A real-time visualized TRSV-based gene silencing method using trichome as a selected marker in cucumber

Area of Science:

• Plant molecular biology
• Genetics
• Agricultural science

Background:

• Cucumber (Cucumis sativus L.) is a globally important vegetable crop.
• While the cucumber genome is sequenced, many gene functions remain uncharacterized.
• Virus-induced gene silencing (VIGS) is effective for gene function analysis, but traditional detection methods are time-consuming.

Purpose of the Study:

• To develop an efficient and visually detectable VIGS system for cucumber.
• To establish a real-time VIGS method for cucumber gene function studies.
• To utilize trichome development as a visual marker for VIGS efficiency.

Main Methods:

• Development of a vacuum-agroinfiltrated Tobacco ringspot virus (TRSV)-based VIGS system in cucumber.
• Cloning of CsGLABROUS3 (CsGL3) into the pTRSV2 vector to create the TRSV-CsGL3 system.
• Validation of the system using CsGL3 (trichome initiation) and CsUFO (flower morphogenesis) genes.

Main Results:

• The TRSV-CsGL3 system enabled visible gene silencing in cucumbers, with glabrous phenotypes observed throughout the plant's life cycle.
• Silencing of the CsUFO gene resulted in expected flower development defects.
• Trichome phenotype served as an effective real-time visual marker for VIGS efficiency.

Conclusions:

• The TRSV-CsGL3 system provides a simplified and real-time visualized VIGS method for cucumber.
• This system facilitates efficient gene function investigation throughout the cucumber life cycle.
• The use of trichomes as a selected marker streamlines VIGS identification procedures.