Screening Of Rice Genes With Highly Endosperm-specific Expression Pattern And Preliminary Analysis Of The Promoter Of Latex-1Gene

Data from biological research can be shared in view of rice functional genomic databases. Employing the research methods in bioinformatics,50seed genes,which maybe endosperm-specific genes, were seeked out from rice databases.Using semi-quantitative RT-PCR assay, the expression pattern of each candidate gene can be understanded. Statistica results indicated that there were12rice endosperm-specific genes. Here we further studied one Gene, named Latex-1for the Latex-related Gene. The RT-PCR analysis showed that Latex-1gene specifically expressed in rice endosperm and did not express in rice embryo and other vegetative organs, such as roots, stems, leaves, flower. Real-time fluorescence quantitative PCR analysis also indicated that the Latex-1gene only expressed in endosperm and its expression level was strong, which was closely related to the grouting process of rice endosperm.For further analysis on the cis-elements of Latex-1promoter, we apply promoter analysis software plantCARE, PLACE and plantpromoterdb2.0. The upstream approximate2.1kb promoter region of Latex-1gene was analyzed, there also existed five significant regulation motifs:AACA motif, GCN4motif, Prolamin-box, ACGT motif, Skn-I like motif, which were necessary for endosperm-specific expression and were found in the corresponding sites of Latex-1promoter. The distribution of these significant regulation elements met endosperm specific promoter sequence characteristic and they intertacted with each other for gene endosperm-specific expression.Then5’end deletion promoter fragments of Latex-1gene were fused into GUS reporter gene and transformed into rice. Meanwhile the approximate2.3kb GluC promoter also was fused into GUS report gene and transformed into rice by agrobacterium-mediated genetic transformation. Molecular detection for transgenic seedlings and the histochemical staining analysis revealed that GUS expression could not be detected in the transformed rice roots, stems, leaves, flower, but clearly observed in seed endosperm. The preliminary results showed that Latex-1promoter was an ideal endosperm specific and strong expression promoter.