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1、安徽農(nóng)業(yè)科學(xué),JournalofAnhuiAgn.Sci.2014,42(25):8584—8586,8647責(zé)任編輯喬利利責(zé)任校對(duì)李巖水稻紋枯病拮抗細(xì)菌的篩選·鑒定及其活性成分的發(fā)酵制備黃河,鄢凱舟,林元山,豐來,羅志威(1.湖南農(nóng)業(yè)大學(xué)東方科技學(xué)院,湖南長(zhǎng)沙410128;2.湖南泰谷生物科技股份有限公司,湖南長(zhǎng)沙410205)摘要[目的]探究水稻紋枯病拮抗細(xì)茵的抑茵活性成分的發(fā)酵制備條件。[方法]通過點(diǎn)接種于平板與水稻紋枯病對(duì)峙生長(zhǎng)對(duì)水稻紋枯病拮抗細(xì)菌進(jìn)行了初篩,通過搖瓶發(fā)酵進(jìn)行了復(fù)篩,并通過形態(tài)鑒定及16SRNA測(cè)序構(gòu)建系統(tǒng)發(fā)育樹對(duì)所得菌株進(jìn)行鑒定;通過單
2、因素試驗(yàn)研究了所得菌株的抑茵活性成分的發(fā)酵制備條件。[結(jié)果]初篩獲得17株水稻紋枯病的拮抗細(xì)菌,復(fù)篩獲得菌株Lys-67,該茵株的發(fā)酵液對(duì)水稻紋枯病生長(zhǎng)具有明顯的抑制作用,菌株Lys-67鑒定為枯草芽孢桿菌(Bacillussubtil~)。經(jīng)單因素試驗(yàn),菌株Lys-67利用麩皮5.0%,于35℃、搖床轉(zhuǎn)速200r/min條件下發(fā)酵3d,經(jīng)牛津杯打孔法檢測(cè),150txl發(fā)酵液中活性成分抑制水稻紋枯病的抑菌圈達(dá)36mm。I結(jié)論l為茵株Lys-67的進(jìn)一步研究及生防應(yīng)用奠定了基礎(chǔ)。關(guān)鍵詞水稻紋枯??;拮抗茵;篩選;發(fā)酵;生物防治中圖分類號(hào)Q93.3;$435.1l1.
3、42文獻(xiàn)標(biāo)識(shí)碼A文章編號(hào)0517—6611(2014)25—08584—03Screening,IdentificationofAntagonisticBacteriaagainstRhizoctoniasolaniandPreparationofItsActiveIngredientbyFermentationHUANGHe,LINYuan-shalletal(Ofien~lScienceandTechnologyCollegeofHunanAgriculturalUniversity,Changsha,Hunan410128)Abstract[Objecti
4、ve]TheaimwastostudythefermentationconditionsofantagonisticbacteriaactiveingredientagainstRhizoctoniasolani.[Method]SomeantagonisticbacteriaagainstR.solaniwerescreenedthroughpointinoculatingcontrarytoR.solanionagarpetriplatesandshakingflaskfermentation;theobtainedstrainwasidentifiedth
5、roughmorphologicalidentificationand16SRNAsequencingtoconstructphylo—genetictree;andfermentationconditionsofactiveingredientoftheobtainedstrainagainstR.solaniwerestudiedthroughsinglefactorexperi—ment.[Result]SeventeenstrainsofbacteriaantagonistR.solaniwereobtained,andstrainLys-67wasob
6、tainedfinally;strainLys-67wasi—dentifiedasBacillussubtil~.Also,150lfermentationbrothofstrainLys-67resultedin36mmofinhibitionzoneagainstR.solani,determi—ningbymethodofdrillingholewithOxfordcuponagarplate,whilestrainLys·67wasculturedinwheatbran5.0%,at35℃,rotationspeed200r/minforthreeda
7、ys.[Conclusion]TheresultslaybasisforfurtherstudyandapplicationofstrainLys-67.KeywordsRhizoctoniasolani;Antagonisticbacteria;Screening;Fermentation;Biologicalcontro1水稻紋枯病是我國(guó)水稻種植區(qū)重要病害,嚴(yán)重時(shí)會(huì)導(dǎo)致1.1.2供試病原真菌。水稻紋枯病菌由湖南農(nóng)業(yè)大學(xué)生物我國(guó)水稻產(chǎn)量年損失在10%以上。目前,在生產(chǎn)上主要用科學(xué)技術(shù)學(xué)院保存。將該菌株接種于WBA中28培養(yǎng),待3%井岡霉素水劑來防治水稻紋枯病,
8、賴開平等研究表明菌絲長(zhǎng)滿