VyDHN1转化‘西拉’愈伤组织遗传体系的建立及其抗逆性鉴定  

Establishment of the Genetic Transformation System Overexpressing VyDHN1 Gene into Callus from Vitis vinifera L.‘Syrah’and Measurement of the Resistance to Abiotic Stresses

作　　者：常贺强 刘牧青 韩国民 代玲敏 CHANG Heqiang;LIU Muqing;HAN Guomin;DAI Lingmin(School of Bioengineering,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250353,China)

机构地区：[1]齐鲁工业大学(山东省科学院)生物工程学院,山东济南250353

出　　处：《中外葡萄与葡萄酒》2025年第2期12-21,共10页Sino-Overseas Grapevine & Wine

基　　金：国家自然科学基金(31901982);齐鲁工业大学(山东省科学院)人才科研项目(2023RCKY227)。

摘　　要：本研究利用农杆菌介导的葡萄转基因方法,对菌液浓度、侵染时长和延迟培养3个条件进行优化,发现农杆菌菌液浓度OD_(600)=0.2、侵染时长10 min、延迟培养21 d时对‘西拉’愈伤组织的转化效率最高,并建立了‘西拉’愈伤组织高效遗传转化体系,获得VyDHN1融合GFP的过表达愈伤组织。正常情况下,转基因株系比野生型愈伤组织生长快,低温、干旱和盐等胁迫处理抑制愈伤组织的长势。低温、干旱和盐胁迫下转基因愈伤组织的细胞膜相对透性、丙二醛含量以及游离脯氨酸含量均低于对照,表明VyDHN1过表达能加快‘西拉’葡萄愈伤的生长速度,在低温、干旱和盐胁迫下保护了细胞膜结构并提高了植物的抗逆境能力。In this study,the Agrobacterium-mediated transgenic method into the callus of Vitis vinifera L.‘Syrah’was established.By optimizing the three basic conditions of transgenic process,namely the bacterial solution concentration,infection duration and delayed culture duration,it was shown that the transformation efficiency of‘Syrah’callus was the highest when the concentration of Agrobacterium solution was OD600=0.2,infection duration was 10 min and delayed culture was 21 d.The‘Syrah’callus overexpressing VyDHN1 fused with GFP was obtained in this study.It was displayed that transgenic lines grew faster than wild-type callus under control condition,however the callus growth was inhibited by low temperature,drought and salt stress.The relative permeability of cell membrane,malondialdehyde content and free proline content of transgenic callus under low temperature,drought and salt stress were lower than those of WT,suggesting that overexpression of VyDHN1 could accelerate the growth rate of callus of‘Syrah’,and could protect the cell membrane structure to improve plant resistance to these abiotic stress.

关 键 词：西拉 愈伤组织 遗传转化 脱水素基因 非生物胁迫抗逆性 

分 类 号：S663.1[农业科学—果树学]