Relation of Manganese to the Carotene and Vitamin Content of Growing Crop Plants

Citation preview

PURDUE UNIVERSITY

THIS IS TO CERTIFY THAT THE THESIS PREPARED UNDER MY SUPERVISION

OTHMAR JOSEPH BURGER

BY

e n title d

RELATION OF. MANGANESE TO THE CAROTENE

AND VITAMIN CONTENT OF GROWING CROP PLANTS

COMPLIES WITH THE UNIVERSITY REGULATIONS ON GRADUATION THESES

AND IS APPROVED BY ME AS FULFILLING THIS PART OF THE REQUIREMENTS

FOR THE DEGREE OF

DOCTOR OF PHILOSOPHY

P rofessor

H

eap of

in

Charge

S c h o o l ,o h D

TO THE LIBRARIAN:----THIS THESIS IS NOT TO BE REGARDED AS CONFIDENTIAL

GRAD. SCHOOL FO R M O—3 - 4 0 —1M

of

Thesis

epartment

RELATION OP MANGANESE TO THE CAROTENE AND VITAMIN CONTENT OF GROWING CROP PLANTS

A Thes is

Submitted to the Faculty

of

Purdue University

by

Othmar Joseph Burger

In Partial Fulfillment of the

Requirements for the Degree

of

Doctor of Philosophy

June, 1950

ProQuest N um ber: 27714095

All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is d e p e n d e n t upon the quality of the copy subm itted. In the unlikely e v e n t that the a u thor did not send a c o m p le te m anuscript and there are missing pages, these will be noted. Also, if m aterial had to be rem oved, a n o te will ind ica te the deletion.

uest ProQuest 27714095 Published by ProQuest LLC (2019). C opyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States C o d e M icroform Edition © ProQuest LLC. ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106 - 1346

ACKNOWLEDGMENTS The author expresses his appreciation and gratefulness to Dr. S. M. Hauge, his major professor, for his interest, encouragement, and assistance in this w o r k .

He is indebted

to Dr. F. W. Quackenbush, and Dr. D. M. Doty for their help and guidance.

Grateful acknowledgment is made of the great

assistance of A . E. Purcell and Katharine Hivon during sampling of the material and to G. Paul Lehman for climatological data. The author expresses his appreciation to Dr. B. R . Bertramson. Dr. A. J. Ohlrogge, Dr. H. H. Kramer, Dr. G. 0. Mott, Pro f . S. R. Miles and Glen Hemstock of the Agronomy Department for their advice and h e l p . The author is grateful to the Tennessee Corporation, Atlanta, Georgia, for the financial aid making possible the fellowship to study this problem. and interest,

For their encouragement

the author is indebted to Dr. J. K. Plummer,

Director of the Products Division of the Tennessee Corpora­ tion and Mr. J. Bergeaux. Agronomist for the Tennessee Manganese Corporation.

TABLE OF CONTENTS Page ABSTRACT ..................................................

i

I.

INTRODUCTION ........................................

1

II.

LITERATURE R E V I E W ....................................2

III.

EXPERIMENTAL P R O C E D U R E .............................. 11

IV.

A.

C o o p é r a t i o n .................................... 11

B.

Location of P l o t s .....................

C.

Minor Element Fertilizers

D.

Soil A p p l i c a t i o n s ..............................15

E.

Spray Applications

F.

Sampling

G.

Methods of Chemical A n a l y s i s ..........

...

11

* ............ 13

............................

20

.................................... . 2 0 21

1.

M o i s t u r e ................................. 21

2.

C a r o t e n e ..................................21

3.

Carotene Lipoxidase Activity ..........

4.

P r o t e i n ..................................21

5.

C h o l i n e ..................................21

6.

Tocopherol

7.

Leaf-Stern R a t i o s ......................... 22

21

......................... 21

H.

Climatological D a t a ........................... 22

I.

Statistical Analysis

. . . . .

EXPERIMENTAL R E S U L T S ........................ A.

Effect of Manganese upon the Carotene Content of Plant Tissues . . . . .

27 27 27

1.

S o y b e a n s .................

27

2.

C o r n ...................................... 41

Page

B.

C.

D.

3.

W h e a t .....................................44

4.

O a t s ....................................... 44

Effect of Manganese on the Activity of the Carotene-Destroying Enzyme . . . . . . . . . 1.

S o y b e a n s .................................. 44

2.

C o r n ..........................

3.

