葡萄SCARECROW Like 14-Like基因的表达特征及胁迫响应研究

Expression pattern and stress response of grape SCARECROW Like 14-Like gene

为探索外源赤霉素(GA3)介导的无核葡萄果实膨大过程中的信号途径的调控基因,寻找验证主要调控元件,本研究以欧亚种无核葡萄品种‘无核白鸡心’(Vitis vinifera L.cv.Centennial Seedless)为材料,通过半定量RT-PCR技术分析属于GRAS基因家族的SCARECROW Like 14-Like(VvSCL14-Like)基因在葡萄不同组织器官和果实发育期的表达。通过启动子克隆、生物信息学分析、promoter∷GUS融合基因和GUS组织染色法对该基因的表达特征进行了研究。半定量RT-PCR结果表明,VvSCL14-Like基因主要在休眠芽中表达,在果实发育过程中无表达。利用GA3对葡萄幼果进行膨大处理,处理后1、3和7d的果实转录组结果表明VvSCL14-Like的转录水平无差异。对VvSCL14-Like启动子进行生物信息学分析,发现多个响应外源激素和逆境胁迫的作用元件。VvSCL14-Like启动子驱动的GUS基因,只在拟南芥萌发初期下胚轴处表达。以GA3和NaCl分别处理阳性拟南芥幼苗,48h后GUS基因均在叶柄和叶脉表达。干旱胁迫处理阳性拟南芥幼苗,15d后GUS基因只发现在根部表达。研究结果表明:VvSCL14-Like基因的表达具有组织特异性,参与外源赤霉素(GA3)和非生物胁迫响应过程,但不是外源赤霉素(GA3)介导的无核葡萄果实膨大过程的主要调控元件。

This paper was concerned on exploring the signal pathway mediated by exogenous gibberellins(GA3)in berry enlargement of seedless grape,and verifying the main regulatory elements.Centennial Seedless(Vitis vinifera L.) was used to detect the expression character of GRAS family gene SCARECROW Like 14-Like(VvSCL14-Like)in different tissues,organs and berry developmental stages by semi-quantitative RT-PCR and grape berry transcriptomic data analysis.The expression pattern of VvSCL14-Like was further studied by promoter cloning,bioinformatic analysis, promoter∷ GUS fusion gene and GUS staining.The semi-quantitative RT-PCR detected VvSCL14-Like expression in dormant buds.No expression in berries was found in three main developmental stages.Berry transcriptomic data revealed that exogenous GA3 application did not lead to significant difference expression on 1,3and 7day after treatment.Bioinformatics analysis showed that VvSCL14-Like promoter has multi-acting elements involving in responses to exogenous hormones and stress resistance.The GUS staining displayed GUS expressions in hypocotyls at the early stage of Arabidopsis seedling development.The GUS expression was found in petioles and leaf veins of transformed young Arabidopsis after 48hGA3 or NaCl treatments.The GUS expression was found in the root of transformed Arabidopsis after 15days stop irrigation.These results indicate that VvSCL14-Like gene is of tissue-specific expression pattern.It might be involved in response to exogenous gibberellins(GA3)and abiotic stress,but not act as a mainregulatory element in berry enlargement process of seedless grape mediated by exogenous gibberellins(GA3).