摘要
连翘既是传统的中药材,又是优良的城市绿化树种,具有重要的经济价值和生态价值.但是连翘基因资料非常匮乏,限制了其分子生物学和基因功能的研究.本研究以连翘根、茎、叶、花和果实等器官的混合样品作为材料,利用Illumina Hi SeqTM 2500测序平台对其进行转录组测序.共获得23164327条干净数据(clean reads),总碱基数为4678791021 bp.Clean reads经de novo组装后获得45112条unigenes.进一步利用五大公共数据库进行同源比对,注释了28699条unigenes.其中,473个基因参与了连翘次生物质的合成和代谢,包括81个与苯丙氨酸和苯丙烷代谢相关的基因.对这81个基因的分析表明,有4个基因编码苯丙氨酸脱氨酶,1个基因编码肉桂酸4-羟化酶,2个基因编码4-香豆酰:辅酶A连接酶.这3个酶催化了连翘中主要药用活性物质苯乙醇苷和木脂素前体肉桂酸衍生物的生物合成.此外,还发现了2个松脂醇-落叶松树脂醇还原酶和1个开环异落叶松脂醇脱氢酶编码基因,这2个酶是木脂素合成的关键酶.最后,分析了长度在1 kb以上的12721个unigenes的基因结构,检测到3199个SSR位点,并对其中40个位点进行了验证.本研究不仅为连翘基因克隆和分子生物学研究提供了丰富的基础数据信息,而且为连翘遗传多样性研究、指纹图谱构建和分子标记辅助选育奠定了分子基础.
F. suspense is not only an important traditional Chinese medicine and but also an excellent urban greening tree species, thus has important economic and ecologic values. However, the lack of gene resources limits the molecular biology and gene functional studies in F. suspensa. In this study, the transcriptome of the mixed samples from root, stem, leaf, flower and fruit of F. suspensa was sequenced using the Illumina HiSeqTM 2500 sequencing platform. After high through-put sequencing, a total of 23164327 clean reads with 4678791021 bp were obtained. The clean reads were then de novo assembled into 45112 unigenes, and 28699 unigenes were annotated by a similarity search against five public databases. Of these annotated unigenes, 473 genes were assigned to second metabolic pathway, including 81 genes associated with phenylalanine and phenylpropanoid metabolism. Further analysis of these 81 genes reveled that there were 4 genes encoding phenylalanine ammonia-lyase, 1 gene encoding cinnamate 4-hydroxylase and 2 genes encoding 4-coumarate： CoA ligase, respectively. These enzymes are involved in the biosynthesis of cinnamic acid derivatives, which are precursors of the mainly pharmaceutically active substances phenylethanoid glycosides and lignan in F. suspensa. In addition, we also identified 2 pinoresinol/lariciresinol reductase encoding genes and 1 secoisolariciresinol dehydrogenase encoding gene, which are the key genes in the lignan biosynthesis. Finally, a total of 3199 SSRs were identified from 12721 unigenes longer than 1 kb, and 40 of them were verified. This work not only provides many valuable basal data for gene cloning and molecular biology research, but also lays the foundation for genetic diversity analysis, finger printing construction and molecular marker assisted breeding in F. suspensa.
出处
《中国科学：生命科学》
CSCD
北大核心
2015年第3期301-310,共10页
Scientia Sinica(Vitae)
基金
山西省科技攻关项目(批准号:20120313015-9)
山西省自然科学基金(批准号:2013011028-1)
山西省卫生厅中药现代化科技产业基地建设项目资助
关键词
RNA-SEQ
转录组
次生代谢
苯乙醇苷
木脂素
SSR分子标记
连翘
RNA-Seq, transcriptome, second metabolism, phenylethanoid glycoside, lignan, SSR molecular marker,Forsythia suspensa
