‘脐红’猕猴桃组培再生和遗传转化体系建立与优化  

Optimization of Regeneration Process in the Genetic Transformation System of‘Qihong’Actinidia chinensis

作　　者：徐长彬 李林 方志蒙 缪星茹 祝小水 雷长生 孙学鹏 胡校粒 XU Changbin;LI Lin;FANG Zhimeng;MIAO Xingru;ZHU Xiaoshui;LEI Changsheng;SUN Xuepeng;HU Xiaoli(College of Horticultural Science,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;Suichang Xiaoshui Family Farm,Suichang 323306,Zhejiang,China;Suichang Beijie People’s Government Agricultural Service Center,Suichang 323306,Zhejiang,China)

机构地区：[1]浙江农林大学园艺科学学院,浙江杭州311300 [2]遂昌县小水家庭农场,浙江遂昌323306 [3]遂昌县北界人民政府农业服务中心,浙江遂昌323306

出　　处：《浙江林业科技》2025年第2期64-70,共7页Journal of Zhejiang Forestry Science and Technology

基　　金：浙江省自然科学基金资助项目(LQ23C150003)。

摘　　要：以‘脐红’猕猴桃Actinidia chinensis‘Qihong’无菌苗的叶片为外植体,探究不同浓度植物生长调节剂组合对猕猴桃愈伤组织诱导及再生的影响,筛选出‘脐红’猕猴桃叶片离体再生体系的最佳配方,并利用根癌农杆菌介导的叶盘法将AcALKBH4超表达载体转入‘脐红’猕猴桃,通过实时荧光定量PCR鉴定出阳性材料,成功通过该体系快速获得转基因植株。结果表明:培养基配方为MS(4.43 g·L^(−1))+蔗糖(30 g·L^(−1))+6-BA(2.0 mg·L^(−1))+NAA(0.5 mg·L^(−1))+TDZ(1.0 mg·L^(−1))+琼脂(8 g·L^(−1))最有利于‘脐红’猕猴桃叶片愈伤组织的诱导和生长,诱导率达85.2%;培养基配方为MS(4.43 g·L^(−1))+蔗糖(30 g·L^(−1))+6-BA(1.0 mg·L^(−1))+NAA(0.1 mg·L^(−1))+琼脂(8 g·L^(−1))时,培养5~6周后,‘脐红’猕猴桃愈伤组织再生出芽率最高可达60.0%。继续培养新生芽,长出叶片后进行转基因阳性植株检测,阳性率为25.0%~30.0%。The effects of different concentrations of plant growth regulator combinations on the callus induction and regeneration of kiwifruit were investigated by using the leaves of aseptic seedlings of Actinidia chinensis‘Qihong’kiwifruit as explant,and the best formula of in vitro regeneration system of‘Qihong’kiwifruit leaves was screened out.AcALKBH4 overexpression vector was transferred into‘Qi hong’kiwifruit by leaf disk method mediated by Agrobacterium tumefaciens,and positive transgenic plants were identified by real-time fluorescence quantitative PCR.We can successfully obtained transgenic plants through this system in a timely manner,which laid a solid foundation for the efficient transformation of kiwifruit.The results showed that adding MS(4.43 mg·L^(−1))+sucrose(30 g·L^(−1))+6-BA(2.0 mg·L^(−1))+NAA(0.5 mg·L^(−1))+TDZ(1.0 mg·L^(−1))+agar(8 g·L^(-1))was the most beneficial to the callus induction and growth of kiwifruit leaves,and the induction rate was 85.2%.The best condition for‘Qi hong’kiwifruit callus in inducing buds was achieved when the medium formulation was MS(4.43 g·L^(−1))+sucrose(30 g·L^(−1))+6-BA(1.0 mg·L^(−1))+NAA(0.1 mg·L^(−1))+agar(8 g·L^(−1))after around 5 to 6 weeks culture.The regeneration rate of callus was up to 60.0%.The new buds were cultured and the positive transgenic plants were detected after the leaves grew out.The positive rate was about 25.0%-30.0%.

关 键 词：猕猴桃 愈伤组织 芽诱导 再生 遗传转化 

分 类 号：S663.4[农业科学—果树学]