摘要
采用Illumina Hi-seq 2500转录组高通量测序,构建黄秋葵花和果荚转录组文库,并利用测序评估、差异基因功能注释等生物信息学方法进行分析。研究结果表明:(1)分别获得黄秋葵花和果荚有效数据7.12Gb和8.14Gb,碱基百分比(Q30)均达到91.0%以上。(2)获得差异表达基因(DEGs)1 336个,其中上调基因319个,下调基因1 017个。(3)获得功能注释的基因有1 131个,GO将455注释基因归成41个功能小类,主要涉及代谢过程、催化活性、单一生物过程和细胞过程等过程;KOG注释了472个DEGs,涉及23个功能分类,其中与次生代谢直接相关过程O和Q类别获得111个注释结果;KEGG将372个DEGs注释到80条代谢通路中,获得F3H、F3′5′H、DFR、ANR、ANS、GT、LAR共10个关键差异基因,其中F3H、DFR在黄秋葵花中表现上调效应,F3′5′H、ANR、LAR在黄秋葵果荚中表现显著上调效应,ANS、GT则分别在花和果荚中均有上调或下调效应。(4)实时定量PCR分析显示,其中7个差异表达基因得到的相对表达量与转录组表达谱分析趋势完全一致。(5)类黄酮代谢途径分析表明,黄秋葵花通过F3H、DFR、ANS、GT途径将NAR催化为生成花青素苷;果荚则将NAR通过F3′5′H将催化为DHM,后在FLS催化下生成黄酮醇类物质等;部分NAR在F3H、DFR催化下,生成无色飞燕草素苷元,再分别在ANS、LAR作用下,进入花青素苷元和原花青素合成途径。该研究结果丰富了黄秋葵转录组信息,为黄秋葵类黄酮物质纯化和功能利用提供参考依据。
The cDNA of okra(flowers and fruits)were sequenced based on Illumina Hi-seq2500and were analyzed by using the bioinformatics methods subsequently,such as sequencing assess and gene function annotation.The result of research shows that:(1)7.12Gb and8.14Gb Clean Data were obtained respectively from the flowers and fruits and the base ratios with quality values higher than30in reads(Q30)were more than91.0%from both.(2)Compared the transcriptome of the flowers and fruits,1131differentially expressed genes(DEGs)were found,including319up-regulated genes and1017down-regulated genes.(3)Annotation analysis indicated that1131genes were annotated.With GO function annotation classification,455functional annotations of these DEGs were divided into41function categories,in which many function categories were mainly involved,such as metabolic process,catalytic activity,single-organism process,cellular process.Through KOG analysis,there were472functional annotations of DEGs,involving23functional classifications.KEGG analysis showed that372DEGs were annotated to80metabolic pathways,and obtained10key DEGs,F3H,F3′5′H,DFR,ANR,ANS,GT,LAR.F3H and DFR showed up-regulated effect in okra flowers;F3′5′H,ANR and LAR showed down-regulated effect in okra fruits;ANS and GT showed up/down-regulated effects in flowers and fruits.(4)The relative expression levels of7DEGS wasin line with that of the transcriptional group by qRT-PCR.(5)Through the flavonoid metabolic pathway,the research showed that anthocyanin were catalyzed by F3H,DFR,ANS(c83240),GT(c82265/c82004)from NAR in okra flowers;The formation of DHM was initiated by F3′5′H,then produced flavonols by FLS;Partly NAR was catalyzed by F3H and DFR to dephinidin,then ultimately went into anthocyanins and proanthocyanidins synthesis pathways.This study enriched transcriptome information of okra and provided reference for the purification and functional utilization of flavonoids in okra.
作者
姚运法
张少平
练冬梅
赖正锋
黄慧明
洪建基
YAO Yunfa;ZHANG Shaoping;LIAN Dongmei;LAI Zhengfeng;HUANG Huiming;HONG Jianji(Subtropical Agriculture Research Institute, Fujian Academy of Agricultural Sciences, Zhangzhou, Fujian 363005, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2018年第11期2000-2009,共10页
Acta Botanica Boreali-Occidentalia Sinica
基金
福建省公益类项目(2016R1012-1)
福建省农业科学院芳香植物创新科技团队(STIT2017-2-11)
