农杆菌介导的黄瓜遗传转化体系优化研究  

Optimization of the Genetic Transformation System of Cucumber Mediated by Agrobacterium tumefaciens

作　　者：陈梅 陈露倩 陈思[1] 杜长霞[1] 樊怀福[1] CHEN Mei;CHEN Luqian;CHEN Si;DU Changxia;FAN Huaifu(College of Horticulture Science,Zhejiang A&F University/Innovation Center of Agricultural Efficient and Green Collaborative Production of Zhejiang Province/Key Laboratory of Quality and Safety Control for Subtropical Fruit and Vegetable,Ministry of Agriculture and Rural Affairs,Hangzhou 311300,China)

机构地区：[1]浙江农林大学园艺科学学院/浙江省山区农业高效绿色生产协同创新中心/农业农村部亚热带果品蔬菜质量安全控制重点实验室,杭州311300

出　　处：《四川农业大学学报》2024年第3期540-545,571,共7页Journal of Sichuan Agricultural University

基　　金：浙江省基础公益研究计划(LGN22C150007);浙江省自然科学基金(LY23C150005);国家自然科学基金(32272700)。

摘　　要：【目的】筛选适用于农杆菌介导的黄瓜遗传转化体系,为通过生物技术进行黄瓜新品种培育奠定基础。【方法】以华北型黄瓜品种‘9930’子叶为试材,探讨卡那霉素浓度(kanamycin, Kan)、预培养时间、预培养基添加乙酰丁香酮(acetosyringone, AS)、分化培养基pH及生根培养基中添加不同外源物质对黄瓜遗传转化的影响。【结果】不定芽分化、生根阶段的Kan临界浓度为150 mg/L;不进行预培养的黄瓜不定芽诱导率最高,为77.17%;当分化培养基pH为5.6时不定芽诱导率最高,为75.26%;外植体在添加0.5 mg/L的IBA生根培养基上生根率达100%,根长也显著增加。【结论】在以黄瓜品种‘9930’子叶节为外植体的遗传转化体系中,150 mg/L Kan可作为不定芽分化和生根阶段的筛选选择压;不进行预培养更有利于其抗性芽诱导;适合不定芽诱导的分化培养基pH为5.6;0.5 mg/L的IBA为诱导外植体生根最佳激素浓度。【Objective】Screening of Agrobacterium-mediated genetic transformation system for cucumber and laying the foundation for the new variety cultivation through biotechnology.【Method】The cotyledon of the North China type cucumber cultivar′9930′was used as the experimental material.Various factors,including kanamycin concentration(Kan),pre-culture time,addition of acetosyringone(AS)to the pre-medium,pH value of the differentiation medium and addition ofdifferent exogenous substances to the rooting medium,were investigated for their effects on the genetic transformation of cucumber.【Re⁃sult】The threshold concentration of Kan at the stages of adventitious bud differentiation and rooting was 150 mg/L.The induction rate of cucumber adventitious buds without preculture treatment was the highest and reached its peak at 77.17%.When the pH value of the differentiation medium was 5.6,the highest in⁃duction rate of adventitious buds was obtained and was 75.26%.The rooting rate of explants on a rooting medium supplemented with 0.5 mg/L of IBA reached 100%,resulting in a significant increase in root length.【Conclusion】In the transgenic transformation system of cucumber cultivar′9930′,a concentra⁃tion of 150 mg/L of Kan can be used as a selection pressure during adventitious bud differentiation and rooting stages.The absence of pre-culture is more favorable for the induction of resistant buds.The suit⁃able pH value for adventitious bud differentiation medium is 5.6,and 0.5 mg/L IBA is the optimal hor⁃ mone concentration for inducing rooting of explants.

关 键 词：根癌农杆菌 黄瓜 遗传转化 

分 类 号：S642.2[农业科学—蔬菜学]