MicroRNA profiling analysis of developing berries for 'Kyoho' and its early-ripening mutant during berry ripening.
BACKGROUND
'Fengzao' is an early-ripening bud mutant of 'Kyoho', which matures nearly 30 days earlier than 'Kyoho'. To gain a better understanding of the regulatory role of miRNAs in early-ripening of grape berry, high-throughput sequencing approach and quantitative RT-PCR validation were employed to identify miRNAs at the genome-wide level and profile the expression patterns of the miRNAs during berry development in 'Kyho' and 'Fengzao', respectively.


RESULTS
Nine independent small RNA libraries were constructed and sequenced in two varieties from key berry development stages. A total of 108 known miRNAs and 61 novel miRNAs were identified. Among that, 159 miRNAs identified in 'Fengzao' all completely expressed in 'Kyoho' and there were 10 miRNAs specifically expressed in 'Kyoho'. The expression profiles of known and novel miRNAs were quite similar between two varieties. As the major differentially expressed miRNAs, novel_144, vvi-miR3626-3p and vvi-miR3626-5p only expressed in 'Kyoho', vvi-miR399b and vvi-miR399e were down-regulated in 'Fengzao', while vvi-miR477b-3p up-regulated in 'Fengzao'. According to the expression analysis and previous reports, miR169-NF-Y subunit, miR398-CSD, miR3626-RNA helicase, miR399- phosphate transporter and miR477-GRAS transcription factor were selected as the candidates for further investigations of miRNA regulation role in the early-ripening of grape. The qRT-PCR analyses validated the contrasting expression patterns for these miRNAs and their target genes.


CONCLUSIONS
The miRNAome of the grape berry development of 'Kyoho', and its early-ripening bud mutant, 'Fengzao' were compared by high-throughput sequencing. The expression pattern of several key miRNAs and their target genes during grape berry development and ripening stages was examined. Our results provide valuable basis towards understanding the regulatory mechanisms of early-ripening of grape berry.
['Guo|Da-Long|DL|http://orcid.org/0000-0001-5379-5255', 'Li|Qiong|Q|', 'Lv|Wen-Qing|WQ|', 'Zhang|Guo-Hai|GH|', 'Yu|Yi-He|YH|']
D005638:Fruit D018507:Gene Expression Regulation, Developmental D018506:Gene Expression Regulation, Plant D018745:Genome, Plant D059014:High-Throughput Nucleotide Sequencing D035683:MicroRNAs D018749:RNA, Plant D017423:Sequence Analysis, RNA D027843:Vitis
miRNA Grape Early ripening ROS