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利用数据库中已发表的拟南芥AtRTE1基因序列,从构建的月季花朵乙烯响应EST数据库中筛选得到一个RTE1同源片段,结合RACE技术,从月季切花品种‘Samantha’花瓣中克隆得到全长为1233bp的RTE1同源基因。该基因包含一个732bp的开放阅读框、339bp的5′非翻译区和162bp的3′非翻译区,编码一个243aa的肽链,其理论分子量为27.6kD,等电点为6.87。该基因推定的氨基酸序列与拟南芥AtRTE1同源性较高,属于RTE家族中的第1组别基因,命名为RhRTE1。该基因在月季花朵自然开放中表达逐步增强,并能够被外源乙烯和切伤处理所显著诱导。将35S::GFP-RhRTE1融合蛋白转入拟南芥,发现RhRTE1蛋白主要定位于细胞膜上,并且转化植株降低了对乙烯的敏感性。以上结果说明RhRTE1可能参与了乙烯介导的月季花朵开放过程,并可能参与了花朵响应伤害的修复过程。
A RTE1 homology fragment was screened from the constructed EST database of rose flowers with the AtRTE1 gene sequence of Arabidopsis thaliana distributed in the database. RTE1 was cloned from rose petal ’Samantha’ by RACE, and the full length was 1233 bp Of RTE1 homologs. The gene contains a 732bp open reading frame, 339bp 5 ’untranslated region and 162bp 3’ untranslated region, encoding a 243aa peptide chain with a theoretical molecular weight of 27.6kD and an isoelectric point of 6.87. The putative amino acid sequence of the gene has high homology with Arabidopsis AtRTE1 and belongs to the first group of genes in the RTE family and is named RhRTE1. This gene is gradually expressed in the natural opening of rose flowers and can be significantly induced by exogenous ethylene and wound healing. The 35S :: GFP-RhRTE1 fusion protein was transferred into Arabidopsis thaliana and RhRTE1 protein was found to be located mainly on the cell membrane, and the transformed plants reduced the sensitivity to ethylene. The above results indicate that RhRTE1 may be involved in the ethylene-mediated rose opening process and may be involved in the flower-healing process.