烟草黑胫病菌（Phytophthora parasitica var.nicotianae）的拮抗内生细菌筛选、鉴定与应用研究
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摘要
2001～2003年，从湖南永州和宁乡等地发病重的田块采集病株共分离到烟草黑胫病菌(Phytophthora parasitica var.nicotianae)菌株6株，经离体叶片致病力筛选，烟草黑胫病菌株phy3为强致病力菌株，病情指数达到89％；并在实验室条件下，研究了几种溶液诱导孢子囊的产生效果，结果证明经0.1％KNO，处理产生孢子囊最多。同时在湖南永州、浏阳和宁乡等地发病重的田块采集健株共分离到内生细菌120株，经室内平皿对峙培养法测定，其中8株内生细菌菌株在室内有较强的拮抗作用。经过内生细菌定殖力的测定，其中6株为烟草内生细菌。将这8株内生细菌与烟草黑胫病菌共培，进一步测定拮抗效果结果显示YN-55、NN-3、YN-4和YN-10 4株拮抗效果最好，均在66％以上，并且这4种细菌均对孢子囊的产生有较强的抑制作用，而菌株发酵液对孢子囊的产生的抑制作用不明显。本文根据形态特征和生理生化性状，初步鉴定内生细菌YN-55为假单胞杆菌属铜绿假单胞菌，NN-3为芽孢杆菌属枯草芽孢杆菌，YN-4、YN-10为芽孢杆菌属凝结芽孢杆菌。
本文还进行了YN-55、NN-3、YN-4和YN-10发酵条件的测定，结果表明YN-55在27.5℃，PH6.5生长最好，生长曲线测定显示在32h内0D值呈逐步上升趋势；NN-3在27.5℃，PH7.5生长最好，在24h 0D值达最高值；YN-4在27.5℃，PH7.0生长最好，在24h OD值达最高值；YN-10在27.5℃，PH7.0生长最好，在26h OD值达最高值。
在温室进行了YN-55、NN-3、YN-4和YN-10 4种拮抗细菌对两个烟草品种苗期烟草黑胫病的防治试验，试验结果显示四种拮抗细菌对烟草黑胫病均有一定的防治效果，防效为30.4％～51.4％，其中以NN-3防治效果最好，防效分别达到51.4％和39.1％。
Six strains of Phytophthora parasitica var. nicotianae were isolated from disease tobaccos in Yongzhou and Ningxiang from heavy disease plots;. Pathogenicity was tested to six strains, the results shows the strain of phy3 have strong pathogenicity. We also studied the effect of several solutions for producing zoosporangium in Artificial culture condition, the result shows the hyphea culture with 0.1% KNO3 produced the most quantity of zoosporangiums. 120 Endophtic bacteria strains were isolated from healthy tobaccos from heavy disease plots in Yongzhou, Ningxiang and Liuyang. By Laboratory dual culture test, seventeen actinomycetes to Phytophthora parasitica var. nicotianae were screened. Among them eight endophytic bacteria showed relatively strong antagonistic actions. By inoculated to tobacco plants, six strains could be re-isolated from the inoculated plants, indicating that they were endophytic bacteria in tobacco. Culture these eight bacteria and Phytophthora parasitica var. nicotianae together, the result showed four endophytic bacteria YN-55, NN-3, YN-4 and YN-10 have the best antagonistic effects. The control effects were above 66%. According morphologic, physiological and biochemical characteries, strain YN-55, NN-3, YN-4 and YN-10 were identified.
Some conditions for culture YN-55, NN-3, YN-4 and YN-10 were studied in shake flask (100ml in 150ml flask). The optimum cultivation PH and temperature for YN-55 were 6.5 and 27.5C respectively, for NN-3 were 7.5 and 27.5C respectively, for YN-4 and YN-10 were 7.0 and 27.5C respectively. The optimum cultivation time for NN-3 and YN-4 were 24h, for YN-10 was 26h.
