龙眼胚性愈伤组织AGO6基因克隆及其在体胚发生过程中的表达分析

Cloning of AGO6 from Embryogenic Callus and Its Expression Analysis by qPCR during Somatic Embryogenesis in Dimocarpus longan Lour.

依据龙眼胚性愈伤组织转录组数据库Unigene序列,利用RT-PCR结合RACE技术,以龙眼胚性愈伤组织c DNA为模板,获得Dl AGO6基因的c DNA全长序列,共3 168 bp(登录号为KF819529),完整开放阅读框2 700 bp,编码900个氨基酸。生物信息学分析表明:Dl AGO6的c DNA所编码的氨基酸序列含有2个高度保守的PAZ和PIWI结构域,具有典型的AGO类蛋白的结构特征;与拟南芥AGO6蛋白序列有较高的同源性。龙眼体胚发生过程中Dl AGO6表达量的q PCR分析表明:Dl AGO6在龙眼松散型胚性愈伤组织时期的表达量最高,在鱼雷形胚时期表达量最低,推测DLAGO6在龙眼松散型胚性愈伤组织阶段的高表达量,可能与细胞的旺盛分裂有关。

Based on the database of longan transcriptome sequences,the Dl AG06 from longan embryogenic callus was isolated by using RACE and PCR. The results showed that the cloned full—length c DNA sequence of Dl AGO6 was 3 168 bp in length（ Genbank： KF819529）,containing a 2 700 bp open reading frame（ ORF） encoding 900 amino acids.Bioinformatics predicted that the deduced Dl AGO6 protein with two conserved PAZ and PIWI domains had the typical characteristics of the AGO family,which had a high identity with the known Arabidopsis thaliana AGO6 protein. The q PCR analysis indicated that the expression level of Dl AGO6 gene was the lowest at the stage of torpedo embryo and reached the maximum at the stage of the friable-embryogenic callus,which suggested that Dl AGO6 may be associated with exuberant cell division.