Comparative expression profiling of miRNAs between the cytoplasmic male sterile line MeixiangA and its maintainer line MeixiangB during rice anther development.

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CONCLUSION: miRNAs are involved in the pollen development during the CMS occurrence in rice. miRNAs are 20-24 nt endogenously expressed small RNAs that play key roles in the regulation of many growth and developmental processes in plants. The knowledge on cytoplasmic male sterility (CMS) regulation by miRNAs in rice is rather limited. In this study, Illumina sequencing was employed to examine the expression profiles of rice anther miRNAs from the CMS line MeixiangA (MxA) and its maintainer line MeixiangB (MxB). A total of 518 known miRNAs and 144 novel miRNAs were identified during rice anther development. Based on the number of sequencing reads, a total of 24 miRNAs were discovered to be differentially expressed between MxA and MxB, and the results were partially validated by qRT-PCR. Among these, 16 miRNAs were decreased and 8 miRNAs were increased in MxA compared with MxB. Target prediction showed that they target genes encoding EF-hand family proteins, F-box domain-containing proteins, MYB transcription factors, PPR-containing proteins and transposons. The expression patterns for targets of osa-miR528, osa-miR5793, osa-miR1432, osa-miR159, osa-miR812d, osa-miR2118c, osa-miR172d and osa-miR5498 were selectively examined, and the results showed that there was a negative correlation on the expression patterns between miRNAs and their targets. These targets have previously been reported to be related with pollen development and male sterility, suggesting that miRNAs might act as regulators of CMS occurrence in rice anthers. Furthermore, miRNA editing events were observed. The U → C and U → A editing phenomenon was validated by molecular cloning and sequencing. These findings contribute to our understanding of the roles of miRNAs during anther development and CMS occurrence in rice.