摘要
草莓离体组织再生和遗传转化体系的研究对草莓分子育种至关重要。为了优化农杆菌介导草莓品种红颜(Fragaria×ananassa Duch.Benihoppe)的遗传转化体系,本研究以红颜草莓组培苗的离体叶盘作为试验材料,首先确定了其离体再生的最佳条件,对影响转化效率的因素进行优化,获得了稳定高效的再生及转化体系。结果表明,红颜草莓离体叶盘最佳暗培养时间为9 d,最佳愈伤诱导培养基为MS+TDZ(2.0 mg/L)+IBA(0.1 mg/L)+2,4-D(0.1 mg/L),最佳愈伤分化培养基为MS+6-BA(0.5 mg/L)+NAA(0.1 mg/L),转化体系的最佳组合为预培养2 d、添加300μmol/L的AS、以OD600nm=0.6的EHA105型的农杆菌浸染5 min、共培养5 d。经优化后的红颜草莓叶盘再生及遗传转化体系对该品种的进一步优化和选育有重要意义。
The study of strawberry in vitro tissue regeneration and genetic transformation system is essential for strawberry variety improvement. In order to optimize the agrobacterium-mediated genetic transformation system of strawberry cultivar Benihope(Fragaria×ananassa Duch. Benihoppe),this research used the leaves as the experimental material. The optimal conditions for the leaf disc regeneration in vitro were determined,and then optimized the factors affecting the transformation efficiency,and obtained a stable and efficient transgene system. The results showed that the optimal dark culture time was 9 days for the leaves of strawberry cultivar Benihope,the best callus induction medium was MS+TDZ(2. 0 mg/L)+IBA(0. 1 mg/L)+2,4-D(0. 1 mg/L). The best callus differentiation medium was MS+6-BA(0. 5 mg/L)+NAA(0. 1 mg/L). The optimal combination of the transformation system was pre-culture for 2 days,addition of 300 μmol/L of AS,agrobacterium from EHA105 with OD600 nm=0. 6 for 5 min,and co-culture for 5 days.The optimized leaf disc regeneration and genetic transformation system in"Benihope"strawberry is of great significance for further optimization and breeding of this variety.
作者
王翠华
刘荇
杜小云
曾弓剑
谭雅慧
周超
刘开鸿
沈祥陵
WANG Cuihua;LIU Xing;DU Xiaoyun;ZENG Gongjian;TAN Yahui;ZHOU Chao;LIU Kaihong;SHEN Xiangling(Biotechnology Research Center,Three Gorges University,Yichang 443000,Hubei,China;Hubei Industry Farmer Ecological Agriculture Co.,Ltd,Yichang 443500,Hubei,China;College of Life Science,Wuhan University,Wuhan 430072,Hubei,China)
出处
《生物资源》
CAS
2020年第2期234-242,共9页
Biotic Resources
基金
三峡大学高层次人才启动基金(2016GCRC02)
三峡大学-勤劳农夫横向合作基金(2017421216000024)。
关键词
草莓
农杆菌
叶盘再生
遗传转化
strawberry
agrobacterium
leaf disc regeneration
genetic transformation