Ferment products of antagonistic bacteria YN-55, NN-3, YN-4 and YN-10 were applied at tobacco seedling in greenhouse in two tobacco cultivars. The greenhouse experimental results indicated that four endophytic bacteria YN-55, NN-3, YN-4 and
YN-10 could control tobacco blank shank, and the control effects were between 30.4% and 51.4%. Among them, NN-3 shows the best control effect, the control effect were 51.4% and 39.1 % respectively.
本文还进行了YN-55、NN-3、YN-4和YN-10发酵条件的测定，结果表明YN-55在27.5℃，PH6.5生长最好，生长曲线测定显示在32h内0D值呈逐步上升趋势；NN-3在27.5℃，PH7.5生长最好，在24h 0D值达最高值；YN-4在27.5℃，PH7.0生长最好，在24h OD值达最高值；YN-10在27.5℃，PH7.0生长最好，在26h OD值达最高值。
在温室进行了YN-55、NN-3、YN-4和YN-10 4种拮抗细菌对两个烟草品种苗期烟草黑胫病的防治试验，试验结果显示四种拮抗细菌对烟草黑胫病均有一定的防治效果，防效为30.4％～51.4％，其中以NN-3防治效果最好，防效分别达到51.4％和39.1％。
Six strains of Phytophthora parasitica var. nicotianae were isolated from disease tobaccos in Yongzhou and Ningxiang from heavy disease plots;. Pathogenicity was tested to six strains, the results shows the strain of phy3 have strong pathogenicity. We also studied the effect of several solutions for producing zoosporangium in Artificial culture condition, the result shows the hyphea culture with 0.1% KNO3 produced the most quantity of zoosporangiums. 120 Endophtic bacteria strains were isolated from healthy tobaccos from heavy disease plots in Yongzhou, Ningxiang and Liuyang. By Laboratory dual culture test, seventeen actinomycetes to Phytophthora parasitica var. nicotianae were screened. Among them eight endophytic bacteria showed relatively strong antagonistic actions. By inoculated to tobacco plants, six strains could be re-isolated from the inoculated plants, indicating that they were endophytic bacteria in tobacco. Culture these eight bacteria and Phytophthora parasitica var. nicotianae together, the result showed four endophytic bacteria YN-55, NN-3, YN-4 and YN-10 have the best antagonistic effects. The control effects were above 66%. According morphologic, physiological and biochemical characteries, strain YN-55, NN-3, YN-4 and YN-10 were identified.
Some conditions for culture YN-55, NN-3, YN-4 and YN-10 were studied in shake flask (100ml in 150ml flask). The optimum cultivation PH and temperature for YN-55 were 6.5 and 27.5C respectively, for NN-3 were 7.5 and 27.5C respectively, for YN-4 and YN-10 were 7.0 and 27.5C respectively. The optimum cultivation time for NN-3 and YN-4 were 24h, for YN-10 was 26h.
Ferment products of antagonistic bacteria YN-55, NN-3, YN-4 and YN-10 were applied at tobacco seedling in greenhouse in two tobacco cultivars. The greenhouse experimental results indicated that four endophytic bacteria YN-55, NN-3, YN-4 and
YN-10 could control tobacco blank shank, and the control effects were between 30.4% and 51.4%. Among them, NN-3 shows the best control effect, the control effect were 51.4% and 39.1 % respectively.
引文
[1] Beauchamp C J, Kloepper J. A review of issues related to meassuring colonization of plant roots bu bacteria[J]. Can. J. Microbiol. 1992,38:1219-1232.
[2] Mcinory J A. Kloepper J W. Plant and soil, 1995,173:337-342.
[3] Gardner J M. Chandler J A. Feldman A W. Plant soil, 1982,86:333-345.
[4] Kluepfel D A. Annu Rev Phytopathol, 1993, 31:441-472.
