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Article|27 Oct 2023|OPEN
Establishment of new convenient two-line system for hybrid production by targeting mutation of OPR3 in allopolyploid Brassica napus 
Hongtao Cheng1 , Mengyu Hao1 , Shifei Sang2 , Yunfei Wen1 , Yating Cai1 , Hui Wang1 , Wenxiang Wang1 , Desheng Mei1 and Qiong Hu,1 ,
1Department of Rapeseed Genetics and Breeding, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences/Key Laboratory for Biological Sciences and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
2College of Life Sciences, Henan Normal University, No. 46 Jianshe East Road, Muye District, Xinxiang, Henan, 453007, China
*Corresponding author. E-mail:

Horticulture Research 11,
Article number: (2024)
Views: 48

Received: 17 Jun 2023
Accepted: 20 Oct 2023
Published online: 27 Oct 2023


The two-line pollination control system, which usually depends on the utilization of thermosensitive or photoperiod genic male-sterile lines, has been widely used in various crops. However, this system is susceptible to instability issues caused by uncontrollable weather fluctuations. A stable and handy two-line pollination control system is highly desirable in many crop species for heterosis exploitation. Oxophytodienoic acid reductase 3 (OPR3) was proven to be involved in jasmonate biosynthesis. In the present study, CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeat) was utilized to mutate two OPR3 homologs in Brassica napus. After two OPR3 homologs were simultaneously mutated, mutants exhibited complete male sterility, and fertility could be easily restored by exogenous MeJA treatment. Hybrids produced from crosses between the opr3 sterile lines and normal varieties exhibited heterosis. This new two-line system based on OPR3 mutation provides higher stability and convenience than traditional systems. By using exogenous MeJA treatment to restore fertility, the system enables more precise control of male fertility transition, which has great potential to significantly contribute to the maneuverable production of hybrid seeds in rapeseed as well as other Brassica species crops.