Physiological responses to saline irrigation in two summer mungbean vigna radiata (l ) wilczek genotypes

P H Y S I O L O G I C A L RE S P O N S E S T O S A L I N E I R R I G A T I O N I N T W O S UM M E R M U N G B E A N [ V I G N A R A D I A T A ( L ) W I L C Z E K ] G E N O T YP E S By DUONG HOANG SON (2010BS100D) Thesis submitted to the Chaudhary Charan Singh Haryana Agricultural University in partial fulfillment of the requirements for the degree of Doctor of Philosophy In Plant Physiology DEPARTMENT OF BOTANY AND PLANT PHYSIOLOGY COLLEGE OF BASIC SCIENCES AND HUMANITIES CCS HARYANA AGRICULTURAL UNIVERSITY HISAR – 125 004 2013 CERTIFICATE–I This is to certify that this thesis entitled, “Physiological responses to saline irrigation in two summer mungbean [Vigna radiata (L.) Wilczek] genotypes” submitted for the degree of Doctor of Philosophy in the subject of Plant Physiology to the CCS Haryana Agricultural University, Hisar, is a bonafide research work carried out by Mr Duong Hoang Son under my supervision and guidance 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 (Dr Neeraj Kumar) Major Advisor Scientist of Plant Physiology Department of Botany and Plant Physiology College of Basic Sciences and Humanities CCS Haryana Agricultural University Hisar-125 004 (Haryana) India CERTIFICATE–II This is to certify that this thesis entitled “Physiological responses to saline irrigation in two summer mungbean [Vigna radiata (L.) Wilczek] genotypes”, submitted by Mr Duong Hoang Son to the CCS Haryana Agricultural University, Hisar, in partial fulfillment of the requirement for the degree of Doctor of Philosophy in the subject of Plant Physiology, has been approved by the Student’s Advisory Committee, after an oral examination on the same, in collaboration with an external examiner MAJOR ADVISOR HEAD OF THE DEPARTMENT DEAN, POST-GRADUATE STUDIES EXTERNAL EXAMINER ACKNOWLEDGEMENTS All above, I want to express my deep sense of indebtedness to my family, especially my parent, my wife Quynh Mai and my sons Nhat Ha and Nhat Minh for their love, understanding and great companion I have no words to express my deep sense of gratitude and indebtedness to my advisor, Dr Neeraj Kumar, Scientist, Department of Botany and Plant Physiology, for his great kindness, constant encouragement and precious time to me in all aspects from the first day I came to India as well as during my study and investigation I owe deep and fervent thanks to Dr A.S Nadwal, Additional Director Research - Directorate of Researh, who spent a lot of time reading and discussing my thesis It gives me immense pleasure to record my sincere gratitude towards the learned members of my advisory committee: Dr S.K Sharma, Sr Scientist (Soil Science), Dr Ramesh Hasija, Sr Scientist (Statistic) and Dr Satish Kumar, Associate Dean PGS, for their intellectual enlightenment, sympathetic interest and pertinent suggestions throughout the pursuit of this study This study could not be completed without their kind help and support It is my profound privilege to express my heartiest thanks to Dr (Mrs.) Sunita Sheokand, Sr Scientist, Dr K.D Sharma, Scientist, Dr Rajiv Angrish, Sr Scientist and Dr H.R Dhingra, Professor of the Department of Botany and Plant Physiology for their timely help and willing cooperation Distinctive words of thanks are due to Dr (Mrs.) Rupa Dhawan, former Head, Dr J.K Sandooja, Head, Department of Botany and Plant Physiology, for providing the necessary facilities and cordial help whenever required I am thanking to all my friends for their help during my study, cheerful company and the research fellows and seniors especially Dr Anita Kumari and Dr Gunjan Geera for her guidance in the analysis of antioxidant enzymes, protein profile and informative discussions during the writing-up of this thesis, and Mr Suraj Bhan and Mr Raghubir Signh for technical assistance I am thankful to Dr Le Van Banh, Director, and Dr Cao Van Phung, Head of Soil Science department, CuuLong Delta Rice Research Institute, Vietnam, for providing me opportunity to study in India Financial help provided by Vietnam International Education Development (VIED), Ministry of Education and Training (MOET) in the form of 322 project fellowship is duly acknowledged (Duong Hoang Son) Date: Hisar ABBREVIATIONS APS - Ammonium per sulphate APX - Ascorbate peroxidase AsA - Ascorbate ATP - Adenosine tri phosphate CAT - Catalase CD - Critical difference CRD - Complete randomized design CSI - Chlorophyll stability index DAT - Days after treatment DMSO - Dimethyl sulphoxide DW - Dry weight EDTA - Ethylene diamine tetra acetic acid FW - Fresh weight G - Genotype G×V - Genotype × variety GR - Glutathione reductase GPX - Glutathione peroxidase GST - Glutathione S-transferase RWC - Relative water content MPa - Mega Pascal MW - Molecular weight NPQ - Non-photo chemical quenching OD - Optical density PS1 - Photosystem I PSII - Photosystem II POX - Peroxidase PVP - Polyvinyl pyrolidone ROS - Reactive oxygen species SDS-PAGE - Sodium dodecyl sulphate- Polyacrylamide gel electrophoresis SOD - Superoxide dismutase TTC - 2,3,5- triphenyl tetrazolium chloride w - Water potential s - Osmotic potential CONTENTS CHAPTERS TITLE PAGE(S) INTRODUCTION 1-4 REVIEW OF LITERATURE 5-19 MATERIAL AND METHODS 20-36 RESULTS 37-72 DISCUSSION 73-83 SUMMARY AND CONCLUSIONS 84-86 BIBLIOGRAPHY i-xiii LIST OF TABLES S No Title Page No Changes in water potential (-MPa) in leaf of mungbean genotypes under saline irrigation 39 Changes in osmotic potential (-MPa) in leaf of mungbean genotypes under saline irrigation 40 Changes in osmotic potential (-MPa) in root of mungbean genotypes under saline irrigation 41 Changes in relative water content (RWC %) in leaf of mungbean genotypes under saline irrigation 41 Changes in relative water content (RWC %) in root of mungbean genotypes under saline irrigation 42 Changes in relative stress injury (RSI %) in leaf of mungbean genotypes under saline irrigation 43 Changes in relative stress injury (RSI %) in root of mungbean genotypes under saline irrigation 44 Changes in Na+/K+ ratio in leaf of mungbean genotypes under saline irrigation 50 Changes in Na+/K+ ratio in root of mungbean genotypes under saline irrigation 50 10 Changes in chloride (Cl-) content (mg g-1 DW) in leaf of mungbean genotypes under saline irrigation 51 11 Changes in chloride (Cl-) content (mg g-1 DW) in root of mungbean genotypes under saline irrigation 52 12 Changes in sulphate (SO42-) content (mg g-1 DW) in leaf of mungbean genotypes under saline irrigation 53 13 Changes in sulphate (SO42-) content (mg g-1 DW) in root of mungbean genotypes under saline irrigation 53 14 Changes in in vitro pollen germination (%) and pollen tuber growth (µm) in two mungbean genotypes under saline irrigation 69 15 Changes