Swida-Barteczka Aleksandra, Kruszka Katarzyna, Grabowska Aleksandra, Pacak Andrzej, Jarmolowski Artur, Kurowska Marzena, Szarejko Iwona, Szweykowska-Kulinska Zofia
Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University in Poznan, Poznań, Poland.
Department of Genetics, Faculty of Biology and Environmental Protection, University of Silesia, Katowice, Poland.
Acta Biochim Pol. 2016;63(4):817-824. doi: 10.18388/abp.2016_1352. Epub 2016 Oct 21.
MicroRNAs are short molecules of 21-24 nt in length. They are present in all eukaryotic organisms and regulate gene expression by guiding posttranscriptional silencing of mRNAs. In plants, they are key players in signal transduction, growth and development, and in response to abiotic and biotic stresses. Barley (Hordeum vulgare) is an economically important monocotyledonous crop plant. Drought is the world's main cause of loss in cereal production. We have constructed a high-throughput Real-Time RT-qPCR platform for parallel determination of 159 barley primary microRNAs' levels. The platform was tested for two drought-and-rehydration-treated barley genotypes (Rolap and Sebastian). We have determined changes in the expression of primary microRNAs responding to mild drought, severe drought, and rehydration. Based on the results obtained, we conclude that alteration in the primary microRNA expression is relative to the stress's intensity. Mild drought and rehydration mostly decrease the pri-miRNA levels in both of the tested genotypes. Severe drought mainly induces the primary microRNA expression. The main difference between the genotypes tested was a much-stronger induction of pri-miRNAs in Rolap encountering severe drought. The primary microRNAs respond dynamically to mild drought, severe drought, and rehydration treatments. We propose that some of the individual pri-miRNAs could be used as drought stress or rehydration markers. The usage of the platform in biotechnology is also postulated.
微小RNA是长度为21 - 24个核苷酸的短分子。它们存在于所有真核生物中,并通过引导mRNA的转录后沉默来调节基因表达。在植物中,它们是信号转导、生长发育以及应对非生物和生物胁迫的关键参与者。大麦(Hordeum vulgare)是一种具有重要经济价值的单子叶作物。干旱是全球谷物产量损失的主要原因。我们构建了一个高通量实时逆转录定量PCR平台,用于平行测定159种大麦初级微小RNA的水平。该平台针对两种经干旱和复水处理的大麦基因型(Rolap和Sebastian)进行了测试。我们确定了初级微小RNA在轻度干旱、重度干旱和复水条件下表达的变化。基于所得结果,我们得出结论,初级微小RNA表达的改变与胁迫强度相关。轻度干旱和复水大多会降低两种测试基因型中初级微小RNA的水平。重度干旱主要诱导初级微小RNA的表达。测试基因型之间的主要差异在于,在遭遇重度干旱时,Rolap中初级微小RNA的诱导作用要强得多。初级微小RNA对轻度干旱、重度干旱和复水处理有动态响应。我们提出,一些单个的初级微小RNA可作为干旱胁迫或复水的标记。还假定了该平台在生物技术中的应用。