OsMYB80下游基因鉴定及其在调控花粉育性中的作用分析

Analysis of the OsMYB80 Downstream Genes in Regulating Pollen Fertility

    摘要
  • 摘要: OsMYB80是水稻花药和花粉发育过程中的重要转录因子，其下游靶标基因的相关研究仍有待补充。该研究从可能受OsMYB80直接调控的基因中筛选出24个可能参与信号转导、泛素化修饰、囊泡运输和未知功能的候选基因，利用qRT-PCR验证其表达模式，并通过CRISPR/Cas9基因编辑技术对其中7个基因(LOC_Os06g11930、LOC_Os09g26470、LOC_Os01g04409、LOC_Os06g35160、LOC_Os06g11135、LOC_Os05g13650、LOC_Os01g14880) 加以敲除。经I2-KI花粉染色分析和结实情况考察发现，上述7个基因敲除后对植株育性影响都不显著，说明上述7个基因并非OsMYB80下游调控花粉育性的关键基因。这些结果也暗示某些受OsMYB80调控的下游基因及其参与的生物化学反应可能在花药花粉发育过程中功能冗余或者只发挥微效作用。

     

    Abstract: OsMYB80 is an important transcription factor for anther and pollen development in rice. However, its downstream target genes need to be further studied. In this study, 24 candidate genes were selected from the genes that may be directly regulated by OsMYB80. The 24 candidate genes may be involved in signal transduction, ubiquitination modification, vesicle transport, and unknown function, and their expression patterns were verified by qRT-PCR. Seven of these genes (LOC_Os06g11930, LOC_Os09g26470, LOC_Os01g04409, LOC_Os06g35160, LOC_Os06g11135, LOC_Os05g13650, LOC_Os01g14880) were mutated using the CRISPR/Cas9 genome editing technology. I2-KI staining of pollen and seed setting analyses were performed on the knockout mutants. None of the seven gene knockout mutants were found to significantly affect plant fertility, indicating that these genes were not key for pollen fertility. These results also suggested that some of the OsMYB80-regulated genes might be functionally redundant or play a minor role in the anther and pollen development.

     

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