火鹤AaSERK基因克隆及其在体细胞胚胎发生中的表达分析

Clone and Expression of AaSERK Gene in Anthurium andraeanum's Somatic Embryogenesis

为了探讨体细胞胚胎发生相关类受体蛋白激酶（SERK）基因在火鹤体细胞胚胎发生中的作用,以火鹤胚性愈伤组织为材料,利用RT-PCR结合RACE技术,克隆了火鹤体细胞胚胎发生相关类受体蛋白激酶基因（AaSERK）,通过实时荧光定量PCR（Real time-PCR）技术分析了AaSERK的相对表达量。结果显示：（1）该基因全长为1 949bp,包含1 866bp开放阅读框,编码蛋白由622个氨基酸组成。（2）预测该基因具有SERK家族典型的结构域：1个信号肽、1个亮氨酸拉链结构域、5个富亮氨酸重复序列结构域、1个SPP基序、1个跨膜结构域、含11个亚区的激酶结构域、1个C端结构域。DNAMAN分析显示,该基因编码的氨基酸序列与其它植物的一致性达到65%-89%。（3）在离体诱导体细胞胚胎发生的各阶段中,AaSERK基因在诱导阶段及发育培养阶段微弱表达或者几乎不表达,在继代培养第30天的胚性愈伤组织中表达量最高。推测该基因可以作为火鹤体细胞胚胎发生的一个标记基因。

To study the function of somatic embryogenesis receptor-like kinase gene （SERK） in Anthurium andraeanum＇ s somatic embryogenesis,we isolated a novel AaSERK gene from the embryogenic callus of A. andraeanum by reverse transeription-polymerase chain reaction （RT-PCR） and rapid amplification of cDNA ends （RACE）. The expression levels of AaSERK were analyzed by Real time-PCR. The results showed that： （1）The full-length cDNA of AaSERK was 1 949 bp with an open reading frame of 1 866 bp encoding a protein of 622 amino acid residues. （2）It was speculated that the AaSERK wag a typical SERK gene with one signal peptide,one leucine zipper domain,five Leu-rich repeat（LRR） domains,one SPP mo- tif,one transmembrane domain,the kinase domain with eleven subdomains, and one C-terminal domain. DNAMAN analysis showed that the putative AaSERK amino acid sequence had 65%-89% identity with those of the other plants. （3）In the different stages of somatic embryogenesis in vitro, the expression of AaSERK was weak or little in the inducing stage and the developmental stage,but was the highest in the embryogenic callus on the 30th day of the subculture stage. It was speculated that the AaSERK might be one o{ the marker genes o{ somatic embryogenesis in A. andraeanum.