抗寒资源枳(Poncirus trifoliata)miR168a克隆、鉴定及遗传转化

Cloning and identification of ptf-miR168a in cold-tolerant Poncirus trifoliata and transformation

基于生物信息学和实验预测,克隆了柑橘抗寒资源枳ptf-miR168a成熟序列和前体序列,并预测了其靶基因。结果表明:ptf-miR168a在植物中高度保守,靶基因是NAC(UCU2)和锌指结构基因(FEZ)家族同源基因;同时,构建了ptf-miR168a前体的超表达载体并在烟草中进行遗传转化,得到了18个转基因系,为进一步鉴定ptf-miR168a在低温胁迫应答中的调控作用奠定基础。

miRNAs(microRNAs)are a type of conserved,endogenous and non-coding small RNAs in plants,and regulate their target genes at post-transcriptional level,which plays important roles in plant.Poncirus trifoliatais a cold-tolerant citrus relative and is often used as rootstock in citrus industry.The mature sequence and precursor sequence were identified in this study.The results showed that ptfmiR168 asequence was highly conserved in plants.The target genes of ptf-miR168 awere analyzed and predicted to be NAC transcription factors and zinc finger genes.The over-expression vector was constructed and transformed to tobacco.A total of 18 transgenic lines were obtained.It will provide valuable materials for further studying the regulatory role of ptf-miR168 ain response to cold stress.