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1、園藝學(xué)報(bào)2014,(4111):2329–2341http://www.ahs.ac.cnActaHorticulturaeSinicaE-mail:yuanyixuebao@126.com鳥(niǎo)巢蕨轉(zhuǎn)錄組高通量測(cè)序及分析*賈新平,孫曉波,鄧衍明,梁麗建,葉曉青(江蘇省農(nóng)業(yè)科學(xué)院農(nóng)業(yè)生物技術(shù)研究所,江蘇省農(nóng)業(yè)生物學(xué)重點(diǎn)實(shí)驗(yàn)室,南京210014)摘要:采用新一代高通量測(cè)序技術(shù)IlluminaHiSeq2000對(duì)鳥(niǎo)巢蕨轉(zhuǎn)錄組(Aspleniumnidus)進(jìn)行測(cè)序,共獲得29254595個(gè)序列讀取片段(reads),包含了5908586517個(gè)堿基序列(bp)信息
2、。對(duì)reads進(jìn)行序列組裝,共獲得42907個(gè)單基因簇(Unigene),平均長(zhǎng)度936bp,序列信息達(dá)到了40.16Mb。數(shù)據(jù)庫(kù)中的序列同源性比較表明,24993個(gè)Unigene與其他物種的已知基因具有不同程度的同源性。鳥(niǎo)巢蕨轉(zhuǎn)錄組中的Unigene根據(jù)GO功能大致可分為細(xì)胞組分、分子功能和生物學(xué)過(guò)程3大類51個(gè)分支,其中有大量的Unigene與代謝進(jìn)程、結(jié)合活性、催化活性和細(xì)胞進(jìn)程相關(guān)。將Unigene與COG數(shù)據(jù)庫(kù)進(jìn)行比對(duì),根據(jù)其功能大致可分為24類。KEGG數(shù)據(jù)庫(kù)作為參考,依據(jù)代謝途徑可將Unigene定位到116個(gè)代謝途徑分支。SSR位點(diǎn)查找發(fā)現(xiàn)
3、,從42907個(gè)Unigene中共找到6067個(gè)SSR位點(diǎn)。SSR不同重復(fù)基序類型中,出現(xiàn)頻率最高的為AG/CT,其次是AC/GT、A/T和AGG/CCT。針對(duì)這些序列,設(shè)計(jì)了20對(duì)引物進(jìn)行了擴(kuò)增效率和多態(tài)性檢測(cè),其中7對(duì)引物在不同蕨類材料中表現(xiàn)出多態(tài)性。關(guān)鍵詞:鳥(niǎo)巢蕨;轉(zhuǎn)錄組;生物信息學(xué);功能注釋;SSR中圖分類號(hào):S68文獻(xiàn)標(biāo)志碼:A文章編號(hào):0513-353X(2014)11-2329-13SequencingandAnalysisoftheTranscriptomeofAspleniumnidus*JIAXin-ping,SUNXiao-bo,DE
4、NGYan-ming,LIANGLi-jian,andYEXiao-qing(ProvincialKeyLaboratoryofAgroBiology,InstituteofAgro-biotechnology,JiangsuAcademyofAgriculturalSciences,Nanjing210014,China)Abstract:ThetranscriptomeofAspleniumniduswassequencedbyIlluminaHiSeq2000platformthatisanewgenerationofhigh-throughputs
5、equencingtechnologytostudytheexpressionprofilingandpredictthefunctionalgenes.Thetargetsamplesequencing,atotalof29254595readsfragmentcontains5908586517bpinsequenceinformationweregenerated.Atotalof42907unigenescontains40.16Mbinsequenceinformationwereformedbyinitialsequencesplicing,w
6、ithanaveragereadlengthof936bp.24993unigeneswereannotatedusingBLASTXsearchesagainsttheNrandSwissProtdatabases.Inthisstudy,allassembledunigenescanbebroadlydividedintobiologicalprocesses,cellularcomponentsandmolecularfunctioncategoriesof51branchesbygeneontology,includingmetabolicproc
7、ess,binding,catalyticactivityandcellularprocess.UnigeneswerefurtherannotatedbasedonCOGcategory,whichcouldbegroupedinto24functionalcategories.KEGGpathwayanalysisshowedthatunigenescanbebroadlydividedinto116classesaccordingtothefunction.Therewere6067SSRin42907unigeneswere收稿日期:2014–07
8、–21;修回日期:2014–09–15基金項(xiàng)目:江蘇省農(nóng)業(yè)科技自主