金柑FcSOC1同源基因的克隆及表达分析

Cloning and Expression Analysing of FcSOC1 Homologous Gene from Fortunella crassifolia Swingle

通过检测这对同源基因的表达水平,来分析这对同源基因与花器官发育的关系。采用同源克隆技术从金柑花中克隆出Fc SOC1基因的c DNA全长序列并进行生物信息分析;利用实时荧光定量PCR技术分析Fc SOC1基因在金柑不同组织、器官的表达特性。序列分析表明获得这对同源基因全长为660 bp和636 bp,分别编码220和212个氨基酸,分别命名他们为Fc SOC1a和Fc SOC1b。同源性分析显示,其与同属于芸香科的克里曼丁桔,相似性达98%,和甜橙的相似性也达到90%以上。实时荧光定量PCR表示Fc SOC1a和Fc SOC1b在营养器官和生殖器官均有表达。在营养器官的表达略有不同,但是在生殖器官的表达均在花蕾发育初期与后期急剧升高,并且在花中的表达量明显大于在其他部位的表达量。这对同源基因的高表达部位为茎、叶、花蕾、花芽、花瓣、花托、子房。Fc SOC1a和Fc SOC1b在花中显著表达,根据其表达特性推测,其可能在调控花器官发育上起着重要作用。根据这两个基因在营养器官的表达不同以及之间同源性较低,说明其功能已经出现分化。

In this paper, the relationship between floral organ development and the expression of the pair of homologous gene was analysied through detecting the expression level of the pair of homologous gene. Using the Fortunella crassifolia Swingle flowers as the experimental material, we cloned the homologous full-length c DNA sequences of Fc SOC1 gene by the homologous cloning technology. The expression characteristics in different organs of Fortunella crassifolia Swingle were analyzed by real-time fluorescence quantitative PCR. Sequence analysis showed that the c DNA of this pair of homologous genes were 660 bp and 636 bp, encoded 220 and 212 amino acids. They were named Fc SOC1 a and Fc SOC1 b respectively. The similarity of the amino acid sequences between the genes and other Rutaceae plants was more than 90. Real-time fluorescent quantitative PCR indicated that Fc SOC1 a and Fc SOC1 b could express in both vegetative and reproductive organs. The expression levels of Fc SOC1 a and Fc SOC1 b in vegetative organs were different, but they both had a high level expression in flowers,and the expresion rised sharply at the eary stage and later stage of the dev elopment of flower buds. These two genes highly expressed in the stems, leaves, flower bud, inforescences, torus and ovary of flowers. Both Fc SOC1 a and Fc SOC1 b were expressed significantly in flowers, and their expression characteristics indicated that they may play an important role in dominating the growth of flower, and this pair of homologous genes may have gene differentiation because they have different function in the growth of vegetative organs.