摘要
【目的】对银耳进行转录组测序分析,分析多糖生物合成途径和挖掘关键功能基因。【方法】采用Illumina Hiseq 2500测序平台对银耳菌丝体进行测序,组装得到unigene序列,并与公共数据库进行对比分析和注释,挖掘果糖和甘露糖代谢通路相关基因。【结果】本研究共得到4.72 Gb raw data,拼接后获得17 008条unigene序列,N50为2073 bp;注释结果表明,76.47%序列能够注释到Uniprot公共数据库,注释到GO分类和KEGG通路的unigene分别为10 152和5671条。进一步分析银耳果糖和甘露糖代谢途径相关基因,挖掘得到74条unigene;其中编码L-iditol 2-dehydrogenase的unigene最多,其次是butanol dehydrogenase。【结论】基于转录组测序数据解析及预测,为研究银耳多糖和其它产物的生物合成途径及分子机制提供了数据支持,也为银耳品质的形成提供理论依据。
【Objective】The study aimed to study on the expression of genes involved in the biosynthesis of fructose and mannose metabolism and explore the functional genes in Tremella fuciformis, the transcriptome was sequenced. 【Methods】 By Illumina Hiseq 2500 platform sequencing technology, the transcriptome of the mycelium was sequenced, assembled and annotated. Moreover, unigene related to the fructose and mannose metabolism pathway were identified.【Results】4.27 Gb raw data was generated, and 17 008 unigenes were obtained after splicing, which the N50 was 2073 bp. 76.47 % unigene was annotated in the Uniprot database, which 10 152 and 5671 unigenes were annotated in the GO classification and KEGG pathways. The fructose and mannose metabolism pathway in Tremella fuciformis involved in 74 unigenes, especially unigene coding L-iditol 2-dehydrogenase and butanol dehydrogenase with the most unigenes. 【Conclusion】This study would be established for revealing the biosynthesis mechanism of polysaccharide and other production. At the same time, this study will provide a theoretical basis for quality formation of Tremella fuciformis.
引文
[1]徐锦堂,兰进.中国药用真菌学[M].北京:北京医科大学、中国协和医科大学联合出版社,1997:421-425.
[2]黎勇,王晓东,高敏.我国银耳的研究历史及现状[J].北方园艺,2014,16:188-191.
[3]梁勤,温文婷,叶小金,等.银耳种质资源遗传多样性及酶学特性分析[J].西南农业学报,2010,23(4):1194-1198.
[4]马素云,贺亮,姚丽芬.银耳多糖结构与生物活性研究进展[J].食品科学,2010,31(23):411-416.
[5]Wu Q,Zheng C,Ning Z X,et al.Modification of low molecular weight polysaccharides from Tremella Fuciformis and their antioxidant activity in vitro[J].International Journal of Molecular Sciences,2007,8(7):670-679.
[6]Wen L R,Gao Q,Ma C W,et al.Effect of polysaccharides from Tremella fuciformis,on UV-induced photoaging[J].Journal of Functional Foods,2016,20:400-410.
[7]Guo H,Yi W,Song J K,et al.Current understanding on biosynthesis of microbial polysaccharides[J].Current Topics in Medicinal Chemistry,2008,8(2):141-151.
[8]陈蕾蕾,王未名,祝清俊,等.细菌多糖的生物合成机制[J].微生物学报,2010,50(12):1583-1589.
[9]王东,宋君,赵树海,等.银耳的分子生物学研究进展[J].北方园艺,2018(4):169-174.
[10]Nagalakshmi U,Wang Z,Waern K,et al.The transcriptional landscape of the yeast genome defined by RNA sequencing[J].Science,2008,320(5881):1344-1349.
[11]Wei Z,Wang W,Hu P,et al.SNVer:a statistical tool for variant calling in analysis of pooled or individual next-generation sequencing data[J].Nucleic Acids Research,2011,39(19):e132.
[12]De W P,Pespeni M H,Ladner J T,et al.The simple fool's guide to population genomics via RNA-Seq:an introduction to high-throughput sequencing data analysis.Molecular[J].Ecology Resources,2012,12(6):1058.
[13]Huang Y T,Wu X Q,Jian D,et al.De novo transcriptome analysis of a medicinal fungi Phellinus linteus and identification of SSR markers[J].Biotechnology & Biotechnological Equipment,2015,29(2):395-403.
[14]Shu S H,Chen B,Zhou M C,et al.De novo sequencing and transcriptome analysis of Wolfiporia cocos to reveal genes related to biosynthesis of triterpenoids[J].Plos One,2013,8(8):e71350.
[15]Lu M Y,Fan W L,Wang W F,et al.Genomic and transcriptomic analyses of the medicinal fungus Antrodia cinnamomea for its metabolite biosynthesis and sexual development[J].Proceedings of the National Academy of Sciences of the United States of America,2014,111(44):e4743-e4752.
[16]聂文强,吴天祥,钟敏,等.真菌灰树花菌丝体转录组测序及分析[J].食品科学,2017,38(20):6-11.
[17]Tang L H,Jian H H,Song C Y,et al.Transcriptome analysis of candidate genes and signaling pathways associated with light-induced brown film formation in Lentinula edodes[J].Applied Microbiology & Biotechnology,2013,97(11):4977-4989.
[18]吴小梅,张昕,李南羿.双孢蘑菇子实体不同发育时期的转录组分析[J].菌物学报,2017,36(2):193-203.
[19]Zhang J X,He C M,Wu K L,et al.Transcriptome Analysis of Dendrobium officinale and its Application to the Identification of Genes Associated with Polysaccharide Synthesis[J].Frontiers in Plant Science,2016,7(5):1-14.