Table Of ContentPURDUE UNIVERSITY
THIS IS TO CERTIFY THAT THE THESIS PREPARED UNDER MY SUPERVISION
Katharine Josephine Hivon
EwiviEffects of Manganese on Ascorbic Acid and
Ascorbic Acid-Oxidizing Enzymes in Growing 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
February 19 50
TO THE LIBRARIAN:----
-$&-
THIS THESIS IS NOT TO BE REGARDED AS CONFIDENTIAL.
GRAD. SCHOOL, FORM 9—3-49—1M
EFFECTS OF MANGANESE ON ASCORBIC ACID
AND
ASCORBIC ACID-OXIDIZING ENZYMES IN GROWING PLANTS
A Thesis
Submitted to the Faculty
of
Purdue University
by
Katharine Josephine Hivon
In Partial Fulfillment of the
Requirements for the Degree
of
Doctor of Philosophy
February, 1950
ProQuest Number: 27712262
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ACKNOWLEDGMENTS
The writer wishes to express sincere appreciation for
the direction and encouragement of the two persons under
whose supervision this research has been conducted: to Dr,
D. M. Doty for interest and advice which has been extended
beyond the time of active staff membership; to Dr. F. W.
Quackenbush for the acceptance of an unfinished project with
undiminished interest and helpfulness.
The writer would like, also, to acknowledge the assis
tance of some of the many persons who have helped in numerous
ways during this research: Dr. S. M. Hauge, for leadership
of the project of which this study was a part; Dr. J. K.
Plummer and Mr. J. Bergeaux of the Tennessee Corporation for
support of the project which furnished the field material;
Drs. B. R. Bertramson, A. J. Ohlrogge, J. E. Steckel, and Mr.
G. A. Hemstock, present and former staff members of the A-
gronomy Department, for cooperation in supplying field mate
rial; Mr. L. C. Shenberger for photographic work, advice and
assistance with the greenhouse research; Prof. S. R. Miles
for instruction in statistical methods; Mr. 0. J. Burger for
assistance with many phases of the research; Drs. Alice P.
Withrow (former member) and G. F. Warren, Profs. N. K. Ellis
and E. R. Honeywell - Horticulture, Dr. G. E. Gould and Mr.
I. C. Brooks - Entomology, and Dr. G. A. Gries - Agricultural
Botany, Departments, for advice and assistance. Gratitude is
also expressed for the technical assistance of Mrs. Katherine
(Henderson) Jackson and Mrs. Burnetta Graham.
VITA
The writer was born in Chicago, Illinois on June 15»
1909* Early education was received in West Lafayette to
which the family had moved in 1914. A diploma was received
from the West Lafayette High School in June, 1927, and the
degree of Bachelor of Science in Home Economics with major
in foods and nutrition from Purdue University in June, 1931.
A student dietitian internship at Indianapolis City Hospital
was completed in June, 1932. Employment as a dietitian oc
cupied the next several years. The positions held were :
assistant dietitian, Indianapolis City Hospital, 1932-1934;
temporary assistant dietitian, Toledo (Ohio) General Hos
pital, summer of 1936; staff, head and acting chief dieti
tian at U. S. Marine Hospitals located at Ellis Island, New
York, New Orleans, Louisiana, and Cleveland, Ohio, respec
tively, as a Civil Service employee of the U. S. Public
Health Service between 1936 and 1943* A position as control
chemist in the Soap Laboratory, Lever Brothers Company, Ham
mond, Indiana was held for a year prior to employment in the
Agricultural Chemistry Department at Purdue University as
one-half time graduate research assistant. This position
has been held from July, 1945 until the present time. The
degree of Master of Science with major in agricultural bio
chemistry was received in February, 1947.
tab le o f contents
Page
ABSTRACT..................................................