W h e a t .....................................47

4.

O a t s ..................................... . 4 7

Effect of Manganese on the Protein Content

Gr.

. . 49

Soybeans

2.

O a t s ....................................... 53

. . . . . .

Effect of Manganese on the Choline Content

49

. . 53

S o y b e a n s .................................. 53

Effect of Manganese on the Tocopherol Content. 53 1.

P.

47

1.

1. E.

44

S o y b e a n s ...................

53

Effect of Manganese on the Leaf-Stern Ratios of P l a n t s ..................................58 1.

S o y b e a n s .................................. 58

2.

W h e a t .................................... 65

Effect of Manganese upon Anatomical Characteristics.... ..........................

65

1.

S o y b e a n s ......................

65

2.

Oats

69

.................

V.

D I S C U S S I O N ........................................ 69

VI.

S U M M A R Y ........................................... 73

VII.

B I B L I O G R A P H Y ...................................... 76

LISTS OF TABLES AND FIG-UBES List of Tables Table

Page

1.

Effect of manganese upon the carotene content of soybean leaves. Test plot on Maumee loam. Walkerton, Indiana, 1946

2.

Location of wheat experiments, soil reaction and description of treatments, 1948 . . . . . . . .

3. 4. 5.

Fertilization treatments on soybeans plots at Nev/land, Indiana on Emil Savage Farm, 1947

14

. . 16

Fertilization treatments on corn plots at PinneyPurdue Farm, Valparaiso, Indiana, 1947 . . . .

17

Fertilizer treatments on soybean plots, Baxter Blosser Farm, Winamac, Indiana, 1948

18

6.

Fertilization treatments on soybean plots on Place Farm, Walkerton, Indiana, 1949. Replicated four t i m e s ...................................... 19

7.

Rainfall data for the minor element field experi­ ment, Blosser Farm, Winamac, Indiana, 1948 . . 23

8.

Temperature data for the minor element field ex­ periment, Blosser Farm, Winamac, Indiana, 1948. 24

9.

Rainfall data for the manganese field experiment. Place Farm, Walkerton, Indiana, 1949

25

10.

Temperature data for the manganese sulfate field experiment. Place Farm, Walkerton, Indiana,, 1949 26

11.

Influence of minor element fertilization on the carotene content and the activity of the carotene-destroying enzyme of soybean leaves, 1947. Average of three replicates . . : ...

29

12.

Influence of minor element treatment on the caro­ tene content and enzyme activity in soybean leaves, Blosser Farm, Winamac, Indiana, 1948. Average of four replicates .............. 30

13.

Influence of minor element treatments on the caro­ tene content and enzyme activity in soybean stems, Blosser Farm, Winamac, Indiana, 1948. Average of four r e p l i c a t e s ..................... 31

Table

Page

14.

Influence of minor element fertilization on the carotene content of soybeans. Blosser Farm, Winamac, Indiana, 1948. Average of four replicates ................................ 32

15.

Influence of manganese on the carotene content and enzyme activity in soybean leaves. Place Farm, Walkerton, Indiana, 1949. Average of four r e p l i c a t e s .......................... 36

16.

Influence of manganese on the carotene content in soybean stems, Place Farm, Walkerton, Indiana, 1949. Average of fourr e p l i c a t e s ................ 37

17.

Influence of manganese on the carotene content of soybeans. Place Farm, Walkerton, Indiana, 1949. Average of fourr e p l i c a t e s ...................... 39

18.

Influence of minor element fertilization on the carotene content and the activity of the caro­ tene destroying enzyme of corn leaves, PinneyPurdue Farm, Valparaiso, Indiana, 1947. One r e p l i c a t e ..................................... 43

19 .

Carotene content, per cent carotene destroyed dur­ ing incubation and leaf-s tern ratio of wheat plants given treatments as listed, located on the Wynkoff, Lewis, and Blosser Farms, respec­ tively, 1948. One replicate on each location . 45

20.

Influence of manganese on weight, carotene content, and crude protein content of Clinton oats, Hayter Fa r m , Walkerton, Indiana, 1949. Average of three replicates .......................... 46

21.

Influence of manganese on the protein content in soybean leaves and stems. Place Farm, Walkerton, Indiana, 1949. Average of four replicates . . 50

22.

Effect on manganese on the choline content of soy­ bean leaves and stems, Place Farm, Walkerton, Indiana, 1949. Average of four replicates . . 55

23.

