目的 人参Panax ginseng中构建烟草脆裂病毒（tobacco rattle virus，TRV）介导的基因沉默体系，为人参中基因功能的验证提供新方法。方法 通过序列比对得到人参中八氢番茄红素脱氢酶（phytoene desaturase，PDS）基因的完整序列，利用保守结构域预测工具得到核心片段，将其整合进pTRV2载体内。重组载体转化农杆菌后侵染人参叶片，观察表型并利用qPCR检测沉默效率。结果 通过序列比对得到了2条PDS基因。PgPDS1基因cDNA序列全长1 746 bp，编码581个氨基酸；PgPDS2基因cDNA序列全长1 299 bp，编码432个氨基酸。2个PgPDS蛋白均具有典型的PDS结构域。将重组载体通过农杆菌转化人参叶片，接种3周后叶片表型无变化，5周后叶片尖端以及边缘出现枯萎现象。经qPCR检测，接种后3周和5周人参内源PgPDS基因表达量分别下降至接种前的70.37%和37.35%，表明体系构建成功。结论 人参中成功构建了TRV介导的基因沉默体系。

Objective Constructing the tobacco rattle virus (TRV)-mediated gene silencing system in Panax ginseng provides a new method for the verification of gene function in ginseng. Methods The complete sequence of phytoene desaturase (PDS) gene was obtained by sequence alignment. Using the conserved domain prediction tool to obtain the core fragment of PDS gene, which was integrated into the pTRV2 vector. The recombinant vector transformed Agrobacterium to infect ginseng leaves, observing the phenotype after a period of time, and detecting the silencing efficiency by qPCR. Results A total of two PDS genes were obtained by sequence alignment. The cDNA sequence of PgPDS1 gene is 1 746bp in length, encoding 581 amino acids; the cDNA sequence of PgPDS2 gene is 1 299 bp in length, encoding 432 amino acids. Both of them have typical PDS conserved domains. The recombinant vector was transformed into ginseng leaf by Agrobacterium, the phenotype of leaves didn’t change after 3 weeks, but the tips and edges of the leaves were wilted after 5 weeks. The results of qPCR showed that the expression of PgPDS gene decreased to 70.37% (3 weeks later) and 37.35% (5 weeks later), respectively, indicating that the system was successfully constructed. Conclusion TRV-mediated gene silencing system was successfully constructed in P. ginseng.

国家重点研发计划（2021YFD1600900）；吉林省科技发展计划项目（20170309007YY）；吉林省自然科学基金项目（20170101011JC）