Abstract: The objective of this work is to clone and identify the seed-specific promoter of Ha ds10 G1 in a LEA（late embryogenesis abundant）protein gene family from oil sunflower. The upstream regulatory region of Ha ds10 G1 gene was isolated from the genomic DNA of sunflower “Aidatou” by PCR method. The cloned region was fused to the GUS reporter gene,and plant expression vector pBI121-PHa ds10 was constructed,which was introduced into Nictiana tabacum NC89 by medicating Agrobacterium tumefaciens. The expression of GUS gene in the transgenic tobacco plants was detected by PCR,RT-PCR and GUS. As results,the gene promoter of Ha ds10 G1 was 1 417 bp,and it had homology with the reported sequence at 89.42%. Cis-acting elements search in the segment revealed that in addition to kernel regulatory sequences of promoter,there was the presence of a number of motifs related to tissue specificity,hormone,and adverse environment,such as RY-repeat elements,ABRE motif and TC-rich element,etc. PCR results showed that the transgenic plants were obtained successfully. GUS activity assays indicated that this promoter drove the expression of GUS gene only in tobacco seed embryo,and not in the other tissues such as roots,stems and leaves. Conclusively,the cloned 1 417 bp upstream regulatory region of Ha ds10 G1 gene from the LEA protein gene family of oil sunflower is seed-specific promoter. The results may provide the tissue-specific promoter for genetic improvement of oilseed crops such as oil sunflower.

Key words: oil sunflower, seed-specific promoter, GUS staining, tobacco