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Characterisation, expression and functional analysis of PAL gene family in Cephalotaxus hainanensis

文献类型： 外文期刊

作者： He, Yuedong 1 ; Zhong, Xiaohong 1 ; Jiang, Xuefei 3 ; Cong, Hanqing 2 ; Sun, Huapeng 1 ; Qiao, Fei 2 ;

作者机构： 1.Hunan Agr Univ, Coll Hort, Changsha, Peoples R China

2.Chinese Acad Trop Agr Sci, Trop Crops Genet Resources Inst, Key Lab Crop Gene Resources & Germplasm Enhanceme, Minist Agr, Haikou, Hainan, Peoples R China

3.Hainan Univ HNU, Coll Hort, Hainan Key Lab Sustainable Utilizat Trop Bioresou, Haikou, Hainan, Peoples R China

关键词： Cephalotaxus hainanensis; Phenylalanine ammonia-lyase; Cephalotaxus alkaloids; Cloning; Expression; Functional analysis

期刊名称：PLANT PHYSIOLOGY AND BIOCHEMISTRY （ 影响因子：4.27； 五年影响因子：4.816 ）

ISSN： 0981-9428

年卷期： 2020 年 156 卷

页码：

收录情况： SCI

摘要： Phenylalanine ammonia lyase (PAL) is the first committed step in the formation of phenylpropanoids, and catalyses the deamination of L-phenylalanine (L-Phe) to yield cinnamic acid. While PALs are common in plants, PAL genes involved in alkaloid biosynthesis in Cephalotaxus hainanensis have never been described. To obtain better knowledge of PAL genes and their number and function involved in Cephalotaxus alkaloid biosynthesis four PAL genes were screened and cloned. In vitro enzymatic analysis showed that all four PAL recombinant proteins could convert L-Phe to product trans-cinnamic acid, and showed strict substrate specificity. Moreover, the expression profiles of four ChPALs were analysed using qRT-PCR, which showed that they had higher transcript levels in roots and stems, and that different ChPALs displayed different response sensitivities and change patterns in response to stimuli. Several metabolic compounds were measured in stimulated leaves using UPLC-MS, and indicating the concentration of Cephalotaxus alkaloids and cinnamic acid in leaves subjected to different conditions. These concentrations increased significantly after treatment with 100 mM NaCl, 100 mM mannitol, 100 mu M SA and 10 mu M ABA. The expression levels of four PAL genes showed indications of upregulation after treatment. These results supply an important foundation for further research on candidate genes involved in the biosynthesis of Cephalotaxus alkaloids.

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