Effect of manganese on the tocopherol content of soybean leaves. Place Farm, Walkerton, Indiana, 1949. Based on composite offour replicates. . 59

24.

Influence of minor elementtreatments on the leafstern ratio of soybean, Blosser Farm, Winamac, Indiana, 1948. Average of four replicates . . 61

Pag

Table 25.

Effect of manganese on ttie leaf-stern ratio of soy­ beans , Place Farm, Walkerton, Indiana, 1949. Average of four r e p l i c a t e s ..................... 62

26.

Effect of manganese on several anatomical charac­ ters of typical soybean plants of each, of the four treatments. Sampled at termination of experiment, August 9, 1949. Place Farm, Walkerton, I n d i a n a ...............

68

List of Figures Figure 1.

Pag

Influence of manganese sulfate and copper sulfate on the carotene content of soybean leaves, 1948

53

2.

Influence of manganese sulfate and copper sulfate on the carotene content of soybean plants,1948. 35

3.

Influence of manganese sulfate on the carotene content of soybean leaves, 1949 ........

4. 5. 6.

Influence of manganese sulfate on the carotene content of soybeans plants, 1949 ........ ..

38 . 40

Influence of manganese sulfate on the carotene content of soybean stems, 1949 Influence of manganese sulfate on the activity of the carotene-destroying enzyme in soybean leaves, 1949

42

48

7.

Influence of manganese sulfate on the protein con­ tent of soybean leaves, 1949 51

8.

Influence of manganese sulfate on the protein con­ tent of soybean stems, 1949 ............. 52

9.

Normal and manganese deficient Clinton oats which were grown on Maumee loam and sampled on May 25, 1949

54

Influence of manganese sulfate on the choline con­ tent of soybean leaves, 1949

56

10.

Figure 11. 12. 13.

Page

Influence of manganese sulfate on the choline con­ tent of soybean s terns, 1949 ...................

57

Influence of manganese sulfate on the tocopherol content of soybean leaves, 1949 ...............

60

Influence of manganese sulfate on the leaf-stern ratio of soybean plants, 1949 .............

63

14.

Normal and manganese deficient Hawkeye soybeans which were grown on Maumee loam. July 21, 1949. 64

15.

Appearance of the terminal leaves of typical trifoliates of the various treatments sampled August 9, 1949

67

RELATION OF MANGANESE TO THE CAROTENE AND VITAMIN CONTENT OF GROWING CROP PLANTSl

-^Abstracted from the doctoral thesis of Othmar Joseph Burger, Purdue University, 1950.___ ______________

0. J. Burger and S. M. Hauge Graduate Fellow in Agricultural Chemistry, Tennessee Corporation Fellowship and Associate in Biochemistry.

AN ABSTRACT

il Manganese deficiencies in plants have been observed on many farms in northern Indiana•

The production of soybean hay

and grain has - been severely reduced in fields exhibiting chlorosis or leaf-yellowing•

Farmers have reported similar

effects in wheat and oats. % Manganese is essential for normal plant growth.

The

concentration of manganese in plant tissues may vary consid­ erably.

Lyon and Beeson (15) reported that the manganese

content of plants increased progressively with increased manganese concentration of the medium in which the plants grew.

Manganese is located mainly in the leaves and actively

growing parts of plants♦ The function of manganese in plant metabolism is not known.

According to McHargue (16, 17) it is closely associ­

ated with vitamins and is responsible in some way for the synthesis of vitamins, probably catalytically.

Manganese

fertilization increases chlorophyll formation (4, 22) and thus functions in forage and seed production (23)♦

Photo­

synthesis is increased by accelerating the oxidation pro­ cesses connected with the photosynthetic reactions in the leaf (5). It has been found that manganese deficiency decreased the carotene content of tomatoes and beets (7).

The lack of

any essential element which reduced the green color of foliage decreased the carotene content (2, 8) .

However, other in­

vestigators have found no consistent correlation between manganese and carotene content (6, 14).

ill Environmental conditions may affect the carotene content of the plants •

Moon (19) found that periods of low rainfall

decreased the carotene content of some grasses.

Other workers

(14, 21) reported that slight environmental differences did not affect the carotene content. Manganese has been found to affect the activity of some enzymes commonly found in plants.

Bailey and McHargue (1)

reported that manganese depressed the activity of catalase, oxidase, and invertase activity.