in yield and yield attributes in two mungbean genotypes under saline irrigation 71 16 Changes in ECe of soil under saline irrigation 72 LIST OF FIGURES S No Title Changes in plant height (cm) in mungbean genotypes under saline irrigation 37 Changes in dry matter (g plant-1) content in leaf of mungbean genotypes under saline irrigation 38 Changes in dry matter (g plant-1) content in root of mungbean genotypes under saline irrigation 38 Changes in lipid peroxidation (MDA) content (nmoles g-1 DW) in leaf of mungbean genotypes under saline irrigation 45 Changes in lipid peroxidation (MDA) content (nmoles g-1 DW) in root of mungbean genotypes under saline irrigation 45 Changes in chlorophyll ‘a’ content (mg g-1 DW) in leaf of mungbean genotypes under saline irrigation 46 Changes in chlorophyll ‘b’ content (mg g-1 DW) in leaf of mungbean genotypes under saline irrigation 47 Changes in total chlorophyll content (mg g-1 DW) in leaf of mungbean genotypes under saline irrigation 47 Changes in chlorophyll stability index (%) in leaf of mungbean genotypes under saline irrigation 48 10 Changes in quantum yield (Fv/Fm) in leaf of mungbean genotypes under saline irrigation 48 11 Changes in hydrogen peroxides (H2O2) content (moles g-1 DW x 10-4) in leaf of mungbean genotypes under saline irrigation 54 12 Changes in hydrogen peroxides (H2O2) content (moles g-1 DW x 10-4) in root of mungbean genotypes under saline irrigation 55 13 Changes in proline content (mg g-1 DW) in leaf of mungbean genotypes under saline irrigation 55 14 Changes in proline content (mg g-1 DW) in root of mungbean genotypes under saline irrigation 56 15 Changes in total soluble carbohydrates content (mg g-1 DW) in leaf of mungbean genotypes under saline irrigation 57 16 Changes in total soluble carbohydrates content (mg g-1 DW) in root of mungbean genotypes under saline irrigation 57 17 Changes in superoxide dismutase (SOD) specific activity (Units mg-1 protein) in leaf of mungbean genotypes under saline irrigation 58 18 Changes in superoxide dismutase (SOD) specific activity (Units mg-1 protein) in root of mungbean genotypes under saline irrigation 59 Page No 19 Changes in catalase (CAT) specific activity (Units mg-1 protein) in leaves of mung bean genotypes under saline irrigation 60 20 Changes in catalase (CAT) specific activity (Units mg-1 protein) in root of mungbean genotypes under saline irrigation 60 21 Changes in peroxidase (POX) specific activity (Units mg-1 protein) in leaf of mungbean genotypes under saline irrigation 61 22 Changes in peroxidise (POX) specific activity (Units mg-1 protein) in root of mungbean genotypes under saline irrigation 61 23 Changes in ascorbate peroxidase (APX) specific activity (Units mg-1 protein) in leaf of mungbean genotypes under saline irrigation 62 24 Changes in ascorbate peroxidase (APX) specific activity (Units mg-1 protein) in root of mungbean genotypes under saline irrigation 63 25 Changes in glutathione reductase (GR) specific activity (Units mg-1 protein) in leaf of mungbean genotypes under saline irrigation 63 26 Changes in glutathione reductase (GR) specific activity (Units mg-1 protein) in root of mungbean genotypes under saline irrigation 64 27 Changes in glutathione S transferase (GST) specific activity (Units mg-1 protein) in leaf of mungbean genotypes under saline irrigation 65 28 Changes in glutathione S transferase (GST) specific activity (Units mg-1 protein) in root of mungbean genotypes under saline irrigation 65 29 Changes in glutathione peroxidase (GPX) specific activity (Units mg-1 protein) in leaf of mungbean genotypes under saline irrigation 66 30 Changes in glutathione peroxidase (GPX) specific activity (Units mg-1 protein) in root of mungbean genotypes under saline irrigation 66 31 Changes in ascorbate (AsA) content (µmoles g-1 DW) in leaf of mungbean genotypes under saline irrigation 67 32 Changes in ascorbate (AsA) content (µmoles g-1 DW) in root of mungbean genotypes under saline irrigation 68 33 Changes in pollen viability in two mungbean genotypes under saline irrigation 68 CHAPTER-I INTRODUCTION Environmental stresses are the most important constraints limiting crop productivity Among these salinity either of soil or water is a serious problem for agriculture all over the world (Majid et al., 2011) Salinity limited the growth and development of plant by altering their morphological, physiological, biochemical attributes and production in most of the arid and semi arid regions of the world (Mudgal et al., 2010; Kandil et al., 2012) There are different causes of the development of soil salinity The major forms are viz (i) natural or primary salinity and (ii) secondary or human-induced salinity Primary salinity is occurred due to the long-term natural accumulation of salts in the soil or surface water Secondary salinity occurs due to anthropogenic activities that disrupt the hydrologic balance of the soil between water applied (irrigation or rainfall) and water used by crops (Geetanjali and Neera, 2008) Salinity created due to high salt concentration in the soil solution is two-fold First, many of salt ions are toxic to plant cells and second, high salt represents a water deficit or osmotic stress Specific ion toxicity is usually associated with excessive intake of sodium, chloride or other ions and causes disrupt plant potassium and calcium nutrition (Zhu, 2007) The deleterious effect of the saline irrigation on plant involve osmotic stress, ion toxicity and mineral deficiency (Ashraf and Harris, 2004) and reduction in growth and alterations in several physiological processes including N2- fixation (Nandwal et al., 2000 a, b; Kukreja et al., 2005) Water potential and osmotic potential become more negative whereas turgor pressure increases with increasing salinity (Mudgal et al., 2010) Osmotic adjustment involves either inorganic ions or low molecules weight organic solutes These play a crucial role in higher plants grown under saline conditions The compatible osmolytes generally found in higher plants are low molecular weight sugars, organic acids, polyols and nitrogen containing compounds such as amino acids, amides, proteins and quaternary ammonium compounds (Dionisio-Sese and Tobita, 1998; Ashraf and Harris, 2004; Mudgal et al., 2010; Sabina and Mehar, 2011) Several physiological and biochemical processes like pigment content and photosynthesis, carbohydrate metabolism, protein synthesis, energy and lipid metabolism are affected by salinity Salt stress disturbs intracellular ion homeostasis in plants, which leads to damage in maintaining cell turgor, enzyme activities, membrane dysfunction, attenuation of Arulbalachandran, D., Ganesh, K.S and Subramani, A 2009 Changes in metabolites and antioxidant enzyme activity of three Vigna species induced by NaCl