Home > Browse Issues > Vol.39 No.7
Bioinformatic Analyses of gma-miR1507a， Construction of Plant Expression Vectors and Genetic Transformation
Ma Ling, Cui Xiaoxia, Huang Yanzhong, Zhao Jinming, Wang Haitang, Guo Na*, Xing Han*
National Center for Soybean Improvement, Key Laboratory of Biology and Genetics and Breeding for Soybean, Ministry of Agriculture, P. R. China, State Key Laboratory of Crop Genetics & Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China
Abstract: miRNA (microRNA) are a class of endogenous and noncoding small molecular RNA with approximately 18-25 nucleotides (nt) in length. They can regulate gene expression through mRNA cleavage or translation inhibition. We analyzed the mature sequence， stem-loop structure and promoter sequence of gmamiR1507a. The level of gma-miR1507a in soybean tissue under stress was tested by stem-loop RT-PCR method. The target genes of gma-miR1507a were predicted by the online software psRNATarget. Then， we constructed artificial miRNA expression vectors-miRNA1507a-pCAMBIA3301. Furthermore， amiRNA1507a gene was overexpressed in soybean cotyledon and the transgenic hairy roots were obtained. The results showed that there were a number of cis-acting elements related to drought stress and pathogen infection in the promoter sequence of precursor gma-miR1507a and 9 target genes of gma-miR1507a were predicted. We obtained the transgenic hairy roots by hairy root-mediated genetic transformation system and GFP staining. The expression level of gmamiR1507a in the transgenic hairy roots significantly increased compared with that in empty vector. The expression of seven predicted target genes significantly reduced in transgenic hairy roots. Based on the results， the transgenic hairy roots of amiRNA1507a overexpression could increase the expression of gma-miR1507a which could be used to analyze the function of gma-miR1507a. In addition， gma-miR1507a may involve in biotic stresses responses in soybean.
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