影响西瓜农杆菌介导的高效遗传转化效率的主要因子.PDF

See discussions, stats, and author profiles for this publication at: /publication/255778851 Establishment of efficient transgenic system mediated by Agrobacterium tumefaciens in watermelon Article · March 2 7 CITATIONS READS 31 1 author: Jian Yong Li Boyce Thompson Institute, USDA, Cornell University 11 PUBLICATIONS 209 CITATIONS SEE PROFILE All content following this page was uploaded by Jian Yong Li on 3 September 2 17. The user has requested enhancement of the downloaded file. 浙江农业学报 ActaAgricuhuraeZhejiangensis19(3)：197～201，2007 影响西瓜农杆菌介导的高效遗传转化效率的主要因子 李建勇 ，丁淑丽 ，孙利祥2，卢 钢 ， (1浙江大学 园艺系，浙江 杭州 310029；杭州市种子技术推广站，浙江 杭州 310021) 摘 要：以西瓜子叶为外植体材料，利用 GUS染色瞬时表达方法研究农杆菌浓度，预培养、共培养时间，预培 养条件以及添加乙酰丁酸酮(As)对西瓜转化效率的影响，优化西瓜遗传转化参数 ，初步建立以Ms+BAP2．0 m L+ 0．2mg／L+Hyg10mg／L+cef300mg／L为分化培养基，黑暗条件下预培养2d，使用OD为0．3的菌 液侵染 10min后，黑暗条件下共培养3d，然后在25 ，光周期为 16h的条件下筛选抗性芽的遗传转化体系。 关键词：西瓜；遗传转化；农杆菌；GUS分析 中图分类号：$651 文献标识码：A 文章编号：1004—1524(2007)03—0197—05 EstablishmentofefficienttransgenicsystemmediatedbyAgrobacteriumtumefaciens inwatermelon LIJian—yong，DINGShu—li，SUNLi—xinag2，LUGang (DepartmentofHorticulture，ZhejiangUniversity，Hangzhou310029，China；HangzhouStationofSeedTechniquePopular— ization，Hangzhou310020，China) A~tract：AnefficientwatermeloncotyledonexplanttransgeniesystemmediatedbyAgrobacteriumtumefacienswasestab— lishedthroughtheexplorationofthefactorsaffectingtransformationefficiencyofAgrobacteriumtumefaciens．Theparameters wereoptimizedbymeasuringhteefficiencyofhistochemiealGUStranssientexpressionincludingpreeuhureperiod，co—culti- vationperiod，additionofASandhtecompositionof regenerationmedia．Th ewatermeloncot)ledonexplantswerepre-eul— turedfor2dinhtedarkness，hteninoculatedwith0．3ofOD~ooAgrobacteriumtume