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Identification of novel microRNAs in Hevea brasiliensis and computational prediction of their targets.

Gébelin V, Argout X, Engchuan W, Pitollat B, Duan C, Montoro P, Leclercq J - BMC Plant Biol. (2012)

Bottom Line: The results showed the most abundant size for miRNAs to be 24 nucleotides, except for seven families.Deep sequencing of small RNAs combined with transcriptomic data is a powerful tool for identifying conserved and novel miRNAs when the complete genome is not yet available.Our study provided additional information for evolutionary studies and revealed potentially specific regulation of the control of redox status in Hevea.

View Article: PubMed Central - HTML - PubMed

Affiliation: CIRAD, UMR AGAP, F-34398 Montpellier, France.

ABSTRACT

Background: Plants respond to external stimuli through fine regulation of gene expression partially ensured by small RNAs. Of these, microRNAs (miRNAs) play a crucial role. They negatively regulate gene expression by targeting the cleavage or translational inhibition of target messenger RNAs (mRNAs). In Hevea brasiliensis, environmental and harvesting stresses are known to affect natural rubber production. This study set out to identify abiotic stress-related miRNAs in Hevea using next-generation sequencing and bioinformatic analysis.

Results: Deep sequencing of small RNAs was carried out on plantlets subjected to severe abiotic stress using the Solexa technique. By combining the LeARN pipeline, data from the Plant microRNA database (PMRD) and Hevea EST sequences, we identified 48 conserved miRNA families already characterized in other plant species, and 10 putatively novel miRNA families. The results showed the most abundant size for miRNAs to be 24 nucleotides, except for seven families. Several MIR genes produced both 20-22 nucleotides and 23-27 nucleotides. The two miRNA class sizes were detected for both conserved and putative novel miRNA families, suggesting their functional duality. The EST databases were scanned with conserved and novel miRNA sequences. MiRNA targets were computationally predicted and analysed. The predicted targets involved in "responses to stimuli" and to "antioxidant" and "transcription activities" are presented.

Conclusions: Deep sequencing of small RNAs combined with transcriptomic data is a powerful tool for identifying conserved and novel miRNAs when the complete genome is not yet available. Our study provided additional information for evolutionary studies and revealed potentially specific regulation of the control of redox status in Hevea.

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Abundance of the 48 conserved miRNA families in Hevea brasiliensis with 0 and 1 mismatch on a minimum length of 19 nucleotides. A) The sequencing frequency of a conserved miRNA family in the small RNA library above 1000 reads. B) The sequencing frequency of a conserved miRNA family in the small RNA library below 1000 reads.
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Figure 3: Abundance of the 48 conserved miRNA families in Hevea brasiliensis with 0 and 1 mismatch on a minimum length of 19 nucleotides. A) The sequencing frequency of a conserved miRNA family in the small RNA library above 1000 reads. B) The sequencing frequency of a conserved miRNA family in the small RNA library below 1000 reads.

Mentions: Of the 48 conserved miRNA families, 12 were predominant with more than 1,000 reads, including 3 families that displayed more than 20,000 reads. The latter corresponded to HbmiR159/319 and HbmiR408 (Figure 3A). By contrast, 36 families were less represented in the dataset with 18 families having fewer than 100 reads: HbmiR1310, HbmiR162, HbmiR168, HbmiR1863, HbmiR2118, HbmiR2910, HbmiR2914, HbmiR2915, HbmiR2916, HbmiR393, HbmiR394, HbmiR395, HbmiR399, HbmiR444, HbmiR476, HbmiR482, HbmiR828 and HbmiR845 (Figure 3B).


Identification of novel microRNAs in Hevea brasiliensis and computational prediction of their targets.

Gébelin V, Argout X, Engchuan W, Pitollat B, Duan C, Montoro P, Leclercq J - BMC Plant Biol. (2012)

Abundance of the 48 conserved miRNA families in Hevea brasiliensis with 0 and 1 mismatch on a minimum length of 19 nucleotides. A) The sequencing frequency of a conserved miRNA family in the small RNA library above 1000 reads. B) The sequencing frequency of a conserved miRNA family in the small RNA library below 1000 reads.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3368772&req=5

Figure 3: Abundance of the 48 conserved miRNA families in Hevea brasiliensis with 0 and 1 mismatch on a minimum length of 19 nucleotides. A) The sequencing frequency of a conserved miRNA family in the small RNA library above 1000 reads. B) The sequencing frequency of a conserved miRNA family in the small RNA library below 1000 reads.
Mentions: Of the 48 conserved miRNA families, 12 were predominant with more than 1,000 reads, including 3 families that displayed more than 20,000 reads. The latter corresponded to HbmiR159/319 and HbmiR408 (Figure 3A). By contrast, 36 families were less represented in the dataset with 18 families having fewer than 100 reads: HbmiR1310, HbmiR162, HbmiR168, HbmiR1863, HbmiR2118, HbmiR2910, HbmiR2914, HbmiR2915, HbmiR2916, HbmiR393, HbmiR394, HbmiR395, HbmiR399, HbmiR444, HbmiR476, HbmiR482, HbmiR828 and HbmiR845 (Figure 3B).

Bottom Line: The results showed the most abundant size for miRNAs to be 24 nucleotides, except for seven families.Deep sequencing of small RNAs combined with transcriptomic data is a powerful tool for identifying conserved and novel miRNAs when the complete genome is not yet available.Our study provided additional information for evolutionary studies and revealed potentially specific regulation of the control of redox status in Hevea.

View Article: PubMed Central - HTML - PubMed

Affiliation: CIRAD, UMR AGAP, F-34398 Montpellier, France.

ABSTRACT

Background: Plants respond to external stimuli through fine regulation of gene expression partially ensured by small RNAs. Of these, microRNAs (miRNAs) play a crucial role. They negatively regulate gene expression by targeting the cleavage or translational inhibition of target messenger RNAs (mRNAs). In Hevea brasiliensis, environmental and harvesting stresses are known to affect natural rubber production. This study set out to identify abiotic stress-related miRNAs in Hevea using next-generation sequencing and bioinformatic analysis.

Results: Deep sequencing of small RNAs was carried out on plantlets subjected to severe abiotic stress using the Solexa technique. By combining the LeARN pipeline, data from the Plant microRNA database (PMRD) and Hevea EST sequences, we identified 48 conserved miRNA families already characterized in other plant species, and 10 putatively novel miRNA families. The results showed the most abundant size for miRNAs to be 24 nucleotides, except for seven families. Several MIR genes produced both 20-22 nucleotides and 23-27 nucleotides. The two miRNA class sizes were detected for both conserved and putative novel miRNA families, suggesting their functional duality. The EST databases were scanned with conserved and novel miRNA sequences. MiRNA targets were computationally predicted and analysed. The predicted targets involved in "responses to stimuli" and to "antioxidant" and "transcription activities" are presented.

Conclusions: Deep sequencing of small RNAs combined with transcriptomic data is a powerful tool for identifying conserved and novel miRNAs when the complete genome is not yet available. Our study provided additional information for evolutionary studies and revealed potentially specific regulation of the control of redox status in Hevea.

Show MeSH