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ALA促进不结球白菜耐盐机理的研究
作 者: Saba Ambreen Memon
导 师: 侯喜林
学 校: 南京农业大学
专 业: 蔬菜学
关键词: 不结球白菜 5-ALA 盐胁迫 矿质含量 抗氧化酶 光合特性 S44基因
分类号: S634.3
类 型: 博士论文
年 份: 2008年
下 载: 33次
引 用: 0次
阅 读: 论文下载
内容摘要
不结球白菜(Brassica campestris ssp.chinensis var. communis Tsen et Lee)又称小白菜、青菜、油菜,原产中国,为十字花科芸薹属芸薹种白菜亚种中的一个变种,以其嫩叶为产品器官的一、二年生蔬菜作物。由于其适应性强、生长周期短、营养价值高,是我国南方的大众化蔬菜,在“菜篮子”工程中占有重要地位,成为衡量市场供应优劣的主要标志。近年来东南亚、日本、美国及欧洲各国也均有引种栽培,不结球白菜日益成为一种世界性的蔬菜。世界上盐碱地面积不断扩大,而有关不结球白菜耐盐性及其机理方面却鲜有研究,本文研究了盐胁迫下ALA对不结球白菜种子萌发、矿质元素含量、光合特性、抗氧化酶活性及耐盐相关基因的影响。1.不结球白菜种子萌发及幼苗期耐盐性的研究试验发现随着盐胁迫的增强,种子的萌发率、根的长度、根、子叶及胚轴的干、鲜重和呼吸强度均显著下降。在50 mmol 1-1 NaCl低盐胁迫下"Za-1", "Su-1", "Ak-1"及"Ha-1"四个品种的种子萌发率均高于对照,其中"Ai-1”表现的呼吸强度最高、根长最长,“Ak-1”表现出最高的下胚轴鲜重(16.8 mg/p),而"Ba-1”则表现出最大的子叶鲜重(25 mg/P)2.盐胁迫下ALA对不结球白菜种子萌发的促进效应盐胁迫下种子的萌发率及幼苗的生长随着ALA浓度的增加而显著增加,在200mmol 1"1 NaCl盐胁迫下添加5Omg1-1 ALA, "Lb-1"和“Ai-1”表现出最高的种子萌发率,而"Qi-1”则表现出最高的下胚轴干、鲜重(25mg/p,1.6mg/P)及子叶干、鲜重(30.4mg/P,2.9mg/P)。在添加:20 mg I-1 ALA’情况下,"Lb-1", "Za-1"及“Su-1”的根长最长。在添加20 mmol I-1 ALA抑制剂LA溶液后,幼苗生长受到显著抑制。ALA可能是通过色素物质影响种子萌发及幼苗生长,而后者在线粒体呼吸途径中又具有重要的作用。3.盐胁迫下ALA对不结球白菜矿质含量的影响盐胁迫下,不结球白菜叶片Mg2+、Ca2+和K+的含量下降,而Nz+和Cl-的含量增加。进一步经ALA处理后,叶片Mg2+、Ca2+、K+的含量增加,Na+和Cl-的含量降低。但在"Qd-1”中Na+含量没有降低,而在“Li-1”和“Ai-1”两品种中Na+明显降低。处理后8天“Ai-1”的Ca2+含量最高(36.08 mg/g);处理后24天,“Li-1”中Mg2+和Cl-含量最高,分别达到4.07 mg/g和7.0 mg/g; "Qd-1"中Na+(47.16 mg/g)和"Ai-1"中K+(43.34 mg/g)的含量在处理24天后达到最高。4.盐胁迫下ALA对不结球白菜色素量、抗氧化酶活性及光合特性的影响试验表明,ALA提高了不结球白菜的光合速率及色素含量,15Ommo11-1 NaCl胁迫下添加100 mg 1-1ALA后,“Qd-1”在1000μmol m-2 s-1光照强度下表现出最大的光合速率15.41μmol m-2 s-1,色素含量也达到最大值,叶绿素a、b及总量分别为1.08 mg g-1FW、0.59 mg/g FW及1.67 mg/g FW,叶绿素a/b为0.54。盐胁迫下耐盐品种“Ai-1”,表现出最大的SOD酶活性(72Ug-1min-1)和POD酶活性(0.08Ug-1min-1)。而"Li-1”中CAT酶活性在50mg 1-1和150mg 1-1盐胁下增加,而添加100mg 1-1ALA后,CAT酶活性分别下降180%和155%。5.盐胁迫下ALA对不结球白菜色素荧光特性的影响盐胁迫下研究了耐盐"Aijiaohuang"、中耐盐’Qingdi"及盐敏感"Lichuandasuo mian"品种的色素荧光特性,结果表明盐胁迫下添加ALA后,中耐盐品种“Qingdi”表现出最高的色素光吸收效率,电子吸收及传递率也有所增加。盐胁迫影响到光合PSⅡ的稳定性,而ALA则显著增加了光合器官的稳定性。6.不结球白菜S44基因的诱导表达特性S44基因有助于增强植株的耐盐性,采用同源克隆的方法,获得了294 bp的S44基因。通过Real-time PCR技术对S44基因的表达研究发现,ALA促进S44基因的表达,其中根的表达量高于叶片中的表达。7.ALA对不结球白菜色素含量、抗氧化酶活性及光合特性的影响非盐胁迫下测定了不同浓度的ALA对不结球白菜色素含量、抗氧化酶活性及光合特性的影响,结果表明250 mg l-1 ALA处理后“Qd-1”叶绿素a含量最高(1.96 mg/g FW),50mgl-1 ALA处理后“Za-1”叶绿素b含量及色素总量最高,分别达到1.74 mg/g FW和2.32 mg/g FW,150 mg I-1 ALA处理后“Ai-1”有最高的Chla比值250 mg-1 ALA处理后,“Zh-1”表现出最高的POD活性1.23 Ug-1.min-1, "Zy-1"有最高的CAT活性0.93Ug-1.min-1,"Qi-1"有最强的SOD活性(287Ug-1.min-1)。同样处理下"Ba-1”具有最高的净光合速率Pn (14.78 (μmol m-2 s-1),而“Qi-1”具有最高的胞内CO2浓度(Ci),50mgl-1 ALA处理“Zh-1"气孔导度((Gs)最高(0.599μmol m-2 s-1)。250mg I-1 ALA也促使“Za-1”达到最大的蒸腾速率11.2μmol m-2 s-1。
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全文目录