stress American European J Agri 2: 109-116 Ashraf, M and Harris, P.J.C 2004 Potential biochemical indicators of salinity tolerance in plants Plant Sci 166: 3–16 Ashraf, M and Rasul, E 1988 Salt tolerance of mung bean plant (Vigna radiata L Wilczek) at two growth stages Plant Soil 110: 63-67 Balasubramanian, V and Sinha, S.K 1976 Effect of salt stress on growth, nodulation and nitrogen fixation in cowpea and mung bean Physiol Plant 36: 1361-1365 Barnett, N.M and Naylor, A.W 1966 Amino acid and protein metabolism in Bermuda grass during water stress Plant Physiol 41: 1222-1230 Bartels, D and Sunkar, R 2005 Drought and salt tolerance in plants Cri Rev Plant Sci 24: 23-58 Bates, L.S., Waldren, R.P and Teare, I.D 1973 Raid determination of free proline for water stress studies Plant Soil 39: 205-207 Beauchamp, C and Fridovich, I 1971 Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels Anal Biochem 44: 276-287 Blokhina, O., Virolainen, E and Fagerstedt, K.V 2003 Antioxidants, oxidative damage and oxygen deprivation stress- a review Ann Bot 91: 179-194 Bor, M.F., Ozdemir, F and Turkan, I 2003 The effect of salt stress on lipid peroxidation and antioxidants in leaves of sugar beet Beta vulgaris L and wild beet Beta maritime L Plant Sci 164: 77-84 Bradford, M M 1976 A rapid and sensitive method for the quantitation ofmicrogram quantities of protein utilizing the principle of protein-dyebinding Anal Biochem 72: 248–254 Cavalcanti, F.R., Lima, J.P.M.S., Ferreira, S.S.L., Viegas, R.A and Silveira, J.A.G 2007 Roots and leaves display contrasting oxidative response during salt stress and recovery in cowpea J Plant Physiol 164: 591-600 Cavalcanti, F.R., Oliveira, J.T.A., Miranda, A.S.M., Ricardo, A.V and Silveira, J.A.G 2004 Superoxide dismutase, catalase and peroxidase activities not confer protection against oxidative damage in salt-stressed cowpea leaves New Phyto 163: 563–571 Chakrabarti, N and Mukherji, S 2002 Growth regulator mediated changes in leaf area and metabolic activity in mung bean under salt stress condition Indian J Plant Physiol 7: 256-263 Chakrabarti, N., Mukherji, S and Chakrabarti, N 2003 a Alleviation of NaCl stress by pretreatment with phytohormones in Vigna radiata Biol Plant 46: 63-66 Chakrabarti, N., Mukherji, S and Chakrabarti, N 2003 b Effect of phytohormone pretreatment on metabolic changes in Vigna radiata under salt stress J Environ Biol 23: 295-300 ii Chesnin, L and Yien, C.H 1950 Turbidmetric determination of available sulphur Soil Sci Soc Am Proc 15: 149-151 Chhabra, R 1973 Kinetics of absorption of chloride and phosphorus, their interaction and effect on growth and composition of tomato plants Ph.D Thesis, KUL, Belgium Corpus, F., Gomez, J.M., Hernandez, J.A and Del Roi, J.A 1993 Metabolism of activated oxygen in leaf peroxisome from two Pisum sativum L cultivars with different sensitivity to sodium chloride J Plant Physiol 141: 160-165 Costa, P.H.A., Neto, A., Bezerra, M., Prisco, J and Filho, E 2005 Antioxidant-enzymatic system of two sorghum genotypes differing in salt tolerance Brazil J Plant Physiol 17: 353-361 Dash, M and Panda, S.K 2001 Salt stress induced change in growth and enzyme activities in germinating Phaseolus mungo seeds Biol Plant 44: 587-589 Demir, Y 2000 Growth and proline content of germination wheat genotypes under ultraviolet light Turkey J Bot 24: 67-70 Dhingra, H.R and Sharma, P.K.1992 Reproductive performance of pea (Pisum sativum L.) under saline condition Indian J Plant Physiol 35: 198-201 Dhingra, H.R and Varghese, T.M 1985 Effect of salt on viability, germination and endogenous level of some metabolites and ions in maize (Zea mays L.) pollen Ann Bot 55: 415-420 Dhingra, H.R and Varghese, T.M 1986 Effect of chloride salts on in vitro germination and tube growth of maize (Zea mays L.) pollen Acta Bot Indica 13: 8-13 Dhingra, H.R and Varghese, T.M 1993 Flowering and male reproductive function of chickpea (Cicer arietinum L.) genotypes as affected by salinity Biol Plant 35: 447-452 Dionisio-Sese, M.L and Tobita, S 1998 Antioxidant responses of rice seedlings to salinity stress Plant Sci 135: 1-9 Dolatabadion, A., Sanavy, S.A.M.M and Chashmi, N.A 2008 The effects of foliar application of ascorbic acid (vitamin C) on antioxidant enzymes activities, lipid peroxidation and proline accumulation of canola (Brassica napus L.) under conditions of salt stress J Agric Crop Sci 194: 206-213 El-baky, A., Hanaa, H., Mohamed, A A and Hussein, M.M 2003 Influence of salinity on lipid peroxidation, antioxidant enzymes and electrophoretic pattern of protein and isozymes in leaves of some onion cultivars J Plant Sci 2: 633-638 Farooq, S and Azam, F 2006 The use of cell membrane stability (CMS) technique to screen for salt tolerant wheat varieties J Plant Physiol 163: 629-637 Farouk, S 2011 Ascorbic acid and α-Tocopherol minimize salt-induced wheat leaf senescence J Stress Physiol Biochem 7: 58-79 iii Foyer, C.H.M., Lelandais, C.G and Kunert, K.J 1991 Effect of elevated cytosolic glutathione pool and photosynthesis in leaves under normal and stress conditions Plant Physiol 97: 863-872 Francois, L.E and Maas, E.V 1999 Hand book of plant and crop stress Chapter 8: Crop response and management of salt-affected soils Marcel Dekker Inc, New York Publisher Pp 169-202 Fusun, E and Mehmet, T.O 2007 Effect of salinity on antioxidant responses of chickpea seedlings Acta Physiol Plant 29: 485–493 Gandhi, A.P and Paliwal, K.V 1975 Effect of different quality irrigation waters and soil texture on the yield and uptake of nutrients by wheat Proc Ind Natl Sci Acad B 41: 440-451 Garg, B.K., Kathju S., Vyas, S.P and Lahiri A.N 1996 Relative effects of sodium salts on growth, yield and metabolism of arid legumes Plant Physiol Biochem 23: 148-152 Geetanjali, M and Neera, G 2008 Salinity and its effects on the functional biology of legumes Acta Physiol Plant 30: 595-618 Guo, Y., Tian, Z., Yan, D., Zhang, Y and Qin, P 2009 Effect of nitric oxide on salt stress tolerance in Kostelezkya virginica Life Sci J 6: 67-75 Gupta, S and Murty, Y.S 1978 Pollen germination in mothbean (Phaseolus aureus) J Indian Bot Soc 57: 53-57 Hafeez, F.Y., Aslam, Z and Malik, K.A 1988 Effect of salinity and inoculation on growth, nitrogen fixation and nutrient uptake of Vigna radiata (L.) Wilczek Plant Soil 106: 3-8 Halliwell, B and Foyer, C.H 1978 Properties and physiological functions of a glutathione reductase purified from spinach leaves by affinity chromatography Planta 139: 9-17 Hameda, E.S and Ralph, C.K 1992 Effects of NaCl salinity and hydrogel polymer treatments on viability, germination and solute contents in maize (Zea meays) pollen Phyton 32: 143-157 Hasegawa, P.M., Bressan, R.A., Zhu, J.K and Bohnert, H.J 2000 Plant cellular and molecular responses to high salinity Annu Rev Plant Mol Plant