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INFLUENCE OF SPRAY MATERIALS

ON THE STRUCTURE OF SOUR
CHERRY LEAVES (PRUNUS CERASJQS
L. VAR. MONTMORENCY)

Thesis for the Degree of M. S.
MICHIGAN STATE COLLEGE

Erik Monradsen Johnson
1949

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THESIS 3 1293 00646 4170

 

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Influence of Spray Materials on the Structure of
Sour Cherry Leaves (Prunus cerasus L. Var.
Montmorency) .

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Erik Monradsen Johnsen

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INFLUENCE OF SPRAI MATERIAIS 0N TIE STRUCTURE OF SOUR CHERRY
LEAVES (PRUNUS CERASUS L. VAR. MONTMORENCY)
—-——-— w t.

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By
Erik Monrodeen J ohneen

A THESIS
Submitted to the School of Graduate Studies of Michigan
State College of Agriculture and Applied Science
in partial fulfillment of the requiremn'be
for the degree of

MASTER OF SCIENCE IN AGRICULTURE

Department of Horticulture

1949

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Table of Contents

Page
Introduction . 1
Review of literature /’ 1
Procedure 4
Results and Discussion 6
Summary 8
Literature Cited _ 10

13 .1758 3

Influence of Spray Materials on the Structure of Sour Cherry
leaves (Prunus cerasus L. var. Montmorency)

By
Erik Monradsen J ohnsen

Introduction

The influence of insecticides and fungicides on the function
of leaves of fruit trees has been a subject of much experimental
work. In general it has been found that certain of these materials
at various concentrations and in various combinations may markedly
affect the size, structure and photosynthetic activity of leaves.
Most studies have dealt, however, with fruits other than the cherry.

The purpose of this study has been to determine the influence
of certain spray materials on the leaf grcnth of the sour cherry
(M oerasus L. var. Montmorenoy), using the longitudinal diameter

or length of the palieade cells as a measure of this influence.

Review of Literature

The concentration at which a chemical compound or spray material
is applied to a plant has been, in many cases, the determining factor
in the response of the plant to the material. Talts (19) demonstrated

that a weak solution of zinc compounds mixed with a growth-substrate

for the germination of spores stimulated spore growth. Stronger
solutions showed either no change in the growth, decreased growth,
arrested growth, or showed death of the spores, depending upcn.the
anion and the concentration. Menzel (11) found no influence on
foliage leaves of Pelargonium from.a weak solution (less than

1 per cent) of a copper compound. However, for stronger solutions

(1 per cent), a.micrcsccpic study of the internal structure showed
shriveling of the cell'wall, decreased length of the palisade cells
in some places, increased size of cells in the epongy'mesophyll;
‘while the intercellular Spaces were much decreased, and in.many places
absent. Although there was no visible influence on the epidermis, the
protoplasm.in the cells, both of palisade and spongy mescphyll,
tended to collect on the inside of the cell wall and showed an arti-
ficially green discoloration. The thickness of the leawes was de-
creased.

These influences an the internal leaf structure of Pelargonium
became more pronounced as the ocncentraticn.of the solution was in-
creased. Furthermore, a relatively high concentration (5.5 per cent)
caused the death of some cells. More cells were killed in.the epi-
dermis than in the spongy mesophyll, and fewer were killed in.the
palisade tissue than in the epongy mesophyll. 0f the plants used by
menzel (11) the influence of chemical compounds on internal structure
was more pronounced with Pelargcnium, but was similar on the leaves
of apple and other plants investigated.

Memel (11) also demonstrated by localized treatments that the

effects of chemical solutions was not restricted to the regions of the

leaf directly below the area treated. From his work there appeared to
be an absorption and. translccation of the chemical, which brought about
physiological changes in untreated areas of the leaf. Roberts, South-
wick and Palmiter (17) showed that the epidermal layer was continuous,
and connected with the vein extensions, but that the cell walls and
cutinised layer was not continuous. Since the outinized layer and cell
walls were not continuous the absorption of spray materials takes

place very readily.

