枸杞苯丙氨酸解氨酶基因的克隆与表达分析

发布时间：2018-11-22 19:06

【摘要】：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。
[Abstract]:In order to study the role of phenylalanine aminolytic enzyme (LcPAL) gene of Lycium barbarum in drought and salt stress, LcPAL gene (Genbank No.KX781247) was cloned by RT-PCR and RACE. The cDNA length of the gene is 2 544 BP, which contains a complete open reading frame of 2 163bp and encodes 720 amino acids. Protein sequence analysis showed that the (GTITASGDLVPLSYIA), containing the typical PAL enzyme activity center sequence had high homology with other plant PAL proteins. Phylogenetic tree showed that the PAL protein of Lycium barbarum LcPAL and Solanaceae plants were clustered together, which indicated that the relationship between them was close. The expression of LcPAL gene was found to be the highest in the leaves of Lycium barbarum and the least in the fruit by real-time fluorescence quantitative PCR. It was induced to express under the stress of polyethylene glycol (PEG), sodium chloride (NaCl), but the expression pattern was different. The results suggested that phenylalanine aminolytic enzyme (LcPAL), was a typical member of the PAL family, which played an important role in the development of Lycium barbarum tissues and played a role in drought resistance and salt stress resistance of Lycium barbarum.
【作者单位】：青海师范大学青藏高原药用动植物资源重点实验室;青海大学农林科学院农业部农产品质量安全风险评估实验室;
【基金】：青海省科技厅项目(2015-ZJ-735);青海省科技厅重点项目(2017-zj-y13) 教育部项目(Z2016098)
【分类号】：Q943.2;S567.19

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