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1、南方農(nóng)業(yè)學(xué)報(bào)JournalofSouthernAgriculture2015,46(3):42l-427ISSN2095—1191;CODENNNXAABhttp://www.nfnyxb.CllDOI:10.39690:issn.2095-1191.2015.3.421一株生姜青枯病拮抗菌的篩選、鑒定及發(fā)酵條件優(yōu)化趙志祥L(zhǎng)2,嚴(yán)婉榮,,一,陳圓1,27肖彤斌·,一,肖敏1,2+(1海南省農(nóng)業(yè)科學(xué)院農(nóng)業(yè)環(huán)境與植物保護(hù)研究所,???71100;2海南省植物病蟲害防控重點(diǎn)實(shí)驗(yàn)室,???71loo)摘要:【目的】篩選對(duì)生姜青枯病菌有較強(qiáng)拮抗
2、活性的細(xì)菌菌株,鑒定并優(yōu)化拮抗菌株的發(fā)酵條件,為拮抗菌株的田間應(yīng)用和活性產(chǎn)物開(kāi)發(fā)提供必要的前期基礎(chǔ)?!痉椒ā繌纳嗫莶“l(fā)病較重的田塊采集17份健康生姜根際根圍土樣,采用稀釋平板分離和室內(nèi)活性測(cè)定,篩選對(duì)生姜青枯病菌有較強(qiáng)拮抗活性的細(xì)菌菌株;對(duì)拮抗菌株進(jìn)行形態(tài)學(xué)、生理生化特性和分子生物學(xué)鑒定,并通過(guò)改變碳源、氮源、發(fā)酵溫度、初始pH和發(fā)酵時(shí)間的長(zhǎng)短等外界條件對(duì)拮抗菌株進(jìn)行發(fā)酵條件優(yōu)化?!窘Y(jié)果】篩選出1株對(duì)生姜青枯病菌有較強(qiáng)拮抗活性的細(xì)菌菌株YA.1。結(jié)合形態(tài)學(xué)、生理生化特性和16SrDNA鑒定,確定YA一1菌株為枯草芽孢桿菌(Baci
3、llussubtilis)。結(jié)合發(fā)酵條件的改變和室內(nèi)活性測(cè)定,確定YA.1菌株的最佳發(fā)酵條件為:發(fā)酵溫度32℃、初始pH7.5、發(fā)酵時(shí)間48h,最佳培養(yǎng)基配方為NaCl1.0%、CaCO,0.2%、酵母粉0.5%、NI-IA-12PO。0.05%、(NH4):HPO。0.05%和蛋白胨1.0%。發(fā)酵條件優(yōu)化后,處理48h,抑菌圈平均直徑由優(yōu)化前的23.0innl提高到29.0mm?!窘Y(jié)論】YA.1菌株對(duì)生姜青枯病菌具有較高的拮抗活性,具有開(kāi)發(fā)成生物農(nóng)藥的潛力。關(guān)鍵詞:生姜青枯??;枯草芽孢桿菌;分離鑒定;拮抗作用;發(fā)酵條件中圖分類號(hào):S
4、476文獻(xiàn)標(biāo)志碼:A文章編號(hào):2095—1191(2015)03—0421—07Screening,identificationandfermentationconditionoptimizationofbiocontrolbacteriaYA一1againstgingerbacterialwiltZHAOZhi—xian91”,YANWan—ron91”,CHENYuanl”,XIAOTong-binl”,XIAOMinl’2+(1InstituteofAgro-Environment&PlantProtectionofHainan
5、AcademyofAgriculturalSciences,Haikou571100,China;2HainanKeyLaboratoryforControlofPlantDiseasesandInsectPests,Haikou571100,China)Abstract:【Objective】BiocontrolbacteriaYA-1withstrongantagonismagainstgingerbacterialwiltwasscreenedandthefermentationconditionswereoptimizedto
6、providenecessarypreliminarybasisforstrainapplicationinfieldandproductdevelopment.【Method】Seventeenheahhygingerrhizospheresoilsampleswerecollectedfromfields,whichwereseverelyhitbygingerbacterialwilt.Diluteplateseparationandindoorbioassaywereusedtoscreenactivestrains.Mor-
7、phologieal,physiologicalandbiochemicalcharacteristicsandbacterial16srRNAgenecharacteristicswereanalyzed.Atthesametime,fortheimprovementofinhibitoryeffect,modificationofCsources,Nsources,fermentationtempera·ture,pHvalueandfermentationtimeweremade.【Result】Inthebioassay,tw
8、elvestrainsprovedtopossessantagonisticeffectagainstRalstoniasolanacearum,amongwhichstrainYA一1hadthebestinhibit