Identification of late-stage pollen-specific promoters for construction of pollen-inactivation system in rice.,Journal of Integrative Plant Biology

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Identification of late-stage pollen-specific promoters for construction of pollen-inactivation system in rice.
Journal of Integrative Plant Biology ( IF 9.3 ) Pub Date : 2020-02-20 , DOI: 10.1111/jipb.12912
Menglong Wang ₁ , Wei Yan _{1,

2} , Xiaoqun Peng _{1,

2} , Zhufeng Chen ₂ , Chunjue Xu ₂ , Jianxin Wu ₁ , Xing Wang Deng _{2,

3} , Xiaoyan Tang _{1,

2}

Affiliation

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.

中文翻译：

鉴定后期花粉特异性启动子以构建水稻花粉灭活系统。

雄性不育种子的大规模生产可通过将与花粉杀伤基因连接的育性恢复基因引入隐性雄性不育突变体中来实现。我们尝试通过使用后期花粉特异性（LSP）启动子驱动玉米α-淀粉酶基因ZM-AA1的表达在水稻中构建该系统。为了在水稻中获得此类启动子，我们对具有减数分裂花药的成熟花粉进行了RNA序列比较分析，并将其与水稻表达数据库中各个组织的转录组数据进行比较，得到269个候选LSP基因。对9个LSP基因的初步测试表明，只有最活跃的OsLSP3启动子才能驱动ZM‐AA1破坏花粉。然后，我们分析了另外22个LSP基因，发现在后期花药中比OsLSP3强12个基因。在第11和第12阶段表达高于OsLSP3的OsLSP5和OsLSP6的启动子可以驱动ZM‐AA1使花粉失活，而在第12阶段显示更高表达的OsLSP4的启动子仅不能驱动ZM‐AA1破坏花粉，表明强第11阶段的启动子活性对于花粉灭活至关重要。在这项研究中确定的强花粉特异性启动子为杂交水稻生产的雄性不育系的基因工程提供了有价值的工具。

更新日期：2020-02-20

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