Limits...
Identification and characterization of microRNAs from Chinese pollination constant non-astringent persimmon using high-throughput sequencing.

Luo Y, Zhang X, Luo Z, Zhang Q, Liu J - BMC Plant Biol. (2015)

Bottom Line: 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.In addition, two transcription factors, a GRF and a bHLH, were experimentally confirmed as the targets of dka-miR396 and dka-miR395, respectively.In addition, dka-miR396g and miR2911a may regulate their target genes associated with glucosylation and insolubilization of tannin precursors.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Horticultural Plant Biology (MOE), College of Horticulture and Forestry Science, Huazhong Agricultural University, Wuhan, 430070, China. luoyujie200701@163.com.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus

Measurement of tannin content in ‘Eshi No. 1’ (CPCNA) fruits at different development stages. A. Representative photos showing the fruits sampled at five stages, 5, 10, 15, 20 and 25 weeks after flowering (WAF) of ‘Eshi No. 1’. B. Analysis of soluble tannin content in the persimmon fruits based on an imprinting method. The red arrows show that the staining became weaker from 15 to 20 WAF. C. Change in the fruit weight at the five sampling stages. D-E. Quantitative measurement of soluble (D) and insoluble (E) tannin in the fruits by folin-ciocalteu method. F. Total tannin content in the fruits.
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Fig1: Measurement of tannin content in ‘Eshi No. 1’ (CPCNA) fruits at different development stages. A. Representative photos showing the fruits sampled at five stages, 5, 10, 15, 20 and 25 weeks after flowering (WAF) of ‘Eshi No. 1’. B. Analysis of soluble tannin content in the persimmon fruits based on an imprinting method. The red arrows show that the staining became weaker from 15 to 20 WAF. C. Change in the fruit weight at the five sampling stages. D-E. Quantitative measurement of soluble (D) and insoluble (E) tannin in the fruits by folin-ciocalteu method. F. Total tannin content in the fruits.

Mentions: Imprinting method was used to determine soluble tannin levels in ‘Eshi No. 1’ fruits. The sections were deeply stained at the beginning of fruit development (5 WAF), when the fruits were small in size. With the progression of development, the fruits grew quickly and became increasingly big until reaching the largest size at 25 WAF (Figure 1A,C). The fruits were still darkly stained until 15 WAF, but the staining began to turn lighter at 20 WAF. At the last experimental stage, 25 WAF, the fruits were only slightly stained (Figure 1B).Figure 1


Identification and characterization of microRNAs from Chinese pollination constant non-astringent persimmon using high-throughput sequencing.

Luo Y, Zhang X, Luo Z, Zhang Q, Liu J - BMC Plant Biol. (2015)

Measurement of tannin content in ‘Eshi No. 1’ (CPCNA) fruits at different development stages. A. Representative photos showing the fruits sampled at five stages, 5, 10, 15, 20 and 25 weeks after flowering (WAF) of ‘Eshi No. 1’. B. Analysis of soluble tannin content in the persimmon fruits based on an imprinting method. The red arrows show that the staining became weaker from 15 to 20 WAF. C. Change in the fruit weight at the five sampling stages. D-E. Quantitative measurement of soluble (D) and insoluble (E) tannin in the fruits by folin-ciocalteu method. F. Total tannin content in the fruits.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4308916&req=5

Fig1: Measurement of tannin content in ‘Eshi No. 1’ (CPCNA) fruits at different development stages. A. Representative photos showing the fruits sampled at five stages, 5, 10, 15, 20 and 25 weeks after flowering (WAF) of ‘Eshi No. 1’. B. Analysis of soluble tannin content in the persimmon fruits based on an imprinting method. The red arrows show that the staining became weaker from 15 to 20 WAF. C. Change in the fruit weight at the five sampling stages. D-E. Quantitative measurement of soluble (D) and insoluble (E) tannin in the fruits by folin-ciocalteu method. F. Total tannin content in the fruits.
Mentions: Imprinting method was used to determine soluble tannin levels in ‘Eshi No. 1’ fruits. The sections were deeply stained at the beginning of fruit development (5 WAF), when the fruits were small in size. With the progression of development, the fruits grew quickly and became increasingly big until reaching the largest size at 25 WAF (Figure 1A,C). The fruits were still darkly stained until 15 WAF, but the staining began to turn lighter at 20 WAF. At the last experimental stage, 25 WAF, the fruits were only slightly stained (Figure 1B).Figure 1

Bottom Line: 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.In addition, two transcription factors, a GRF and a bHLH, were experimentally confirmed as the targets of dka-miR396 and dka-miR395, respectively.In addition, dka-miR396g and miR2911a may regulate their target genes associated with glucosylation and insolubilization of tannin precursors.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Horticultural Plant Biology (MOE), College of Horticulture and Forestry Science, Huazhong Agricultural University, Wuhan, 430070, China. luoyujie200701@163.com.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus