ANALYSIS OF ABA-DEPENDENT ANTIOXIDANT RESPONSE OF WHEAT SEEDLINGS UNDER THE INFLUENCE OF EXOGENOUS SUGARS

Thesis

Submitted to the Punjab Agricultural University in partial fulfillment of the requirements for the degree of

MASTER OF SCIENCE in BIOCHEMISTRY (Minor Subject: Biotechnology) By Sandeep Kaur (L-2010/11-BS-185-M)

Department of Biochemistry College of Basic Sciences and Humanities PUNJAB AGRICULTURAL UNIVERSITY LUDHIANA-141004 2013

CERTIFICATE-I This is to certify that this thesis entitled ANALYSIS OF ABA-DEPENDENT ANTIOXIDANT RESPONSE OF WHEAT SEEDLINGS UNDER THE INFLUENCE OF EXOGENOUS SUGARS submitted for the degree of Master of Science in the subject of Biochemistry (Minor subject: Biotechnology) of Punjab Agricultural University, Ludhiana, is a bonafide research work carried out by Sandeep Kaur (L-2010/11-BS-185-M) under my 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.

Major Advisor Dr. Vikramjit Kaur Assistant Biochemist Deptt. of Biochemistry Punjab Agricultural University Ludhiana- 141004.

CERTIFICATE-II

This is to certify that the thesis report entitled, ANALYSIS OF ABA-DEPENDENT ANTIOXIDANT RESPONSE OF WHEAT SEEDLINGS UNDER THE INFLUENCE OF EXOGENOUS SUGARS submitted by Sandeep Kaur (L-2010/11-BS-185-M) to the Punjab Agricultural University, Ludhiana, in partial fulfillment of the requirements for the degree of Master of Science in the subject of Biochemistry (Minor subject: Biotechnology) has been approved by the Students Advisory Committee along with Head of the Department after an oral examination of the same, in collaboration with an external examiner.

Major Advisor (Dr. Vikramjit Kaur)

External Examiner Dr. S.K Singla Professor of Biochemistry Deptt. of Biochemistry Panjab University, Chandigarh

Head of the Department (Dr. A.K. Gupta)

Dean Postgraduate Studies (Dr. Gursharan Singh)

ACKNOWLEDGEMENT To everyone who helped me complete my manuscript, thank you. Here's the acknowledgement I wrote in my thesis for all my angels. First, I would like to thank God for all He has given me. Without Him, none of this would have been possible. Thanks God, the merciful and the passionate, for providing me the opportunity to step in the excellent world of science. I dedicate my gratitude to my God, whose grace and blessings in crunch situations allowed me to reign in odd crisis hours and to complete this endeavour. It is a matter of great pleasure for me to acknowledge my indebtness to my Major Advisor Dr. Vikramjit Kaur, Assistant Biochemist, Department of Biochemistry for her judicious guidance, motivation, enthusiasm, constant supervision and untiring help throughout and for all the patience and support she gave me since day first of my thesis completion. Her guidance helped me in all the time of research and writing of this thesis. I could not have imagined having a better advisor and mentor for my Masters study. Besides my advisor, I would like to thank the rest of members of my advisory committee Dr. A.K. Gupta, Professor-cum-Head, Department of Biochemistry, Dr. S.S. Gosal, Director of Research, Panjab Agricultural university and Dr. Bavita Asthir , Senior Biochemist (Dean PGS Nominee), Department of Biochemistry, for their encouragement, insightful comments, support and concrete suggestions during the tenure of my work. Last but not the least, I would like to thank my loving parents, for giving birth to me at the first place and supporting me morally and spiritually throughout my life. I dedicate this golden chapter of my life to my family. No choice of words will suffice to adequately register my gratitude to my sister and brother, loveable and close friendsGagan, Pallavi, Manpreet, my junior Lovelace and our lab attendant Mrs Inderpal Kaur for their care, help and moral advocacy. The successful completion of this manuscript was made possible through the invaluable contribution of a number of people. I would like to say thank you to all the direct and indirect help received from various quarters, but not mentioned here. Date: Place: Ludhiana. Sandeep Kaur

Title of the Thesis

Name of the Student and Admission No. Major Subject Minor Subject Name and Designation of Major Advisor Degree to be Awarded Year of award of Degree Total Pages in Thesis Name of University ABSTRACT

Analysis of ABA-dependent antioxidant response of wheat seedling under the influence of exogenous sugars SandeepKaur L-2010/11-BS-185-M Biochemistry Biotechnology Dr. VikramjitKaur Assistant Biochemist M.Sc. 2013 68 + Annexure + Vita Punjab Agricultural University Ludhiana- 141004, Punjab (India)

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Sugar(Glucose; G and Sucrose; S) regulation of seed germination, growth and antioxidant potential, and involvement of ABA biosynthesis in these regulations were studied in two wheat cultivars varying in ABA sensitivity, where C306 was ABA-higher sensitive and PBW343 was ABA-lesser sensitive. Two concentrations (1.5% and 3%) of both sugars were used for germination and growth experiments where sodium tungstate was used as inhibitor of ABA biosynthesis. Germination inhibition was observed under both sugars supply but ABA biosynthesis was involved only under 3% S in PBW343 and 1.5% S in C306 otherwise not involved in such inhibition. Growth of seedlings was inhibited under both concentration of G and under 3% S in both cultivars where ABA biosynthesis contributed in such inhibition under 1.5% G in both cultivars and 3% S in PBW343. Growth was stimulated under 1.5% S, it involved ABA biosynthesis in both cultivars. Increases in antioxidant enzymes were higher under G than under S in both cultivars and were higher in C306 than PBW343 under both sugar supply. These increases involved ABA biosynthesis in both cultivars. Reduced state of ascorbate was affected under G while increased under S in roots and shoots of PBW343. Reduced state of ascorbate was increased under both G and S in shoots but decreased in roots of C306. Maintainence of reduced state of ascorbate under S involved ABA biosynthesis in both cultivars. Roots of C306 showed toxicity symptoms under G and S where there were concomitant increases in dehydroascorbate, malondialdehyde, H2O2 and decreases reduced state of ascorbate. Keywords: ABA, Glucose, Sucrose, Antioxidant, Wheat. ______________________ Signature of Major Advisor ___________________ Signature of the Student

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CHAPTER

TOPIC

PAGE

I II III IV V

INTRODUCTION REVIEW OF LITERATURE MATERIAL AND METHODS RESULTS AND DISCUSSION SUMMARY REFERENCES

1-2 3-20 21-26 27-56 57-59 60-68

VITA

69