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Published on: August 21, 2019
Identification and characterization of microRNAs from Chinese pollination constant non-astringent persimmon using
Yujie Luo1, Xiaona Zhang2, Zhengrong Luo3
1Key Laboratory of Horticultural Plant Biology (MOE), College of Horticulture and Forestry Science, Huazhong Agricultural University, Wuhan, 430070, China. luoyujie200701@163.com.
Background:
microRNAs (miRNAs) have been shown to play key roles in regulating gene expression at post-transcriptional level, but miRNAs associated with natural deastringency of Chinese pollination-constant nonastringent persimmon (CPCNA) have never been identified.
Results:
In this study, two small RNA libraries established using 'Eshi No. 1' persimmon (Diospyros kaki Thunb.; CPCNA) fruits collected at 15 and 20 weeks after flowering (WAF) were sequenced through Solexa platform in order to identify miRNAs involved in deastringency of persimmon. A total of 6,258,487 and 7,634,169 reads were generated for the libraries at 15 and 20 WAF, respectively. Based on sequence similarity and hairpin structure prediction, 236 known miRNAs belonging to 65 miRNA families and 33 novel miRNAs were identified using persimmon transcriptome data. Sixty one of the characterized miRNAs exhibited pronounced difference in the expression levels between 15 and 20 WAF, 17 up-regulated and 44 down-regulated. Expression profiles of 12 conserved and 10 novel miRNAs were validated by stem loop qRT-PCR. A total of 198 target genes were predicted for the differentially expressed miRNAs, including several genes that have been reported to be implicated in proanthocyanidins (PAs, or called tannin) accumulation. In addition, two transcription factors, a GRF and a bHLH, were experimentally confirmed as the targets of dka-miR396 and dka-miR395, respectively.
Conclusions:
Taken together, the present data unraveled several important miRNAs in persimmon. Among them, miR395p-3p and miR858b may regulate bHLH and MYB, respectively, which are influenced by SPL under the control of miR156j-5p and in turn regulate the structural genes involved in PA biosynthesis. In addition, dka-miR396g and miR2911a may regulate their target genes associated with glucosylation and insolubilization of tannin precursors. All of these miRNAs might play key roles in the regulation of (de)astringency in persimmon fruits under normal development conditions.
Insights
This study identified key microRNAs (miRNAs) involved in the natural deastringency of Chinese pollination-constant nonastringent persimmon (CPCNA). These miRNAs regulate genes controlling tannin accumulation, crucial for fruit flavor development.
Area of Science:
- Plant Molecular Biology
- Fruit Ripening and Quality
- Post-transcriptional Gene Regulation
Background:
- MicroRNAs (miRNAs) are critical regulators of gene expression.
- The specific miRNAs involved in the natural deastringency of Chinese pollination-constant nonastringent persimmon (CPCNA) remain unidentified.
Purpose of the Study:
- To identify and characterize miRNAs associated with the natural deastringency process in CPCNA persimmon.
- To investigate the roles of these miRNAs in regulating genes related to tannin accumulation and fruit astringency.
Main Methods:
- Small RNA sequencing of persimmon fruits at two developmental stages (15 and 20 weeks after flowering).
- Bioinformatic analysis for miRNA identification, including known and novel miRNAs.
- Prediction and validation of miRNA target genes using transcriptome data and stem-loop qRT-PCR.
Main Results:
- Identified 236 known and 33 novel miRNAs in persimmon fruits.
- Found 61 differentially expressed miRNAs between the two developmental stages.
- Predicted 198 target genes, including those involved in proanthocyanidin (tannin) accumulation and transcription factors (GRF, bHLH).
Conclusions:
- Several miRNAs, including miR395p-3p, miR858b, miR156j-5p, dka-miR396g, and miR2911a, are implicated in persimmon deastringency.
- These miRNAs likely regulate tannin biosynthesis and modification through transcription factors and other target genes.
- The findings provide insights into the molecular mechanisms underlying fruit astringency in persimmon.
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