Physiol 51: 463-499 Hauser, E.J.P and Morrison, J.H 1964 The cytochemical reduction of nitroblue tetrazolium as an index of pollen viability Am J Bot 51: 748-752 He, Y and Zhu, Z.J 2008 Exogenous salicylic acid alleviates NaCl toxicity and increases antioxidative enzyme activity in Lycopersicon esculenttum Biol Plant 52: 792-795 Heath, R.L and Packer, L 1968 Photoperoxidation in isolated chloroplasts I Kinetics and stoichiometry of fatty acid peroxidation Arch Biochem Biophy 125: 189-198 Hernandez, J.A., Campillo, A., Jimenez, A., Alarcon, J J and Sevilla, F 1999 Response of antioxidant systems and leaf water relations to NaCl stress in pea plants New Phytol 141: 241-251 iv Hernandez, J.A., Jimenez, A., Mullineaux, P and Sevilla, F 2000 Tolerance of pea (Pisum sativum L.) to long term salt stress is associated with induction of antioxidant defenses Plant Cell Eviron 23: 853-862 Hiscox, J.D and Isrealstam, G.F 1979 A method for the extraction of chlorophyll from leaf tissue without maceration Canadian J Bot 57: 1332-1334 Hossain, M.A and Fujita M 2010 Evidence for a role of exogenous glycinebetaine and proline in antioxidant defense and methylglyoxal detoxification systems in mung bean seedlings under salt stress Physiol Mol Biol Plants 16: 19-29 Hossain, M.A., Hasanuzzaman, M and Fujita, M 2011 Coordinate induction of antioxidant defense and glyoxalase system by exogenous proline and glycinebetain is correlated with salt tolerance in mungbean Front Agric China 5: 1-14 Humaira, G and Rafiq, A 2006 Effect of salintity on pollen viability of different canola (Brassica napus L.) cultivars as reflected by the formation of fruits and seeds Pak J Bot 38: 237-247 Hussain, T.M., Chandrasekhar, T., Hazara, M., Sultan, Z., Saleh, B.Z and Gopal, G.R 2008 Recent advances in salt stress biology - a review Biotechnol Mol Biol 3: 8-13 Jain, R.K., Singh, N and Jain, S 2003 Engineering salinity tolerance in crop plants: A Reality Physiol Mol Biol Plant 9: 29-39 Jaleel, C.A., Gopi, R., Manivannan, E and Panneerseevam, R 2007 Response of antioxidant defence system of Catharantnus roseus (L.) G Don to poclobutrazol treatment under salinity Acta Physiol Plant 29: 205-209 Joseph, B and Jini, D 2011 Development of salt stress-tolerant plants by gene manipulation of antioxidant enzymes Asian J Agric Res 5: 17-27 Kabir, M.E., Karim, M.A and Azad, M.A.K 2004 Effect of potassium on salinity tolerance of mung bean (Vigna radiata L Wilczek) J Biol Sci 4: 103-110 Kandil, A.A., Arafa, A.A., Sharief, A.E and Ramadan, A.N 2012 Genotypes differences between two mungbean varieties in response to salinity stress at seedling stage Int J Agric Sci 4: 278283 Katsuhara, M., Otsuka, T and Ezaki, B 2005 Salt stress induced lipid peroxidation is reduced by glutathione S transferase, but this reduction of lipid peroxides is not enough for recovery of root growth in Arabidopsis Plant Sci 169: 369-373 Kestwal, R.M., Kestwal, D.B and Chiang, B.H 2012 Analysis and enhancement of nutritional and antioxidant properties of Vigna aconitifolia sprouts Plant Foods Hum Nutr 67: 136-141 Khan, M.H and Patra, H.K 2007 Sodium chloride and cadmium induced oxidative stress and antioxidant response in Calamus tenuis leaves Indian J Plant Physiol 12: 34-40 v Khan, M.I and Khatoon, A 1997 Effect of seawater and NaCl salinity on the in vitro germination of pollen and tube growth of catharanthus roseus (L) Pak J Bot 29: 351-354 Khatun, S and Flowers, T.J 1995 The estimation of pollen viability in rice J Exp Bot 46: 151-151 Koleyoreas., S.A 1958 A new method for determining drought resistance Plant Physiol 33: 232-233 Kukreja, S., Nandwal, A.S., Kumar, N., Sharma, S.K., Kundu, B.S., Unvi, V and Sharma, P.K 2006 Responses of chickpea roots to short term salinization and desalinization: plant water status, ethylene evolution, antioxidant activity and membrane integrity Physiol Mol Biol Plants 12: 67-73 Kukreja, S., Nandwal, A.S., Kumar, N., Sharma, S.K., Sharma, S.K and Unvi, V 2005 Plant water status, H2O2 scavenging enzymes, ethylene evolution and membrane integrity of Cicer arietinum roots as affected by salinity Biol Plant 49: 305-308 Kukreja, S., Nandwal, A.S., Kumar, N., Sharma, S.K., Sunder, S., Sharma, S.K., Sharma, S.K., Devi, S and Kumar, A 2010 Ethylene evolution and modification of antioxidant defense mechanism as indices of salinity stress tolerance in Cicer arietinum L nodules Indian J Plant Physiol 15: 203-212 Laemmli, U.K 1970 Cleavage of structural proteins during the assembly of the head bacteriophage T4 Nature 227: 680-685 Lee, D.H., Kim, Y.S and Lee, C.B 2000 Chilling stress induced changes of antioxidant enzymes in the leaves of cucumber: in gel enzyme activity assay Plant Sci 159: 75-85 Lee, D.H., Kim, Y.S and Lee, C.B 2001 The inductive responses of antioxidant enzymes by salt stress in the rice (Oryza sativa) J Plant Physiol 158: 737-745 Lee, G., Carrow, R.N., and Duncan, R.R 2004 Photosynthetic responses to salinity stress halophyte seashore paspalum ecotypes Plant Sci 166: 1417-1425 Levine, A., Tenhaken, R., Dixon, R and Lamb, C 1994 H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response Cell 79: 583-593 Levitt, J 1980 Responses of plants to environmental stresses Volume II Water, radiation, salt, and other stresses 2nd Eds Academic Press, Berlin and New York Publisher Pp 365-454 Li, S.W and Xue, L 2010 The interaction between H2O2 and NO, Ca2+, cGMP and MAPKs during adventitious rooting in mung bean seedling In Vitro Cell Dev Biol Plant 46: 142-148 Li, Y 2008 Kinetics of the antioxidant response to salinity in the halophyte limonium bicolor Plant Soil Environ 54: 493-497 Liang, Y.C., Chen, O., Liu, Q., Zhang, W.H and Ding, R.X 2003 Exogenous silicon (Si) increases antioxidant enzyme activity and reduces lipid peroxidation in roots of salt-stressed barley (Hordeum vulgare L.) J Plant Physiol 160: 1157-1164 vi Maia, J.M., Macedo, C.E.C., Voigt, E.L., Freitas, E.L and Silveira, J.A.G 2010 Antioxidative enzymatic protection in leaves of two contrasting cowpea cultivars under salinity Biol Plant 54: 159-163 Maity, S., Bannjee, G., Roy, M., Pal, C., Pal, B., Chak, D and Bhattacharjee, A 2000 Chemical induced prolongation of seed viability and stress tolerance capacity of mung bean seedling Seed Sci Technol 28: 155-162 Majid, G J., Ali, S., Foad, M., Seyed Ali, M M S and Iraj, A 2011 The role of phytohormones in alleviating salt stress in crop plants Australia J Crop Sci 5: 726-734 Maliwal, G.L and Paliwal KV 1982 Salt tolerance of some mungbean (Vigna radiata), urdbean (Vigna mungo) and gaur (Cyamopsis tetragonoloba) varieties at germination and early stages Legume Res 5: 23-30 Mandhania, S., Madan, S and Sawhney, V 2006 Antioxidant defense mechanism under salt stress in wheat seedlings Biol Plant 50: 227-231 