日本结缕草滞绿基因ZjSGR对烟草的转化及功能分析

发布时间：2018-06-01 11:02

本文选题：日本结缕草 + 滞绿基因　；参考：《草业学报》2017年05期

【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。
[Abstract]:SGR gene is one of the key genes regulating chlorophyll degradation in plants. It also plays an important role in regulating plant senescence and responding to abiotic stress. In this study, DNA recombinant technology was used to construct 35S::ZjSGR:YFP plant expression vector, and the results of Agrobacterium tumefaciens mediated transformation of tobacco. PCR and qRT-PCR showed that ZjSGR gene of Zoysia japonica was integrated into tobacco genome. And can be expressed in transgenic tobacco. Transgenic tobacco showed a yellow phenotype in normal growing environment. Compared with the control, the chlorophyll content was lower and the photosynthetic rate decreased. Subcellular localization experiment showed that ZjSGR was located in chloroplast. Transmission electron microscopy showed that overexpression of ZjSGR gene resulted in abnormal chloroplast development. The results of QRT-PCR showed that the expression levels of SAG113, SAG12NCED and AAO_3 in tobacco transgenic with ZjSGR gene were significantly higher than those in wild type, and the senescence process was accelerated obviously. This study demonstrated that ZjSGR may play an important role in the regulation of chlorophyll degradation and senescence.
【作者单位】：北京林业大学草坪研究所;深圳市国艺园林建设有限公司;
【基金】：深圳市科技计划项目(JCYJ20160331151245672,JSGG20160229155434792)资助
【分类号】：S688.4

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