Ulyksa Esa Nqx/K Thuksa Esa Vkuqokaf'kd Fofh Ns'kh Xksoa'kh
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Study on Genetic Variation in Milk Genes and Their Association with Milk Performance Traits in Indigenous Cattle Breeds ns'kh xkSoa'kh; uLyksa esa nqX/k thuksa esa vkuqokaf'kd fofHfofHkUurkvksakUurkvksa ,oe~ nqXnqX/k/k mRiknu lEcaflEcaf/k/k xq.kksa ij muds izHkko dk v/;;u VIJAY KUMAR AGRAWAL M.V.Sc THESIS DOCTOR OF PHILOSOPHY (Animal Genetics and Breeding) 2017 Department of Animal Genetics and Breeding College of Veterinary and Animal Science Rajasthan University of Veterinary & Animal Sciences, Bikaner-334001 Rajasthan Study on Genetic Variation in Milk Genes and Their Association with Milk Performance Traits in Indigenous Cattle Breeds ns'kh xkSoa'kh; uLyksa esa nqX/k thuksa esa vkuqokaf'kd fofHkUurkvksa ,oe~ nqX/k mRiknu lEcaf/k xq.kksa ij muds izHkko dk v/;;u THESIS Submitted to the Rajasthan University of Veterinary & Animal Sciences, Bikaner in partial fulfillment of the requirement for the degree of DOCTOR OF PHILOSOPHY (Animal Genetics and Breeding) By VIJAY KUMAR AGRAWAL M.V.Sc 2017 Rajasthan University of Veterinary & Animal Sciences, Bikaner College of Veterinary and Animal Science, Bikaner CERTIFICATE - I Date…………… This is to certify that Vijay Kumar Agrawal had successfully completed the comprehensive examination held on 12/11/2016 as required under the regulation for PhD degree. (G.C. Gahlot) Head Department of Animal Genetics and Breeding College of Veterinary and Animal Science, Bikaner Rajasthan University of Veterinary & Animal Sciences, Bikaner College of Veterinary and Animal Science, Bikaner CERTIFICATE - II Date………...…… This is to certify that this thesis entitled STUDY ON GENETIC VARIATION IN MILK GENES AND THEIR ASSOCIATION WITH MILK PERFORMANCE TRAITS IN INDIGENOUS CATTLE BREEDS submitted for the degree of DOCTOR OF PHILOSOPHY in the subject of Animal Genetics and Breeding embodies bonafide research work carried out by Mr. Vijay Kumar Agrawal , under my guidance and supervision and that no part of this thesis has been submitted for any other degree. The assistance and help received during the course of investigation have been fully acknowledged. The draft of the thesis was also approved by the advisory committee on 16/06/2017. (G.C. Gahlot) (G.C. Gahlot) Head Major Advisor Department of Animal Genetics and Breeding College of Veterinary and Animal Science, Bikaner (Tribhuwan Sharma) Dean College of Veterinary and Animal Science Bikaner Rajasthan University of Veterinary & Animal Sciences, Bikaner College of Veterinary and Animal Science, Bikaner CERTIFICATE – III Date………….... This is to certify that the thesis entitled STUDY ON GENETIC VARIATION IN MILK GENES AND THEIR ASSOCIATION WITH MILK PERFORMANCE TRAITS IN INDIGENOUS CATTLE BREEDS submitted by Mr. Vijay Kumar Agrawal to Rajasthan University of Veterinary & Animal Sciences, Bikaner, in partial fulfillment the requirements for the degree of DOCTOR OF PHILOSOPHY in the subject of Animal Genetics and Breeding, after recommendation by the external examiner, was defended by the candidate before the following members of the examination committee. The performance of the candidate in the oral examination on his thesis has been found satisfactory, we therefore, recommend that the thesis be approved. (G.C. Gahlot) Major Advisor (Rajeev Kumar Joshi) (S.K. Kashyap) Advisor Advisor (Rajesh Nehra) (R.K.Singh) Advisor Dean P.G. Nominee (G.C. Gahlot) Head Department of Animal Nutrition APPROVED (S.K. Kashyap) DEAN POST GRADUATE STUDIES Rajasthan University of Veterinary & Animal Sciences, Bikaner College of Veterinary and Animal Science, Bikaner CERTIFICATE – IV Date………….... This is to certify that Mr. Vijay Kumar Agrawal of the Department of Animal Genetics and Breeding, College of Veterinary and Animal Science, Bikaner has made all corrections/modifications in the thesis entitled , STUDY ON GENETIC VARIATION IN MILK GENES AND THEIR ASSOCIATION WITH MILK PERFORMANCE TRAITS IN INDIGENOUS CATTLE BREEDS which were suggested by the external examiner and the advisory committee in the oral examination held on……………….. The final copies of the theses duly bound and corrected were submitted on……………………, are forwarded herewith for approval. (G.C. Gahlot) Major Advisor Enclosed one original and three copies of bound thesis. Forwarded to the Dean, Post Graduate Studies, Rajasthan University of Veterinary & Animal Sciences, Bikaner, through the Dean. (G.C. Gahlot) Head Department of Animal Genetics and Breeding (Tribhuwan Sharma) DEAN College of Veterinary and Animal Science, Bikaner APPROVED (S.K. Kashyap) DEAN POST GRADUATE STUDIES ACKNOWLEDGEMENTS First of all, I would like to thank the almighty GOD for his endless mercies and all the blessings he bestowed in my life. With an overwhelming sense of legitimate