ABSTRACT 11-15 摘要 15-18 ABBREVIATIONS 18-19 GENERAL INTRODUCTION 19-42 Origins and Distribution 19-20 Plant Habit 20 Flowers 20-21 Commercial Production 21 Climatic and Soil Requirements 21-22 Seeding and Spacing 22 Cultural Practices 22 Fertilizer Requirements 22-23 Nutrients for Vegetables 23 Importance and Uses 23-26 Uses 23-24 Nutritional Value 24-25 Medicinal Value 25-26 Factors Affecting Pak choi production 26-27 Abiotic Factors 26 Biotic Factors 26-27 Environmental Fluctuation 27-29 Plant Responses to Salinity 29-33 Morphological Effects 30 Physiological Effects 30-32 Biochemical Effects 32 Effects of Salinity on the Enzyme Activities 32-33 5.Aminolevulinic Acid Application in Plant Growth 33-35 Aims of Thesis 35-36 Literatures Cited 36-42 Chapter 1 Salt Tolerance of Pak choi at Germination and Seedling Growth 42-55 Abstract 42 Introduction 42-43 1 Materials and Methods 43-45 1.1 Plant Materials 43 1.2 Seed Germination 43-44 1.3 Seed Respiration 44 1.4 Data Collection and Statistical Analysis 44-45 2 Results 45-51 2.1 Different NaCl Treatments on the Seed Germination of Pak choi 45-51 2.2 NaCl Treatment on the Seed Respiration 51 3 Discussion 51-53 4. Conclusion 53 Literature Cited 53-55 Chapter 2 Promotion by 5-ALA of Seed Germination of Pak choi under Salt Stress 55-85 Abstract 55 Introduction 55-56 1. Materials and Methods 56-59 1.1 Plant Material 56-57 1.2 Seed Germination 57 1.3 Determination of ALA 57-58 1.4 Method of Protoheme 58 1.5 Reagents 58 1.6 Determination of Protoheme 58-59 1.7 Data Collection and Statistical Analysis 59 2. Regults 59-81 2.1 Effect of ALA on Seed Germination under Salt Stress 59-67 2.2 Inhibition of Levulinic Acid(LA)on ALA-Enhanced Seed Germination 67-76 2.3 Effect of ALA on the Heme Content of Pak choi under Salt Stress 76-79 2.4 Determine the Endogenous ALA under Salt Stress 79 2.5 Effect of ALA on Respiration of Pak choi under Salt Condition 79-81 3 Discussion 81-82 4. Conclusion 82-83 Literature Cited 83-85 Chapter 3 Effect of ALA on the Mineral Element Content in Pak choi under Salt Stress 85-95 Abstract 85 Introduction 85-86 1. Materials and Methods 86-88 1.1 Seed Material 86 1.2 Preparation of Plant Materials 86-87 1.3 Preparation of Hoagland Solution 87 1.4 Determination of Elements 87 1.5 Data Collection and Statistical Analysis 87-88 2. Results 88-91 2.1 Effect of ALA on Mineral Element Content of "Qingdi" under Salt Stress 88-89 2.2 Effect of ALA on Mineral Element Content of "Aijiaohuang" under Salt Stress 89-90 2.3 Effect of ALA on Mineral Element Content of "Lichuandasuomian" under Salt Stress 90-91 3. Disussion 91-92 4. Conclusion 92 Literature Cited 92-95 Chapter 4 Role of ALA on Antioxidative Enzymes,Chlorophyll Content and Photosynthesis ofPak choi under Salt Stress 95-113 Abstract 95 Introduction 95-97 