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。
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.
出处
《园艺学报》
CAS
CSCD
北大核心
2015年第8期1495-1504,共10页
Acta Horticulturae Sinica
基金
国家‘863’计划项目(2012AA100202)
中国农业科学院科技创新工程项目(CAAS-ASTIP-IVFCAAS)
科技部国际合作项目(2011DFR31180)
农业部园艺作物生物学与种质创制重点实验室项目
