ARUN SHARMA College of Agriculture

131 Pages·2011·11.45 MB·English

MARKER-ASSISTED IMPROVEMENT OF PEARL MILLET
(Pennlsetum glaucum) DOWNY MILDEW RESISTANCE IN ELITE
HYBRID PARENTAL LINE H 771833-2
BY
ARUN SHARMA
Thesls submitted to the Chaudhary Charan Slngh
Haryana Agricultural University in partial fulfillment
of the requirements for the degree of:
DOCTOR OF PHILOSOPHY
In
PLANT BREEDING
College of Agriculture
Chaudhary Charan Singh Haryana Agricultural University,
Hlsar
DEDICATED
TO MY
REVERED PARENTS
AND
LOVING TANYA
CERTIFICATE I
This is to certify that this dissertation entitled, "Marker-assisted
improvement of pearl millet (Pennisetum glaucum) downy mlldew
resistance in elite hybrid parental line H 771833-2", submitted forthe
degree of Doctor of Philosophy in the subject of Plant Breeding, of
Chaudhary Charan Singh Haryana Agricultural University, Hisar, is a
bonafide research work carried out by Mr. Arun Sharma under my
supervision and that no part of this thesis has been submitted for any other
degree.
The assistance and help received during the courseof investigation
have been fully acknowledged.
Head, Oil Seed Section
Department of Plant Breeding
CCS Haryana Agricultural University
Hisar - 125004
[C.T. HASH]
CO-MAJOR ADVISOR
Principal Scientist
Molecular Breeding
ICRiSAT, Patancheru-502 324
Hyderabad, A.P.
CERTIFICATE II
This Is to certlfy that this dissertation entitled, "Marker-asslsted
Improvement of pearl mlllet (Pennlsetum glaucum) downy mildew
resistance In elite hybrid parental line H771833-2" submitted by
Mr. Arun Sharma to the Chaudhafy Charan Singh Haryana Agricultural
University in partial fulfillment of the requirements for the degree of Doctor
of Philosophy in the subject of Plant Breeding has been approved by the
Student's Advisory Committee after an oral examination on the same, in
collaboration wlth an External Examiner.
0 ~ 2 ~ ' ~
@M' EXTERNAL EXAMINE
'
CO-MAJOR ADVISOR
&bl"
HEAD OF THE DEPARTMENT
Acknowledgements
Fervently and modestly, I do1t he genuine cooperation and inspiration offered to me by the
Chairman o f my Advisory commiffee, Dr. D.C. Nijhawan, Professor & Head, OIIseeds section,
Department o f Plant Breeding, CCS HAU, Hisar, for hi.s valuable counsel and meticulous
guidance during my study at HAU and kind encouragement during the process o f achieving
thefinul form o f this thesis.
It give.^ me immense pleasure in exlending my deep sense o f reverence and gratitude to
Dr. C.T. Hash, Principal Scientisl, Molecular Breeding, ICRISAT, Patancheru and Co-
chairman o/ my Advisory cornmiltee, for providing research facilities, suggestion o f the
research problem, immnsely valuable guidance and constructive criticism throughout the
period of this invesfigalion andpreparation o f the manuscript.
I am sincerely obliged and indebted to Dr. R.K. Behl, Professor and Dr. Ashok Chhabra,
Associate Professor, Department o f Plant Breeding, for their invaluable consultations,
sagacious advice and able direction throughout the course o f study and bringing out the
manuscript.
I am also thankful to other members o f my advisory committee, Dr. (Mrs.) Suntosh DhiNon,
Professor, Department o f Chemistry & Biochemistry, Dr. R.K. Join, Professor, Department of
Biotechnolou & Molecular Biolow and Dr. K.K. Saxena, Senior Scientist, Department o f
Mathemotics & Statistics,f or constructive suggestions and guidance for completion o f thi.s
research project.
I am grateful to Dr. B.P.S. Latlrer, Pr(/essor & Head, Department o f Plant Breeding and aho
to the former Head, Dr. G.P. Lodhi for extending all possible help and support during the
course o f my study.
fi gives me pleasure in expressing my sincere thanks lo Dr. S. Sivaramakrishnan, Senior
Scientist (Radio Isutope Labnratury), Dr. N. Seetharama, Senior Scientist (Molecular Biology),
Dr. R.P. Thokur, Senior Scientist (Putholou), Dr. K.N. Rai, Principul Scientist (Pearl millet
Breeding), Dr. F. R. Biilinger, Principal Scientist (Cereal Physiologv) and Dr. H.D. Upadbyaya,
Scientist (Genetic Resources Divisiun) for their kind help which enabled me lo accomplish this
task with ease.
With respectful regnrds and immense pleasure, I wish to acknowledge and express my thanks
to Dr. S. Chandru (Senior Statistician) and Mr. Hori (Scientflc Officer, Statistics unit) for
vuluuble advice and help rendered during analyses.
I express my sincere gratitude lo Dr. S. N. Nigam, Principal Scientist, Groundnut Breeding, for
boosting my morale and giving me timef or scientijir discucsions.
Dr. CJ. Howarth, Scientist, IGER, Aberyslwyth, UK, deserves special mention to permit me to
reproduce linkage and QTL maps used in the present study. I am also grateful to Dr. Rattan
Yailuv, Molecular Geneticist, IGER, Abeyslwylh, UK, for being supportive and encouraging
throughout this research project.
Assistance rendered by Ms. Seetha and Ms. Santuslt helped me in ~~omplelinmgy lab work
effrentiy on11 smoothly.
I feel privileged to express heartfelt words of appreciation to all PMB stafl especially Mr.
