Optimization of CGMMV-VIGS System in Cucurbits

doi:10.11924/j.issn.1000-6850.casb2024-0328

Abstract

Abstract:

This study aims to improve the efficiency of cucumber green mottle mosaic virus (CGMMV)-mediated virus-induced gene silencing (VIGS) in cucurbit crops. Using germplasms from cucumber, melon and watermelon, effectiveness in gene silencing was tested by using various inoculation methods with CGMMV-VIGS vector, cultivation temperatures, and relative humidity in incubator. The results demonstrated that vacuum infiltration and seed imbibition were the most effective inoculation methods, achieving the highest frequency of gene silencing (FGS) in cucurbit crops. Cucumber, melon, and watermelon showed high effectiveness of PDS silencing EGSL (100%) at the temperatures of 22℃ and 25℃ after one month of growth, significantly higher than the EGSL at 30℃. After three months, the EGSL for melon and watermelon were 89.7% and 95%, respectively at 22℃ and 85.6% and 86.1% at 25℃, both significantly higher than that at 30℃. With 30% and 50% relative humidity in incubator, the EGSL for cucumber were 70% and 72%, respectively, which were significantly higher than the EGSL at 80% humidity. For watermelon, the EGSL at 30% relative humidity was 69%, significantly higher than the EGSL at 50% humidity (38%) and 80% humidity (33%). Overall, by optimizing the inoculation methods and environmental parameters in the CGMMV-VIGS, we have improved the gene silencing rate and effectiveness in cucurbit crops. This method will facilitate rapid acquisition of VIGS germplasm resources for research about crop quality and abiotic stress resistance.

Key words: cucumber green mottle mosaic virus (CGMMV-VIGS), gene silencing, PDS silencing EGSL, cucurbits, seed inoculation, temperature, relative humidity

WAN Lili, WANG Zhuanrong, TANG Mi, ZHANG Xuejun, REN Jian, ZENG Hongxia, ZHANG Na, WEI Jiaqi, XIONG Jianshun. Optimization of CGMMV-VIGS System in Cucurbits[J]. Chinese Agricultural Science Bulletin, 2024, 40(30): 40-47.

Figures/Tables 5

References 35

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