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为了研究夜来香生物钟和花香代谢机制,本研究以夜来香花瓣为材料,利用PCR技术,从中获得了夜来香LIS基因的全长cDNA,并命名为CnLIS。生物信息学分析结果表明:该cDNA序列全长为1 800 bp,编码560个氨基酸,其中ORF为1 683 bp,5’UTR为56 bp,3’UTR为61 bp;该序列编码的氨基酸序列与软枣猕猴桃、温州蜜柑LIS基因的同源性分别为55%、50%,属于类异戊二烯合成C1超级家族。实时荧光定量PCR分析结果表明,0~12 h CnLIS表达水平较低,16 h后表达量明显上调,20 h表达量达最大,随后表达量急剧下降,其表达量随花瓣开放与闭合呈节律性变化。推测CnLIS可能参与夜来香香气形成,调控花香形成过程,同时,该基因的表达可能受生物钟基因所调控。
In order to study the biological clock and floral metabolism mechanism, the full-length cDNA of LIS was obtained from Caryophylla trifoliata using PCR technique. The full-length cDNA was named as CnLIS. The results of bioinformatics analysis showed that the full length of this cDNA was 1 800 bp, encoding 560 amino acids. The ORF was 1 683 bp, the 5 ’UTR was 56 bp, and the 3’ UTR was 61 bp. The amino acid sequence The homology of the LIS genes in jujube and mandarin was 55% and 50% respectively, belonging to the C1 superfamily of isoprenoid synthesis. Real-time PCR analysis showed that CnLIS expression was low at 0-12 h, significantly up-regulated after 16 h, and reached the maximum at 20 h, then decreased sharply. The expression of CnLIS was decreased with the opening and closing of petals Variety. It is speculated that CnLIS may participate in the formation of the nocturnal aroma and regulate the floral formation process. Meanwhile, the expression of CnLIS may be regulated by the circadian clock gene.