日本结缕草滞绿基因ZjSGR对烟草的转化及功能分析  

Functional analysis of the stay-green gene ZjSGR from Zoysia japonica using transgenic Nicotiana tabacum

作　　者：檀鹏辉 袁丽丽 樊波 于安东[1] 董笛 滕珂[1] 晁跃辉[1] 

机构地区：[1]北京林业大学草坪研究所,北京100083 [2]深圳市国艺园林建设有限公司,广东深圳518038

出　　处：《草业学报》2017年第5期155-162,共8页Acta Prataculturae Sinica

基　　金：深圳市科技计划项目(JCYJ20160331151245672;JSGG20160229155434792)资助

摘　　要：SGR基因是植物体内调控叶绿素降解的关键基因之一,在调控植物衰老以及响应非生物胁迫等方面也发挥着重要的作用。本研究利用DNA重组技术,构建35S::ZjSGR:YFP植物表达载体,并通过农杆菌介导的方法成功转化烟草。PCR和qRT-PCR结果显示日本结缕草滞绿基因ZjSGR已整合到烟草基因组中,并能够在转基因烟草中高效表达。在正常生长环境下,转基因烟草表现出黄化的表型。与对照相比叶绿素含量较低,光合速率下降。亚细胞定位实验证明ZjSGR定位在叶绿体,透射电镜观察结果表明过表达ZjSGR基因使叶绿体发育异常,且有加速降解的趋势。qRT-PCR结果发现转ZjSGR基因的烟草中SAG113,SAG12,NCED和AAO_3的表达量明显高于野生型,衰老进程明显加快。本研究证明ZjSGR可在调控叶绿素降解和衰老的进程中发挥重要作用。In green plants, SGR is a key gene with roles in chlorophyll degradation, plant development, senes-cence ,and responses to abiotic stresses. In this study, the coding domain sequence of ZjSGR from Zoysia ja -ponica was inserted into the 3302Y vector, yielding the plant expression vector 35S ：： ZjSGR ：YFP , by DNA recombination technology. The construct was introduced into tobacco plants by Agrobac terium tume fac iens- mediated transformation. The transgenic plants were confirmed to harbor the construct by PCR, and qRT - PCR analyses confirmed that Zj SGR was successfully heterologously expressed in the transgenic tobacco plants. The transgenic plants showed a rapid yellowing phenotype corresponding to a lower chlorophyll content and lower photosynthetic rate. Subcellular localization analyses showed that ZjSGR was localized in the chloro- plasts in stably transformed tobacco plants. Transmission electron microscope analyses revealed that the chlo- roplasts in transgenic plants showed retarded development and decomposition processes. Analyses of gene tran-script levels by qRT -PCR showed that the transcript levels of SAG113, SAG12, NECDy and AA03 were higher in ZjSGR-overexpressing tobacco plants than in control plants. The results of this study highlight the important roles of ZjSGR in controlling chlorophyll degradation and senescence in plants.

关 键 词：日本结缕草 滞绿基因 转基因烟草 叶绿素 衰老 

分 类 号：S688.4[农业科学—观赏园艺]