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A spontaneous thermo-sensitive female sterility mutation in rice enables fully mechanized hybrid breeding

Cell Research volume 32, pages 931–945 (2022)Cite this article

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Abstract

Male sterility enables hybrid crop breeding to increase yields and has been extensively studied. But thermo-sensitive female sterility, which is an ideal property that may enable full mechanization in hybrid rice breeding, has rarely been investigated due to the absence of such germplasm. Here we identify the spontaneous thermo-sensitive female sterility 1 (tfs1) mutation that confers complete sterility under regular/high temperature and partial fertility under low temperature as a point mutation in ARGONAUTE7 (AGO7). AGO7 associates with miR390 to form an RNA-Induced Silencing Complex (RISC), which triggers the biogenesis of small interfering RNAs (siRNAs) from TRANS-ACTING3 (TAS3) loci by recruiting SUPPRESSOR OF GENE SILENCING (SGS3) and RNA-DEPENDENT RNA POLYMERASE6 (RDR6) to TAS3 transcripts. These siRNAs are known as tasiR-ARFs as they act in trans to repress auxin response factor genes. The mutant TFS1 (mTFS1) protein is compromised in its ability to load the miR390/miR390* duplex and eject miR390* during RISC formation. Furthermore, tasiR-ARF levels are reduced in tfs1 due to the deficiency in RDR6 but not SGS3 recruitment by mTFS1 RISC under regular/high temperature, while low temperature partially restores mTFS1 function in RDR6 recruitment and tasiR-ARF biogenesis. A miR390 mutant also exhibits female sterility, suggesting that female fertility is controlled by the miR390-AGO7 module. Notably, the tfs1 allele introduced into various elite rice cultivars endows thermo-sensitive female sterility. Moreover, field trials confirm the utility of tfs1 as a restorer line in fully mechanized hybrid rice breeding.

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Fig. 1: Phenotypes of the tfs1 mutant and 4266.
Fig. 2: TFS1 is AGO7.
Fig. 3: Defective recruitment of RDR6 by mTFS1.
Fig. 4: The function of mTFS1 is partially recovered by low temperature.
Fig. 5: The utility of tfs1 as a restorer line in fully mechanized hybrid rice breeding.
Fig. 6: Field trials of hybrid seed production under regular/high temperature.
Fig. 7: A proposed model of the molecular defects of mTFS1, which confer thermo-sensitive female sterility in rice.

Data availability

All DNA sequencing and sRNA sequencing data from this study have been deposited in the National Center for Biotechnology Information (NCBI) with the accession number PRJNA827282. Source data for all graphs have been provided. All other data are available from the corresponding authors upon reasonable request.

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Acknowledgements

We thank Biogle GeneTech for providing a gene-editing vector and transgenic plants. This work was funded by the National Natural Science Foundation of China (NSFC31971924), Hong Kong Research Grant Council (GRF14122415), and JSPS KAKENHI (18H02380).

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Authors and Affiliations

  1. Department of Biology, Hong Kong Baptist University, Hong Kong, China

    Haoxuan Li, Jianhua Zhang & Guanqun Wang

  2. School of Life Sciences and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Hong Kong, China

    Haoxuan Li, Jianhua Zhang & Guanqun Wang

  3. Southern Regional Collaborative Innovation Center for Grain and Oil Crops in China, College of Agriculture, Hunan Agricultural University, Changsha, Hunan, China

    Haoxuan Li & Nenghui Ye

  4. Department of Botany and Plant Sciences, Institute of Integrative Genome Biology, University of California, Riverside, CA, USA

    Chenjiang You & Xuemei Chen

  5. Division of Plant and Microbial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 2-1-2 Kannondai Tsukuba, Ibaraki, Japan

    Manabu Yoshikawa

  6. Guangdong Provincial Key Laboratory for Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, Guangdong, China

    Xiaoyu Yang & Jie Cui

  7. College of Horticulture Science and Engineering, Shandong Agricultural University, Taian, Shandong, China

    Xiaoyu Yang

  8. The Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong, China

    Haiyong Gu & Chuanguo Li

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Contributions

J.Z. led and managed the project. J.Z., X.C., G.W., and N.Y. conceived the study. H.L. and G.W. performed the research. C.Y. analyzed the sequencing data. M.Y. performed the BYL in vitro experiments. H.G. performed the field planting. C.L. provided the mutant lines. J.C. instructed the RNA gel blotting. H.L., G.W., X.C., and X.Y. wrote the manuscript. All authors read and approved the manuscript.

Corresponding authors

Correspondence to Xuemei Chen, Nenghui Ye, Jianhua Zhang or Guanqun Wang.

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Li, H., You, C., Yoshikawa, M. et al. A spontaneous thermo-sensitive female sterility mutation in rice enables fully mechanized hybrid breeding. Cell Res 32, 931–945 (2022). https://doi.org/10.1038/s41422-022-00711-0

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