Bhaskar Raj, Mr. Canopalhi, Mr. OmParkash, Mr. Murali and Mr. Izaz Ahmed for their kind
cooperation and help rendered
I thank Mr. K,P rabhakar, Senior Secretary (PMB) and former secretary Mr. S.B. Stanleyfor
being very nice tu me by pmmptly completing any assignment given to them
Special thanks are mended lo Library sta& Learning Systems Unit staflespeciully Mr. Prasad
Rao and Mr. Vidya Sagar (Photographer)f or their excellent asshtance during my mearch
work at ICRISAT.
It was with the utmost blessings and good wishes of my parents and aflection of my brothers
Siddhartha and Rajeshwar, sisler Sujata and brother-in-law Ashwini that I could anain
academic heights by undergoing doctorate studies.
I shall befuiling in my du@ i/ I do not express my cordial thanks to all my colleagues and
friends, Maria, Azhqu, Ranjana, Lava, Gopala, Dewart, Kulkarni, Sarosh, Raghu, Naveen,
Gaurav, Atul, AMK Raju, Dinesh, Manoj, Rupesh, Anil and Deepak for their untiring help,
whole-bearled cooperation and nice company during my stay both, at ICRISAT and Hi.~ar.
I take BB upportuni@m thank Dr. C.L.L. Gowda, Program Leader, Learning System. Unit,
ICRISAT and Dr. B. Diwaker, former Program Leader, for their kind couperution to work in
collaboration with PMB divi.vion of ICRISAT on Memorandum of Understanding.
Financial usvistance received in form of Merit stipend awarded by CCS HAU and the grant of
Resenrch Schol~irshipb y ICRISATis also acknowledged
Special thanks is also extended to Be Department for International Development (DFID),
Plunt Sciences, UK,J urpwviding fund7 to carry out this investigation.
HISAR
Mflrc11 07, 2001
CONTENTS
Chapter Description Page
Number Number
1 INTRODUCTION 1-5
2 REVIEW OF LITERATURE 6-22
3 MATERIALS AND METHODS 23-44
4 RESULTS 45-63
5 DISCUSSION 64-76
6 SUMMARY AND CONCLUSION 77-79
LITERATURE CITED 80-10 3
APPENDICES i-vi
LIST OF TABLES
Table No. Particulars Page No.
Table 1. Details of target traits and probes-enzyme combinations used for 23
marker-assisted downy mildew resistance improvement of pearl
millet pollinator H 771833-2
Table 2. MAS operations and results in each generation 35
Table 3. Agronomic performance of DMR testcross entries compared to 46
mean of control HHB 67 and the mean of the trial as a whole
Table 4. Agronomic performance of bristled testcross entries compared to 50
mean of control HHB 67 and the mean of the trial as a whole
Table 5. Chi-square estimates for goodness of fit to a range of classical 54
Mendelian segregation ratios for greenhouse screen downy mildew
reaction against Patancheru isolate among 193 testcrosses of pearl
millet cross ICMP 451 x H 771833-2
Table 6. Marker genotype data for 38 BC4F2D MR entries that had lower 55
disease incidence during their testcross downy mildew screening
Table 7. Marker genotypes of selected BC4F2p lants and selected plants 56
in their progenitor generations along with DM1 (%).
Table 8. Summary of QTLs identified using MapmakerlQTL for the response 57
to downy mildew in pearl millet cross ICMP 451 x H 771833-2.
-
LIST OF FIGURES
Figure No. Particulars Page No.
Figure 1. Genetic linkage map of pearl millet cross ICMP 451 x H 771833-2 29
Figure QTL map for downy mildew resistance in pearl millet cross
ICMP 451 x H 771833-2
Figure 3. QTLs associated with morphological characters in pearl millet 31
cross ICMP 451 x H 771833-2
Figure 4. Donor parent ICMP 451-P6
Figure 5. Recurrent parent H 771833-2
Figure 6. MAS-improved DMR version of H 771833-2
Figure 7. Schematic for transfer of downy mildew resistance by 36
marker-assisted backcrossing in pearl millet
Figure 8. Testcross trial at flowering stage 38
Figure 9. Testcross trial post-anthesis measurement of plant height 38
Figure 10. Testcross trial at harvesting stage 38
Figure 11. Distribution of 193 testcross progenies for disease incidence (%) 53
when screened under greenhouse conditions at ICRISAT against
a downy mildew pathogen population from Patancheru, India
Figure 12. Panicles of the original non-bristled HHB 67, moderately 66
resistant to downy mildew; and bristled DMR version of HHB 67,
pollinator improved by MAS
Figure 13. Original HHB 67 66
Figure 14. Bristled version of HHB 67 66
Figure 15. Autorad~ographso btained from screening of BC~FIp rogenies 58.
based on donor ICMP 451 and recurrent parent H 771833-2
Figure 16. Autoradiographs obtained from screening of BCjFq progenies 59
based on donor ICMP 451 and recurrent parent H 771833-2
Figure 17. Autoradiographs obtained from screening of BC4F, progenies 60
based on donor ICMP 451 and recwrrent parent H 771833-2
Flgure No. Particulars Page No,
Figure 18. Autoradiograph obtained from screening of BC4F, progenies 61
based on donor ICMP 451 and recurrent parent H 771833-2
Figure 19. Autoradiograph obtained from screening of BC4F, progenies 61
based on donor ICMP 451 and recurrent parent H 771833-2
Figure 20. Autoradiographs obtained from screening of BC4F2progenies 62
based on donor ICMP 451 and recurrent parent H 771833-2
Figure 21. Autoradiographs obtained from screening of BC4F2p rogenies 63
based on donor ICMP 451 and recurrent parent H 771833-2
Figure 22. Modified three-way hybrid seed production scheme 75