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当前位置: 首页 > 文章 > 茄子SmMsrA基因VIGS表达载体的构建及表达分析 园艺学报 2015,42 (8) 1495-1504
Position: Home > Articles > VIGS Expression Vector Construction and Expression Analyses of SmMsrA Gene in Eggplant Acta Horticulturae Sinica 2015,42 (8) 1495-1504

茄子SmMsrA基因VIGS表达载体的构建及表达分析

作  者：
赵祯;刘富中;张映;齐东霞;陈钰辉;连勇
单  位：
中国农业科学院蔬菜花卉研究所
关键词：
茄子;SmMsrA基因;VIGS;Gateway技术;果实发育
摘  要：
利用RT-PCR从茄子单性结实品系D-10中获得甲硫氨酸亚砜还原酶A基因(Sm Msr A)的编码区。通过Gateway同源重组技术,构建了3个分别含Sm Msr A基因不同片段的烟草脆裂病毒(Tobacco rattle virus,TRV)介导的基因沉默(VIGS)表达载体p TRV2-Sm Msr Ai。表达载体转入农杆菌GV3101后,用注射压迫法侵染茄子叶片。采用表型观察、TRV病毒分子检测和q RT-PCR技术,评价构建的VIGS体系对Sm Msr A基因的沉默效果。结果表明,报告基因PDS沉默后叶片呈现明显的光漂白现象,Sm Msr A基因沉默后叶片呈花叶状,果实变小,叶片和果实中的Sm Msr A基因的表达量明显降低,表明Sm Msr A基因是正向调控茄子果实发育的相关基因。
译  名：
VIGS Expression Vector Construction and Expression Analyses of SmMsrA Gene in Eggplant
作  者：
ZHAO Zhen;LIU Fu-zhong;ZHANG Ying;QI Dong-xia;CHEN Yu-hui;LIAN Yong;Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences;
关键词：
eggplant;;SmMsrA gene;;VIGS;;Gateway technology;;fruit development
摘  要：
The Coding sequence of methionine sulfoXide reductase A(SmMsrA) was amplified from eggplant(Solanum melongena L.)parthenocarpic line D-10 using reverse transcription polymerase chain reaction(RT-PCR)method.Through the Gateway recombination technology,three TRV(Tobacco rattle virus)-mediated VIGS expression vectors were constructed,pTRV2-SmMsrAi,containing different fragments of SmMsrA gene.The VIGS expression vectors were transformed into Agrobacterium strain GV3101,and then inoculated eggplant leaves by Agro-Syringe.Phenotypes observation,TRV molecular detection and qRT-PCR thechnology were used to observe and evaluate the VIGS System for silencing the SmMsrA gene effection.The results showed that the positive control of silencing reporter gene PDS showed photo-bleaching phenotype in eggplant leaves;Silencing SmMsrA gene were observed pale yellow(mosaic)leaves and bore smaller fruits;The expression of SmMsrA gene was decreased obviously in the leaves and fruits of the infected eggplant plants.These results suggested that the SmMsrA gene was a related gene in positive regulation of fruit development in eggplant.

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