According to Jones (13)

large amounts of manganese are found in tissues which contain large amounts of oxidizing enzymes • There is an enzyme system in plants which is responsible for the destruction of carotene during the field curing pro­ cess .

This was shown by Mitchell and Hauge (18) to be a

lipoxidase and that soil fertility or additions of fertilizers containing N, P, and K had no effect on the carotene-destroying system of alfalfa. The lipoxidase activity may be inhibited by alphatocopherol (11, 12).

The autoxidation of oils and fats may

be inhibited by the presence of carotene and vitamin A be­ cause of the greater lability of these substances to oxida­ tion (10).

According to Heftmann (9) tocopherol appears to

be responsible for protection of carotene and other autoxidizable substrates. It was the purpose of this study to determine the effect of manganese on the carotene content of corn, wheat, oats, and soybeans, on the carotene-destroying enzyme system, on

iv the protein, choline, and tocopherol contents of soybeans.

MATERIALS AND METHODS Field experiments were conducted on a Maumee loam soil in the Kankakee River Valley of northern Indiana.

The sur­

face layer of this soil is dark gray and the M horizon is a yellowish gray clay loam (23).

This soil has good internal

drainage because of artificial drainage by open ditches. Experimental plots were located on areas where severe man­ ganese deficiency in plants had been observed during the previous year. < ■

The. soil had a pH of about 6.5.

After 100 pounds of PgOg and KgO were drilled into the

soil, manganese fertility plots were laid out.

On June 1,

1949, Hawkeye soybeans were planted about 2 inches deep in rows 3 feet apart *

On June 25, M12SO4 (Tecmangam, 65$ MnSO^.)

was applied either to the soil or as a spray to the foliage of the soybean plants.

It was applied to the soil at the

rates of 75 and 150 pounds per acre about 1^ inches to each side of the seed row and about 2 inches deep with a Planet Junior drill.

Treatments were replicated four times in

individual 16-foot rows. Samples of soybeans were taken periodically throughout the major portion of the growing season by harvesting the above-ground portion of the plants from a 2-foot section of each plot from all four replicates.

The dates of sampling

were July 5, July 13, July 21, August 1, and August 9.

V The effect of manganese fertilization on the carotene content of corn, wheat, and

oats was studied on selected

plots in northern Indiana*An area of wheat

and oats which

were growing in a manganese deficient area was set off and a 10-foot square portion was treated with a spray application of MnS04 at the rate of 5 pounds per acre,

The corn was

given a soil treatment of 29*5 pounds per acre of MhS04 (67% MnS04) •

This was applied about 1^ inches to each side

of the seed row and about 2 inches deep with a Planet Junior drill. The wheat and oats were sampled on June 1, 25, 1949, respectively; the July 30, 1947 * ground level*

corn was sampled on

1947 and May July 7 and

When sampled, the plants were cut off at the Carotene analyses were made on the leaves of

corn, wheat, and oats* The plots were located from 60-100 miles from the laboratory.

To minimize deterioration, the samples were

collected during the early morning hours, placed into insu-* lated chests which contained about 15 pounds of crushed ice, brought to the laboratory as soon as possible, and unpacked immediately into a refrigerator for temporary storage.

The

leaves and stems were separated, finely chopped, and thor­ oughly mixed.

Samples of 5 grams each were placed in small

glass bottles and stored in a freezing room, -15°C., for subsequent carotene analyses.

Portions of the leaves and

stems were taken for moisture determinations♦

A 1-gram

sample of oven-dry material was used for protein analyses

vi and two gi*suns were taken for the choline déterminations •

A

10-gram sample of fresh material was used for tocopherol analyses. Carotene determinations were made by the method of Zscheile and Whitmore (24)•

The carotene-destroying enzyme

activity was determined by measuring the loss of carotene which took place during the incubation of plant material at 37° C • for a period of 40 h o urs.

The protein content was

calculated by multiplying the Kjeldahl nitrogen by the factor 6.25,

The method of Engel (3) was used for choline

analyses and the modified method of Parker and MacFarland (20) was employed for tocopherol analyses.

RESULTS AND DISCUSSION In a preliminary study made in 1946, leaves of soybean plants from the minor element experimental plots of Steckel (23) were analyzed for carotene.

The characteristic chlo­

rosis and necrosis appeared during the growing period on the untreated plots, and was markedly decreased as the quan­ tity of MhSO^ was increased with complete visible recovery on the higher levels. Carotene determinations on the fresh leaf tissue indi­ cated a definite relationship between carotene content and manganese fertilization.