INTRODUCTION.............................................. 1
REVIEW OF THE LITERATURE................................. 6
Suggested Functions of Manganese...................... 6
General......... 6
Effects of Manganese on Ascorbic Acid............. 9
Effects of Manganese on Ascorbic Acid-oxidizing
Enzyme s........... 11
Suggested Functions of Ascorbic Acid and Ascorbic
Acid-oxidizing Enzymes............................. 12
Effect of Light and Other Factors on Ascorbic Acid
Synthesis................ 13
FIELD CULTURE............................................. 17
Soy Bean Plants........................................ 17
Experimental...................... 17
Materials. ................ ................ .... 17
Methods......... 17
Preliminary Studies.......................... 17
Sampling...................................... 23
Methods of Chemical Analysis................ 23
Moisture................................. 23
Ascorbic acid............................ 23
Enzymatic ascorbic acid-oxidizing activi
ty...................................... 25
Experimental Results............................... 26
First Season. ................................. 26
Second Season. ........... 29
Third Season .............................. 35
Corn and Wheat Plants................................. 39
Experimental ............ 39
Materials........................................ 39
Methods.......................................... 42
TABLE OF CONTENTS (Continued)
Page
Sampling ............... 42
Methods of Chemical Analysis............... 42
Experimental Results......... 42
Discussion of Results of Field Experiments........... 44
Effects of Manganese on Ascorbic Acid .... 45
Effects of Manganese on Ascorbic Acid-oxidizing
Enzymes.............. 45
Effects of Growth and Other Factors on Ascorbic
Acid and Ascorbic Acid-oxidizing Enzymes...... 47
SOLUTION CULTURE OF GREEN BEAN PLANTS. .............. 51
Experimental.................... 51
Materials........................................... 51
Methods ....... 52
Solution Culture of Plants.......... 52
Sampling......................................... 59
Methods of Chemical Analysis.......... 60
Moisture....................... 60
Ascorbic Acid........ 60
Enzymatic Ascorbic Acid-oxidizing Activity.. 60
Manganese. ....... 60
Study of the Effects of Parathion (Pot Culture) 6l
Study of the Effects of Sunlight (Pot Culture) 61
Experimental Results..................... 62
Discussion of Results of Experiments with Green Bean
Plants ..... 89
Appearance of Plants....................... 89
Ascorbic Acid and Enzyme Activity of Leaves and
stems.......... 92
Ascorbic Acid-oxidizing Enzyme Activity........... 93
Ascorbic Acid Content.... ................. 94
Effect of Sunlight on Ascorbic Acid Content....... 96
SUMMARY.................................................... 98
LITERATURE CITED........................... 101
LIST OF TABLES
Table Page
1. Description of soy bean plants used in field
experiments........................... 18
2• Effect of manganese and other micro-elements on
reduced ascorbic acid content of leaves of
soy bean plants grown in the field (1947)....* 2?
3* Effect of manganese and other micro-elements on
enzymatic ascorbic acid-oxidizing activity of
leaves of soy bean plants grown in the field
(1947) 28
4» Effect of manganese and copper on reduced ascor
bic acid content of leaves of soy bean plants
grown in the field (1948)...................... 31
3. Effect of manganese and copper on enzymatic as
corbic acid-oxidizing activity of leaves of
soy bean plants grown in the field (1948)..... 32
6. Effect of manganese on a single harvest of
leaves of soy bean plants grown in the field
(1948).............................. 36
7. Effect of manganese on reduced ascorbic acid
content of leaves of soy bean plants grown
in the field (1949)............................ 37
8. Effect of manganese on enzymatic ascorbic acid-
oxidizing activity of leaves of soy bean
plants grown in the field (1949)............ 38
9. Effect of three iron-manganese ratios on reduced
ascorbic acid content of green bean plants
grown in cultural solution ( 1948)........... 63
10. Effect of three iron-manganese ratios on en
zymatic ascorbic acid-oxidizing activity of
green bean plants grown in cultural solution
(1948).......................................... 64
11* Effect of three iron-manganese ratios on reduced
ascorbic acid content of green bean plants
grown in cultural solution (1948-1949)....... * 83
12. Effect of three iron-manganese ratios on en
zymatic ascorbic acid-oxidizing activity of
freen bean plants grown in cultural solution
1948-1949 )........................ 84
LIST OF TABLES (Continued)
Table Page
13# Effect of parathion insecticide on reduced as
corbic acid content of leaves of green bean
plants grown in pot culture (1949).............. 86
14* Effect of sunlight on reduced ascorbic acid con
tent of leaves of green bean plants grown in
pot culture (1949).......... .................. 88
