转录组测序法研究草珊瑚叶和根的基因差异表达  

Profiling Differential Gene Expressions in Leaves and Roots of Sarcandra glabra Based on Transcriptome

作　　者：车苏容[1] 张家源 卢伟 祁克明 魏艺聪[1] CHE Surong;ZHANG Jiayuan;LU Wei;QI Keming;WEI Yicong(Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China)

机构地区：[1]福建中医药大学,福建福州350122

出　　处：《福建农业学报》2020年第6期598-610,共13页Fujian Journal of Agricultural Sciences

基　　金：福建省自然科学基金项目(2017J01841)。

摘　　要：【目的】从基因表达的水平,初步分析草珊瑚叶和根之间次生代谢差异的分子机制,为二者之间临床疗效差异形成的分子机制分析提供信息。【方法】以福建省福州市的草珊瑚作为样品,采用Illumina HiSeqTM高通量测序技术测定草珊瑚叶和根的转录组,然后经过滤和Trinity组装,得到的unigenes再通过blast与Nr、Nt、Pfam、KOG、Swiss-Prot、Kegg和GO进行比对注释,并对叶和根的基因差异表达进行分析,尤其是对KEGG代谢通路富集的差异基因进行分析。【结果】转录组测序结果共获得0.4亿多个clean reads,经Trinity组装后共得到508271个unigenes,其平均长度为740 bp,最大长度为17.3 kb。基于blast分析,共有148561个unigenes在七大功能注释数据库中得到成功注释,占总基因数的58.80%。在分析基因表达水平差异时,发现草珊瑚叶和根的共同基因有93127个,叶和根的差异基因分别为36327个和52268个;同时还发现在29732种不同表达的unigenes中,有12511个上调基因和17221个下调基因;代谢相关KEGG具有显著差异的通路有淀粉和蔗糖代谢、苯丙烷类生物合成、乙醛酸和二羧酸代谢、光合生物的固碳作用、吞噬体、谷胱甘肽代谢、光合作用、丙氨酸、天冬氨酸和谷氨酸代谢、倍半萜类和三萜类生物合成、卟啉和叶绿素代谢、氮素代谢、昼夜节律-植物、光合作用-天线蛋白、芪类、二芳基庚酸和姜酚生物合成、不饱和脂肪酸生物合成、柠檬烯和蒎烯降解、类胡萝卜素生物合成、二萜类生物合成、类黄酮生物合成、脂肪酸延伸等。其中与药效密切相关的次生代谢通路苯丙烷类、倍半萜类和三萜类、二萜类、类黄酮类生物合成等途径分别有193个、82个、40个、35个差异表达基因,而上调倍半萜合酶、ent-kaur-16-烯合酶、黄酮醇合酶/黄烷酮3-羟化酶等基因和下调8-羟基香叶醇脱氢酶、vinorine合酶、角鲨烯合酶等关键酶基【Objective】Base on transcriptome sequencing,the molecular mechanisms that caused the secondary metabolic differences between the leaves and roots of Sarcandra glabra were studied for clinic applications of the two parts,as well as for determination of the effective components in the medicinal herb.【Method】Specimens of S.glabra were collected from Fuzhou,Fujian for a transcriptome analysis on the leaves and roots using the Illumina HiSeq platform.After filtration and the Trinity assembly,the unigenes were compared with Nr,Nt,Pfam,KOG,Swiss-Prot,KEGG,and GO by BLAST,and the differentially expressed genes analyzed.A special attention was paid on the differentially enriched genes in the KEGG metabolic pathway.【Result】More than 40 million clean reads were obtained from the sequencing.The Trinity assembly yielded 508271 unigenes with an average length of 740 bp.Based on BLAST,148561 unigenes,accounting for 58.80% of the total,were successfully annotated using 7 functional annotation databases.There were 29732 unigenes identified with differential gene expressions between the leaves and the roots.Of which,12511 were up-regulated and 17221 down-regulated.Dozens of significantly different KEGG metabolic pathways were found that associated with functions such as starch/sucrose metabolism,phenylpropanoid biosynthesis,glyoxylate/dicarboxylate metabolism,carbon fixation in photosynthetic organisms,phagosome/glutathione metabolism,photosynthesis,alanine/aspartate/glutamate metabolism,sesquiterpenoid/triterpenoid biosynthesis,porphyrin/chlorophyll metabolism,nitrogen metabolism,circadian rhythm-plant/photosynthesis-antenna proteins,stilbenoid/diarylheptanoid/gingerol biosynthesis,unsaturated fatty acids biosynthesis,limonene/pinene degradation,carotenoid biosynthesis,diterpenoid biosynthesis,flavonoid biosynthesis,fatty acid elongation,etc.Insofar as pharmacodynamics is concerned,the secondary metabolic pathway of the phenylpropanoid biosynthesis had 193 differentially expressed genes between the leaves and the root

关 键 词：草珊瑚 叶 根 Illumina测序 基因差异表达分析 次生代谢 

分 类 号：R932[医药卫生—生药学]