中文摘要：

本研究通过PCR方法从拟南芥基因组序列中分离了3个理论推断与耐寒相关的miRNA片段（miRNA402,miRNA319和miRNA393）和1个与植物生长相关的片段miRNA172。利用重叠延伸PCR技术将miRNA172小分子,以及3个与耐寒相关的miRNA402,miRNA319和miRNA393小分子串连在一起分别导入植物表达载体pVKH-35S-GUS-pA,取代表达载体中的GUS基因,构建了pVKH-35S-mir172-pA和pVKH-35S-mir31＋402＋393-pA融合表达载体。经PCR和双酶切及测序鉴定,pVKH-35S-mir172-pA和pVKH-35S-mir319＋402＋393-pA构建成功,为后续利用基因工程手段,miRNA转化木薯,提高木薯生长和木薯耐寒相关研究奠定基础。

英文摘要：

In this article,we have isolated one small molecule miRNA172 related with plant growth,3 cold-resistant related miRNA402,miRNA319 and miRNA393 from Arabidopsis genomic DNA by PCR method.Then miRNA172 and the fused miRNAs with miRNA402,miRNA319 and miRNA393 have been replaced the GUS gene in pVKH-35S-GUS-pA by splicing overlap extension PCR and constructed plant botanic expression vectors of pVKH-35S-mir172-pA and pVKH-35S-mir319＋402＋393-pA,respectively.The expressing vectors of pVKH-35S-mir172-pA,pVKH-35S-mir319＋402＋393-pA have been identified to be successfully constructed by PCR,restriction enzyme digestion and sequence analysis.The result would be the preliminary work for use of miRNAs to improve the cassava growth and cold tolerance by gene engineering method.