John Wiley & Sons, Ltd.

Genome-Wide Identification Of Chromium Stress-Responsive Micrornas And Their Target Genes In Tobacco (Nicotiana Tabacum) Roots

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Tobacco can easily accumulate certain heavy metals in leaves and thus poses a potential threat to human health. To systematically dissect Cr-responsive miRNAs and their targets at the global level, four small RNA libraries were constructed from the roots of Cr-treated (Cr) and Cr-free (control) two contrasting tobacco genotypes,Yunyan2 (Cr-sensitive) and Guiyan1 (Cr-tolerant). Using high-throughput-sequencing-technology, we identified 53 conserved and 29 novel miRNA families. Comparative genomic analysis of 41 conserved Cr-responsive miRNA families revealed that 11 miRNA families showed up-regulation in Guiyan1 but unaltered in Yunyan2, and 17 miRNA families were up-regulated only in Yunyan2 under Cr stress. Only 1 family, miR6149, was down-regulated in Yunyan2 but unchanged in Guiyan1. Of the 29 novel miRNA families, 14 expressed differently in the two genotypes under Cr stress. Based on a high-throughput degradome sequencing homology search, potential targets were predicted for the 41 conserved and 14 novel Cr-responsive miRNA families. COG functional category analysis revealed that some of these predicted target transcripts of miRNAs are responsive to biotic and abiotic stresses. Furthermore, the expression patterns of many Cr-responsive miRNAs were validated by stem-loop RT-PCR. The results of the present study provide valuable information and a framework for understanding the function of miRNAs in Cr tolerance.

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