The leaves of soybeans which re­

ceived no manganese contained 258 micrograms of carotene per gram dry matter while the carotene content of the leaves of

vil the soybeans which received a soil treatment of 212 pounds of MhS04 per acre was 535 micrograms per gram.

The leaves

of those plants which received 10 pounds MnS04 per acre as a spray contained 544 micrograms per gram.

When heavy appli­

cations of N, P, and K were applied in addition to MnS04 , the carotene content was increased to 660 micrograms per gram. Copper and zinc sulfate appeared to be of little value in correcting the leaf chlorosis or increasing the carotene content. The carotene contents of the leaves and stems of soy­ beans grown on manganese-deficient muck soil during 1949 are summarized in figures 1 and 2.

The 5 pounds of MhS04 per

acre applied as a spray to the foliage was very effective as an immediate corrective treatment for manganese deficient plants as manifested by increased carotene content.

The

greatest increase in carotene content was obtained by the soil application of 150 pounds of MnS04 per acre. effect persisted throughout the entire season.

This

This in­

crease was significantly higher than the soil treatment of MnS04 at the rate of 75 pounds per acre.

Apparently, the

manganese from the spray treatment applied at the beginning of the season was dissipated toward the end of the season as manifested by evidence of chlorosis and lowered carotene content. The effects of various manganese treatments on the carotene content of the stems were different from those on the leaves, as shown in figure 2.

It is apparent that the

viii

380

LU

S O I L - 150

S O IL - 7 5

Q 300

Ld CL 220

Ld

O I 80 4 was generally higher than the protein content of the stems of plants re­ ceiving manganese treatments.

This might be related to the

apparent maturity of the plants.

The reproduction period of

the plants of the no treatment plots were delayed considerably. After the initial period, the MnSO^ treatments produced a definite reduction of choline content of leaves of soybeans, but an apparent increase in the stems, as shown in Table 3,

PLANTS

xii

CD

-P

cd

ON THE CAROTENE

CONTENT

IN THE LEAVES

OF VARIOUS

CD

-p

-P

Üo •H CD

CD O i —t H

GO

O

GO

'sh 02

CO

02

—I o !> i « —I to

O i —l

!> i —I i —I

CO

CD P cd o 1— 1

1 —I 04 X X 'i'

LO • p CD f i P CD02 CD P fi fi CD sh O fi O S O cd * o & -p cd id * LO CD ^ CQ CD fi ^ fi Eh H CD P O fi cd

SULFATE OF MANGANESE

02

CD CD CQ 0 p cd CD 0 CD 0 P CD F-.•H O P o 0, fi (d O a 1—1 o o

O

INFLUENCE

H

oa So

T) 5 I

cd fi 04 ta cd

as i to

Z 60

vb ^1» LU 5 0

45

LU

CL

35

I0 DAYS JU LY 5

Pig. vity of leaves, L.S.D. L.S.D.

18

26

AFTER T R E A T M E N T JU L Y 13

JU LY 2 I

37

45

A P P L IC A T IO N AUG I

AUG 9

3. Influence of manganese sulfate on tlie acti­ th.e carotene-des troying enzyme in soybean 1949. Average of four replicates. (5%): July 5, 8; July 13, 3; July21, 8; August 1, 8; August 9, 7. (1%): July 5, 12; July 13, 5; July21, 11; August 1, 11; August 9, 10.

xiv

• CD ▻

M m M 0 GO O EH !s»

* -P

EH

CD

02

to

1— 1

LO r— 1

LO 1— 1

CO

H

CO 02 «T>

!m

&

rH tO 0 rH f-0

CD

O O

CO rH

-P H M EH O Ph

CD ^ CD 1— 1

1— 1 ir.

10

O H>

CO 1

1 1 1 1

* ❖

X *

03 CD

-p cd o •H

I—I A CD Fh Î

g $

02

H

CO

O

co

02 02

O to

0 to

00 02

CO

s

CD

CO H

cd

CO

I 1I I 0

H1 o

CD -P

CO

O




CO

cd CD

Ï

0

tO

02 02

rH to

0 to

02 02

rH O to

r'a

o

rH

$ 5;

}> )

u

*

LO

00 0 to

to 0 to

C-

*

X =; * *

O to

to

GO

00

CD 02

02 02

£> 02

00 02

02

00

CO

0

rH to

O to

0 to

02 to

02

tO

0 Pk

o rH

I

LO

3

H

CD - P CQ t i CD CD

bû oS *h ti

m

CD

-p

CD ^ O

• CO

43

%

LO

1 0

LO o

o 10 rH

0 hO cd -p C 0 0

U O)

