洛阳烟区烟株根围抗烟草疫霉放线菌的筛选与鉴定
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摘要
为筛选对烟草疫霉具有拮抗作用的放线菌菌株,有效防治烟草黑胫病,对洛阳烟区烟株根围土壤中分离的放线菌采用改良平板对峙法与菌丝生长速率法测定了供试菌株及其发酵液对烟草疫霉的拮抗作用,并通过形态学观察、生理生化测定及16S rDNA序列分析鉴定了优势拮抗放线菌的种类。结果表明:在获得的227个放线菌单菌落中,8个菌株的发酵液对烟草疫霉菌丝生长的抑制率达100%,被确定为优势拮抗放线菌。根据这些菌株的形态学观察、生理生化反应与16S rDNA序列分析结果,将菌株YY75鉴定为鲁萨链霉菌(Streptomyces lucensis),菌株YY124、YY135鉴定为高贵链霉菌(S. nobilis),菌株LN204、LN221、LN247鉴定为娄彻链霉菌(S. rochei),菌株SX92鉴定为珊瑚链霉菌(S. coralus),菌株SX59鉴定为金黄回旋链霉(S. aureoverticillatus),均属于放线菌门、放线菌纲、链霉菌目、链霉菌科、链霉菌属。
To screen the antagonistic actinomycetes against Phytophthora parasitica from tobacco rhizospheric soils in order to control tobacco black shank,the antagonistic effects of actinomycetes from rhizospheric soils in Luoyang tobacco-growing areas and their fermentation broths against P. parasitica were determined by improved plate confrontation test and mycelial growth rate method. Dominant antagonistic strains were identified by morphological and biochemical features and 16 S rDNA analysis. The results showed that 227 strains of actinomycetes were isolated,in which the inhibition rates of the fermentation broths of 8 strains against P. parasitica reached 100%,therefore the 8 strains were determined as the dominant antagonistic strains.The results of morphological and biochemical features and 16 S rDNA analysis indicated that YY75,SX92 and SX59 were identified as Streptomyces lucensis,S. coralus and S. aureoverticillatus,respectively. YY124 and YY135 were identified as S. nobilis. LN204,LN221 and LN247 were identified as S. rochei. The 8 dominant strains all belonged to Actinobacteria,Actinobacteria,Stretomytales,Streptomycetaceae,Streptomyces.
To screen the antagonistic actinomycetes against Phytophthora parasitica from tobacco rhizospheric soils in order to control tobacco black shank,the antagonistic effects of actinomycetes from rhizospheric soils in Luoyang tobacco-growing areas and their fermentation broths against P. parasitica were determined by improved plate confrontation test and mycelial growth rate method. Dominant antagonistic strains were identified by morphological and biochemical features and 16 S rDNA analysis. The results showed that 227 strains of actinomycetes were isolated,in which the inhibition rates of the fermentation broths of 8 strains against P. parasitica reached 100%,therefore the 8 strains were determined as the dominant antagonistic strains.The results of morphological and biochemical features and 16 S rDNA analysis indicated that YY75,SX92 and SX59 were identified as Streptomyces lucensis,S. coralus and S. aureoverticillatus,respectively. YY124 and YY135 were identified as S. nobilis. LN204,LN221 and LN247 were identified as S. rochei. The 8 dominant strains all belonged to Actinobacteria,Actinobacteria,Stretomytales,Streptomycetaceae,Streptomyces.
引文
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[17]边传红.洛阳市烟田腐霉菌与烟草根际拮抗放线菌的鉴定[D].洛阳:河南科技大学,2016.BIAN Chuanhong.Identification of Pythium in tobacco fields and antagonistic actinomycetes in tobacco rhizosphere soils[D].Luoyang:Henan University of Science and Technology,2016.
[18]廖振林,刘菁,陈建宏,等.广西北海红树林土壤放线菌的分离与鉴定[J].安徽农业科学,2010,38(23):12693-12694,12702.LIAO Zhenlin,LIU Jing,CHEN Jianhong,et al.Isolation and identification of actinobacteria from Mangrove Forest in Beihai,Guangxi Province[J].Journal of Anhui Agricultural Sciences,2010,38(23):12693-12694,12702.
[19]Singh S B,Zink D L,Herath K B,et al.Discovery and antibacterial activity of lucensimycin C from Streptomyces lucensis[J].2008,49(16):2616-2619.
[20]Anukool U,Gaze W H,Wellington E M H.In situ monitoring of streptothricin production by Streptomyces rochei F20 in soil and rhizosphere[J].Applied and Environmental Microbiology,2004,70(9):5222-5228.
[2]周喜新,周倩,胡日生,等.烟草黑胫病生物防治研究进展[J].江西农业学报,2011,23(7):124-126.ZHOU Xixin,ZHOU Qian,HU Risheng,et al.Research advance in biological control of tobacco black shank[J].Acta Agriculturae Jiangxi,2011,23(7):124-126.
[3]李斌.烟草黑胫病菌拮抗放线菌的筛选[J].西南农业学报,2012,25(5):1708-1713.LI Bin.Screening of antagonistic actinomyce against Phytophthora nicotianae[J].Southwest China Journal of Agricultural Sciences,2012,25(5):1708-1713.