Manivannan, P., Jaleel, C.A., Sankar, B., Somasundaram, R., Murali, P.V., Sridharan, R and Panneerselvam, R 2007 Salt stress mitigation by calcium chloride in Vigna radiata L Wilczek Acta Bio Cra Ser Bot 49: 105-109 Mannan, M.A., Karim, M.A., Khaliq, Q.A., Haque, M.M., Mian, M.A.K and Ahmed, J.U 2009 Proline accumulation, water status and chlorophyll content in leaf in relation to salt tolerance in soybean Indian J Plant Physiol 14: 130-134 María, P.B., Patricia, L.M., Susana, M.G., María, E.C and María, L.T 2000 Relationship between antioxidant defence systems and salt tolerance in Solanum tuberosum Australia J Plant Physiol 27: 273–278 Marler, T.E and Mickelbart, M.V 1993 Growth and chlorophyll fluorescence of Spondias purpurea L as influenced by salinity Trop Agric 70: 245-247 Mass, E.V and Grieve, C.M 1987 Sodium induced calcium deficiency in salt stress corn Plant Cell Eviron 10: 559-564 Maxwell, K and Johnson, G.N 2000 Chlorophyll fluorescence a practical guide J Exp Bot 51: 659668 McCord, J.M and Fridovichi, I 1969 Superoxide dismutase An enzymic function for erythocuprein (hemocuprein) J Biol Chem 244: 6049-6055 Meloni, D.A., Oliva, M.A., Martinez, C.A and Cambraia, J 2002 Photosynthesis and activity of superoxide dismutase and glutathione reductase in cotton under salt stress Environ Exp Bot 49: 69-76 Mensah, J.K and Ihenyen, J 2009 Effect of salinity on germination, seedling establishment and yield of three genotypes of mung bean (Vigna mungo L Hepper) in Edo state, Nigeria Nigerian Ann Nat Sci 8: 17-24 vii Minhas, P.S., Sharma, D.R and Khosla, B.K.1990 Mung bean response to irrigation with waters of different salinities Irrig Sci 11: 57-62 Misra, N and Gupta, A.K 2006 Effect of salinity and different nitrogen sources on the activity of antioxidant enzymes and indole alkaloid content in Catharanthus roseous seedlings J Plant Physiol 163: 11-18 Misra, N and Dwivedi, U.N 2004 Genotypic difference in salinity tolerance of green gram cultivars Plant Sci 166: 1135-1142 Mitchell, J.P and Shennan, C 1991 Tomato fruit yields and quality under water deficit and salinity J Am Soc Hort Sci 116: 215-221 Mittova, V., Tal, M., Volokita, M and Guy, M 2003 a Up-regulation of the leaf mitochondrial and peroxisomal antioxidative systems in response to salt-induced oxidative stress in the wild salttolerant tomato species Lycopersicon pennellii Plant Cell Environ 26: 845-856 Mittova, V., Frederica, L.T., Guy K., Leonardo, G., Micha, V., Moshe, T., Christine, H.F and Micha, G 2003 b Coordinate induction of glutathione biosynthesis and glutathione metabolizing enzymes is correlated with salt tolerance in tomato FEBS Letters 554: 417-421 Mohamed, M.H and EL Kramany, M.F 2005 Salinity tolerance of some mungbean varieties J App Sci Res 1: 78-84 Mohammed, A.H.M.A 2007 Physiological aspects of mung bean plant (Vigna radiata L Wilczek) in response to salt stress and gibberellic acid treatment Res J Agric Biol Sci 3: 200-213 Mudgal, V., Madaan, N and Mudgal, A 2010 Biochemical mechanisms of salt tolerance in plants: a review Int J Bot 6: 136-143 Munns, R 2002 Comparative physiology of salt and water stress Plant Cell Environ 25: 239-250 Munns, R 2005 Genes and salt tolerance: bringing them together New Phytol 167: 645-663 Nafees, A.K., Shabina, S., Asim, M., Rahat, N and Noushina, I 2010 Application of salicylic acid increases contents of nutrients and antioxidative metabolism in mung bean and alleviates adverse effects of salinity stress Int J Plant Bio 1: 1-8 Nakano, Y and Asada, K 1981 Hydrogen peroxide is scavenged by ascorbate specific hydrogen peroxidase in spinach chloroplast Plant Cell Physiol 28: 131-140 Nandwal, A S., Godara, M., Kamboj, D V., Kundu, B S., Mann A., Kumar, B and Sharma, S K 2000 a Nodule functioning in trifoliate and pentafoliate mungbean genotypes as influenced by salinity Biol Plant 43: 459-462 Nandwal, A.S., Godara, M., Sheokand, S., Kamboj, D.V., Kundu, B.S., Kuhad, M.S., Kumar, B and Sharma, S.K 2000 b Salinity induced changes in plant water status, nodule functioning and ionic distribution in phenotypically differing genotype of Vigna radiata L J Plant Physiol 156: 352-359 viii Navneet, K.Y., Sarika, M.A.I and Mohd, A 2011 In-silico analysis and homology modelling of coatprotein of mungbean yellow mosaic India virus J Food Leg 24: 138-141 Neelam, S 2013 Enhancement of catalase activity under salt stress in germinating seeds of Vigna radiata Asian J Biomed Pharma Sci 3: 6-8 Kumar, N., Sunder, S., Nadwal, A.S., Waldia, R.S and Sharma, M.K 2008 Genotypic differences in water status, membrane integrity, ionic content, N2-fixing efficiency and dry matter of mung bean nodules under saline irrigation Physiol Mol Biol Plants 14: 363-368 Neto, A.D.A., Prisco, J.T., Filho, J.E., Medeiros, J.V.R and Filho, E.G 2005 Hydrogen peroxide pretreatment induces salt stress acclimation in maize plants J Plant Physiol 162: 1114-1122 Ottaviano, E., Gorla, M.S., Mulcahi, D.L 1975 Gamatic and intergamatophytic infuence on pollen tube growth In: Gamete Competition in Plants and Animals (Ed D.L Mulcahi), pp 125-130 North Holland Publishing Co, Amsterdam Publisher Panda, S.K 2001 Oxidative response of green gram under salinity stress Indian J Plant Physiol 6: 438-440 Pandey, S.K., Bahuguna, R.N., Pal, M., Trivedi, A.K., Hemantaranjan, A and Srivastava, J.P 2010 Effects of pre-treatment and foliar application of zinc on growth and yield components of mungbean (Vigna radiata L.) under induced salinity Indian J Plant Physiol 15: 164-167 Parida, A.K., and Das, A.B 2005 Salt tolerance and salinity effects on plants- a review Ecotox Environ Saf 60: 324-349 Parida, A.K., Das, A.B and Das, P 2002 NaCl stress causes changes in photosynthetic pigments, proteins and other metabolic components in the leaves of a true mangrove, Bruguiera parviflora, in hydroponic cultures J Plant Biol 45: 28-36 Parveen, R., Karmoker, J.L., Sabanando, C and Bimal, C.S 2004 The effect of salinity on ion accumulation and anatomical attributes in mungbean (Phaseolus radiatus L cv BARI-3) seedlings Int J Agric Biol 6: 495-498 Passardi, F., Penel, C and Dunand, C 2004 Performing the paradoxical: how plant peroxidases modify the cell wall Trends Plant Sci 9: 534-540 Patterson, B.D., Machae, E.A and Ferguson, I.G 1984 Estimation of hydrogen peroxide in plant extracts using titanium (IV) Anal Biochem 139: 487-492 Parvaiz, A and Satyawati, S 2008 Salt stress and phyto-biochemical responses of plants Plant Soil Environ 54: 89-99 Picchioni, G A and Miyamoto, S 1990 Salt effects on growth and ion uptake of Pistachio rootstock seedlings J Am Soc Hort Sci 115: 647-653 ix Pooja, R and Sharma, K.D 2010 Salicylic acid induced amelioration in growth, biochemical metabolites and yield of mungbean (Vigna radiata L.) under salinity stress Indian J Plant Physiol 15: 219-225 