Anand Asha, Chauhan Shailja, Chodon Aparna, Vimala Kumar Kavitha Velayudha, Saravanakumar S, Pandi Gopal
Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, India.
Department of Life Sciences, CHRIST (Deemed to be University), Bengaluru, Karnataka, India.
BioTechnologia (Pozn). 2024 Mar 29;105(1):19-32. doi: 10.5114/bta.2024.135639. eCollection 2024.
The microRNAs are endogenous, regulating gene expression either at the DNA or RNA level. Despite the availability of extensive studies on microRNA generation in plants, reports on their abundance, biogenesis, and consequent gene regulation in plant organelles remain naVve. Building on previous studies involving pre-miRNA sequencing in , we demonstrated that three putative microRNAs were raised from the chloroplast genome. In the current study, we have characterized the genesis of these three microRNAs through a combination of bioinformatics and experimental approaches. The gene sequence for a miRNA, designated as AecpmiRNA1 ( chloroplast miRNA), is potentially located in both the genomic DNA, i.e., nuclear and chloroplast genome. In contrast, the gene sequences for the other two miRNAs (AecpmiRNA2 and AecpmiRNA3) are exclusively present in the chloroplast genome. Target prediction revealed many potential mRNAs as targets for AecpmiRNAs. Further analysis using 5' RACE-PCR determined the AecpmiRNA3 binding and cleavage site at the photosystem II protein N (psbN). These results indicate that AecpmiRNAs are generated from the chloroplast genome, possessing the potential to regulate mRNAs arising from chloroplast gene(s). On the other side, the possibility of nuclear genome-derived mRNA regulation by AecpmiRNAs cannot be ruled out.
微小RNA是内源性的，可在DNA或RNA水平上调节基因表达。尽管对植物中微小RNA的生成已有广泛研究，但关于它们在植物细胞器中的丰度、生物合成以及随之而来的基因调控的报道仍然很少。基于之前涉及[具体植物名称]中前体微小RNA测序的研究，我们证明了三个假定的微小RNA来自叶绿体基因组。在当前研究中，我们通过生物信息学和实验方法相结合的方式对这三个微小RNA的起源进行了表征。一种名为AecpmiRNA1（叶绿体微小RNA）的微小RNA的基因序列可能位于基因组DNA中，即核基因组和叶绿体基因组中。相比之下，另外两个微小RNA（AecpmiRNA2和AecpmiRNA3）的基因序列仅存在于叶绿体基因组中。靶标预测揭示了许多潜在的mRNA作为AecpmiRNAs的靶标。使用5' RACE-PCR的进一步分析确定了AecpmiRNA3在光系统II蛋白N（psbN）处的结合和切割位点。这些结果表明AecpmiRNAs是从叶绿体基因组产生的，具有调节叶绿体基因产生的mRNA的潜力。另一方面，不能排除AecpmiRNAs对核基因组衍生的mRNA进行调控的可能性。
