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Background  As one of the main crops in the world, sterility of rice ( Oryza sativa  L.) significantly affects the production and leads to yield decrease. Our previous research showed that  OsPUB73 , which encodes U-box domain-containing protein 73, may be associated with male sterility. However, little information is available on this gene that is required for anther development. In the present study, we knocked out  OsPUB73  by using the CRISPR/Cas9 system and studied the cytological and transcriptome of the gene-defect associated with pollen development and sterility in the rice variety (Taichung 65).    Results  The sequence analysis indicated that  OsPUB73  was comprised of 3 exons and 2 introns, of which CDS encoded 586 amino acids including a U-box domain. The expression pattern of  OsPUB73  showed that it was highly expressed in the anther during meiosis stage. The  ospub73  displayed low pollen fertility (19.45%), which was significantly lower than wild type (WT) (85.37%). Cytological observation showed tapetum vacuolated at the meiosis stage and pollen exine was abnormal at the bi-cellular pollen stage of  ospub73 . RNA-seq analysis detected 2240 down and 571 up-regulated genes in anther of  ospub73  compared with WT during meiosis stage. Among of 2240 down-regulated genes, seven known genes were associated with tapetal cell death or pollen exine development, including  CYP703A3  ( Cytochrome P450 Hydroxylase703A3 ),  CYP704B2  ( Cytochrome P450 Hydroxylase704B2 ),  DPW  ( Defective Pollen Wall ),  PTC1  ( Persistant Tapetal Cell1 ),  UDT1  ( Undeveloped Tapetum1 ),  OsAP37  ( Aspartic protease37 ) and  OsABCG15  ( ATP binding cassette G15 ), which were validated by quantitative real-time polymerase chain reaction (qRT-PCR). These results suggested  OsPUB73  may play an important role in tapetal or pollen exine development and resulted in pollen partial sterility.    Conclusion  Our results revealed that  OsPUB73  plays an important role in rice male reproductive development, which provides valuable information about the molecular mechanisms of the U-box in rice male reproductive development.

Cytological and transcriptome analyses reveal OsPUB73 defect affects the gene expression associated with tapetum or pollen exine abnormality in rice