A convenient and high-efficiency Agrobacterium rhizogenes-mediated hairy root transformation for CRISPR/Cas9-based gene editing efficiency evaluation in different Citrullus species

ISHS Secretariat
A convenient and high-efficiency Agrobacterium rhizogenes-mediated hairy root transformation for CRISPR/Cas9-based gene editing efficiency evaluation in different Citrullus species

The CRISPR/Cas9 genome-editing system has been widely used in crop genetic improvement. To rapidly select efficient single guide RNAs (sgRNAs) in the genus watermelon, which is difficult to genetically transform, the Agrobacterium rhizogenes-mediated hairy root method is often used to detect efficient target sites in different Citrullus species. In this study, hairy roots were induced in different plant tissues of Citrullus lanatus, Citrullus mucosospermus and Citrullus amarus, in a non-sterile environment. Using this method, targeted mutations in ClCIPK17 could be detected in 90.9% of watermelon accessions. A total of 31 different mutation types in ClCIPK17 were recorded, where deletion of nucleotides was most common. Among all examined hairy roots, 73.9% contained mutations at the sgRNA1 site. Using sgRNA1 of ClCIPK17, stable transgenic watermelon plants were obtained from explants and targeted mutations in ClCIPK17 were confirmed. The results demonstrate that the hairy root transformation system can be used to evaluate the efficiency editing of sgRNA targets in watermelon. Yong Zhao is enrolled as a PhD candidate at Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences. He devotes himself to research on the natural variation and genetic basis of metabolites of watermelon.

Yong Zhao won the ISHS Young Minds Award for the best poster presentation at the VII International Symposium on Cucurbits in China in June 2023.

Yong Zhao, National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou Henan, 450009, China, e-mail: 1690539046@qq.com

The article is available in Chronica Horticulturae

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CRISPR/Cas9
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Young Minds Award Winners