论文部分内容阅读
笔者课题组首次发现可可毛色二孢菌(Lasiodiplodia theobromae)可导致贵州惠水茶树(Camellia sinensis)发生茶叶斑病。采用RNA-seq和miRNA-seq技术分析侵染茶树后病原菌的基因表达谱,可为L.theobromae和茶树的互作提供研究启示。本研究采用RNA-seq对侵染茶树后L.theobromae菌株GZHS-2017-010的mRNA进行测序,共获得41.82GB reads (样本数据量为6.29~7.90GB)。将reads拼接得到179 383条转录本,最终获得87 468个Unigenes,总长度为58.14Mb。52 840个Unigenes在至少一个数据库中被注释(占总数的60.41%)。采用eggNOG对Unigenes进行注释,10个COG分别为(O) Posttranslational modification,protein turnover,chaperones、(T) Signal transduction mechanisms、(K) Transcription、(G) Carbohydrate transport and metabolism、(L) Replication,recombination and repair、(U) Intracellular trafficking,secretion,and vesicular transport、(J)Translation,ribosomal structure and biogenesis、(E) Amino acid transport and metabolism、(I) Lipid transport and metabolism、(C) Energy production and conversion。Nr对Unigenes的分析表明,Diplodia corticola数据可注释4 942个Unigenes,Diplodia seriata数据可注释3 814个Unigenes,Macrophomina phaseolina数据可注释1 699个Unigenes,Rhizophagus irregularis数据可注释1 285个Unigenes,Neofusicoccum parvum数据可注释1 251个Unigenes,Spizellomyces punctatus数据可注释1 163个Unigenes,其他物种的数据可注释14 641个Unigenes。KEGG分析表明,大量的mRNA可富集在carbohydrate metabolism、amino acid metabolism等代谢通路中。miRNA-seq分析检测出547个known或conservative miRNA和100个novel miRNA。靶基因KEGG富集性分析表明,ptc-miR6427-3pL-1R-21ss16AT、sly-MIR319c-p52ss16CT18TC、cas-MIR157b-p5、mtr-miR168b、mtr-miR396b-3pL+2R-2、hbr-MIR6173-p32、aly-MIR157c-p31ss6GA、aly-MIR157c-p51ss1TG被富集在Starch and sucrose metabolism通路中,并调控TRINITYDN44125-c0g1、TRINITYDN33727c0g2、TRINITYDN44986c2g3、TRINITYDN44889c0g2、TRINITYDN46030c0g1等靶基因的表达。