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Genetic Studies of Salinity Tolerance in Wheat

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Genetic Studies of Salinity Tolerance in Wheat Genetic Studies of Salinity Tolerance in Wheat By Nawar Jalal Shamaya B.Sc., M.Sc. A thesis submitted for the degree of Doctor of Philosophy School of Agriculture, Food and Wine Discipline of Plant Breeding and Genetics Australian Centre for Plant Functional Genomics March 2013 Table of Contents ABSTRACT ............................................................................. I DECLARATION ..................................................................... III ACKNOWLEDGEMENT ................................................... IV LIST OF FIGURES ............................................................. VI LIST OF TABLES ............................................................... XI CHAPTER 1: GENERAL INTRODUCTION .................... 1 1.1 INTRODUCTION ....................................................................... 1 1.2 LITERATURE REVIEW ............................................................. 2 1.2.1 Salinity ................................................................................................. 2 1.2.1.1 What is salinity? ................................................................................... 3 1.2.1.2 Phases of plant response and tolerance to salinity ............................ 4 1.2.1.2.1 First phase - osmotic effect ............................................................. 4 1.2.1.2.2 Second phase - ion-specific effect ................................................... 5 1.2.1.3 Key conclusions ..................................................................................... 6 1.2.2 Physiological traits associated with salinity tolerance in plants .... 7 1.2.2.1 Osmotic stress tolerance ....................................................................... 7 1.2.2.1.1 Growth rate ...................................................................................... 8 1.2.2.1.2 Stomatal conductance ..................................................................... 9 1.2.2.1.3 Measurement of osmotic stress tolerance ...................................... 9 + 1.2.2.2 Na exclusion ....................................................................................... 12 + 1.2.2.2.1 Na exclusion in durum wheat ..................................................... 14 + 1.2.2.3 Na tissue tolerance ............................................................................ 16 1.2.2.4 Plant tolerance to two phases of salt stress (osmotic and ionic stress) 17 1.2.3 Wheat ................................................................................................ 18 1.2.4 Detection of the genetic loci associated with salinity tolerance ... 20 1.2.4.1 QTL analysis ....................................................................................... 21 1.2.4.1.1 Mapping population ...................................................................... 21 1.2.4.1.2 Molecular markers ........................................................................ 26 1.2.4.1.3 Linkage map .................................................................................. 27 1.2.4.1.4 QTL analysis methods ................................................................... 27 1.2.4.2 Selective genotyping analysis (SGA) ................................................. 30 1.2.4.3 Bulked segregant analysis (BSA) ...................................................... 31 1.2.4.4 QTL for salinity tolerance traits ....................................................... 31 1.3 AIM ........................................................................................ 37 CHAPTER 2 IMPROVEMENT OF SCREENING TECHNIQUES FOR SALINITY TOLERANCE ............. 38 2.1 INTRODUCTION ..................................................................... 38 2.1.1 Gladius and Drysdale ...................................................................... 39 2.2 MATERIALS AND METHODS .................................................. 41 2.2.1 Plant Materials: ................................................................................ 41 2.2.2 Growth conditions ............................................................................ 41 2.2.2.1 Field capacity ...................................................................................... 42 2.2.3 Phenotypic characterisation ............................................................ 44 2.2.4 Experimental Design ........................................................................ 46 2.2.5 Statistical analysis ............................................................................ 47 2.3 RESULTS ................................................................................ 48 2.3.1 Shoot dry weight (SDW) .................................................................. 48 2.3.2 Salt tolerance index (STISDW) .......................................................... 49 th + 2.3.3 4 leaf Na accumulation ................................................................. 51 th + 2.3.4 4 leaf K accumulation .................................................................. 52 2.3.5 Plant height ....................................................................................... 54 2.3.6 A comparison of projected shoot area (PSA) with shoot fresh weight (SFW) and with shoot dry weight (SDW) ..................................... 55 2.4 DISCUSSION ........................................................................... 57 CHAPTER 3: ENHANCING SALINITY TOLERANCE IN ELITE BREAD WHEAT CULTIVARS ...................... 61 3.1 INTRODUCTION ..................................................................... 61 3.2 MATERIALS AND METHODS .................................................. 62 3.2.1 Plant materials ................................................................................. 62 3.2.2 Growth conditions ............................................................................ 63 3.2.2.1 Soil based experiment ......................................................................... 63 3.2.2.2 Hydroponics experiment .................................................................... 63 3.2.3 Experimental design ........................................................................ 65 3.2.3.1 Soil based experiment ......................................................................... 65 3.2.3.2 Hydroponics experiment .................................................................... 68 3.2.4 Linkage analysis ............................................................................... 70 3.2.5 Phenotypic characterisation ............................................................ 70 3.2.6 Statistical analysis ............................................................................ 72 3.2.7 QTL analysis ..................................................................................... 73 3.3 RESULTS ................................................................................ 74 3.3.1 Linkage analysis ............................................................................... 74 3.3.2 Soil based experiment ...................................................................... 76 3.3.2.1 Phenotyping ......................................................................................... 76 3.3.2.2 Trait correlation ................................................................................. 82 3.3.2.3 QTL analysis ....................................................................................... 84 3.3.3 Hydroponics experiment ................................................................. 86 3.3.3.1 Phenotyping ......................................................................................... 86 3.3.3.2 Trait correlation ................................................................................. 90 3.3.3.3 QTL analysis ....................................................................................... 93 3.3.4 The response of RILs to saline conditions varies between soil based and hydroponics experiments .......................................................... 99 3.4 DISCUSSION ......................................................................... 102 3.4.1 Soil based experiment .................................................................... 102 3.4.2 Hydroponics experiment ............................................................... 105 3.4.3 The response of RILs to saline conditions varies between soil based and hydroponics experiments ........................................................ 108 3.5 CONCLUSION ....................................................................... 110 CHAPTER 4 GENETIC BEHAVIOUR OF NA+ EXCLUSION IN AFGHANI LANDRACES OF DURUM WHEAT .............................................................................. 112 4.1 INTRODUCTION ................................................................... 112 4.2 MATERIALS AND METHODS ................................................ 114 4.2.1 Plant materials ............................................................................... 114 4.2.2 Crossing ........................................................................................... 114 4.2.3 Growth conditions .......................................................................... 115 4.2.4 Experimental design ...................................................................... 116 rd + rd + 4.2.5 Measurement of 3 leaf Na and 3 leaf K accumulation ....... 116 4.2.6 Genotyping characterising ............................................................ 116 4.2.6.1 DNA extraction ................................................................................
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