Heinicke (s), Hoffmaml (4,5), Eyre (9), and others have reported
findings of reduced photosynthesis following spray applications, and
greater reduction from strong spray materials than for mild spray
materials. Mills (12), Rassnmssen, Toenjes and Strong (16) have
shown that the cumulative effect of reduced yield is greater for lime
sulfur than for elemental sulfur sprays. Dutton (2) showed that a
spray injurious to the foliage leaves of the apple reduced growth of
wood, and the rate of reduction was different for lime sulfur and
Bordeaux mixture. Horsfall and Turner (8), Horsfall, Harvey, and
Suit (7), Bonds, Folsom, and Tobey (l), and others have shown that spray
materials may reduce the growth of plants. According to Southwick and
Childers (18) the influence of Bordeaux mixture on the photosynthetic
process of apple leaves was primarily physiological rather than
mechanical.

Pickett and Birlceland (13, 14, 15) believed that the reduction
in photosynthetic activity brought about by some spray materials on

apple foliage may be explained, in part, by the reduction of the

amount of the internally exposed cell wall surface. Kenworthy (10)
found a highly significant correlation between the total length or
depth of the palisade mesophyll and the ratio of the internally ex-
posed cell wall surface to the external surface. This was later
confirmed by Pickett and Birkeland (14) who found that the so-called
”stronger" or more caustic spray materials, as lime sulfur, inhibited
the development of the palisade cells more than the so-called “mild"
or less caustic spray materials such as elemental sulfur. Also, re-
peated applications inhibited the development of palisade cells more
than single applications. According to these reports a measurement
of the depth of the palieade tissue or longitudinal diameter of pali-
sade cells becomes a practical test of the physiological effects of
spray materials that should save investigators considerable time in

evaluating new spray materials .

Procedure

Four-year-old cherry trees grown on the Horticultural farm of
Michigan State College, at East Lansing, Michigan, were used in this
study. The trees were grafted on seedlings of m mahaleb L. Eight
different spray treatments were used. There were four replications

for each treatment; each replication having two trees.

The treatments were applied five times during the season, using
4 preharvest and l after-harvest application. The treatments used
may be outlined as follows (amounts listed are for 100 gallons of
water):
1. Basic copper sulfate-~15 lbs.*
Hydrated lime-4 lbs .
Arsenate of lead-~2 lbs.
2. Basic copper sulfate-"3 lbs.*
Hydrated lime-«.4. lbs.
Benzene hexachlcrideul’: lbs .1”
3. Basic copper sulfate-79 lbs.‘I
Hydrated lime-m4 lbs .
Parathion (15%)-1 lb.

4. Ferric dimethyldithiocarbamate-n-l 1/2 lbs.
Arsenate of lead--2 lbs.

5. Ferric dimethyldithiccarbamate--l 1/2 lbs.
Benzene hexachlcride--3 lbs .1»

6. Ferric dimethyldithiccarbamate—l 1/2 lbs.
Parathion (l5%)--1 lb.

7. Liquid lime sulfur--2 gals.
Arsenate of lead-22 lbs.

8. Liquid lime sulfur-~22 gals.
Parathion (15%)«1 1b.

On August 7, 1948 sixteen leaves were collected from each treat-
ment, except for treatment No. 6 where only 12 leaves were collected.
Two leaves were collected from each tree. The leaves selected were
removed from the southern periphery of the tree and near the middle of

shoots of average vigor.

*The basic copper sulfate was known as Tennessee 26 which contains
a metallic equivalent of 28% copper.

IM'The benzene hexachlcride contained 6% game. isomer.

The portion of the leaf used was located midway between the base
and the apex of the leaf. From each leaf, one 5/4 centimeter square
piece of leaf, including half of the midrib, was placed in a formalin-
acetic acid-alcohol killing and fixing solution, dehydrated and embedded
in paraffin.

The sections were out eight microns thick and were stained with
haemotoaqlin and safranin. One slide of several sections was made
from each leaf piece, with the sections being made across the midrib
as a means of orientation. The total length of the three layers of
palisade cells was measured at ten different places near the midrib

and in areas devoid of smaller veins.

Results and Discussion

The results of the measurements are shown in Table 1. Table 2
shows the occurrence of significant differences between the mean
values for treatments. The slides showed dark spots of materials
that were formed in the epidermal cells, and in some sections this
material moved out over other parts of the leaf sections during the
staining process (see Figure 1). These dark spots could not be
associated with any certain treatment, and may be the gum-like sub-
stance fcund consncnly on cherry leaves.

