摘要
SGR(stay green regulation)基因是植物体内叶绿素代谢的关键调控基因,在调控植物衰老进程中发挥着重要的作用。本研究利用从日本结缕草(Zoysia japonica)中分离的ZjSGR基因,通过农杆菌介导的方法,完成对匍匐剪股颖(Agrostis stolonifera)的转基因操作。经过诱导、分化、生根、炼苗及土培过程,最终获得了19株再生匍匐剪股颖植株。PCR方法检测证实:其中15株再生匍匐剪股颖植物为转化阳性;RT-PCR分析表明:15株转基因匍匐剪股颖能够转录表达ZjSGR基因;利用荧光定量RT-PCR技术筛选ZjSGR基因高水平表达的转基因植株,结果显示:10号、13号、15号表达水平最高。选择这3株表达水平最高的转基因植株,进行生理生化分析,结果显示:转基因匍匐剪股颖叶绿素含量明显降低,净光合速率显著下降,可溶性糖及丙二醛含量与野生型植株相比有所提高。以上结果表明,ZjSGR基因在匍匐剪股颖中过量表达,能够通过促进叶绿素的降解加速植物的衰老。
SGR(stay green regulation)are key regulatory genes of chlorophyll metabolism,which play criticial roles in senescence process in plants.In this work,the ZjSGR gene isolated from Zoysia japonica was used to complete the transgenic creeping bentgrass(Agrostis stolonifera)by Agrobacterium-mediated method.A total of 19 regenerated creeping bentgrass plants were obtained after inducing callus,differentiation,rooting,seedling adaptation and planting in soil.By PCR technology,15 seedlings were confirmed as transgenic plants,with ZjSGR gene inserted into genome successfully.After analyzing with RT-PCR,the ZjSGR gene were proved to be transcribed into mRNA in those plants.The Real-Time Fluorescence Quantitative RT-PCR(qRT-PCR)was used to select transgenic plant with a high expression level of ZjSGR gene,and the result showed that line 10,13 and 15 had the top three expression levels of ZjSGR,which were selected for further experiments.The analyses of bio-physiology and chemistry in transgenic and WT plants showed that the chlorophyll contents and net photosynthetic rate were drastically decreased in three transgenic plants,while the contents of MDA and soluble sugar were obviously increased.Those results demonstrated that overexpression of ZjSGR gene in creeping bentgrass accelerated plant senescence by degradation of chlorophyll.
作者
周志湘
韩烈保
晁跃辉
张铁军
ZHOU Zhixiang;HAN Liebao;CHAO Yuehui;ZHANG Tiejun(College of Grassland Science,Beijing Forestry University,Beijing 100083,China)
出处
《草业科学》
CAS
CSCD
2020年第3期444-450,共7页
Pratacultural Science
基金
国家自然科学基金面上项目“紫花苜蓿产量性状配合力的分子遗传机理与预测模型分析”(31772656)。
作者简介
第一作者:周志湘(1981-),男,湖南安仁人,在读博士生,主要从事草坪草抗逆生理研究。E-mail:zhxzhou2008@aliyun.com;通信作者:张铁军(1981-),男,山东莱州人,副教授,博士,主要从事草地植物遗传育种研究。E-mail:zhangtiejun@bjfu.edu.cn。
