Biological Sciences and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, Turkey.
Funct Integr Genomics. 2010 Nov;10(4):493-507. doi: 10.1007/s10142-010-0181-4. Epub 2010 Jul 31.
We aim to identify conserved and dehydration responsive microRNAs (miRNAs) in Hordeum vulgare (barley). A total of 28 new barley miRNAs belonging to 18 distinct miRNA families were identified. Detailed nucleotide analyses revealed that barley pre-miRNAs are in the range of 46-114 nucleotides with average of 77.14. Using 28 newly detected miRNAs as queries, 445 potential target mRNAs were predicted. The predicted miRNAs were differentially expressed and some of them behaved similarly in leaf and root tissues upon stress treatment. Hvu-MIR156, Hvu-MIR166, Hvu-MIR171, and Hvu-MIR408 were detected as dehydration stress-responsive barley miRNAs. To discover target transcripts of barley miRNAs a modified 5' RLM-RACE was performed and seven cleaved miRNA transcripts were retrieved from drought stressed leaf samples. In silico analysis indicated 15 potential EST targets. Measurement of expression levels showed a positive correlation between levels of miRNA expression and suppression of their target mRNA transcripts in dehydration-stress-treated barley.
我们旨在鉴定大麦(Hordeum vulgare)中保守和脱水响应的 microRNAs(miRNAs)。共鉴定出 28 种属于 18 个不同 miRNA 家族的新大麦 miRNAs。详细的核苷酸分析表明,大麦前体 miRNA 的长度在 46-114 个核苷酸之间,平均长度为 77.14。使用 28 个新检测到的 miRNA 作为查询,预测了 445 个潜在的靶 mRNA。预测的 miRNAs 表现出差异表达,其中一些在受到胁迫处理时在叶片和根组织中的表现相似。Hvu-MIR156、Hvu-MIR166、Hvu-MIR171 和 Hvu-MIR408 被检测为对脱水胁迫有反应的大麦 miRNAs。为了发现大麦 miRNAs 的靶转录本,进行了改良的 5' RLM-RACE,并从干旱胁迫的叶片样本中回收了 7 个切割的 miRNA 转录本。计算机分析表明有 15 个潜在的 EST 靶标。表达水平的测量显示,在脱水胁迫处理的大麦中,miRNA 表达水平与它们的靶 mRNA 转录本的抑制之间呈正相关。