摘要
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.
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.
关键词
冬小麦
促进剂
春化作用
人工染色体
GFP, pool-PCR, promoter, TAC, transient expression, Triticum aestivum L., vernalization.