Hi • rH

a •H nd 0 GO GO 0

rH •

10

Q

P

co

CO •

J

CO

T5 £ 0

»>5 r— I rH 0 02 H

00

Ph X w

• y--^ H* O

CO d g

LO CO '— '

cd bO P cd S 0 0 EH •N -p 0 s

cd bO P cd S 0 0 EH

cd 0 A -P

ra 5 cd Ph Pk GO

:|c

T X; xc

. -p p 0 p. 0 i rH -P P cd 0 •H

co

g

Ë.

PH 0 H

1

I

• o

to LO • o

e bO ^ CT!
i— 1 DO

03 t> to

1 1 1

CO LO 02

02 O to

03 co 03

LO 02 to

02 'xh to

1 1 1

i— I o

o CO ti

'xh ^ O O CO to d d s o

o CO d

-p d o o

o CO d o

LO o

LO 02

LO LO t> 02

o to rH

o co t>3 cd s u A to CO

i— !

02

to

1xM

lO

CO

02 co © O ti to >nO t-4 d 02 ro

to

CO

sH CT> i —I

© > d d lo •H O -P O CD < d !>s CD r-j î> 5 W r H tO (p d H o f-o

S

❖ © d © -p o d cd o

V © a d to 60 d 02 V'' f-o

r>a

3 CQ

S o

GO

V

l>* iH tO

sVO

I

.60

02 -P d

LO

LO LO

t> 02

o LO 1—I

02

CO

xji

iO

02

0 e rf rP

1 1

5

xh

g

O CO ti 'M i^>

q q

À LO to

to

Trf

0

m 0 0 q

1

INFLUENCE OF MINOR ELEMENT FERTILIZATION ON THE CAROTENE CONTENT OF SOYBEANS.

32

—1 cd p o p

I 1 0 f-3 M

1

pH pH O §

00

'sh CT5

r~i

g

.5 a d

m o 3

CO

g ti © Pi Pi •rH

io

cd

0

•H

a

t0 a

-P

i —Ii —I0002 nP LQ " nP O^ 02 H 02

DLO

to

1

Oh

INFLUENCE

O to CD H I I to LO lO I

CO

-P

02 02 H 02

tO H

'th

Cd .£2

02i00 tO'sMtO'sh'>js totO"^h

$>a E to O bO ti to to

_ GOsMOE-OOOOOO0O^^COGOO-O5O2

CO ti o

E i>a bo -1 1 ti i>'\ to V

COtotoOE^LOCOin ^cOOaOLOO-ODOc 0 ^ h t 0 C 0 ’=*, t 0 0 2 C 0

to

05 O

o

CO

I—I

o

g

COOO-OGOOOSM05 OjLOCOto^tocOtO

O)

0 > -< * © ti 0 to o ti cd o

?S&,

I IH

to ti 0 E 0 1 -1 to

0

CO

C0

05

0

i to to co

to o H to

*»

44 contained less carotene tlian the corn from the other treated plots. 3.

Wheat■

The effect of manganese sulfate on the caro­

tene content of the leaves of wheat is given in Table 19,

Al­

though the carotene values of the wheat leaves from treated plots were higher than the controls 9 the increase was not great.

The carotene content of the stems of the deficient

plants was higher than that of the stems of the sprayed wheat. The application of sodium chloride, sodium sulfate, or calcium sulfate did not affect the carotene content appre­ ciably.

This lack of effect might have been caused by the

lack of rainfall which prevented the particular salt from being effective. 4.

Oats.

Samples of manganese deficient and normal

oat plants were taken on May 25, 1949.

There was a marked

difference in the carotene content. Table 20.

The carotene

content of the leaves of the normal plants was about 40 per cent higher than that of the deficient plants.

There was

also a difference in the carotene content of the stems.

The

stems of the deficient plants, however, had a higher carotene content than the stems of the normal plants. B.

1.

Effect of Manganese on the Activity of the Carotene-Destroying Enzyme . Soybeans.

Mb satisfactory data were obtained in the

45 Table 19 CAROTENE CONTENT, PER CENT CAROTENE DESTROYED DURING INCUBA­ TION AND LEAF-STEM RATIO OF WHEAT PLANTS GIVEN TREATMENTS AS LISTED, LOCATED ON THE WYNKOFF, LEWIS, AND BLOSSER FARMS, RESPECTIVELY, 1948.

