栝楼黄酮类生物合成关键酶基因的挖掘  

Mining of Key Enzyme Genes for Flavonoid Biosynthesis of Trichosanthes kirilowii

作　　者：胡秀芹 梁秀坤 张文玉 刘言娟[2] 辛杰 张永清[1] HU Xiu-qin;LIANG Xiu-kun;ZHANG Wen-yu;LIU Yan-juan;XIN Jie;ZHANG Yong-qing(College of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Medical College,Linyi University,Linyi 276000,China;Binzhou Inspection and Testing Center,Binzhou 256600,China)

机构地区：[1]山东中医药大学药学院,山东济南250355 [2]临沂大学医学院,山东临沂276000 [3]滨州市检验检测中心,山东滨州256600

出　　处：《中药材》2024年第7期1655-1660,共6页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金项目(81803651)。

摘　　要：目的:深入挖掘栝楼黄酮类次生代谢产物生物合成途径相关酶基因,以便解析栝楼黄酮类化合物生物合成的调控机制。方法:以栝楼雌雄株花芽为材料,采用转录组测序、生物信息学及功能注释等方法筛选差异表达基因(DEGs);对关键的酶基因进行qRT-PCR验证;利用HPLC法测定栝楼雌雄株叶、茎、花三个部位的总黄酮含量;应用Pearson法分析栝楼雌雄株不同部位基因表达量与总黄酮含量之间的相关性。结果:分别从雌雄株花芽文库中获得了17619567和16699544条高质量的reads,组装并去冗余后得到156032条Unigenes,经筛选得到25435条DEGs;KEGG代谢通路发现,有80条DEGs参与了栝楼黄酮类生物合成,其中包括ANR、CHS、HCT等关键酶基因;栝楼雄株各部位的总黄酮含量均高于雌株(P≤0.001),且黄酮生物合成途径排名前三的关键基因在雄株中的表达量显著高于雌株(P≤0.001);ANR、CHS和HCT基因与总黄酮含量在栝楼雌雄植株中呈现出不同的相关性。结论:该结果为进一步研究栝楼功能基因在黄酮类成分代谢路径中的调控作用以及雌雄株黄酮类成分差异的遗传机制奠定了基础。Objective:To explore the enzyme genes associated with the biosynthesis pathway of secondary metabolites of flavonoids from Trichosanthes kirilowii,so as to understand the regulatory mechanism of the biosynthesis of flavonoids of Trichosanthes kirilowii.Methods:The flower buds of female and male Trichosanthes kirilowii were used as materials,the differentially expressed genes(DEGs)by transcriptome sequencing,bioinformatics and gene function annotation were screened,qRT-PCR assay was used to verify the key en-zyme genes.The contents of total flavonoids in leaves,stems and flowers of female and male Trichosanthes kirilowii were determined by HPLC.The correlations between the expressions of genes and the contents of total flavonoids in different parts of Trichosanthes kirilowii were analyzed by Pearson method.Results:17619567 and 16699544 high quality reads were obtained from the female and male flow-er bud libraries,respectively.156032 Unigenes were obtained after assembly and redundancy removal,and 25435 DEGs were ob-tained.The KEGG metabolic pathway showed that 80 DEGs were involved in the flavonoid biosynthesis of Trichosanthes kirilowii,inclu-ding key enzyme genes such as ANR,CHS and HCT.The total flavonoid content of male Trichosanthes kirilowii was higher than that of female Trichosanthes kirilowii(P≤0.001),and the expressions of the top three key genes of flavonoid biosynthesis pathway in male Tri-chosanthes kirilowii were significantly higher than in female Trichosanthes kirilowii(P≤0.001).ANR,CHS and HCT genes showed dif-ferent correlations with total flavonoid content in male and female Trichosanthes kirilowii plants.Conclusion:The results lay a foundation for further study the regulatory role of functional genes in the metabolic pathway of Trichosanthes kirilowii and the genetic mechanism of the difference of flavonoids between male and female strains.

关 键 词：栝楼 次生代谢 生物合成 关键酶基因 黄酮类成分 

分 类 号：R282.71[医药卫生—中药学]