目的 克隆灰毡毛忍冬苯丙氨酸解氨酶（LmPAL1）基因全长序列，并进行生物信息学和表达模式分析。方法 提取灰毡毛忍冬花总RNA，通过实时荧光定量PCR（qRT-PCR）和RACE技术克隆LmPAL1基因的全长cDNA序列；运用相关软件对该基因序列进行生物信息学分析；利用qRT-PCR测定灰毡毛忍冬茎、叶和不同花期花中该基因的相对表达量。结果 克隆的LmPAL1基因开放阅读框（ORF）长度为2 145 bp，编码714个氨基酸，生物信息学分析预测为亲水性蛋白，定位于叶绿体中，包含PAL shielding结构域（527~641 aa），并且含有PAL/HAL活性中心序列GTITASGDLVPLSYIAG（196~212 aa），与其他LmPAL1具有较高的相似度；LmPAL1基因qRT-PCR结果显示，7个花期材料以金黄色开花期相对表达量较高；与茎、叶比，白色花蕾期花的相对表达量最高，叶的最低。结论 成功克隆了LmPAL1基因，为进一步研究该基因的功能以及遗传改良灰毡毛忍冬品质奠定基础，同时为探究灰毡毛忍冬绿原酸生物合成和调节机制提供了依据。

Objective To clone the full length of LmPAL1 gene and analyze bioinformatics and expression patterns from Lonicera macranthoides.Methods The total RNA of L. macranthoides was extracted. The full-length cDNA sequence of LmPAL1 gene was cloned by RT-PCR and RACE technique; The genome sequence in bioinformatics was analyzed by using the relevant software; The relative expression of the gene in stem, leaf, and different flower period was determined by using real-time PCR.Results The cloned LmPAL1 gene open reading frame (ORF) was 2 145 bp, encoding 714 amino acids. It was predicted by bioinformatics analysis as hydrophilic protein, being located in the chloroplasts, containing PAL shielding structure domain (527-641 aa). This gene contained PAL/HAL active center sequence GTITASGDLVPLSYIAG (196-212 aa), which was highly similar to other phenylalanine ammonia-lyase. Real-time PCR results showed that the relative expression level of golden yellow flowering flower was higher in seven florescence periods. When comparing the stem, leaf, and white flower bud period, the relative expression of flower was the highest and the leaf was the lowest.Conclusion In this study, PAL1 gene of L. macranthoides was cloned successfully, laying a foundation for further study of the function of this gene and genetic improvement of L. macranthoides quality and providing the research basis for exploring the biosynthesis and regulation of chlorogenic acid in L. macranthoides.

国家自然科学基金资助项目（81673546）；国家自然科学基金资助项目（81203007）；中央引导地方科技发展专项资金项目（2017XF5044）；湖南省自然科学基金项目（2017JJ3237）；湖南省教育厅资助科研项目（16B193）；湖南省科技厅重点研发项目（2017SK2124）；湖湘中药资源保护与利用2011协同创新中心（湘教通[2015]351）；国家中医药管理局中药标准化项目（ZYBZH-Y-HUN-24）；湖南中医药大学中药学一流学科项目（校行科字[2018]3号）