[5] Lynch J M., Fletcher M, Latham M J. In "Mycrolial Elology A Conceptual approach". Lynch J M, Poole N J (eds) Black well Scientific Publications, 1979,171-189.
[6] Chanway C P. Endophytes: they're not just fungi[J]. Can. J. Bot.,1996,74:321-322.
[7] Chen C E. Bauske M, Musson G, et al. Biological control of Fusarium wilt on contton by use of endophytic bacteria[J]. Biological Control ,1995,5:83-91.
[8] Andrews J H , Ann Rev Phytopathol, 1992,30:603-635.
[9] Chen C, Banske E M, Myssen G, Kloepper J W. In "Improving plant productivity with rhizosphere bacteria". Ryder M H, Stephens P M, Bbowen G D(eds). CSIRO, Australia, 1994, 191-193.
[10] Sayonara M P Assis, Rosa L R Mariano, Sami J Michereff et al. In Advances in Biological Control of P:lant Diseases Tang Wenhua, R James Cook., Albest Rovira (Eds). China Agricultural University Pressing, 1996,347-353.
[11] 崔林、孙振、孔福在等．马铃薯内生细菌的分离及环腐病拮抗菌的筛选鉴定[J]．植物病理学报，2003，33(4)：353～358．
[12] 何红、蔡学清、洪永聪等．辣椒内生细菌的分离及拮抗菌的筛选[J]．中国生物防治，2002，18(4)：171～175．
[13] 黎起秦、罗宽、林纬等．番茄青枯病内生拮抗细菌的筛选[J]．植物病理学报，2003，33(4)：364～367．
[14] Brdder U S, Reis V D. In Dolsinelli M Marerrassi R, Vincenzini M (Eds). Mtrogen Fixationkluwer Academic Pubishers Dordsecht, 1991:105-111.
[15] Shishido M, Loeb B M, Chanway C P. External and internal root colonization of lodgepole pine seedings by two growth-promoting Bacillus strains originated from different root microsites[J]. Can.
J. Microbiol.,1989, 35:456-463.
[16] Van Peer R. Schippers B. Plant growth responses to bacterization with selected Pseudomonas spp. strains and rhizosphere microbial development in hydroponic culture[J]. Can. J. Microbiol., 1989,35:456-463.
[17] Brooks D S, Gonzalez, C F, Appel D N. (abstract) Phytopathol, 1988,78:1626.
[18] 何红，蔡学清，关雄等．辣椒内生枯草芽孢杆菌(Bacillus subtilis)BS-2和BS-1防治辣椒炭疽病研究[J]．植物病理学报，2003，33(2)：170～173．
[19] Chanway C P. Endophytes: they're not just fungil[J].Can. J. Bot, 1996,74:321-322.
[20] Buchenaver H. Biological control of soil-borne disease by rhizobacteris[J]. J. Plant Disease and Protection, 1995,44:40-50.
[21] Velazhahan R., Samiyappan R. Relationship between antagonistic activties of Pseudomonas fluarescens isolates against Rhizochonic solani and their production of lyric enzymes[J]. J. Plant Disease and Protection, 1999,106(3):244-250.
[22] 孔庆科，丁爱云．内生细菌作为生防因子的研究进展[J]．山东农业大学学报(自然科学版)，2001，32(2)：256～260．
[23] Lemanceau P, Lantour X. Effects of the plant on the soil-borne microflofta; application to the fluorescent pseudomonas[R], the 7th international Cogress of Plant Pathology(ICPP),2.7.4a.
[24] 金玲、巴峰、计平生等．小麦内生有害细菌的发现和作用研究[J]．植物病理学报，1996，26(2)：123～126．
[25] 中国农业科学院植物研究所．中国农作物病虫害(下)[M]．1996：575～580．
[26] Broadbent P. Bactetia and actinomyctes antagonistic to fungal root pathogens in Australian siol[J].Aust. J. Biol. Sci.,1971,24:925～944.
