Establishment of new convenient two-line system for hybrid production by targeting mutation of OPR3 in allopolyploid Brassica napus

Abstract

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.

Acknowledgements

This work was supported by the Agricultural Science and Technology Innovation Project (CAAS-ZDRW202105), the Sci-Tech Innovation 2030 Agenda (2022ZD04009), the National Key Research and Development Program of China (2022YFD1200804), Key Research Projects of Hubei Province (No. 2021EHB026 and 2022BBA0039), and the Fundamental Research Funds for Central Non-profit Scientific Institution (No. 1610172020001).

Author contributions

H.T.C. and Q.H. designed and conducted the study. M.Y.H. and S.F.S. performed most of the experiments; Y.F.W., Y.T.C., H.W., W.X.W., and D.S.M. provided the materials and performed some experiments. H.T.C. wrote the manuscript. Q.H. revised the manuscript. All of the authors read and approved the final manuscript.

Conflict of interest

The authors have declared that no competing interests exist in relation to this manuscript.

Data availability

All data supporting the findings of this study are available within the paper and within the supplementary data.

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