse may have been due to
higher P fertilization rates. Higher yields on soils with
both 7.5 and 15.0 pp2m Zn applications may also have been
due to higher P rates. A gradual increase in Zn uptake by
beans was generally obtained at each Zn treatment on unlimed
soils from the first to third crOps, and the increase in Up-
take may have been due to a gradual decrease in pH values
from the first to third crOps. Uptake was inversely corre-
lated with soil pH values. Similar quantities of Zn ferti-
lizer were extracted from unlimed soils by pea bean crops
(0.1 to 3 percent at both 7.5 and 15.0 pp2m Zn application).

Conclusions from this investigation considered to be
most important were:

1. Pea beans generally did not reSpond to Zn applica-

tion on soils with pH values below 7.0.

71

2. On all soils, highest yields were obtained when the
Zn concentration in plants was between 25 and 34 ppm.

3. On the basis of both greenhouse and field studies,
a 0.1 N_HCl extraction procedure was found to be a
good soil test for plant available Zn in Michigan.

4. Generally, yields were increased when Zn fertilizers
were applied after liming soils which tested pH 6.3

or below.

1.

10.

11.

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Bingham, F. T., J. P. Martin, and J. A. Chastain. 1958.
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Boawn, L. C., F. G. Viets Jr., and C. L. Crawford. 1954.
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Nelson. 1960. Effect of nitrogen carrier, nitrogen rate,
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Bonner, J., and J. E. Varner. 1965. Plant Biochemistry.
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Bould, C. 1963. Mineral nutrition of plants in soils.
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Bould, C., D. J. D. Nicholas, J. A. H. Tolhurst, and
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Brechley, W. E. 1914. Inorganic plant poisons and
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Brinkerhoff, F., B. Ellis, J. Davis, and J. Melton. 1967.
Field and laboratory studies with zinc fertilization of
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Sta., East Lansing, Mich. 49:NO. 3, 262-275.

Broadbent, F. E., and G. R. Bradford. 1952. Cation-
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Burd, J. S., and J. C. Martin. 1923. Water diSplacement
Of soils and the soil solution. J. Agr. Sci. 13:265-295.

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74

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81

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APPENDIX

82

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SOumm $.wv 0.N «.5 NHIO EmoH H SOumxooum
"aaosv<
m.~m m.o a.m mwumm emoH awao
o.O¢ m.o o.m mm-dm EmoH meu
m.mm 5.0 o.m amuaa smoa
Nam m.¢m a.m m.n aauo smoH HHH> umcmflz
«.mm ¢.o m.m qumm EmoH xmflo
o.mm m.o m.m mmumm EmoH wade
o.mm m.o 5.5 mmnaa awed mmao
Nam o.mn m.¢ m.” aauo emoa mmHo >wcmm HH> umcmflz
m.mm m.o m.m fimnon Emoa mafia
¢.mm n.o H.m omuwm awed Swan
0.NN m.o o.m flmnfia eon wade
«.mm 5.0 5.5 ¢aum amoH mmau
xmm m.mn m.¢ m.> m-o amoH H> umcmflz
"ummsvd
qu300 SN Hauoe SN manmuomuuxm mm AmmSUSHV musuxme mmflumm USM umvuonsm
Aflom SH SN Ema Spawn
UmDCHUCOU ll 4d GHQMB

89

 

UwDCHUCOU

$.ma m.o m.m ow-mm EmoH ancmm
m.mm m.a m.~ mmumm emoH
m.mm o.m o.m mmnfla emoH
m.mm m.¢ m.h ¢a-m amoH

acumm N.mn «.m m.~ muo emoH HH noozaou
m.am m.o o.m m¢u¢m emoH
N.mn $.o m.fi «m-mm EmoH
N.nm m.« m.~ mm-m amoH

wcmawflz o.mm ¢.¢ 5.5 m-o EmoH H coozaoo
$.m¢ N.0 m.> m¢-0¢ EmoH Swan
m.~« >.« h.» o¢u¢m EmoH Swan
m.a¢ m.m m.> «mumfi amoH
o.¢« m.m m.“ ma-oa emoH

:Oumnm «.0d Nfiv 0.N. OHIO EMOH > COumeOHm
m.mm m.o m.m m¢u¢m EmoH mmau
o.mm m.o 5.5 «mnmm Emoa undo
m.mm m.o m.> mmuma awed
o.m« m.« m.~ mfium amoH

