Abstract：
CONCLUSIONS: We have developed and optimized a rapid, versatile Agrobacterium-mediated transient expression system for cannabis seedlings that can be used in functional genomics studies of both hemp-type and drug-type cannabis. Cannabis (Cannabis sativa L.) holds great promise in the medical and food industries due to its diverse chemical composition, including specialized cannabinoids. However, the study of key genes involved in various biological processes, including secondary metabolite biosynthesis, has been hampered by the lack of efficient in vivo functional analysis methods. Here, we present a novel, short-cycle, high-efficiency transformation method for cannabis seedlings using Agrobacterium tumefaciens. We used the RUBY reporter system to monitor transformation results without the need for chemical treatments or specialized equipment. Four strains of A. tumefaciens (GV3101, EHA105, LBA4404, and AGL1) were evaluated for transformation efficiency, with LBA4404 and AGL1 showing superior performance. The versatility of the system was further demonstrated by successful transformation with GFP and GUS reporter genes. In addition, syringe infiltration was explored as an alternative to vacuum infiltration, offering simplicity and efficiency for high-throughput applications. Our method allows rapid and efficient in vivo transformation of cannabis seedlings, facilitating large-scale protein expression and high-throughput characterization studies.
摘要：
结论：我们开发并优化了一种快速，用于大麻幼苗的多功能农杆菌介导的瞬时表达系统，可用于大麻型和药物型大麻的功能基因组学研究。大麻（CannabissativaL.）由于其多样化的化学成分，在医疗和食品工业中具有广阔的前景，包括专门的大麻素。然而,研究涉及各种生物过程的关键基因，包括次级代谢产物的生物合成，由于缺乏有效的体内功能分析方法而受到阻碍。这里,我们提出了一部小说，短周期,利用根癌农杆菌高效转化大麻幼苗的方法。我们使用RUBY报告系统来监测转化结果，而不需要化学处理或专用设备。对4株根癌农杆菌（GV3101、EHA105、LBA4404和AGL1）的转化效率进行了评价，LBA4404和AGL1表现出卓越的性能。通过用GFP和GUS报告基因的成功转化进一步证明了系统的多功能性。此外,研究了注射器渗透作为真空渗透的替代方法，为高通量应用提供简单性和效率。我们的方法可以快速有效地在体内转化大麻幼苗，促进大规模蛋白质表达和高通量表征研究。
