摘要
【目的】建立香椿中快速高效的遗传转化系统,为基因功能研究及分子育种提供技术支持。【方法】以香椿组培苗为材料,采用农杆菌介导真空渗透法进行瞬时转化植物叶片,并利用甜菜红素RUBY报告基因载体评估转化效率和目的基因表达水平,针对农杆菌菌株类型、侵染浓度、真空渗透时间等条件进行优化研究,并采用来自不同地区的5个香椿组培苗材料进行验证与评估。【结果】建立了适用于香椿的瞬时转化体系,筛选出了适用于香椿瞬时转化的农杆菌菌株为K599和GV3101菌株,通过比较转化效率获得的最适转化条件为150μmol/L乙酰丁香酮、菌液浓度OD_(600)为0.5,真空渗透30 min,共培养3-4 d。结果表明该转化体系具有高效的转化效率。【结论】成功建立了适合香椿的瞬时基因转化体系,基于RUBY报告基因表达情况优化了转化条件,并在不同香椿材料转化验证,为分子育种奠定基础。
【Objective】Establishing a fast and efficient genetic transformation system in Toona sinensis may provide technical support for gene function research and molecular breeding.【Method】This study utilized tissue-cultured seedlings of T.sinensis as the experimental materials,employed Agrobacterium-mediated vacuum infiltration,and adopted the red RUBY reporter gene to monitor transformation events and gene expression in the plants.Research was conducted to optimize conditions such as the type of Agrobacterium strain,infection concentration,and vacuum infiltration duration.Finally,validation and evaluation were conducted using five tissue-cultured seedlings from different provenances of T.sinensis.【Result】Simultaneously,a transient transformation system suitable for T.sinensis has been established.The Agrobacterium strains suitable for transient transformation in T.sinensis were screened out,namely K599 and GV3101 strains.The optimal transformation conditions obtained by comparing transformation efficiency were 150μmol/L acetosyringone,OD_(600) at 0.5,vacuum infiltration for 30 min,and co-cultivation for 3‒4 d.Ultimately,the results indicated that the transformation system has high transformation efficiency.【Conclusion】This system provides technical support for rapidly and effectively conducting functional identification of genes in T.sinensis.
作者
张瑶毅
殷恒福
刘军
韩小娇
范艳如
王民炎
曹受金
ZHANG Yao-yi;YIN Heng-fu;LIU Jun;HAN Xiao-jiao;FAN Yan-ru;WANG Min-yan;CAO Shou-jin(Central South University of Forestry and Technology,Changsha 410000;Research Institute of Subtropical Forestry,Chinese Academy of Forestry,Hangzhou 310000)
出处
《生物技术通报》
北大核心
2025年第11期221-227,共7页
Biotechnology Bulletin
基金
国家自然科学基金项目(32301637)。
关键词
香椿
农杆菌
瞬时转化
体系建立
体系优化
Toona sinensis
Agrobacterium
transient transformation
system establishment
system optimization
