Karimi Marzieh, Ghazanfari Farahnaz, Fadaei Adeleh, Ahmadi Laleh, Shiran Behrouz, Rabei Mohammad, Fallahi Hossein
Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Shahrekord University, Shahrekord, P.O.Box 115, Iran.
Institute of Biotechnology, Shahrekord University, Shahrekord, P.O.Box 115, Iran.
PLoS One. 2016 Jun 2;11(6):e0156519. doi: 10.1371/journal.pone.0156519. eCollection 2016.
Spring frost is an important environmental stress that threatens the production of Prunus trees. However, little information is available regarding molecular response of these plants to the frost stress. Using high throughput sequencing, this study was conducted to identify differentially expressed miRNAs, both the conserved and the non-conserved ones, in the reproductive tissues of almond tolerant H genotype under cold stress. Analysis of 50 to 58 million raw reads led to identification of 174 unique conserved and 59 novel microRNAs (miRNAs). Differential expression pattern analysis showed that 50 miRNA families were expressed differentially in one or both of almond reproductive tissues (anther and ovary). Out of these 50 miRNA families, 12 and 15 displayed up-regulation and down-regulation, respectively. The distribution of conserved miRNA families indicated that miR482f harbor the highest number of members. Confirmation of miRNAs expression patterns by quantitative real- time PCR (qPCR) was performed in cold tolerant (H genotype) alongside a sensitive variety (Sh12 genotype). Our analysis revealed differential expression for 9 miRNAs in anther and 3 miRNAs in ovary between these two varieties. Target prediction of miRNAs followed by differential expression analysis resulted in identification of 83 target genes, mostly transcription factors. This study comprehensively catalogued expressed miRNAs under different temperatures in two reproductive tissues (anther and ovary). Results of current study and the previous RNA-seq study, which was conducted in the same tissues by our group, provide a unique opportunity to understand the molecular basis of responses of almond to cold stress. The results can also enhance the possibility for gene manipulation to develop cold tolerant plants.
春霜是一种重要的环境胁迫，威胁着李属树木的产量。然而，关于这些植物对霜冻胁迫的分子反应的信息却很少。本研究利用高通量测序技术，鉴定了耐冷H基因型杏仁生殖组织中差异表达的miRNA，包括保守型和非保守型。对5000万至5800万条原始读数的分析，共鉴定出174个独特的保守型和59个新的微小RNA（miRNA）。差异表达模式分析表明，50个miRNA家族在杏仁的一种或两种生殖组织（花药和子房）中差异表达。在这50个miRNA家族中，分别有12个和15个呈现上调和下调。保守miRNA家族的分布表明，miR482f的成员数量最多。通过实时定量PCR（qPCR）对耐冷（H基因型）和敏感品种（Sh12基因型）的miRNA表达模式进行了验证。我们的分析揭示了这两个品种在花药中有9个miRNA和在子房中3个miRNA差异表达。对miRNA进行靶标预测并随后进行差异表达分析，共鉴定出83个靶基因，其中大多数是转录因子。本研究全面编目了两种生殖组织（花药和子房）在不同温度下表达的miRNA。本研究结果与我们团队之前在相同组织中进行的RNA测序研究结果，为理解杏仁对冷胁迫反应的分子基础提供了独特的机会。这些结果也增加了通过基因操作培育耐寒植物的可能性。
