【摘要】 为构建烟草脆裂病毒（tobacco rattle virus,TRV）介导的十字花科植物的病毒诱导的基因沉默（virusinduced gene silencing,VIGS）体系，以十字花科蔬菜上海青和芥菜为研究对象，以内源八氢番茄红素脱氢酶（phytoene desaturase,PDS）基因为标记基因，构建pTRV2-BrPDS、pTRV2-BjuPDS-g和pTRV2-BjuPDS-c载体，并进一步对农杆菌的侵染水平进行优化，构建上海青和芥菜的高效VIGS系统。结果显示，pTRV2-BrPDS、pTRV2-BjuPDS-g和pTRV2-BjuPDS-c载体构建成功。当农杆菌OD₆₀₀=1.0时，上海青植株白化表型最明显，当农杆菌OD₆₀₀=0.8时，芥菜的沉默效果最佳。通过qRT-PCR证实，白化表型是由TRV重组病毒侵染植株后，内源性PDS基因发生沉默而引起的。以上研究结果为利用VIGS技术研究十字花科植物的基因功能提供了理论依据。更多还原

【Abstract】 To establish an efficient virus-induced gene silencing（VIGS） system in important Brassicaceae vegetables, Brassica rapa subsp. chinensis and Brassica juncea were chosen as the material and the endogenous phytoene desaturase（PDS） gene was selected as the marker gene. The pTRV2-BrPDS, pTRV2-BjuPDS-g and pTRV2-BjuPDS-c vectors were constructed, and the infection efficiency of Agrobacterium was optimized to construst VIGS system with high efficient. The results showed that the pTRV2-BrPDS, pTRV2-BjuPDS-g and pTRV2-BjuPDSc vectors were successfully constructed. The albinism phenotype of B. rapa subsp. chinensis plants was the most obvious with OD₆₀₀=1.0 of Agrobacterium, while the silencing efficiency of B. juncea was optimal with OD₆₀₀=0.8 of Agrobacterium. The qRT-PCR analysis confirmed that the observed albinism phenotype was resulted from the silencing of the endogenous PDS gene in plant infected with the TRV-based recombinant virus. Above results provided theoretical basis for studing the gene functions of B. rapa subsp. chinensis and B. juncea using VIGS technology.更多还原