Treatment

ONE REPLICATE ON EACH LOCATION.

Description of Plants

Enzyme

Carotene^ Leaves Stems r/gm r/gm

Le af— Stem Activity Ratio % loss

————

Normal plants

242

60

60

0.56

None

Chlorotic

203

119

80

1.42

M11SO 4**

Clilorosis corrected

245

41

58

0.56

None

Clover with, wheat, chlorotic

493

20Q

*■—

2.73

Clover with wheat, chlorosis corrected 524

129

——

1.49

MnSO^**

None

Chlorotic

165

69

70

1.51

MnS 04^

Chlorosis corrected

242

49

58

0.70

None

Chlorotic

234

35

32

0.76

ME1SO 4**

Chlorosis corrected

306

27

41

0.27

NaCl*~x"*

Partly chlorotic

272

24

39

0.41

Na 2S 04'ÎHMî-

Partly chlorotic

247

34

45

0.37

CaS04««*

Partly chlorotic

263

45

32

0.60

*

Expressed on dry basis.

****

Spray, 5 lbs. per acre.

*** Soil treatment, 5 lbs. per acre.

46

Table 20 INFLUENCE OF MANGANESE ON WEIGHT, CAROTENE CONTENT, AND CRUDE PROTEIN CONTENT OF CLINTON OATS, HAYTER FARM, WALKERTON, INDIANA, 19 49.

Category

AVERAGE OF THREE REPLICATES. Clinton Oats Normal Deficient 50 Plants 50 Plants 10 inches tall 6 inches tall Leaves Stems Leaves Stems 47.00

26.00

34.30

17.00

8.00

2.90

4.30

1.60

Dry matter, per cent

14.98

10.92

12.44

9.42

Crude Protein, per cent

28.88

21.62

32.44

25.25

555.00

84.00

383.0 0

100.00

Carotene mgm/lOO gram plant material

38.20

2.40

28.00

2.70

Enzyme activity, carotene loss on incubation, per cent

20.00

78.00

55.00

91.00

Green weight, grams Dry weight, grams

Carotene content, Y/gnu , dry basis

47 study of tlie carotene-destroying enzyme in 1947 or 1948, Tables 11, 12, and 13.

The enzymatic activity of the Es-Min-

E 1 minus copper treatment was significantly lower than the other minor element soil treatment: that is, less carotene was destroyed in the tissues during incubation. In 1949, however, highly significant data were ob­ tained, Table 15, and Figure 6 .

The soil treatments of 75

and 150 pounds of manganese sulfate per acre produced sig­ nificant increases in enzyme activity in soybean leaves. The spray treatment of 5 pounds per acre also had the same effect, but for only part of the season.

In this experiment,

manganese apparently acted as an activator for this lipoxidase enzyme.

Such effects are not unusual because manganese

serves as an activator for other enzymes such as arginase, carboxylase, some phosphatases, cholinesterase, and others (89) . 2. Table 18.

Corn.

The carotene-destroying activity is shown in

The values are quite erratic and no conclusions

can be made.

Considering the averages, there is an indication

that the treatments increase the enzyme activity. 3.

W he at .

The carotene-destroying activity of the

wheat tissue appears to be unusually high, as shown in Table 19.

This was especially true in the manganese deficient

plant material.

When manganese was supplied, the activity

was reduced. 4.

Oats.

The carotene-destroying enzyme activity of

48

75

70

LU LU LU 5 0

| NO

Z CH LU
0> 'sh

© > C

g g 0 0 © ©

r-4

05

©

© ©

o r-ne£

O i— ! 03 •H M

I— t I— I I— I I— I

0

02 02

g g © © bO bO d d

o • o

o o

o o o o

xh O

CO

â ê

o o o o

00 LO 00 M CT> CO CO

d>o Ha

©

xh 02 r-4 O

Th co co co

0

d A

co co ’-ch CO to to co •

d

&

o o

o o o o

r-4 CO d r-4 h>

O

o co r-4 i—I

i—f 02 i—I i—I

d ©

to io "sh m

o o o o

e bO . © d h a t— I 1 —1 d H 43 tO O d 10 $> LO 0 r-4 0

u

© A

r-4 02 CO Xh

©

Ph H

1 —1

"r4

5 d

0