The data given in Table 1 show that sprays containing copper are
more injurious to leaves of cherry trees than sprays containing ferric
dimethyldithiocarbamate, and that sprays containing liquid lime

sulfur were less injurious than sprays containing ferric dimethyldi-

 

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Table 2. The Occurrence of Significant Differences between

 

 

 

 

Treatments '

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thiocarbamate (Figures l, 2, and 3). The different insecticides
likewise showed different influences on the leaves. Sprays containing
parathion were less injurious than sprays containing benzene hexa-
chloride. Benzene hexachloride in combination with ferric dimethyl-
dithiocarbamate, or parathion was less injurious than arsenate of lead
with the same fungicides. However, benzene hexachloride in com-
bination with basic copper sulfate was slightly more injurious than
when basic copper sulfate was used with arsenate of lead. Apparently
benzene hexachloride has an undesirable reaction with c0pper compounds.
The greatest thickness of the palisade layers was associated with the
use of parach in combination with liquid lime sulfur, while the
smallest thickness of the palisade layers was associated with the use
of benzene hexachloride in combination with basic copper sulfate.

The results obtained in this study show that all of the spray
materials used had an effect upon the development of palisade cells,
and that certain combinations may have a greater influence than sprays
containing only one insecticide or fungicide. Since the development
of the palisade cells is correlated with the photosynthetic activity
of the leaves , the changes in their development brought about by the
use of spray materials should eventually be reflected in the growth
and fruitfulness of the tree. The length of time required for this
influence to become measureable in the field will depend upon the
magnitude of the influence. Many of the differences reported here
may appear small in numerical value, but are statistically significant,

and, although their influence on tree growth could not be measured

within a single year, the trees should eventually show a correspondingly

significant difference in growth and production.

Summary

Three commonly used fungicides and three insecticides were
studied in combination with one another, such as may be used in com-
mercial orchards. Their effect upon the structure of sour cherry
leaves (m cerasus L. var. Montmorency), using the depth of the
palisade mesophyll as an index, was studied.

The fungicides were found to have a very marked effect upon the
palisade development. The palisade development associated with the
use of basic copper sulfate was less than that associated with either
ferric dimethyldithiocarbamate or liquid lime sulfur when the same
insecticide was used. Likewise the palisade development associated
with the use of ferric dimethyldithiocarbamate was less than that
associated with the use of liquid lime sulfur.

The insecticides were found also to have a marked effect upon
palisade development. Arsenate of lead resulted in less palisade de-
velopment than parathion when the same fungicide was used.

Benzene hexachloride with basic 00pper sulfate resulted in less
palisade development than either arsenate of lead or parathion.

When benzene hexachloride was used in combination with ferric dimethyl-
dithiocarbamate the palisade development was greater than when the
latter was used with arsenate of lead, but less than when parathion

was used in combination with ferric dimethyldithiccarbamate.

The magnitude of the physiological effects that spray materials
may have is illustrated by comparing the palisade development asso-
ciated with the mixture of liquid lime sulfur and parathion with
that associated with the use of benzene hexachloride and basic copper
sulfate. There was 28.7 per cent less palisade development in the
leaves sprayed with benzene hexachloride and basic copper sulfate than

when the leaves were sprayed with liquid lime sulfur and parathion.

l.

2.

3.

4.

5.

6.

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8.

9.

10.

ll.

10
Literature Cited

Bonds, D. Folsom, and E. R. Tobey. 1929
Potato spraying and dusting experiments. 1926-1928.
Maine Agr. Exp. Sta. Bul. 552: 97-140.

Dutton, W. C. 1952
Spray injuries II. Secondary effects of spray injury to
apple foliage. Mich. Agr. Exp. Sta. Spec. Bul. 219. 58 pp.

Heinicke, A. J. 1958 7
How lime sulfur spray affects the photosynthesis of an
entire apple tree. Amer. Soc. Hort. Sci. Proc. 55: 256-259.

Hoffman, M. B. 1955
The effect of certain spray materials on the carbon dioxide
assimilation by McIntosh apple leaves. Amer. Soc. Hort.
Sci. Proc. 29: 589-595. A

Hoffman, M. B. 1954
Carbon dioxide assimilation by apple leaves as affected
by line sulfur sprays II: Field experiments. Amer.
Soc. Hort. Sci. Proc. 50: 169-175.