COumm m .Ouv m . N N . N. mlO EMOH >H COumeOHm
m.mm m.o o.m N¢upm EmoH xmao
o.mm m.o m.m pmnmm Emoa mmflo
m.0¢ «.o m.~ mmuwm amoH mmao
«.mw o.m m.~ ¢muma emoH
m.mm m.a «.5 maum emoH

SOumm O.¢¢ H.m O.m mIO EmoH HHH SOumeOHm

"Haosvd
>uSSOU SN HMuOB SN mHQMuUMHuxm mm AmmSUSHV musuxwe mmflumm OSm umouonsm
HHom :H cw 8mm gamma

 

 

UmSSHuSOU II

4H magma

USSSHuSOU

 

 

N.5m N.0 H.m m¢-~m Emofl uSHm
o.mw m.o m.> 5m-mm awed uHHm
o.¢m m.o 5.» mmumd amoa uHSm
ncmauflz m.m« m.m N.N nauo emoH uHHm H Smasflum
"UOSUS
m.mm «.5 N.5 o«-mm awed Smao
m.m¢ m.m N.5 mmumfi amoH Swan
m.m¢ ¢.m H.5 waned emoH Smao
mmmwSBMASm m.mm m.m m.m HHIO Emoa >mHo H mmBSSwA
o m.mm N.0 m.~ mwuon amflo
av «.om H.m m.» omuoa amoS Sada
mm3mcwfl m.®m N.® $.w O.HIO EMOH HHH wHHH>uKAOm
m.mm w.o H.m mwumm mafia
o.¢m m.m m.~ mmuma Swan
0.0m m.m 0.5 manm amao
mm3mSmA m.mm m.Oa 0.5 mIO ESOH wmao HH wHHH>u>om
o.mm 0.0 m.~ N«-om Smao
m.m> m.aa 5.5 omuma Smao
0.0m o.ma 0.N ma-oa amau
moucoz ¢.mm mum m.~ oauo EmoH undo H mSSH>u>om
”Haosvd
qu300 SN HMuOB SN wHQMuumuuxm mm AmwSUSflv muzuxma wmflumm USS vauonsm
HHom SH SN 8mm Spawn
nmscflucoo -- SS magma

91

USSSHSSOU

 

m.mm m.o o.m m¢um¢ Smao
o.mm ¢.o m.~ m«-mm Smao
¢.mo o.m >.m mmuma SmSu
¢.m¢ m.¢ m.m manma emoS Swag
omcmua o.mm a.¢ m.m NH-o amoH Swan xuflxSmm
N.Nw «.o m.m mwumm Smao
m.mm o.m N.5 mm-mfi Smao
m.¢m H.0 o.m maum Swan
Scmflwflz «.mm m.o o.m muo EmoS Smao Sumxusm
SomSSOSSO ¢.Nm m.mH m.m NHIO SSHU SomSSOSSO
w.m¢ m.o m.m m¢-0¢ emoS Smao
o.mm ¢.o o.m 0¢uom awed Swan
«.o« «.o m.> omu¢m emoS Smau
m.mm 0.5 ¢.m Smnm amoS SmHo
omcmua $.«m m.m m.m muo emoS Smao ucmx
“Samuom
«.mw ¢.o m.m mwuam EmoS uaflm
«.SN “.0 N.m «mumm amoS uHSm
w.>m m.o o.m mmum amoS uHSm
Samanflz m.«m m.m m.> muo amoS SSHm HH Smaeflum
"UODUS
>uSSOU SN HSuOB SN manSuoSnuxm mm ASSSUSHV musuxma mwflumm USS HmUuonsm
aflom ca SN SSS SuSmS

 

 

Smscflucoo n- «a manna

USSSHuSOU

 

 

 

