首页|褐角苔遗传转化体系的建立

褐角苔遗传转化体系的建立

扫码查看
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pCAMBIA1305.2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100 μmol/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-qPCR)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.
Establishment of a Genetic Transformation System for Foliocer-os fuciformis
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 hygro-mycin as a screening marker,the pre-culture time of explants,the OD value and addition amount of the bacte-rial suspension,the concentration of acetosyringone(AS),and the co-culture time were optimized for the es-tablishment of the genetic transformation system.The results showed that the optimum concentration of hy-gromycin was 15 mg/L for the screening of resistant seedlings,and the state of the explants was the best af-ter 4 d of pre-culture.The optimal infection conditions were addition of 80 μL Agrobacterium suspension with OD600=0.8 in the co-culture medium,AS concentration of 100 μmol/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 horn-worts.

Folioceros fuciformisexplantsAgrobacterium-mediated methodgenetic transformation system

黄丹、彭涛、姜山、王顺莉、李旭强、马怀富、盛福源

展开 >

贵州师范大学生命科学学院,贵州贵阳 550025

贵州师范大学国际教育学院,贵州贵阳 550025

褐角苔 外植体 农杆菌介导法 遗传转化体系

2024

广西科学
广西科学院 广西壮族自治区科学技术协会

广西科学

CSTPCD北大核心
影响因子:0.516
ISSN:1005-9164
年,卷(期):2024.31(3)