Cloning and identification of novel miRNAs in the flower organs of Korla fragrant pear at anthesis

ABSTRACTThe objective of the study was to learn more about some newly identified miRNAs related to calyx persistence in Korla fragrant pear (Pyrus sinkiangensis Yu). Small RNAs were subjected to high-throughput sequencing after extraction from the ovaries and sepals of flowers with either a deciduous or a persistent calyx. Differentially expressed miRNAs were screened, and 73 new miRNAs were obtained. Twenty of these new miRNAs were selected to further validate all of the new miRNAs. Their mature miRNAs were cloned and identified, the secondary structures of the precursor miRNAs (pre-miRNAs) were analysed, and then qRT-PCR analysis was conducted. The results showed that the mature sequences of nine new miRNAs (novel_miRNA) in different samples were consistent with the results of high-throughput sequencing. Overall, this study improved current methods for the molecular cloning and identification of fruit tree miRNA. Nine new miRNA types were identified. This study laid a good foundation for elucidating the...