褐角苔遗传转化体系的建立  

Establishment of a Genetic Transformation System for Folioceros fuciformis

作　　者：黄丹 彭涛[1] 姜山 王顺莉 李旭强 马怀富 盛福源 HUANG Dan;PENG Tao;JIANG Shan;WANG Shunli;LI Xuqiang;MA Huaifu;SHENG Fuyuan(School of Life Sciences,Guizhou Normal University,Guiyang,Guizhou,550025,China;School of International Education,Guizhou Normal University,Guiyang,Guizhou,550025,China)

机构地区：[1]贵州师范大学生命科学学院,贵州贵阳550025 [2]贵州师范大学国际教育学院,贵州贵阳550025

出　　处：《广西科学》2024年第3期469-477,共9页Guangxi Sciences

基　　金：国家自然科学基金项目(32060611,31560508)资助。

摘　　要：本研究旨在筛选适用于农杆菌(Agrobacterium)介导的角苔植物遗传转化体系,为今后研究早期陆生植物的进化提供理论基础。实验使用携带p CAMBIA1305.2质粒的农杆菌AGL1菌株对褐角苔(Folioceros fuciformis)外植体进行侵染,将潮霉素作为筛选标记,通过比较外植体预培养时间、菌液OD值、菌液添加量、乙酰丁香酮(AS)浓度和共培养时间来构建该物种的遗传转化体系。结果表明,褐角苔抗性植株筛选所用潮霉素的最适浓度为15 mg/L,预培养4 d的外植体侵染后状态最好。最佳侵染条件为共培养基中添加80μL OD600为0.8的农杆菌菌液,AS浓度为100μmol/L,共培养时间为3 d。通过潮霉素筛选、PCR、实时荧光定量PCR(RT-q PCR)和β-葡萄糖苷酸酶(GUS)染色鉴定获得转基因植株褐角苔。研究结果可为后续角苔植物基因功能的研究提供技术支持。The selection of Agrobacterium mediated genetic transformation system for hornworts provides a theoretical basis for the future study of early terrestrial plant evolution.In this study,Agrobacterium AGL1 carrying the plasmid pCAMBIA1305.2 was used to infect the explants of Folioceros fuciformis.With hygromycin as a screening marker,the pre culture time of explants,the OD value and addition amount of the bacterial suspension,the concentration of acetosyringone(AS),and the co culture time were optimized for the establishment of the genetic transformation system.The results showed that the optimum concentration of hygromycin was 15 mg/L for the screening of resistant seedlings,and the state of the explants was the best after 4 d of pre culture.The optimal infection conditions were addition of 80μL Agrobacterium suspension with OD 600=0.8 in the co culture medium,AS concentration of 100μmol/L,and co culture time of 3 d.The transgenic plants were identified by hygromycin screening,PCR,RT qPCR and beta glucuronidase(GUS)staining.The findings provide technical support for the subsequent research on the gene functions of hornworts.

关 键 词：褐角苔 外植体 农杆菌介导法 遗传转化体系 

分 类 号：Q37[生物学—遗传学]