[4]杨兵,刘慧美,龙章富.微生物防治烟草黑胫病的研究进展[J].农药,2015,54(9):629-634.YANG Bing,LIU Huimei,LONG Zhangfu.Recent of advances in microbiological control of Phytophthora parasitica var.nicotianae[J].Agrochemicals,2015,54(9):629-634.
[5]Lukic A,Welty R E,Lucas G B.Antifungal spectra of actinomycetes isolated from tobacco[J].Antimicrobial Agents and Chemotherapy,1972,1(4):363-366.
[6]Kortemaa H,Pennanen T,Smolander A,et al.Distribution of antagonistic Streptomyces griseoviridis in rhizosphere and nonrhizosphere sand[J].Phytopathology,1997,145(4):137-143.
[7]Loqman S,Barka E A,Clément C,et al.Antagonistic actinomycetes from Moroccan soil to control the grapevine gray mold[J].World Journal of Microbiology and Biotechnology,2009,25(1):81-91.
[8]周喜新,周倩,胡日生,等.烟草黑胫病菌拮抗放线菌LY18的分离筛选与鉴定[J].农学学报,2011,1(7):18-22.ZHOU Xixin,ZHOU Qian,HU Risheng,et al.Isolation and identification of antagonistic actinomyces LY18 against Phytophthora parasitica var.nicotianae[J].Journal of Agriculture,2011,1(7):18-22.
[9]王静,孔凡玉,张成省,等.放线菌F8对烟草黑胫病的拮抗作用及其产酶活性[J].中国烟草科学,2013,34(2):49-53.WANG Jing,KONG Fanyu,ZHANG Chengsheng,et al.Antagonism effect and enzyme-producing comparison of Actinomycetes F8 against tobacco black shank[J].Chinese Tobacco Science,2013,34(2):49-53.
[10]郑雅楠,杨宇,吕国忠,等.土壤放线菌分离方法研究[J].安徽农业科学,2006,34(6):1167-1168,1170.ZHENG Yanan,YANG Yu,LüGuozhong,et al.Study on the method of Actinomycetes isolation from soil[J].Journal of Anhui Agricultural Sciences,2006,34(6):1167-1168,1170.
[11]丁钥琪.烟田拮抗微生物的互作及其应用效果研究[D].洛阳:河南科技大学,2017.DING Yueqi.Interactions and applications of antagonistic microorganisms in tobacco field[D].Luoyang:Henan University of Science and Technology,2017.
[12]方中达.植病研究方法[M].北京:中国农业出版社,1998.FANG Zhongda.Methodology for plant pathology[M].Beijing:China Agriculture Press,1998.
[13]周德庆.微生物学实验手册[M].上海:上海科学技术出版社,1986.ZHOU Deqing.Medical microbiology laboratory[M].Shanghai:Shanghai Scientific&Technical Publishers,1986.
[14]中国科学院微生物研究所放线菌分类组.链霉菌鉴定手册[M].北京:科学出版社,1975.Division of Actinomycetes in the Microorganism Institute of the Chinese Academy of Science.Identification manual of Streptomyces[M].Beijing:Science Press,1975.
[15]阎逊初.放线菌的分类和鉴定[M].北京:科学出版社,1992.YAN Xunchu.Classification and identification of actinomycetes[M].Beijing:Science Press,1992.
[16]宋喜乐,赵世民,赵云波,等.洛阳地区烟草根际土壤中多粘类芽孢杆菌的分离与鉴定[J].烟草科技,2016,49(12):13-20.SONG Xile,ZHAO Shimin,ZHAO Yunbo,et al.Isolation and identification of Paenibacillus polymyxa from rhizosphere soil of tobacco plantation in Luoyang[J].Tobacco Science&Technology,2016,49(12):13-20.
[17]边传红.洛阳市烟田腐霉菌与烟草根际拮抗放线菌的鉴定[D].洛阳:河南科技大学,2016.BIAN Chuanhong.Identification of Pythium in tobacco fields and antagonistic actinomycetes in tobacco rhizosphere soils[D].Luoyang:Henan University of Science and Technology,2016.
[18]廖振林,刘菁,陈建宏,等.广西北海红树林土壤放线菌的分离与鉴定[J].安徽农业科学,2010,38(23):12693-12694,12702.LIAO Zhenlin,LIU Jing,CHEN Jianhong,et al.Isolation and identification of actinobacteria from Mangrove Forest in Beihai,Guangxi Province[J].Journal of Anhui Agricultural Sciences,2010,38(23):12693-12694,12702.
[19]Singh S B,Zink D L,Herath K B,et al.Discovery and antibacterial activity of lucensimycin C from Streptomyces lucensis[J].2008,49(16):2616-2619.
[20]Anukool U,Gaze W H,Wellington E M H.In situ monitoring of streptothricin production by Streptomyces rochei F20 in soil and rhizosphere[J].Applied and Environmental Microbiology,2004,70(9):5222-5228.