STUDIES ON MOLECULAR DIVERSITY, HETEROSIS, COMBINING ABILITY AND HIGH ...

235 Pages·2013·9.06 MB·English

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“STUDIES ON MOLECULAR DIVERSITY, HETEROSIS,
COMBINING ABILITY AND HIGH TEMPERATURE
TOLERANCE IN Brassica juncea L. Czern & Coss”

THESIS

Submitted to the

G.B. PANT UNIVERSITY OF AGRICULTURE & TECHNOLOGY,
PANTNAGAR–263145, (U.S. NAGAR), Uttarakhand, INDIA

By

VIKRANT SINGH
M.Sc. Ag. (Genetics and Plant Breeding)

IN PARTIAL FULFILMENT OF THE REQUIREMENTS
FOR THE DEGREE OF

DOCTOR OF PHILOSOPHY
(Genetics and Plant Breeding)

May, 2013
ACKNOWLEDGEMENT

All glory goes to the almighty whose blessing is behind the success. Praise, pride and
perfection belong to almighty, so first of all I would like to express my deepest sense of gratitude to
the omniscient and omnipresent power of the universe, the Almighty God.
Words fail to express my sense of indebtedness and profound gratitude towards my
favorite teacher since U.G. program and Major Advisor as well as Chairman Dr. Ram Bhajan,
Professor/Senior Scientist of Genetic and Plant Breeding for his noble guidance, constructive
criticism, innovative ideas and valuable suggestions during the course of this research program.
With profound sense of gratitude, I express my sincere thanks to the esteemed members of
my advisory committee Dr. J.S. Verma, Professor, Department of Genetics & Plant Breeding; Dr.
P.K. Shrotria, Professor, Department of Genetics & Plant Breeding; Dr. R.P. Awasthi, Professor,
Department of Plant Pathology and Dr. S.K. Guru, Associate Professor, Department of Plant
Physiology for providing advice and all kinds of help during the course of the research.
I owe my indebtedness to Dr. H.S. Chawla, Professor and Head of the Department of
Genetics & Plant Breeding; the Joint Director, N.E. Borlaug Crop Research Centre; the Dean,
Post Graduate Studies and the Dean, College of Agriculture, G.B.P.U.A. & T. Pantnagar for
providing opportunity and necessary facilities during the whole research program
My sincere thanks go to Dr. R.K. Khulbe, Assistant Professor, Department of Genetics &
Plant Breeding for providing laboratory facilities at PCPGR Lab. A lot of thanks to my senior Dr.
Arif Khan, Dr. Manjeet Singh and Dr. Amrita Dey for proper guidance of molecular diversity
analysis. I wish thanks to all the members of Brassica division for their timely-untimely prestigious
as well as valuable help.
All the words in the lexicon will be futile and meaningless if I fail to express the obligation
from the fathom of my heart for my beloved and venerable family (nani Kansa Devi, grandfather
Sri Ram Badan Singh, father Sri Lal Bijendra Singh, uncle Sri Ashok Singh and Sri Bajarang
Bahadur Singh, mother Smt. Asha Singh, aunty Smt. Puspa Singh and Smt. Savitri Singh as
well as to my all brothers and sisters) for their blessing, affection, sacrifice and moral as well as
financial support.
I am extremely thankful to my God gifted elder brother Dr. Prashant Vikram, Visiting
Research Fellow, IRRI, Philippines, as an ideal person for their valuable guidance and moral
support. It is a great pleasure to acknowledge the affection rendered by my seniors and batch mates
(Dr. Ranjeet Kumar, Dr. Ram Lakhan, Dr. Chandra Mohan, Dr. Vineet Kumar, Dr. Ajaya
Paliwal, Mr. Devender Singh, Gunjan, Bhim jyoti, Priyanka, Reena, Kalyan and Uma
Maheswar). The words fail to express my deep sense of indebtedness to my heartiest loving
juniors Nitin alias Katiyar, Naveen, Surandra, Vipin, Pankaj, Sanjeev, Girish, Madhav,
Himanshu, Praveen, Puri, Narolia, Renu and Sweta for their affection as well as help in field as
well as lab works.
Fellowship support, provided by ICAR-SRF (PGS) and DST-INSPIRE programme for
pursuing Ph.D. on the basis of my talent, is duly acknowledged.
Last but not the least, I express my thanks to each and every person who directly or
indirectly aided me in completion of this work.

