摘要
丝裂原活化蛋白激酶MAPK级联反应在植物响应生物与非生物胁迫中发挥着重要作用。为了获得转基因马铃薯,进一步探索StMKK1在信号转导和胁迫响应中的功能,从四倍体栽培品种马铃薯Desiree的cDNA中克隆到StMKK1基因302 bp的目的片段,并将目的基因的正向片段利用Eco RⅠ和KpnⅠ,反向片段利用HindⅢ和XbaⅠ与重组载体35s-pART27进行连接,构建成35s-StMKK1-pART27-RNAi表达载体;利用农杆菌介导的遗传转化方法,将35s-StMKK1-pART27-RNAi载体导入马铃薯品种Desiree中,得到7株沉默的阳性马铃薯转基因植株,不同的转基因植株中StMKK1基因的沉默效率经检测均在92%以上。
The mitogen-activated protein kinase(MAPK)cascades play important roles in plant response to biotic and abiotic stresses.The aim of this study is to develop StMKK1 knock-down transgenic potato lines for further investigating the functions of StMKK1 in signal transduction and stress response in potato.In this study,a 302 bp DNA fragment of StMKK1 gene was amplified from the cDNA of tetraploid potato cultivar Desiree,and the sense DNA fragment of StMKK1 was cloned into 35s-pART27 using restriction enzymes Eco RI and KpnⅠwhile the antisense DNA fragment of StMKK1 was ligated to 35s-pART27 by using restriction enzymes HindⅢand XbaⅠto generate a destination RNA interference(RNAi)vector,namely 35s-StMKK1-pART27-RNAi.The 35s-StMKK1-pART27-RNAi vector was transformed into potato cultivar Desiree by Agrobacterium-mediated gene transformation.A total 7 transgenic potato lines were obtained,and assessment of the silencing efficiency was more than 92%reduction in StMKK1 expression in most transgenic potato lines compared with wild-type potato cultivar Desiree.The results of this study will further contribute to understanding of the role of StMKK1 in signal transduction and stress response in potato.
作者
陈小康
李芳芳
王文斌
单卫星
杜羽
CHEN Xiaokang;LI Fangfang;WANG Wenbin;SHAN Weixing;DU Yu(College of Horticulture,Northwest A&F University,Yangling Shaanxi 712100,China;College of Agronomy,Northwest A&F University,Yangling Shaanxi 712100,China)
出处
《西北农业学报》
CAS
CSCD
北大核心
2020年第12期1822-1830,共9页
Acta Agriculturae Boreali-occidentalis Sinica
基金
陕西省博士后基金(2017BSHYDZZ63)
国家现代农业产业技术体系项目(CARS-09)。