pride and genuine obligation which gives me exuberant pleasure and privilege to express my eternal gratitude of my learned and revered advisor Professor (Dr) G.C. GahlotGahlot,, for his excellent and praiseworthy guidance, pertinent criticism, juvenile encouragement and parental affection during whole course of my study. I am grateful to him for being with me throughout the course of my research programme. Your insatiable hunger for quality work has inspired me to produce something up to your expectations for me. I feel highly obliged to DeanDean, College of Veterinary and Animal Science, Bikaner for their official help in providing me all the necessary facilities during the entire period of my research work. I am extremely thankful to State Animal Husbandry DepartmentDepartment, Government of Rajasthan for providing me study leave and financial assistance during the course of doctoral programme. I gratefully acknowledge learned members of my advisory committee, Professor Rajeev K. JoshiJoshi, Professor S.S.S.KS. KKK.. KashyapKashyap, Asst. Professor Rajesh Nehra and Professor R.K. Singh, for their understanding, encouragement and personal attention which have provided good and smooth basis for my PhD tenure. I warmly thanks for their valuable advice, constructive criticism and extensive discussion around my work. I would like express my gratitude and thanks to Professor S.B.S. Yadav, Professor V.K. ChoudharyChoudhary, Professor Urmila PannuPannu, Professor Tribhuwan SharmaSharma, Professor Basant BaisBais, Assistant Professor Manju Nehra, Assistant Professor Seetaram GuptaGupta, Assistant Professor Sumit Prakash YadavYadav,Yadav Assistant Professor Virendra Kumar for providing me technical support to carry out part of my research work. The constant affection and moral support given to me by colleagues Drs. Amitosh KumarKumar, Harvinder SinghSingh, Kritika GahlotGahlot, JaiprakashJaiprakash, PrakashPrakash, Ajay DewnaDewna, Govind DhakadDhakad, Manoj NetraNetra, Hemlata and all the juniors had pushed me and motivated me during the happy and hard moments. The completion of this study was not possible without the support of non teaching staff. I would like to express my sincere thanks to all of them especially Pradeep JiJiJi,Ji Deepak JiJi, MMahaveerahaveer JiJiJi,Ji Vijay JiJiJi,Ji and Shyam Ji and staffs of dairy farmsfarms. No words could be traced in the lexicon of my heart for the constant and unconditional support in the form of technical advice and digital support and inspiration and encouragement given to me by my best friends Dr. Mohammed Ashraf and Rituraj BhatiBhati. It seems impossible to express my heartiest feelings to my wife, colleague and best friend Dr Sonal Thakur whose unconditional love, patience and continuous support made me successful in all the aspects related with my life and doctoral research programme. A special thanks to my family. Words are unable to express my cardinal regards and love of my respected father Sh. R.K. AgrawalAgrawal and mother Smt. Bimla Devi who laid foundation stone of my education and constantly supported throughout my life. I deeply expressed my sincere regards to my brother DineshDinesh, Manmohan and AjayAjay. Especially I would like to pay my sincere affection and love to younger kids Utkarsh and Siddhika for sparing their valuable time to me. The affection and moral support of my in laws Dr. DheerajDheeraj,Dheeraj Shikha and little Ryan in the form of ever smiling face are graciously and lovingly acknowledged. Date: Vijay Kumar Agrawal Place: Bikaner LIST OF CONTENTS CHAPTER TITLE PAGE No. 1. INTRODUCTION 1-6 2. REVIEW OF LITERATURE 7-18 3. MATERIALS AND METHODS 19-28 4. RESULTS AND DISCUSSION 29-84 5. SUMMARY AND CONCLUSION 85-92 6. LITERATURE CITED 93-108 APPENDICES I-IV LIST OF TABLES Table No. Particulars Page No. 2.1 K232A polymorphism of DGAT1 gene in different cattle breeds 15 Primer sequences and e xpected fragment sizes of PCR 3.1 22 products of selected genomic regions 3.2 PCR programming for amplification of exon-3 of PRL gene 23 3.3 PCR programming for amplification of exon-4 of PRL gene 23 3.4 PCR programming for amplification of exon-8 of DGAT1 gene 24 4.1 Gene and genotypic frequencies of exon-3 of PRL gene 31 detected through RFLP analysis 4.2 Hardy-Weinberg equilibrium for exon-3 of PRL gene 33 4.3 Genotypic frequencies of SSCP patterns detected under 35 restriction patterns for exon-3 of PRL gene 4.4 Within-population heterozygosity estimates, PIC and F IS values 36 of Rathi, Sahiwal and Kankrej cattle for exon-3 of PRL gene 4.5 Observed and effective number of alleles; and Shannon 38 information index for exon-3 of PRL gene 4.6 Pair wise genetic distance matrix between studied cattle breeds 39 for exon-3 of PRL gene 4.7 List of accession numbers obtained for different SSCP patterns 39 of exon-3