Abstract

BackgroundMicroRNAs (miRNAs) are small RNAs (sRNA) ~21 nucleotides in length that negatively control gene expression by cleaving or inhibiting the translation of target gene transcripts. miRNAs have been extensively analyzed in Arabidopsis and rice and partially investigated in other non-model plant species. To date, 109 and 62 miRNA families have been identified in Arabidopsis and rice respectively. However, only 33 miRNAs have been identified from the genome of the model tree species (Populus trichocarpa), of which 11 are Populus specific. The low number of miRNA families previously identified in Populus, compared with the number of families identified in Arabidopsis and rice, suggests that many miRNAs still remain to be discovered in Populus. In this study, we analyzed expressed small RNAs from leaves and vegetative buds of Populus using high throughput pyrosequencing.ResultsAnalysis of almost eighty thousand small RNA reads allowed us to identify 123 new sequences belonging to previously identified miRNA families as well as 48 new miRNA families that could be Populus-specific. Comparison of the organization of miRNA families in Populus, Arabidopsis and rice showed that miRNA family sizes were generally expanded in Populus. The putative targets of non-conserved miRNA include both previously identified targets as well as several new putative target genes involved in development, resistance to stress, and other cellular processes. Moreover, almost half of the genes predicted to be targeted by non-conserved miRNAs appear to be Populus-specific. Comparative analyses showed that genes targeted by conserved and non-conserved miRNAs are biased mainly towards development, electron transport and signal transduction processes. Similar results were found for non-conserved miRNAs from Arabidopsis.ConclusionOur results suggest that while there is a conserved set of miRNAs among plant species, a large fraction of miRNAs vary among species. The non-conserved miRNAs may regulate cellular, physiological or developmental processes specific to the taxa that produce them, as appears likely to be the case for those miRNAs that have only been observed in Populus. Non-conserved and conserved miRNAs seem to target genes with similar biological functions indicating that similar selection pressures are acting on both types of miRNAs. The expansion in the number of most conserved miRNAs in Populus relative to Arabidopsis, may be linked to the recent genome duplication in Populus, the slow evolution of the Populus genome, or to differences in the selection pressure on duplicated miRNAs in these species.

Highlights

  • MicroRNAs are small RNAs ~21 nucleotides in length that negatively control gene expression by cleaving or inhibiting the translation of target gene transcripts. miRNA* units [8]. One strand (miRNA) have been extensively analyzed in Arabidopsis and rice and partially investigated in other non-model plant species

  • By removing redundant sequences from these two data sets, we identified 5,998 (Additional file 1) and 6,339 (Additional file 2) unique small RNAs (sRNA) sequences from leaf and vegetative buds, respectively

  • A total of 2,607 and 2,167 unique sRNA sequences matched the Populus genome assembly; these were considered for further analysis. sRNAs of 21 nt in length were the most abundant class among the 15–30 nucleotide sequences (Fig. 1) suggesting that most of the small RNAs identified are processed by the Populus DCL1 homolog

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Summary

Introduction

MicroRNAs (miRNAs) are small RNAs (sRNA) ~21 nucleotides in length that negatively control gene expression by cleaving or inhibiting the translation of target gene transcripts. miRNAs have been extensively analyzed in Arabidopsis and rice and partially investigated in other non-model plant species. MiRNAs have been extensively analyzed in Arabidopsis and rice and partially investigated in other non-model plant species. Only 33 miRNAs have been identified from the genome of the model tree species (Populus trichocarpa), of which 11 are Populus specific. Gene sequences among the different Populus species show high similarity (>95%) [2] and as close as 99% between P. balsamifera and P. trichocarpa for the few cDNAs sequenced in P. balsamifera. In addition to its economic and ecological importance [3], Populus was chosen as a model for trees because it has a relatively small genome (500 MB), just four times the size of the Arabidopsis genome. The genome sequence for Populus trichocarpa facilitates functional analyses of genes in Populus as well as comparative and functional genomics with closely related species, especially within the Salicaceae

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