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Article|20 Jan 2022|OPEN
Targeted creation of new mutants with compact plant architecture using CRISPR/Cas9 genome editing by an optimized genetic transformation procedure in cucurbit plants
Tongxu Xin1 ,† , Haojie Tian1 ,† , Yalin Ma1,2 ,† , Shenhao Wang3 , Li Yang4 , Xutong Li1 , Mengzhuo Zhang1,2 , Chen Chen5,6 , Huaisong Wang1 and Haizhen Li5,6 , Jieting Xu7 , Sanwen Huang2 , Xueyong Yang,1 ,
1Key Laboratory of Biology and Genetic Improvement of Horticultural Crops of Ministry of Agriculture, Sino-Dutch Joint Lab of Horticultural Genomics, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
2Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518124, China
3College of Horticulture, Northwest A&F University, Yangling 712100, China
4Key Laboratory of Horticultural Plant Biology, Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, China
5Hunan Vegetable Research Institute, Hunan Academy of Agricultural Sciences, Changsha 410125, China
6Hunan Vegetable Research Institute, Hunan Academy of Agricultural Sciences, Changsha 410125, China
7Wimi Biotechnology (Jiangsu) Co., Ltd, Changzhou, 213000, China
*Corresponding author. E-mail: yangxueyong@caas.cn
Tongxu Xin,Haojie Tian and Yalin Ma contributed equally to the study.

Horticulture Research 9,
Article number: uhab086 (2022)
doi: https://doi.org/10.1093/hr/uhab086
Views: 23

Received: 21 Oct 2021
Accepted: 12 Dec 2021
Published online: 20 Jan 2022

Abstract

Fruits and vegetables in the Cucurbitaceae family, such as cucumber, melon, watermelon, and squash, contribute greatly to the human diet. The widespread use of genome editing technologies has greatly accelerated gene functional characterization and crop improvement. However, most economically important cucurbit plants, including melon and squash, remain recalcitrant to standard Agrobacterium tumefaciens-mediated transformation, limiting the effective use of genome editing technology. In this study, we used an “optimal infiltration intensity” strategy to establish an efficient genetic transformation system for melon and squash. We harnessed the power of this method to target homologs of the ERECTA family of receptor kinase genes and created alleles that resulted in a compact plant architecture with shorter internodes in melon, squash, and cucumber. The optimized transformation method presented here enables stable CRISPR/Cas9-mediated mutagenesis and provides a solid foundation for functional gene manipulation in cucurbit crops.