Previously, we isolated a vernalization-related gene, VER2, from winter wheat (Triticum aestivum L.) and its expression was restricted in the immature leaves of vernal-ized wheat seedlings. To further investigate the ...Previously, we isolated a vernalization-related gene, VER2, from winter wheat (Triticum aestivum L.) and its expression was restricted in the immature leaves of vernal-ized wheat seedlings. To further investigate the regulation of VER2 expression and the function of its promoter, we iso-lated a 41.7 kb genomic clone containing VER2 gene from a transformation-competent artificial chromosome (TAC) li-brary of wheat (Triticum aestivum-Haynaldia villosa). The sequence analysis showed that there were eleven predicted genes in the TAC. The exons of gene 3 corresponded to the cDNA sequence of VER2 gene. Analysis of VER2 promoter structure showed that there were three small repeat se-quences divided by two large repeat sequences. The putative response elements, such as abscisic acid response elements (ABRE), MeJA-response elements (Me-JARE), low-tempe- rature response elements (LTR), endosperm expression ele-ments, MYB binding sites and similar elements to GA re-sponse elements (GARE), were involved in the VER2 pro-moter region. Construct containing the VER2 promoter (-5895 to +73) driving GFP reporter gene was bombarded into vernalized or non-vernalized immature leaves in wheat. The vernalized immature leaves showed bright green fluo-rescence after incubation for 24 h, however, the green fluo-rescence was not observed in the non-vernalization leaves under the same condition. These results suggested that vernalization was essential for the function of VER2 promoter in the immature leaves of winter wheat.展开更多
文摘Previously, we isolated a vernalization-related gene, VER2, from winter wheat (Triticum aestivum L.) and its expression was restricted in the immature leaves of vernal-ized wheat seedlings. To further investigate the regulation of VER2 expression and the function of its promoter, we iso-lated a 41.7 kb genomic clone containing VER2 gene from a transformation-competent artificial chromosome (TAC) li-brary of wheat (Triticum aestivum-Haynaldia villosa). The sequence analysis showed that there were eleven predicted genes in the TAC. The exons of gene 3 corresponded to the cDNA sequence of VER2 gene. Analysis of VER2 promoter structure showed that there were three small repeat se-quences divided by two large repeat sequences. The putative response elements, such as abscisic acid response elements (ABRE), MeJA-response elements (Me-JARE), low-tempe- rature response elements (LTR), endosperm expression ele-ments, MYB binding sites and similar elements to GA re-sponse elements (GARE), were involved in the VER2 pro-moter region. Construct containing the VER2 promoter (-5895 to +73) driving GFP reporter gene was bombarded into vernalized or non-vernalized immature leaves in wheat. The vernalized immature leaves showed bright green fluo-rescence after incubation for 24 h, however, the green fluo-rescence was not observed in the non-vernalization leaves under the same condition. These results suggested that vernalization was essential for the function of VER2 promoter in the immature leaves of winter wheat.
