黄瓜遗传转化体系优化的研究  

Study on Optimization of Agrobacterium-mediated Transformation System of Cucumber

作　　者：李蕾[1] 孟永娇[1] 张璐[1] 娄群峰[1] 李季[1] 钱春桃[1] 陈劲枫[1] 

机构地区：[1]南京农业大学园艺学院,作物遗传与种质创新国家重点实验室,江苏南京210095

出　　处：《华北农学报》2015年第5期115-121,共7页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金重点项目(314330075);国家重点基础研究发展计划“973”项目(2012CB113904);国家“863”项目(2012AA100202);农业部行业专项(201403032);国家自然基金青年基金项目(31301781)

摘　　要：为了提高农杆菌介导的黄瓜遗传转化效率,以亚洲生态型长春密刺、欧洲生态型Poinset76和Marketmore763种黄瓜为材料,筛选了适合遗传转化最佳的基因型,并通过改变培养基固化剂、添加抗氧化剂和有机添加物的方式对遗传转化体系进行优化。结果发现,3种基因型黄瓜中,长春密刺的抗性芽诱导分化表现最佳,而Poinset76和Marketmore76稍差;在脱乙酰吉兰糖胶为固化剂的培养基上,3种基因型的抗性芽诱导率比以琼脂粉为固化剂的培养基诱导率分别高28.21,19.71,15.39个百分点;共培养基中添加50 mg/L抗氧化剂α-硫辛酸时,长春密刺抗性芽诱导率最高达66.67%,平均芽分化数最高达1.32;在选择培养基中添加1.5 g/L的有机添加物水解酪蛋白,长春密刺抗性芽诱导率和平均芽分化数最高,分别为67.15%和1.42。此外,在前人建立的农杆菌介导的遗传转化体系的基础上,得到了以长春密刺为遗传转化的最适基因型,脱乙酰吉兰糖胶为培养基固化剂,共培养基中添加50 mg/Lα-硫辛酸或选择培养基中添加1.5 g/L水解酪蛋白的优化遗传转化体系,极显著提高了外植体抗性芽诱导率和平均芽分化数,推动了黄瓜转基因进程。For the purpose of improving the efficiency of Agrobacterium-mediated transformation of cucumber,the most suitable genotype was selected from Asian ecotype Changchunmici and European ecotype Poinset76 and Marketmore76. And then the Agrobacterium-mediated transformation system of cucumber was optimized with changing medium curing agent and adding antioxidant and organic additive into medium in different culture stage. The results indicated that the resistant buds differentiation of Changchunmici was the best of the three cucumber genotypes,Poinset76 and Marketmore76 were poorer. The resistant buds induction rates of three genotypes on medium gelled with gellan gum were increased by 28. 21,19. 71 and 15. 39 percentage points respectively compared with those on medium gelled with agar. Adding 50 mg / L α-lipoic acid into co-culture medium could make the resistant buds induction rates of Changchunmici up to 66. 67% and number of shoots per explants up to 1. 32. Similarly,when adding 1. 5 g / L casein hydrolysate into selection culture medium,the resistant buds induction rate and number of shoots per explants of Changchunmici were the highest,67. 15% and 1. 42 respectively. In conclusion,this article gained the optimization of transformation system that was using Changchunmici as appropriate genotype and gellan gam as medium curing agent,then adding 50 mg / L α-lipoic acid into co-culture medium or 1. 5 g / L casein hydrolysate into selection culture medium based on previous Agrobacterium-mediated transformation system. This optimizedtransformation system very significantly improved the resistant buds induction rate and number of shoots per explants of cucumber and promoted the process of cucumber transformation.

关 键 词：黄瓜 遗传转化 基因型 固化剂 抗氧化剂 有机添加物 

分 类 号：S642.03[农业科学—蔬菜学]