Date: May, 2013
Place: Pantnagar
Dr. Ram Bhajan Department of Genetics and Plant Breeding
Professor College of Post Graduate Studies
G.B. Pant University of Agriculture &
Technology, Pantnagar–263145,
Distt.-U.S. Nagar,
Uttarakhand, INDIA.

CERTIFICATE
This is to certify that the thesis entitled “STUDIES ON MOLECULAR DIVERSITY,
HETEROSIS, COMBINING ABILITY AND HIGH TEMPERATURE
TOLERANCE IN Brassica juncea L. Czern & Coss” submitted in partial fulfillment of
the requirements for the degree of DOCTOR OF PHILOSPHY with major in Genetics
and Plant Breeding and minor in Plant Pathology of the college of Post Graduate
Studies, G.B. Pant University Of Agriculture & Technology, Pantnagar, is a record of bona
fide research carried out by Mr. Vikrant Singh, Id. No. 39352, under my supervision, and
no part of the thesis has been submitted for any other degree or diploma.
The assistance and help received during the course of this investigation has been
duly acknowledged.

Pantnagar
May,2013
CERTIFICATE
We, the undersigned members of the advisory committee of Mr. Vikrant Singh, Id. No.
39352, a candidate for the degree of DOCTOR OF PHILOSOPHY with major in Genetics
and Plant Breeding and minor in Plant Pathology, agree that the thesis entitled “STUDIES
ON MOLECULAR DIVERSITY, HETEROSIS, COMBINING ABILITY AND HIGH
TEMPERATURE TOLERANCE IN Brassica juncea L. Czern & Coss” may be
submitted in partial fulfillment of the requirements for the degree.
CONTENTS
S. No. CHAPTER PAGE No.
1 INTRODUCTON 1-5
2 REVIEW OF LITERATURE 7-46
3 MATERIALS AND METHODS 47-79
4 RESULTS AND DISCUSSION 81-164
5 SUMMARY AND CONCLUSIONS 165-170
LITERATURE CITED
APPENDICES
VITA
ABSTRACT
LIST OF TABLES
Table No. Title Page
3.3.1 Pedigree/parentage and origin/source of the parental lines including 48
check cultivars
3.3.2 Details of the crosses with code for cross identification number as 4 9
mentioned in the text
3.6.4.4.1.1 Detail description of primer sequences of SSRs marker 62
3.7.1.1.1 Data on Lines (L.), Testers (T.) and Crosses (L.×T.) in one way table 65
3.7.1.1.2 Analysis of variance for simple RBD 66
3.7.1.1.3 Data on Line×Tester in a two way table 67
3.7.1.1.4 Analysis of variance for line×tester analysis including parents 67
3.7.1.1.5 Analysis of Variance for Line×Tester pooled over environments 68
3.7.1.2.1 Analysis of variance for combining ability in Line×Tester 70
3.7.1.2.2 Analysis of variance for combining ability pooled over environments 71
4.1.1 Analysis of variance for different heat stress related traits in Indian 84
mustard
4.1.2 Selection parameters for different heat stress related traits in Indian 84
mustard
4.1.3 Categorization of Indian mustard genotypes in heat stress tolerant and 94
susceptible groups based on physiological traits
4.1.4 Correlation coefficients between seed yield and stress related traits in 94
different environments in Indian mustard
4.1.5 Correlation coefficient between physiological traits and yield based 94
stress related parameters in B. juncea (L.)
4.2.1.1 Range of SSR loci scored, number and size of exclusive loci 97
amplified in Indian mustard genotypes
4.2.2.1 Similarity matrix for Jaccard’s coefficient of 45 genotypes (G) in 105
Indian mustard
4.3.1.1 Analysis of variance for simple RBD of yield and its contributing 111
characters in Indian mustard in early (E1), timely (E2) and late (E3)
sown environments (E)
4.3.1.2 Combined analysis of variance for simple RBD of different 112
characters over environments (E1, E2 and E3)
4.4.1.1 Analysis of variance for line×tester of different characters with 115
respect to early (E1), timely (E2) and late (E3) sown environment (E)
4.4.1.2 Combined analysis of variance for line×tester of different characters 116
over environments (E1, E2 and E3)
4.4.2.1 General combining ability effects of lines (L.) in early (E1), timely 117
(E2), late (E3) sown and pooled (P) over environments (E)
4.4.2.2 General combining ability effects of testers (T.) in early (E1), timely 120
(E2), late (E3) sown and pooled (P) over environments (E)
4.4.3.1 Specific combining ability effects of crosses (L.×T.) for yield 128
contributing traits in early (E1), timely (E2), late (E3) sown and
pooled (P) over environments (E)
4.4.4.1 Components of variance for various characters with respect to early 140
(E1), timely (E2), late (E3) sown and pooled (P) over environments
(E)
4.4.5.1 Proportional contribution of lines, testers and their interactions to 142
total variance with respect to early (E1), timely (E2), late (E3) sown
and pooled (P) over environments (E)
4.5.1 Magnitude of heterosis over better parent (BP) and standard check 145
(SV) of crosses for yield contributing traits in early (E1), timely (E2),
late (E3) sown and pooled (P) over environments
4.5.2 Prospective cross combinations based on heterosis (SV & BP) vis-a- 163
vis per se performance for seed yield and its contributing traits
LIST OF FIGURES

