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为研究枸杞苯丙氨酸解氨酶基因LcPAL在干旱和盐逆境中的作用,采用RT-PCR和RACE方法克隆了LcPAL基因(Genbank No.KX781247)。该基因cDNA长度为2 544bp,含有2 163bp的完整开放阅读框,编码720个氨基酸。蛋白序列分析表明,其包含典型的PAL酶活性中心序列(GTITASGDLVPLSYIA),与其他植物的PAL蛋白有很高的同源性。系统进化树表明,枸杞LcPAL与茄科植物的PAL蛋白聚为一类,说明两者的亲缘关系较近。利用实时荧光定量PCR方法检测发现,LcPAL基因在枸杞的叶中表达量最高,果实中表达量最少。其在聚乙二醇(PEG)、氯化钠(NaCl)胁迫下诱导表达,但表达模式不同。研究结果推测从枸杞中克隆获得苯丙氨酸解氨酶(LcPAL),是典型的PAL家族成员,在枸杞各组织发育过程中具有重要功能,且在枸杞抗干旱和抗盐逆境过程也起一定作用。
In order to study the role of Lycium phenylalanine ammonia lyase gene LcPAL in drought and salt stress, LcPAL gene (Genbank No. KX781247) was cloned by RT-PCR and RACE. The gene cDNA length of 2 544bp, contains 2 163bp complete open reading frame, encoding 720 amino acids. Protein sequence analysis showed that it contained a typical GERTASGDLVPLSYIA, which had high homology with PAL proteins of other plants. Phylogenetic tree showed that Lycium barbarum LcPAL and Solanaceae PAL protein clustered together, indicating that the two are relatively close genetic relationship. Real-time PCR detection showed that the expression level of LcPAL gene was the highest in the leaves of Lycium barbarum and the lowest in the fruit. The expression was induced by polyethylene glycol (PEG) and sodium chloride (NaCl), but the expression patterns were different. The results presumed that the phenylalanine ammonia-lyase (LcPAL) cloned from Lycium chinense L. is a typical PAL family member and plays an important role in the development of Lycium barbarum, and plays an important role in the anti-drought and salt-tolerance stress of Lycium barbarum effect.