Promila, K and Kumar, S 2000 Vigna radiata seed germination under salinity Biol Plant 43: 423426 Rai, V K 2002 Role of amino acids in plant responses to stress Biol Plant 45: 481-487 Rinallo, C 1992 Effects of acidity of stimulate rain on the fruiting “summered” apple trees J Environ Qual 21: 61-68 Rodriguez, R.M.P., Kerkeb, L., Bueno, P and Donaire, J.P 1999 Changes induced by NaCl in lipid content and composition, lipoxygenase, plasma membrane H+-ATPase and antioxidant enzyme activities of tomato (Lycopersicon esculentum Mill) calli Plant Sci 143: 143-150 Rolfe, B.G and Gresshoff, P.M 1988 Genetic analysis of legume nodule initiation Annu Rev Plant Physiol 39: 297-319 Roxas, V.P., Sundus, A.L., Daniel, K.G., James, R.M and Allen, R.D 2000 Stress tolerance in transgenic tobacco seedlings that overexpress glutathione S-transferase/glutathione peroxidase Plant Cell Physiol 41: 1229-1234 Saffan, S.E.S 2008 Effect of salinity and osmotic stresses on some economic plants Res J Agric Biol Sci 4: 159-166 Saha, P., Chatterjee, P and Biswas, A.K 2010 NaCl pretreatment alleviates salt stress by enhancement of antioxidant defense system and osmolyte accumulation in mungbean (Vigna radiata L Wilczek) Indian J Exp Biol 48: 593-600 Sairam, R.K and Tyagi, A 2004 Physiology and molecular biology of salinity stress tolerance in plants Curr Sci 86: 407-421 Sairam, R.K., Rao, K.V and Srivastava, G.C 2002 Differential responses of wheat genotypes to long term salinity stress in relation to oxidative stress, antioxidant activity and osmolytic concentration Plant Sci 163: 1037-1046 Sass, J.E 1964 Botanical microtechnique Oxford and IBH Publishing Co, New Delhi Publisher Pp 12-22 Shabala, S 2000 Ionic and osmotic components of salt stress specifically modulate net ion fluxes from bean leaf mesophyll Plant Cell Environ 23: 825-837 Shabina, S and Khan, N.A 2004 Activities of carbonic anhydrase, catalase and ACC oxidase of mung bean (Vigna radiata) are differentially affected by salinity stress Food Agric Environ 2: 241-249 Shabina, S and Mehar, F 2011 Salt tolerance in mung bean Vigna radiata L genotypes: role of proline and glycinebetaine J Func Environ Bot 1: 139-147 x Sharma, S.K and Khajanchi, L 2009 Raising crops using saline waters: management strategies including nutrient management Enhancing nutrition use efficiency in problem soil ICAR Azad Hind Stores (P) Ltd Publisher Pp 186-207 Sheldon, A., Menzies, N.W., So, B.H and Dalal, R 2004 The effect of salinity on plant available water In: Supersoil: 3rd Australian New Zealand Soils Conference (Ed B Sing), pp 1-5 Sydney, Australia, December 5-9 2004 Sheokand, S., Kumari, A and Sawhney, V 2008 Effect of nitric oxide and putrescine on antioxidative responses under NaCl stress in chickpea plants Physiol Mol Biol Plants 14: 355-362 Sheoran, I.S and Garg, O.P 1979 Quantitative and qualitative changes in peroxidase during germination of mung bean under salt stress Physiol Plant 46: 147-150 Siegel, B.S and Siegel, S.M 1969 Anomalous substrate specificities among the algal peroxidases American J Bot 57: 285-287 Shigeoka, S., Ishikawa, T., Tamoi, M., Miyagawa, Y., Takeda T., Yabuta, Y and Yoshimura, K 2002 Regulation and function of ascorbate peroxidase isoenzymes J Exp Bot 53: 1305-1319 Singh D.P and Singh B.B 2011 Breeding for tolerance to abiotic stresses in mungbean J Food Legumes 24: 83-90 Singh, S.P., Singh, B.B and Singh, M 1994 Effect of kinetin on chlorophyll, nitrogen and proline in mungbean (Vigna radiata) under saline conditions Indian J Plant Physiol 37: 37-39 Siyag, R.S., Rao, R.V.G.K., Agarwal, M.C., Singh, O.P and Oosterban, R.J 1995 Salinization of soil and yield of crops in relation to crop rotation and conjunctive use of canal water and saline water In: Reclamation and Management of Waterlogged Saline Soil (Ed R.S Siyag), pp 205-212 Department of Soil Science CCS Haryana Agricultural University, Hisar, India Smillie, R.M and Nott, R 1982 Salt tolerance in crop plants monitored by chlorophyll fluorescence in vivo Plant Physiol 70: 1049-1054 Smirnoff, N 2000 Ascorbic acid, metabolism and functions of a multifaceted molecule Curr Opin Plant Biol 3: 229-235 Song, J.D., Choi, K.H., Eu, Y.J., Rhew, T.H., Lee, D.H., and Lee, C.H 2001 Differential expression of ACC synthase and ACC oxidase genes in mung bean leaves under saline and oxidative stresses J Plant Biol 44: 118-126 Soomro, A., Mirjat, M.S., Oad, F.C., Soomro, H., Samo, M.A and Oad, N.L 2001 Effect of irrigation intervals on soil salinity and cotton yield Online J Biol Sci 1: 472-474 Stone, J.R and Yang, S 2006 Hydrogen peroxide: A signaling messenger Antioxit Redox Signal 8: 243-270 xi Sullivan, C.Y and Ross, W.M., 1979 Selecting for drought and heat resistance in grain shorgum In: Stress Physiology in Crop Plant (Eds H Mussel and R.C Staples), pp 263-281 R.C John Wiley and Sons, Ltd Publisher New York Sumithra, K., Jutur, P.P., Carmel, B.D., Reddy, A.R 2006 Salinity-induced changes in two cultivars of Vigna radiata: responses of antioxidative and proline metabolism Plant Growth Regul 50: 11-22 Suriyan, C.U., Chalermpol, K 2010 Effect of glycinebetaine on proline, water use, and photosynthetic efficiencies, and growth of rice seedlings under salt stress Turkish J Agric For 34: 517-527 Taiz, L and Zeiger, E 1998 Plant Physiology Chapter 4: Water balance of plants Sinauer Associates Inc, Massachusetts, Publisher Pp 47-65 Takahama, U and Oniki, T 1992 Regulation of peroxidase-dependent oxidation of phenolic in the apoplast of spinach leaves by ascorbate Plant Cell Physiol 33: 379-387 Talukdar, D 2011 Isolation and characterization of NaCl-tolerant mutations in two important legumes, Clitoria ternatea L and Lathyrus sativus L.: induced mutagenesis and selection by salt stress J Med Plant Res 5: 3619-3628 Uchida, A., Jagendorf, A.T., Hibino, T and Takabe, T 2002 Effect of hydrogen peroxide and nitric oxide on both salt and heat stress tolerance in rice Plant Sci 163: 515-523 Vaidyanathan, H., Sivakumar, P., Chakrobarty, R and Thomas, G 2003 Scavenging of reactive oxygen species in NaCl- stressed rice (Oryza sativa L.) differential responses in salt tolerant and salt sensitive varieties Plant Sci 165: 1411-1418 Venkateshan, A and Chellappan, K.P 1999 Salinity effect on the activities of certain antioxidant enzymes in Ipomoea pes-capraesweet, a halophyte Indian J Plant Physiol 4: 40-42 Verma, S and Mishra, S.N 2005 Putrescine alleviation of growth in salt stressed Brassica juncea by inducing anti-oxidative defence system J Plant Physiol 162: 669-677 Volkmar, K M., Hu, Y and Steppuhn, H 1998 Physiological responses of plants to salinity: a review Can J Plant Sci 78: 19-27 Wahid, A., Hameed, M and Rasul, E 2004 Salt-induced injury symptom, changes in nutrient and pigment composition, and yield characteristics