调控马铃薯干旱胁迫响应相关转录因子的miRNA功能研究

MicroRNAs(miRNAs)是一类非蛋白质编码的内源小分子RNA，通过与靶mRNA的互补配对而在转录后水平调控基因的表达，在植物的生长发育和抗逆性中发挥着非常重要的调控功能。本研究将利用miRNA高通量测序技术和生物信息学方法分离出马铃薯响应干旱胁迫的miRNA，用软件psRNATarget、GO和KEGG对其靶基因进行预测和功能注释，筛选出调控干旱胁迫响应相关转录因子的miRNA，然后用降解组测序技术和RACE方法对筛选出的miRNA靶基因进行验证；构建干旱胁迫响应相关转录因子的miRNA的过表达和抑制表达载体，转化马铃薯，对获得的转基因植株进行表型和抗旱性鉴定，阐明miRNA通过调控干旱胁迫响应相关转录因子来调控功能基因的机理，为利用miRNA改良马铃薯抗旱性奠定理论基础。

MicroRNAs (miRNAs) are a group of small, endogenous noncoding RNAs, which regulates gene expression at posttranscriptional level through complementary pairing with the target mRNA, and play very important role in plant growth and development, and in response to abiotic stress. In the research, the potato miRNA responding to drought stresses will be isolated and identified by high throughput sequencing and bioinformatics method. The target genes will be predicted by online software psRNATarget, and annotated function using GO and KEGG. The miRNA regulating transcription factor related to drought stress response will be screened out. The miRNA target genes will be verified using degradome sequencing and RACE method. Some potato miRNAs regulating transcription factor related to drought stress response will be cloned and constructed their overexpression and RNA interferencing (RNAi) vectors, and tansferred them into potato genome and obtained transgenic potato plants. Phenotype and drought resistance of the transgenic potato plants will be analyzed to elucidate mechanism of miRNA regulating function gene by controlling transcription factor related to drought stress response, and provide theoretical foundation for improvrment drought resistance of potato by miRNA.