日本结缕草滞绿基因 ZjSGR 对烟草的转化及功能分析

doi:10.11686/cyxb2016492

草业学报 ›› 2017, Vol. 26 ›› Issue (5): 155-162.DOI: 10.11686/cyxb2016492

日本结缕草滞绿基因 ZjSGR 对烟草的转化及功能分析

檀鹏辉¹, 袁丽丽², 樊波², 于安东¹, 董笛¹, 滕珂^{1, *}, 晁跃辉^{1*, *}

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

收稿日期:2016-12-26 修回日期:2017-02-10 出版日期:2017-05-20 发布日期:2017-05-20
作者简介:檀鹏辉(1993-),女,河北邢台人,在读硕士。E-mail:penghuitan@bjfu.edu.cn
基金资助:
深圳市科技计划项目(JCYJ20160331151245672,JSGG20160229155434792)资助

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

TAN Peng-Hui¹, YUAN Li-Li², FAN Bo², YU An-Dong¹, DONG Di¹, TENG Ke^{1, *}, CHAO Yue-Hui^{1, *}

1.Institute of Turfgrass Science, Beijing Forestry University, Beijing 100083, China;
2.Shenzhen Guoyi Park Developments CO., LTD, Shenzhen 518038, China

Received:2016-12-26 Revised:2017-02-10 Online:2017-05-20 Published:2017-05-20

摘要/Abstract

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AAO3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: In green plants, SGR is a key gene with roles in chlorophyll degradation, plant development, senescence, and responses to abiotic stresses. In this study, the coding domain sequence of ZjSGR from Zoysia japonica 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 Agrobacterium tumefaciens-mediated transformation. The transgenic plants were confirmed to harbor the construct by PCR, and qRT-PCR analyses confirmed that ZjSGR 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 chloroplasts in stably transformed tobacco plants. Transmission electron microscope analyses revealed that the chloroplasts in transgenic plants showed retarded development and decomposition processes. Analyses of gene transcript levels by qRT-PCR showed that the transcript levels of SAG113, SAG12, NECD, and AAO3 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.

中图分类号:

檀鹏辉, 袁丽丽, 樊波, 于安东, 董笛, 滕珂, 晁跃辉. 日本结缕草滞绿基因 ZjSGR 对烟草的转化及功能分析[J]. 草业学报, 2017, 26(5): 155-162.

TAN Peng-Hui, YUAN Li-Li, FAN Bo, YU An-Dong, DONG Di, TENG Ke, CHAO Yue-Hui. Functional analysis of the stay-green gene ZjSGR from Zoysia japonica using transgenic Nicotiana tabacum[J]. Acta Prataculturae Sinica, 2017, 26(5): 155-162.

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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

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