农杆菌介导的苜蓿遗传转化及PDH45转基因苜蓿耐盐性  

Agrobacterium-mediated Genetic Transformation of Alfalfa and Salt-tolerant of PDH45 Transgenic Alfalfa

作　　者：金太成[1] 刘雨晴[1] 许亚男 孟大伟[1] 周晓梅[1] 杨丽萍 Jin Taicheng;Liu Yuqing;Xu Yanan;Meng Dawei;Zhou Xiaomei;Yang Liping(School of Life Science, Jilin Normal University, Siping, 136000;Key Laboratory for Plant Resources Science and Green Production, Siping, 136000)

机构地区：[1]吉林师范大学生命科学学院,四平136000 [2]吉林省植物资源科学与绿色生产重点实验室,四平136000

出　　处：《分子植物育种》2018年第10期3190-3195,共6页Molecular Plant Breeding

基　　金：吉林省教育厅"十三五"科研规划项目(JJKH20180794KJ)资助

摘　　要：耐盐性基因的遗传转化能够提高植物的抗盐胁迫能力,有效解决盐胁迫抑制植物生长的问题。本研究中紫花苜蓿(Medicago sativa)被作为植物材料,利用农杆菌介导的叶盘转化法将PDH45基因转入紫花苜蓿中。影响遗传转化效率的各种条件被优化,包括农杆菌菌液浓度的优化、侵染时间的优化和选择培养基中卡那霉素浓度的优化。采用半定量(reverse transcription-PCR,RT-PCR)对PDH45转基因苜蓿进行了检测,结果表明转基因植株中PDH45基因均表达。在Na Cl浓度为50 mmol/L、100 mmol/L、150 mmol/L、200 mmol/L条件下,对PDH45转基因植株进行了耐盐性检测实验,结果表明中度盐胁迫150 mmol/L NaCl条件下,PDH45转基因植株表现出较强的耐受能力。本研究通过实验条件的优化,我们快速地获得了PDH45转基因苜蓿,为耐盐性转基因苜蓿的应用和推广提供了研究基础。The genetic transformation o f salt-tolerant genes can improve the ability of plants to resist salt stress and effectively solve the inhibition of salt stress to plants. In this research, Medicago sativa was used as the plant materials, and PDH45 gene was transformed into Medicago sativa by using the method of Agrobacterium-mediated leaf disc transformation. A variety of conditions influencing efficiency of genetic transformation were optimized,including Agrobacterium bacteria concentration, Agrobacterium infection time, kanamycin concentration of the selective medium. PDH45 transgenic alfalfa was tested by using the semi-quantitative RT-PCR, and the results showed that PDH45 genes were all expressed in the transgenic plants. Under the Na Cl concentration of 50 mmol/L,100 mmol/L, 150 mmol/L and 200 mmol/L, the study detected the salt-tolerant of PDH45 transgenic alfalfa, and the experimental results proved that PDH45 transgenic plants had strong tolerance to moderate salt stress of150 mmol/L Na Cl. This study rapidly obtained PDH45 transgenic alfalfa by optimizing experiment condition, and would provide research base for the application and spread of salt-tolerant transgenic alfalfa.

关 键 词：紫花苜蓿 叶盘转化法 PDH45 盐胁迫 

分 类 号：S541.9[农业科学—作物学]