1. Materials and Methods 97-98 1.1 Seed Materials 97 1.2 Preparation of Plant Material 97 1.3 Determination of Chlorophyll 97 1.4 Measurement of Antioxidative Enzymes 97-98 1.5 Measurement of Photosynthesis Parameter 98 1.6 Data Collection and Statistical Analysis 98 2 Results 98-104 2.1 Effect of ALA on Antioxidative Enzyme under Salt Stress 98-100 2.2 Effect of ALA on Chlorophyll Content under S alt Stress 100-101 2.3 Effect of ALA Treatment on Photosynthesis under Salt Stress of Pak choi Leaves 101-104 3 Discussion 104-107 4.Conclusion 107-108 Literature cited 108-113 Chapter 5 Changes of Chlorophyll Fluorescence Parameters in Pak choi by ALA under Salt Stress 113-128 Abstract 113 Introduction 113-114 1. Materials and Methods 114-115 1.1 Seed Materials 114 1.2 Preparation of plant material 114-115 1.3 Measurement of Chlorophyll Fluorescence in Modulated Light 115 1.4 Statistical Analysis 115 2 Results 115-123 3. Discussion 123-124 4. Conclusion 124-125 Literature cited 125-128 Chapter 6 Expression of S44 Gene Induced by ALA in Pak choi under Salt Stress 128-141 Abstract 128 Introduction 128-130 1 Materials and Methods 130-131 1.1 Plant Materials and Nucleic Acid Extraction 130 1.2 Primer Designing 130 1.3 Cloning and DNA Sequencing 130 1.4 Construction of cDNA and Sequence Analysis 130-131 1.5 Real-time PCR analyses 131 1.6 Real-time PCR product sequence 131 2 Results 131-135 2.1 RT-PCR analysis under salt stress 131-133 2.2 Gene expression of S44 gene by ALA under Salt Stress 133-135 3 Discussion 135-137 4.Conclusion 137-138 Literature Cited 138-141 Chapter 7 Promotive Effect of ALA on Chlorophyll,Anti-oxidative Enzyme and Photosynthesison Pak choi under hot stress conditions 141-203 Abstract 141 Introduction 141-143 1. Materials and methods 143-145 1.1 Seed Materials 143 1.2 Preparation of Plant Materials 143-144 1.3 Determination of Chlorophyll 144 1.4 Determination of Antioxidative Enzymes 144 1.5 Measurement of Photosynthetic Parameter 144-145 1.6 Data Collection and Statistical Analysis 145 2 Results 145-193 2.1 Effect of ALA Treatment on Activity of POD of Pak choi Leaves 145-153 2.2 Effect of ALA Treatment on Activity of CAT of Pak choi Leaves 153-161 2.3 Effect of ALA Treatment on Activity of SOD of Pak choileaves 161-168 2.4 Effect of ALA Treatment on Chlorophyll Content of Pak choi 168-178 2.5 Effect of ALA Treatment on Photosynthesis of Pak choi 178-193 3. Discussion 193-197 4. Conclusion 197 Literature Cited 197-203 Summary and Conclusion 203-205 Innovation 205-207 Publications 207-209 Acknowledgements 209
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中图分类: > 农业科学 > 园艺 > 蔬菜园艺 > 白菜类 > 白菜(小白菜)
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