高产抗真菌抗生素ustilagoesculenta菌株的选育-breeding of high-yielding antifungal antibiotic ustilagogulenta strain.docxVIP

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高产抗真菌抗生素ustilagoesculenta菌株的选育-breeding of high-yielding antifungal antibiotic ustilagogulenta strain.docx

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高产抗真菌抗生素ustilagoesculenta菌株的选育-breeding of high-yielding antifungal antibiotic ustilagogulenta strain

5.2展望·······························································································-42-主要参考文献······························································································-44-致谢·······································································································-47-附录Ⅰ···································································································-48-附录Ⅱ···································································································-49-摘要传统食用担子菌Ustilagoeseulenta菌株可代谢产生一种对部分人体病原真菌、部分植物病原真菌等具有较强拮抗作用的抗真菌抗生素。该抗生素具有安全、稳定、抗菌谱广等特点，为开发和生产新型农用抗真菌抗生素提供了可能。自茭白分离38株菌株。综合形态学和分子生物学方法，共鉴定了33株菌株为U.esculenta菌株。以Aspergillesnigersp.为拮抗菌，琼脂块法测定分离菌株的抑菌活性，筛选到抑菌活性较强的分离菌株UEZC。为筛选高产抗真菌抗生素菌株，以UEZC作为出发菌株，经60Co-γ射线辐射诱变，以拮抗菌对峙法初筛，琼脂块法、管碟法复筛，筛选到较高产抗真菌抗生素突变株MUE-4，其抑菌活性较UEZC菌株提高了近1倍，遗传稳定性良好。经ITS1-5.8SrDNA-ITS2基因序列分析，高产突变株MUE-4较其出发菌株UEZC在ITS1-5.8SrDNA-ITS2区段发生了较为明显的变异。采用响应面（RSA）法对诱变选育得到的高产菌株MUE-4进行发酵工艺的初步研究，在所获得的最优发酵条件下，抑菌活性较优化前提高了46%，较诱变出发菌株UEZC提高了194%。关键词：Ustilagoesculenta；抗真菌抗生素；鉴定；ITS1-5.8SrDNA-ITS2；60Co-γ射线；诱变育种AbstractThetraditionalediblebasidiomycetesUstilagoeseulentastrainwasanantifungalantibioticsproducingstrain.TheantifungalantibioticsthatUEZCproducedhadstrongantagonismonpartofhuman,plantspathogenicfungiandsoon,andtheantifungalantibioticswasstableandhadabroadantifungalspectrum.Allofthesecharacteristicsoftheantifungalantibioticsprovidedthepossibilityforthenewantifungalagriculturalantibioticsdevelopmentandproduction.The38isolatesfromZizanialatifoliawereobtained.Studycolligatedthemorphologyandthemolecularbiologicalmethodswasused,theresultsofwhichshowedthat33strainsamongthemwereU.eseulentastrains.Aspergillesnigersp.asantagonisticbacteria,anddetectforantifungalactivitybyagarblockmethodmethod,thestrainUEZCwhichhavestrongerantifungalactivitywasscreened.Toselecthigh-yieldantifungalantibioticsproducingstrains,thestartingstrainUEZCwastreatedwith60Co-γray.Throughtheconfrontculturemethodasprimaryscreening;agarblo