Figure No. Title Page
4.1.1 Maximum temperature during 2010-11 87
4.1.2 Minimum temperature during 2010-11 87
4.1.3 Mean temperature during 2010-11 87
4.1.4 Cell Membrane Stability (%) of Indian mustard genotypes 90
4.1.5 % Reduction in Relative Water Content from Leaf of Indian 90
mustard genotypes
4.1.6 Acquired Thermal Tolerance (%) of Indian mustard genotypes 90
4.1.7 Chlorophyll a (mg/g fresh leaf weight) of Indian mustard 91
genotypes
4.1.8 Chlorophyll b (mg/g fresh leaf weight) of Indian mustard 91
genotypes
4.1.9 Chlorophyll a/b ratio of Indian mustard genotypes 91
4.1.10 Total Chlorophyll Content (mg/g fresh Leaf) of Indian mustard 92
genotypes
4.1.11 % Reduction of Seed Yield due to Seedling (First column) and 92
Terminal Heat Stress (Adjacent second column) of Indian mustard
genotypes
4.1.12 Mean Productivity to Seedling (First column) and Terminal 92
(Adjacent second column) Heat Stress (g/plant) of Indian mustard
genotypes
4.1.13 Tolerance to Seedling (First column) and Terminal (Adjacent 93
second column) Heat Stress of Indian mustard genotypes
4.1.14 Stress Tolerance Efficiency to Seedling (First column) and 93
Terminal (Adjacent second column) Heat Stress (%) of Indian
mustard genotypes
4.1.15 Stress Susceptibility Index to Seedling (First column) and 93
Terminal (Adjacent second column) Heat Stress of Indian mustard
genotypes
4.2.1.1 PCR amplification profile of 45 Indian mustard genotypes with 100
SSR primer Na10-Co1a. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.1.2 PCR amplification profile of 45 Indian mustard genotypes with 100
SSR primer Na10-Co1c. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.1.3 PCR amplification profile of 45 Indian mustard genotypes with 101
SSR primer Ni02-D08a. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.1.4 PCR amplification profile of 45 Indian mustard genotypes with 101
SSR primer Ni03-H07a. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.1.5 PCR amplification profile of 45 Indian mustard genotypes with 102
SSR primer Ol10-A03a. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.1.6 PCR amplification profile of 45 Indian mustard genotypes with 102
SSR primer Ol10-F11a. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.1.7 PCR amplification profile of 45 Indian mustard genotypes with 103
SSR primer Ra02-A04a. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.1.8 PCR amplification profile of 45 Indian mustard genotypes with 103
SSR primer Ra02-A04c. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.1.9 PCR amplification profile of 45 Indian mustard genotypes with 104
SSR primer Ra02-E01a. M, 3 Kb ladder; Lanes 1 to 45, genotypes
4.2.2.1 Dendrogram of 45 Indian mustard genotypes constructed by using 109
UPGMA based on Jaccard’s similarity coefficient
4.4.2.1 General combining ability effects for days to flowering 121
4.4.2.2 General combining ability effects for day to marturity 121
4.4.2.3 General combining ability effects for plant height (cm) 121
4.4.2.4 General combining ability effects for main shoot length (cm) 122
4.4.2.5 General combining ability effects for primary branches number 122
4.4.2.6 General combining ability effects for secodary branches number 122
4.4.2.7 General combining ability effects for siliqua length (cm) 123
4.4.2.8 General combining ability effects for seed number per siliqua 123
4.4.2.9 General combining ability effects for seed yield per plant (g/plant) 123
4.4.2.10 General combining ability effects for harvest index (%) 124
4.4.2.11 General combining ability effects for 1000s seed weight (g) 124
4.4.2.12 General combining ability effects for oil content (%) 124