0.0 m.o m.m m¢nom Scam
o.oa a.a o.m omnom Scam
m.a« m.a m.m omu¢a Scam
m.ma m.a >.m «aum Scam SamoS
wwumHSSz m.md m.a >.m OIO ESOH mUSSm HHH SHHHSN
o.ma m.o m.m m¢n>n Scam
o.¢m m.o m.m pmumm Scam SamoS
0.0m m.o m.m mm-0m amoH Sncmm
N.5m ¢.a ¢.m omum amoH Socmm
efluucc m.on «.m m.m muo amoa Socmm HH cHHumx
¢.m «.a m.m ¢¢nmm Scam
m.m $.a p.m mmumm Sumo
«.SN m.a m.m mmu¢a Scam
S. m.m« m.a o.m «Hum Scam Samoa
9 SSHS>SHB USSHU O.Nm m.m a.o mnO ESOa >USSm H SHHqu
¢.m O.H m.m pfiumm Scam
¢.m m.o m.m mm->a Scam
S.ma a.m m.m Nana Scam
wmum>mufi UCMHU N.m.fi wlv m.m mIO UCMm >EMOH MXmMXHSVm
m.om m.o m.m m¢uom amoH Sucmm
«.ma 5.0 m.~ omuma amoH Sucmm
m.am m.m m.> md-m amoH accmm
m.m¢ >.m N.5 m-n emoH Socmm
mHmH mswmmum $.Nd m.¢ «.5 >10 ESOH mUSSm umEEm
"Sonuuo
muSSoo SN HSuoa SN SHQSuUSuuxm mm ASSSUSNV Susuxme mmflumm USS HSUHonsm
HSom ca an ESS nuSmS
UmSCHUCOU ll Aw.“ MHQMB

USSSHuSOU

 

 

 

o.mm 0.N o.m SSISS SmoH Swan Sccmm
m.~n m.m m.m SmISN SmoH Sada Sncmm
«.SN o.a m.m SNISH SmoH Sncmm
N.fin m.o m.m fiduma SmoH
szmcH m.om S.H m.m Nfiuo Smoa Socmm xcscom
«.mm a.d m.m mmumn SmoS Sncmm
o.ow 0.N m.m mmumm SmoS Sncmm
o.mm m.H S.m mmuma SmoH
o.mm O.H m.m SHIm SmoH Sncmm
SmsmcH m.mm ¢.N m.m mlo Smoa macaw mSmSmHHSm
m.n¢ m.o m.m NSIHS Sada
0.0m N.0 N.m Hmumm Swan
xv m.mm N.N m.~ mmuhm SmoH Swan
9 N.mm m.w m.“ SNISH SmoH Swan
m.¢m m.¢ N.5 SHIQH SmoS mmao
mmmmcmo m.>m m.m S.S OHIO Smofi mcflamo
“Samoa
SOuSmsom m.>H m.m m.m OHIO ESOH >USSm mSHmHSSZ
EHSUuSoz w.mm H.N m.w OHIO USSm >ESOH EHSUuSoz
m.mw m.o «.S SSINS SmoH
m.«m m.¢ S.m NSISN SmoH Swan macaw
«.mm H.N H.m «mImH SmoS Svcmm
m.mm O.H m.m SHINH SmoH SSSmm
S.Hm m.o m.m Nanm Smoa Socmm
HSSSSA O.mm m.m H.m mIO ESOH >USSm SUHumuz
"nonuuo
qu900 SN HSuOB SN mHnSuoSuuxm mm ASSSUSHV musuxwa mmHuSm USS umUuonsm
HHom SS SN SSS SuSmS
USSSHuSoo II 4H SHQSB

94

 

 

m.m m.o H.m SSISS Scam
N.SH m.o o.m mmnan Scam SSmoH
m.am m.o m.¢ anumm Scam SSmoH
¢.mw S.H m.¢ SNISH Smoa
o.m¢ ¢.a o.m SHIOH Smoa
oonmSmSmx m.«¢ m.n m.m oauo SmoS aflcflao>
"HHOSD
o.mH H.N m.> NSISN SmoH
m.mm 0.N m.m SNINH SmoH Swan
¢.mm o.a m.m NHIS SmoH Smau
acumm N.5N H.m N.5 mno Smoa SSmSz
m.m m.o H.m mwnmm Scam
m.mm S.H m.m mmumm SmoS Sccmm
m.¢m m.m m.m mmnmm SmoS macaw
o.mH S.H m.m NNIHH SmoH chmm
choH m.mH m.m m.m aHuo SmoH mxooq
«.SN m.o 0.N mmumm SmoS Socmm
N.5m m.m H.5 mmuwH Smoa chmm
3SSmuSmS3 N.mN O.O w.w ¢HIO ESOH MSSQSA
m.mm «.H o.m SSImm Scam
«.SN m.a m.m mmuom SmoH Socmm
m.mm S.H ¢.m OSISH SmoH
m.~m m.a o.m maum SmoH
OONSESHSx 0.0m O.m H.m mIO ESOH CONSESHSX
"Samoa
muSSOU SN HSuOB SN SHQSuoSuuxm mm AmmSoSHV muSuxmB mmHumm USS umUuonsm
HSom SS cu SSS nuSmS

 

 

USSSHuSOU II dH SHQSB

HICHIGQN STQTE UNIV. LIBRMTIES
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