Characterization Of Tissue-specific Promotors In Soybean

Soybean is an important grain and oil crop around the world.The seed contains abundant fat,vegetable protein and multiple nutritive elements.However,biotic stresses such as pathogens,pests and abiotic stresses such as high temperature and drought in the growing environment often impair the growth,which lead to decline of production and deterioration of quality.Thus,an economical and effective methods need to be delivered for multiple improvements of soybean.The genome of soybean is large and the mechanism of gene expression is complicated.With more and more resistance genes are discovered,the research for regulatory elements of gene expression,such as promoter,admits of no delay.However,the promoters we often use in present transgenic technology are constitutive,which may cause extra metabolism burden or toxic effect in transgenic plants.So,for deeper research in spatio-temporal expression of gene,we should pay more attention to the tissue-specific promoter which can guide target genes to express in specific tissues and organs.In our study,we select tissue-specific genes from published soybean transcriptome data,and then search the promoter sequence from the gene's upstream in Phytozome.The promoter sequence was cloned from Glycine max,cultivar Williams 82,and the gene expression vectors of promoter:GUS were constructed.We transform the vectors into Arabidopsis using Agrobacterium tumefaciens mediated method,respectively.The promoters' expression patterns were identified by GUS staining and RT-PCR.The promoter:GUS expression vectors were transformed into Agrobacterium rhizogenes and then transformed into soybean where the hairy root grow from.The expressions of promoters in hairy roots were verified by GUS staining.The study have acquired 2 leaf-specific promoters and 8 root-specific promoters,and the expression specificities were verified in Arabidopsis transgenic plants and soybean hairy roots.