Horsfall', James G. 1945
Fungicides and their action. Chronica Botanica Company.
Waltham, Mass.

Horsfall, Jo Go, Go Ea Re Hervey, and Re Fe Suit. 1939
Dwarfing of cucurbits sprayed with Bordeaux mixture.
Jour. Agr. Res. 58: 911-928.

Horsfall, Jo Go, and No Turner. 1943
Injuriousnoss of Bordeaux mixture. Amer. Pot. Jour.
20: 508-320.

The effect of sulfur fungicides on the photosynthesis
and respiration of apple leaves. Cornell Agr. Exp. Sta.
Mom. 222. 40 PP.

Kenworthy, A. L. 1959
A study of the internally exposed surface of foliage
loaves of five varieties of apples. Unpublished Thesis.
Kansas State College.

Menzel, Klaus-0hr. 1955
Unto rsuchungen der schadigenden Wirkengen kupforhaltiger
Spritzmittel. Angewante Botanik. Berlin 17: 225-255.

12.

15.

14.

15.

16.

17.

18.

19.

11

Mills, W. D. 1947
Effect of sprays of lime sulfur and elemental sulfur
on apple in relation to yield. Cornell Agr. Exp. Sta.
Mom. 275. 58 pp.

PiCkBtt, We Fe. and Birkeland, C. J. 1941
Common spray materials alter the internal structure
of apple leaves. Amer. Soc. Hort. Sci. Proc. 58: 158-162.

, and . 1942

 

The influence of some spray materials on the internal

structure and chlorophyll content of apple leaves.
Kansas Agr. Exp. Sta. Tech. Bul. 55. 54 pp.

, and . 1942

 

 

Further studies on the effect of common spray materials on
the internal structure of apple leaves. Amer. Soc. Hort.
305.. PI‘OO. 403 371-3730

Rasmussen, E. J., W. Toenjes, and F. C. Strong. 1948
Effect of spray treatment on foliage injury, pest control,
and yield and quality of apples. Mich. Agr. Exp. Sta.
Spec. Bul. 347. 26 pp.

Roberts, E. A., M. D. Southwick and D. H. Palmiter. 1948
A microchemical examination of McIntosh apple leaves showing
relationship of cell wall constituents to penetration of
spray solutions. Plant Physiol. 25: 557-559.

SOUth'iOk, Fe ‘30. and Ne Fe Childers. 1940
The influence of Bordeaux mixture on the rate of photo-
synthesis and respiration of apple leaves. Amer. Soc.
Hort. Sci. Proc. 57: 574.

Talts, von J. 1952
Einfluss der Schwermetallsalze auf Penicillium Glaucum
(mit besonderer Berucksichtighung der anionenwirhmg).
Protoplasma, Leipzig 15: 188-258.

Acknowledgment

The author wishes to acknowledge the assistance
of Dr. A. L. Kenmorthy (principal adviser), Mr. A. E.
Mitchell, Dr. D. P. Watson, Dr. H. B. Tubey, and Dr.
I. M. Felber (deceased) for their assistance in plan-
ning and conducting these investigations, and for aid

in the preparation of the manuscript.

Figure 1. Comparative effects of spraying leaves with basic copper
sulfate in combination with:

A. Arsenate of lead. The total length of palisade
cells-406.4 microns

B. Lime ,and benzene hexachloride. The total
length of palisade cells-105.1 microns

C. Lime, and parathion (15%). The total length
of palisade cells—125.2 microns

 

 

 

Figure 2. Comparative effects of spraying leaves'with ferric
dimethyldithiocarbamate in combination with:

A. .Arsenate of lead. The total length of
palisade cells--ll5.5 microns

B. Benzene hexachloride. The total length of
palisade cells--l56.2 microns

C. Parathion. Total length of palisade cells--
138 09 microns

 

 

Figure 5. Comparative effects of spraying leaves with liquid
lime sulfur in combination with:

A. Arsenate of lead. The total length of

palisade cells--155.5 microns

B. Parathion. The total length of palisade cells-
144.6 microns

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