J Integr Plant Biol. ›› 2020, Vol. 62 ›› Issue (8): 1246-1263.DOI: 10.1111/jipb.12912

• Plant Reproduction Biology • Previous Articles    

Identification of late-stage pollen-specific promoters for construction of pollen-inactivation system in rice

Menglong Wang1†, Wei Yan1, 2†, Xiaoqun Peng1, 2†, Zhufeng Chen2, Chunjue Xu2, Jianxin Wu1, Xing Wang Deng2, 3* and Xiaoyan Tang1, 2*   

  1. 1Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou 510631, China
    2Shenzhen Institute of Molecular Crop Design, Shenzhen 518107, China
    3School of Life Sciences, Southern University of Science and Technology, Shenzhen 518055, China

    These authors contributed equally to this work.
    *Correspondence: Xing Wang Deng (deng@pku.edu.cn); Xiaoyan Tang (txy@frontier-ag.com, Dr. Tang is fully responsible for the distribution of all materials associated with this article)
  • Received:2019-10-10 Accepted:2020-01-16 Online:2020-01-22 Published:2020-08-01

Abstract:

Large‐scale production of male sterile seeds can be achieved by introducing a fertility‐restoration gene linked with a pollen‐killer gene into a recessive male sterile mutant. We attempted to construct this system in rice by using a late‐stage pollen‐specific (LSP ) promoter driving the expression of maize α‐amylase gene ZM‐AA1 . To obtain such promoters in rice, we conducted comparative RNA‐seq analysis of mature pollen with meiosis anther, and compared this with the transcriptomic data of various tissues in the Rice Expression Database, resulting in 269 candidate LSP genes. Initial test of nine LSP genes showed that only the most active OsLSP3 promoter could drive ZM‐AA1 to disrupt pollen. We then analyzed an additional 22 LSP genes and found 12 genes stronger than OsLSP3 in late‐stage anthers. The promoters of OsLSP5 and OsLSP6 showing higher expression than OsLSP3 at stages 11 and 12 could drive ZM‐AA1 to inactivate pollen, while the promoter of OsLSP4 showing higher expression at stage 12 only could not drive ZM‐AA1 to disrupt pollen, suggesting that strong promoter activity at stage 11 was critical for pollen inactivation. The strong pollen‐specific promoters identified in this study provided valuable tools for genetic engineering of rice male sterile system for hybrid rice production.

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