of mung bean Int J Agric Biol 6: 11431152 Weatherley, P.E 1950 Studies on the water relations of the cotton plant I The field measurement of water deficit in leaves New Phytol 40: 81-97 Welburn, R.W 1994 The spectral determination of chlorophyll a and b as well as total carotenoids using varios solvents with spectrophotometer of different resolution J Plant Physiol 144: 307-313 Willekens, H., Montagu, M and Camp, W 1995 Catalase in plants Mol Breed 1: 207-228 xii Wilson, D.A and Reisenaur, H.M 1963 Cobalt requirements of symbiotically grown alfalfa Plant Soil 19: 364-373 Yasar, F., Ellialtioglu, S and Yildiz, K 2008 Effect of salt stress on antioxidant defense systems, lipid peroxidation, and chlorophyll content in green bean Russian J Plant Physiol 55: 782-786 Yasar, F., Uzal, O., Tufenkci, S and Yildiz, K 2006 Ion accumulation in different organs of green bean genotypes grown under salt stress Plant Soil Eviron 52: 476–480 Yemm, E.W and Willis, A.J 1954 The estimation of carbohydrates in plant extract by anthrone Biochem J 57: 508-514 Yeo, A 1999 Predicting the interaction between the effects of salinity and climate change on crop plants Scientia Hort 78: 159-174 Yousif, Y.H., Bingham, F.F and Yermanos, D.M 1972 Growth, mineral composition and seed oil of seasame (Sesamun indicum L.) as affected by NaCl Soil Sci Soc Am Proc 36: 450-453 Zaffar, M.D., Hemantaranjan, A and Sunil, K.P 2007 Antioxidatice response of mung bean (Vigna radiata L.) to salt stress Legume Res 30: 57-60 Zaidi, P.H and Singh, B.B 1995 Modulation of adverse effect of salinity by growth regulators in soybean photosynthetic area, pigment, efficiency and plant growth Plant Physiol Biochem 22: 135-142 Zayed, M.A and Zeid, I.M 1998 Effect of water and salt stresses on growth, chlorophyll, mineral ions and organic solutes contents, and enzymes activity in mung bean seedlings Biol Plant 40: 351-356 Zhu, J K 2001 Plant salt tolerance Trends Plant Sci 6: 66-71 Zhu, J.K Plant salt stress 2007 Handbook of Plant Science John Wiley and Sons Ltd, New York Publisher Pp 1304-1307 xiii ABSTRACT Title of thesis : Physiological responses to saline irrigation in two summer mungbean [Vigna radiata (L.) Wilczek] genotypes Name of the degree holder : Duong Hoang Son Title of the degree : Doctor of Philosophy Admission No : 2010BS100D Name and address of Major advisor : Dr Neeraj Kumar Scientist of Plant Physiology Department of Botany & Plant Physiology, College of Basic Sciences & Humanities CCS Haryana Agricultural University, Hisar-125004, Haryana, (India) Degree awarding University/Institute : CCS HAU, Hisar-125004 (Haryana) India Year of award of degree : 2013 Major subject : Plant Physiology Total number of pages in the thesis : 86 + xiii Number of words in the abstract : Approximate 590 Key words: Salinity tolerance; Mungbean; Growth; Physiology; Seed yield; SDS-PAGE The effects of saline irrigation on morpho-physiological traits, antioxidant defense mechanism and protein profile (SDS-PAGE) in two summer mungbean (Vigna radiata L.) genotypes viz MH 421 and SML 668 differing in their sensitivity to salinity was investigated under natural conditions of screen house At the flowering stage (30-35 DAS), plants were exposed to single saline irrigation (Cl-dominated) of levels 2.5, 5.0 and 7.5 dS m-1 Sampling was done after and 14 days of treatment (DAT) The control plants were irrigated with distilled water The water potential (Ψw) of leaf and osmotic potential (Ψs) of leaf and root decreases in both the genotypes MH 421 showed more negative values Ψw of leaf i.e from 0.20 to -0.46 MPa as compared to -0.18 to -0.40 MPa in SML 668, respectively The Ψs of leaf decreased from -1.36 to -1.89 MPa in MH 421 and -1.36 MPa to -1.80 MPa in SML 668 with increase in salinity level from control to 7.5 dS m-1 at DAT A similar trend of change in the osmotic potential (Ψs) of root was observed The proline content of leaf increased significantly from 11.4 to 144.5 % and 15.7 to 219.4 % and the total soluble carbohydrate (TSC) content from 16.6 to 79.8 % and 22.5 to 81.9 % in SML 668 and MH 421, respectively, with increasing level of saline irrigation from control to 7.5 dS m-1 A marked increase in hydrogen peroxide H2O2, lipid peroxidation (MDA content) and relative stress injury (RSI %) was noticed in leaf and root which was much higher in SML 668 than MH 421 A similar trend was observed at 14 DAT Saline irrigation increased the Cl- and SO42- concentration in leaf by 2.8 and 3.1 fold in MH 421 and 3.0 and 3.6 fold in SML 668, respectively, at 7.5 dS m-1 Moreover, MH 421 had overall lower Na+/K+ ratio than SML 668 More negative values of Ψw of leaf, Ψs of leaf and root and better accumulation of osmotically active solutes, i.e proline, TSC and low Na+/K+ ratio in leaf and root of MH 421, helped in maintaining the higher RWC % of these organs than noticed in SML 668 ROS scavenging enzyme of leaf i.e SOD, CAT, POD, APX, GR, GPX and GST increased by 296.7, 158.5, 146.3, 159.1, 125.8, 95.9, 81.0 % in SML 668 and by 806.8, 180.3, 231.4, 174.9, 190.2, 112.6, 172.6 % in MH 421, respectively, upon increasing levels of saline irrigation from control to 7.5 dS m-1 after days of treatment (DAT) Despite the increase in the activity of these enzymes AsA content decreased by 52.5 and 41.3% Higher activities of antioxidant enzymes, lower accumulation H2O2, MDA and AsA content in MH 421 than in SML 668 indicated those enzymes play a key role in removal of ROS in MH 421 better than in SML 668, thus minimizing the cellular damage caused by ROS under saline irrigation Pollen viability (%), in vitro pollen germination and tube growth were also adversely affected with increasing salinity; the effect being most pronounced in SML 668 SDS-PAGE revealed more deletion than addition of polypeptide bands in SML 668 than MH 421 with increasing levels of saline irrigation in leaf and root at both the stages The yield parameters like number of pods plant-1, number of seeds plant-1, 100 seed weight and seed weight plant-1 decreased more in SML 668 with increasing salinity Hence, the mechanism of salt tolerance was better in MH 421 than in SML 668 as found from physiological traits studied MAJOR ADVISOR HEAD OF THE DEPARTMENT SIGNATURE OF STUDENT CURRICULUM VITAE Name Date of birth Place of birth Mother’s name Father’s name Permanent address : : : : : : Telephone E-mail Academic qualifications : : Degree University/ Board B.Sc University of Natural Sciences, Vietnam M.Sc Ph.D CCS HAU, Hisar, India CCS HAU, Hisar, India DUONG HOANG SON June 21, 1976 HaNoi (VietNam) Smt Hoang Thi Kim Quy Sh Duong Thanh Tu 27/10 LeBinh Street, NinhKieu Distric, CanTho City - VietNam 00847103-839451 dhoangsonvn@yahoo.com Year of passing 2001 %age of marks/OGPA 61.2 2007 2013 71.2 71.2 Co-curricular