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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。
By examining the expression levels of these homologous genes, we analyzed the relationship between the homologous genes and flower organ development. The full-length cDNA sequence of Fc SOC1 gene was cloned from the kiwifruit by homologous cloning technology and bioinformatics analysis was performed. The expression of Fc SOC1 gene in different tissues and organs of kumquat was analyzed by real-time fluorescence quantitative PCR. Sequence analysis showed that the full length of the homologous genes was 660 bp and 636 bp, encoding 220 and 212 amino acids, respectively, which were named Fc SOC1a and Fc SOC1b respectively. Homology analysis showed that the similarity with kremlin, belonging to the same genus Rutaceae was 98%, and the similarity with orange was more than 90%. Real-time fluorescence quantitative PCR showed that both Fc SOC1a and Fc SOC1b were expressed in vegetative and reproductive organs. The expression in vegetative organs was slightly different, but the expression in reproductive organs increased sharply in the early and late stages of flower bud development, and the expression level in flowers was significantly greater than that in other parts. The high expression of these homologous genes stems, leaves, buds, flower buds, petals, receptacle, ovary. Fc SOC1a and Fc SOC1b are highly expressed in flowers, suggesting that they may play important roles in the regulation of floral organs development. According to the different expression of these two genes in vegetative organs and the low homology between them, indicating that their function has been differentiated.