摘要摘 要 新蚜虫疠霉(Pandora neoaphidis)是虫霉门(Entomophthoromycota)中极具代表性的蚜虫专化性病原真菌，分布广泛，可侵染多种蚜虫。适宜条件下可在较短的时期内形成流行病，在蚜虫自然防治中起主导作用。为了系统分析了两个阶段的基因表达和功能相关的转录组信息，本研究通过二代测序平台Illumina Hiseq 2500对新蚜虫疠霉的两种发育形态(菌丝和分生孢子)进行转录组测序。得到90 639个不含N(模糊碱基)的组装片段(contigs)和49 138条单基因簇(unigenes)。49 138条单基因簇经基因本体论(Gene Ontology, GO)功能显著性富集分类后归到47种功能分支中，相关的基因功能主要归类至分子功能、细胞组分和生化过程三大类中。根据每百万reads中来自于某基因每千碱基长度的reads数(reads per kilobase per million of mapped reads, RPKM)的表达量算法一共得到3 555个差异表达的基因，其中上调的基因有2 294个，下调的基因有1 261个。利用京都基因及基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)对差异表达基因进行途径显著性富集分析后发现与之相关的代谢路径有22条，其中糖胺聚糖降解和类胡萝卜素生物合成两条途径显著上调。本研究为进一步挖掘和分析与新蚜虫疠霉生长发育相关的功能基因提供了理论依据。
Abstract：Abstract Pandora neoaphidis, one of important Entomophthoromycota fungi, is a representative obligated aphid pathogenic fungi that can infect a variety of aphids and are widely distributed. Under suitable conditions, P. neoaphidis can cause a strong epidemic in the aphids in short term and plays a leading role in biological control of aphids. Here, the RNA-seq data of hyphae and primary conidia were carried out using second-generation sequencing platform Illumina Hiseq 2500, and the differentially expressed genes were assessed systematically. Totally, 90 639 contigs and 49 138 unigenes were detected among the RNA-seq data. Results based on Gene Ontology (GO) functional enrichment analysis showed that 49 138 unigenes were annotated to 47 function categories, covering 3 ontologies including molecular function, cellular component and biological processes. Besides, according to reads per kilobase per million of mapped reads(RPKM), a total of 3 555 genes expressed differentially, involving in 2 294 up-regulated genes and 1 261 down-regulated genes. Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis involved 22 metabolic pathways during the conidia stage, of which carotenoid biosynthesis and glycosaminoglycan degradation were significantly up-regulated against the hyphae stage. These basis data would provide a solid foundation for further excavation and analysis of functional genes related to the growth and development of P. neoaphidis.
[1]陈春.蚜虫专化性病原真菌(虫霉目)随寄主迁飞扩散而传播流行的生物学实证研究[J].浙江大学博士论文论文, 2005, 1(1):6-11[2]陈相波, 陈春, 沈金慧, 等.利用改良的方法快速调查田间新蚜虫疠霉的流行[J].中国生物防治学报, 2014, 30(6):731-735[3]李增智, 主编.中国真菌志[J].虫霉目, 2000, 1(1):1-24[4]薛承美, 解廷娜, 叶素丹, 等.利用实时荧光定量筛选新蚜虫疠霉内参基因[J].农业生物技术学报, 2014, 22(12):1575-1583[5]吴书峰, 王志, 乔治军, 等.大豆抗胞囊线虫号小种转录组测序及分析[J].农业生物技术学报, 2016, 24(4):519-529[6]Bayram ?, Krappmann S, Ni M, et al.VelBVeALaeA complex coordinates light signal with fungal development and secondary metabolism[J].Science, 2008, 320(5882):1504-1506[7]Chen C, Xie T N, Ye S D.Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis[J].Brazilian Journal of Microbiology, 2016, 47(1):259-265[8]Chen C H, De May B S, Gladfelter A S, et al.Physical interaction between VIVID and white collar complex regulates photoadaptation in Neurospora[J].Proceedings of the National Academy of Sciences of the United States of America, 2010, 107(38):16715-16720[9]Flaherty J E，Dunkle L D.Identification and expression analysis of regulatory genes induced during conidiation in Exserohilum turcicum[J].Fungal Genetics and Biology, 2005, 42(1):471-481[10]Ganem S, Lu S, Lee B, et al.G-Protein subunit of Cochliobolus heterostrophus involved in virulence,asexual and sexual reproductive ability,and morphogenesis[J].Eukaryotic Cell, 2004, 3(1):1653-1663[11]James T Y, Kauff F, Schoch C L, et al.Reconstructing the early evolution of Fungi using a six-gene phylogeny[J].Nature, 2006, 443(1):818-822[12]Krystofova S, Borkovich K A.The heterotrimeric G-Protein subunits GNG-1 and GNB-1 form a G dimer required for normal female fertility,asexual development and G protein levels in Neurospora crassa[J].Eukaryotic Cell, 2005, 4(1):365-378[13]Liu ZQ, Lin S, Baker PJ, et al.Transcriptome sequencing and analysis of the entomopathogenic fungus Hirsutella sinensis isolated from Ophiocordyceps sinensis[J].BMC Genomics, 2015, 16(1):106-123[14]Lauter F R, Yamashiro C T, Yanofsky C.Light stimulation of conidiation in Neuro sporacrassa: studies with the wild-type strain and mutants wc-1,wc-2 and acon-2[J].Journal of Photochemistry and Photobiology B: Biology, 1997, 37(3):203-211[15]Ma?agocka J, Grell M N, Lange L, et al.Transcriptome of an entomophthoralean fungus (Pandora formicae) shows molecular machinery adjusted for successful host exploitation and transmission[J].Journal of Invertebrate Pathology, 2015, 128(1):47-56[16]Pietro A D, Garcia F, Meglecz E, et al.A MAP kinase of the vascular wilt fungus Fusarium oxysporum is essential for root penetration and pathogenesis[J].Molecular Microbiology, 2001, 39(1):1140-1152[17]Qin L, Gong X Y, Xie J T, et al.Phosphoribosylamidotransferase,the first enzyme for purine de novo synthesis,is required for conidiation in the sclerotial mycoparasite Coniothyrium minitans[J].Fungal Genetics and Biology, 2011, 48(1):956-965[18]Xu J H, Feng M G.The time-concentration-mortality modeling and virulence indices for two entomophthoralean species,Pandora delphacis and Pneoaphidis,against the green peach aphid,Myzus persicae[J].Biological Control, 2000, 17(1):29-34[19]Zhang W, Liu H T.MAPK signal pathways in the regulation of cell proliferation in mammalian cells[J].Cell Research, 2002, 12(1):9-18