activities : - Medals/Honours received : - Subjects Biology (Major subject: Plant Physiology) Plant Physiology Plant Physiology (submitted) List of publications: (i) Research Papers - Published: 1 Son, D.H., Kumar, N., Nandwal, A.S., Kumar, S and Sharma, S.K 2013 Comparative physiology of two summer mungbean genotypes to salt stress International Journal of Biotechnology and Bioengineering Research (6): 603-608 (ii) Research paper presented and published in proceedings of Conference/Seminar: Son, D.H., Kumar, N., Nandwal, A.S., Bhasker, P., Kumar, S and Sharma, S.K 2013 Plant water status, osmolyte accumulation, membrane integrity and ionic distribution in two summer Vigna radiata L cultivars under saline irrigation Summary published in National Conference of Plant Physiology on: “Current trends in plant biology research”, pp 494-495, organised at Directorate of Groundnut Research, Junagadh, Gujarat, India, December 12-16, 2013 Kumar, N., Son, D.H., Nandwal, A.S., Devi, S., Yadav, R and Sharma, S.K 2013 Effect of saline irrigation on antioxidant responses in two summer mungbean (Vigna radiata L.) cultivars Summary published in National Conference of Plant Physiology on: “Current trends in plant biology research”, pp 599-600, organised at Directorate of Groundnut Research, Junagadh, Gujarat, India, December 12-16, 2013 Son, D.H., Kumar, N., Nandwal, A.S and Yadav, R 2012 Physiological responses to saline irrigation in two summer mungbean [Vigna radiata (L.) Wilczek] genotypes Abstract in National Seminar of Plant Physiology on: “Physiological and molecular approaches for development of climate resilient crops”, pp 84-85, organised at Directorate of Sorghum Research, Rajender Nagar, Hyderabad, India, December 12-14, 2012 Son, D.H., Kumar, N., Nandwal, A.S., Devi, S., Kumar, S., Sharma, M.K., Sharma, S.K and Yadav, R 2014 Activities of antioxidant enzymes in two summer mungbean Vigna radiata (L.) cultivars in response to saline irrigation Abstract in National Seminar on: “Reorientation of Agricultural Research to Ensure National Food Security”, pp 32-33, organised at Chaudhary Charan Singh, Haryana Agricultural University, Hisar, India, January 6-7, 2014 UNDERTAKING OF COPYRIGHT I, Duong Hoang Son, Admission No 2010BS100D undertake that I give copyright to the CCS HAU, Hisar of my thesis entitled, “Physiological responses to saline irrigation in two summer mungbean [Vigna radiata (L.) Wilczek] genotypes” I also undertake that patent, if any, arising out of the research work conducted during the programme shall be filed by me only with due permission of the competent authority of CCS HAU, Hisar (Haryana) Signature of the Student [...]... marked in roots GST increased by 95% was reported at 10 dS m-1 salinity level in chickpea roots (Kukreja et al., 200 5) GST activity was reported to increase in salinity stress in mungbean (Hossain et al., 201 1) 2.6.6 Glutathione peroxidase (GPX) GPX is ubiquitously occurring enzymes in plant cells that involved in scavenging of H2O2 and sever to detoxify products of lipid peroxidation formed due to activity... thus important in providing protection against oxidative damage in plants by maintaining the reduced form of glutathione (Foyer et al., 199 1) GR activity has been reported to increase in B juncea seedlings with increase in salinity level (Verma and Mishra, 200 5) Similarly, Abbaspour (201 2) reported an increased GR activity with increase in NaCl concentration in Pistacia vera Salt tolerance in the leaves... 201 0) A rapid accumulation of proline under salt stress has been observed in mungbean crop (Singh et al., 199 4) Arora and Saradhi (200 2) studied Vigna radiata were exposed to different concentrations of NaCl in light and dark Proline accumulation in the shoots was higher in light than in dark, the increased in proline content upto 286% as compared to control in light under 200 mM NaCl Saffan (200 8) observed... in addition to reacting with H 2O2 may react 13 with superoxide anion (O- 2), singlet oxygen (1O 2) and hydroxyl radical (OH -) (Shigeoka et al., 200 2) Hernandez et al (199 9) reported rapid increased in APX activity in response to 90 mM and 110 mM NaCl concentrations in Pisum sativum leaves Sheokand et al (200 8) also reported an increase in APX activity under salt stress in chickpea Salinity induced increase... sustainable agricultural (Hussain et al., 200 8) Mungbean may also be sown as an inter crop or as a green manure or cover crop In order to overcome these problems, genotypes which are resistant to salinity are to be identify Selection and breeding programmed to increase salt tolerance will be more successful if selection is based directly on the physiological mechanism (s) or character (s) confirming tolerance... et al., 200 1) or to differentiate between salt tolerant and sensitive genotypes (Suriyan and Chalermpol, 201 0) The Fv/Fm ratio can be used to detect damage to photosymtem II and possible photo-inhibition (Ahmed et al., 200 2) Lee et al (200 4) observed in Paspalum vaginatum 9 Swartz ecotypes that with the increase of salinity level (1.1-49.7 dS m- 1) initial chlorophyll fluorescence (Fo) increased while... observed increasing with increasing saline irrigation concentration and days interval (Soomro et al., 200 1) 19 CHAPTER-III MATERIAL AND METHODS The present investigations were conducted on two released genotypes of mungbean [Vigna radiata (L. ) Wilczek] namely MH 421 and SML 668 The seeds were procured from the Pulses Section, Department of Genetic and Plant Breeding and the experiment was conducted in the... mutant genotypes Similarly, in tomato, it has been detected that GST activity increased in leaves and roots of salt tolerant but unchanged in control tomato cultivars under 100m M NaCl (Mittova et al., 2003 b) Misra and Gupta (200 6) studied wild type and mutant of Canthranthus roseus subjected to salinity A higher GST activity was observed in leaf pairs (apical, middle, and basal) and roots under saline. .. the proline content increased in all plants (wheat, barley, mungbean and kidney bean) under effect of 200 mM NaCl Similarly, proline content increased with increasing salt treatments in cowpea, black gram and green gram compared to control Accumulation of proline was more in root compared to shoot (Arulbalachandran et al., 200 9) Shabina and Mehar (201 1) subjected the seven varieties of mungbean to 50... al., 200 5) In contrast, Kukreja et al (200 6) observed two- fold increase in SOD activity in chickpea roots under shortterm salinization SOD activity increased under salt stress was observed in both leaves and roots of mungbean (Chakrabarti and Mukherji, 2003 a), mungbean seedling (Saha et al., 201 0) Manivannan et al (200 7) reported SOD activity was increased in leaves, stems and roots with increased