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为探讨肌动蛋白解聚因子(actin depolymerizing factor,ADF)是否参与乙烯对月季花瓣细胞扩展的调节,以切花月季‘Samantha’为试材,从花瓣中分离到响应乙烯的ADF基因,并将其命名为Rh-ADF1,在GenBank中的登录号为JN048105。推定的氨基酸序列分析表明,Rh-ADF1的开放阅读框长度为423 bp,编码140个氨基酸。半定量RT-PCR分析结果表明,Rh-ADF1在花朵的各个器官中均有表达,在花瓣中略高于其他器官;该基因表达能够被乙烯处理强烈诱导升高,被乙烯作用抑制剂1-MCP(1-Methylcyclopropene)处理明显降低。在洋葱表皮细胞中进行的瞬时表达分析表明,Rh-ADF1蛋白分布在整个细胞中,主要与actin组成的微丝网络结合。据此推测,Rh-ADF1基因在转录水平上参与乙烯对花瓣扩展的调节,乙烯可能通过Rh-ADF1基因调控了花瓣细胞骨架的动态变化,进而影响花瓣细胞扩展和形态变化。
In order to investigate whether ADF is involved in the ethylene regulation of rose petal cell expansion, the ADF gene responsive to ethylene was isolated from the petals of Samantha ’Cut rose’ It was named Rh-ADF1 and its accession number in GenBank was JN048105. The putative amino acid sequence analysis showed that the open reading frame of Rh-ADF1 was 423 bp, encoding 140 amino acids. Semi-quantitative RT-PCR analysis showed that Rh-ADF1 was expressed in all organs of flowers and slightly higher than other organs in the petals. The expression of Rh-ADF1 was strongly induced by ethylene treatment and was inhibited by ethylene inhibitor 1-MCP (1-Methylcyclopropene) treatment was significantly reduced. Transient expression analysis in onion epidermal cells showed that the Rh-ADF1 protein was distributed throughout the cell and bound mainly to actin filaments. It is speculated that Rh-ADF1 gene is involved in ethylene regulation of petal expansion at the transcriptional level. Ethylene may regulate the dynamic changes of petal cytoskeleton via Rh-ADF1 gene, which may affect the expansion and morphological changes of petal cells.