‘红颜’草莓短缩茎再生及遗传转化体系建立  

Establishment of regeneration and genetic transformation system of‘Benihoppe’strawberry shortened stem

作　　者：甘珂珂 刘宪夺 贾燕 沈元月 黄芸 GAN Keke;LIU Xianduo;JIA Yan;SHEN Yuanyue;HUANG Yun(College of Plant Science and Technology/Beijing Key Laboratory of New Technology in Agricultural Application,Beijing University of Agriculture,Beijing 102206,China;Beijing Jiaming Weiye Technology Development Co.,LTD.,Beijing 102206,China)

机构地区：[1]北京农学院植物科学技术学院/农业应用新技术北京市重点实验室,北京102206 [2]北京佳铭伟业科技发展有限公司,北京102206

出　　处：《北京农学院学报》2024年第3期11-15,共5页Journal of Beijing University of Agriculture

基　　金：北京农学院科技创新“火花行动”支持计划。

摘　　要：【目的】为了获得‘红颜’草莓再生及遗传转化植株。【方法】以八倍体‘红颜’草莓(Fragaria×ananassa Duch.‘Benihoppe’)组培苗短缩茎为试验材料,将草莓长日照开花抑制基因FaTFL1和FaSOC1的靶点构建至CRISPR/Cas9基因编辑载体pDIRECT-22C中。通过建立‘红颜’草莓短缩茎再生及遗传转化体系获得FaTFL1和FaSOC1基因编辑植株。【结果】‘红颜’草莓短缩茎不经过愈伤诱导,可直接诱导形成不定芽,整个再生周期2个月左右。通过优化培养基条件,建立了农杆菌介导的短缩茎遗传转化体系。通过扩增CRISPR/Cas9载体中的Cas9基因对遗传转化获得的草莓植株进行鉴定,阳性率18.62%。【结论】成功建立了一种以八倍体‘红颜’草莓组培苗短缩茎为外植体的高效再生体系以及农杆菌介导的稳定遗传转化体系。[Objective]In order to obtain regenerated and genetically transformed plants of the‘Benihoppe’strawberry.[Methods]The shortened stems of octoploid strawberry(Fragaria×ananassa Duch.‘Benihoppe’)were used as experimental materials.The targeted sequences of FaTFL1 and FaSOC1 known to suppress flowering process of‘Benihoppe’strawberries under long-day conditions,were incorporated into the CRISPR/Cas9 vector pDIRECT-22C.FaTFL1 and FaSOC1 gene edited plants were obtained by establishing shortened stem regeneration and genetic transformation system of‘Benihoppe’strawberry.[Results]The shortened stem is directly induced to form adventitious buds without callus generated,and the whole regeneration cycle is about two months.By optimizing the medium conditions,a genetic transformation system mediated by Agrobacterium tumefaciens was established.The Cas9 elements of the CRISPR/Cas9 vector of genetically transformed strawberry plant was identified,resulting in a positivity rate of 18.62%.[Conclusion]A highly efficient regeneration system utilizing the shortened stem bases of octoploid‘Benihoppe’strawberry tissue culture seedlings as experimental materials,along with an Agrobacterium-mediated genetic transformation system,has been established.

关 键 词：‘红颜’草莓 短缩茎 再生 遗传转化 

分 类 号：S668.03[农业科学—果树学]