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) 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.

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)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。本项目利用miRNA高通量测序技术和生物信息学方法分离鉴定出了59个马铃薯已知的miRNA和1279个新的miRNA。靶基因预测获得已知miRNA的165个靶基因，新miRNA的2981个靶基因；对预测的靶基因进行了功能注释，并用降解组测序鉴定了马铃薯miRNA的靶基因。鉴定出了分别调控马铃薯转录因子MYB、NF-YA、HD-ZIP和NAC的miR159、miR169、miR166和miRNA164及其靶基因StGAMyb-like1、StGAMyb-like2.1、StGAMyb-like2.2；StNF-YA1、StNF-YA3、StNF-YA4、StNF-YA5、StNF-YA6；StHB14-lik、StHB14、StHB15、StHB15-like、StREV、ATHB12和StNAC083、StNAC262、StNAC280。从马铃薯中克隆出了miR159的3个靶基因StGAMyb-like1、StGAMyb-like2.1、StGAMyb-like2.2；miR166的4个StHB14、StHB15、StREV、ATHB12和miRNA164的靶基因NAC262，并对其进行了功能鉴定。阐明了在干旱胁迫下miR159、miR169、miR166和miRNA164通过调节转录因子来调控马铃薯抗旱性的分子机理。取得的研究结果为利用miRNA改良马铃薯的抗旱性奠定了理论基础。