[27] English J. T., Mitchell D. J. relationship between the development of root systems of tobacco and infection by Phytophthora parasitica var. nicotianae[J]. Phytopathology, 1988,78:1478-1483.
[28] 沈其益．棉花病害基础研究与防治[M]．北京：科学出版社，1995．69—187．
[29] 王远山，王平，胡正嘉．绿针假单胞菌PL9菌株对烟草黑胫病菌的拮抗作用研究[J]．华中农业大学学报，2002，21(3)：248～251．
[30] 方敦煌，顾金刚，李江涛等．烟草黑胫病菌的拮抗根际细菌的筛选[J]．云南农业大学学报，2001，16(2)：93～97．
[31] 王万能，肖崇刚．烟草内生细菌118防治黑胫病的机理研究[J]．西南农业大学学报，2003，25(1)：28—31．
[32] English J. T., Mitchell D. J. Influence of an introduced composite of microorganism on infection of tobacco by Phytophthora parasitica var. nicotianae[J]. Phytopathology, 1988,78:1484-1490.
[33] Cartwright D.K., Spurr H.W. Biological control of Phytophthora parasitica var. nicotianae on Tobacco Seedlings with Non-pathogenic Binucleate Rhizoctonia fung[J]. Soil Biology & Biochemistry, 1998, 30(14):1879-1884.
[34] Patel D.N., Patel B.N. Evaluation of plant extracts and Trichoderma harzianum rifai against Phytophthora parasitica var. nicotianae[J]. Tobacco Research, 1999, 25(1):4-8.
[35] Sreeramulu K.R., Onkarappa T., Swamy H.N. Biocontrol of damping-off and blanck shank disease in tobacco nursey[J]. Tobacco Research, 1998,24(1):1-4.
[36] 薛宝娣，李娟，陈永萱．木霉(TR-5)对病原真菌的拮抗机制和防病效果研究[J]．南京农业大学学报，1995，18(1)：31～36．
[37] 徐同，柳良好．木霉几丁质酶及其对植物病原真菌的拮抗作用[J]．植物病理学报，2002，32(2)：97～103．
[38] 王革，李梅云，段玉琪等．木霉菌对烟草黑胫病菌的拮抗机制及其生物防治研究[J]．云南农业大学学报，2001，23(3)：222～226．
[39] 李梅云，李天飞，刘开启等．烟草黑胫病木霉生防菌株的筛选[J]．中国烟草科学，2001(2)：43～46．
[40] Kuc J. Expression of latent mechanisms for resistance to blue mold andother disease in tobacco, In CORESTA Inf. Bull., 9th Tob. Sci. cong., 198,25～43.
[41] Chawla H S. In vitro selection of barley and wheat for resitance against Helminthosporium sativum. Their. Appl. Genet, 1987, 74:841.
[42] 王荔，杨艳琼，杨德等．运用致病毒素筛选抗烟草黑胫病细胞突变体Ⅱ抗毒素愈伤组织的再生植株及其后代的抗病性鉴定[J]．云南农业大学学报，1999，14(3)：273～279．
[43] 王荔，罗文富，杨艳琼等．运用致病毒素筛选抗烟草黑胫病细胞突变体Ⅰ抗毒素愈伤组织的有效筛选[J]．云南农业大学学报，1999，14(1)：16～21．
[44] 黄丽华，陈廷俊，宋军．烟草黑胫病粗毒素高产培养条件的筛选[J]．安徽农业科学，1999，27(4)：368～370．
[45] 赵蕾，梁元存，刘延荣．壳聚糖对烟草抗黑胫病的作用[J]．应用与环境生物学报，2000，6(5)：436～439．
[46] 徐后娟，张军，慕立义等．二氢茉莉酸丙酯诱导烟草抗黑胫病作用研究[J]．农药学报，2003，5(1)：73～76．
[47] 杨献营．烟草病害防治的新途径—诱导抗病性的研究进展[J]．中国烟草，1993(4)：29～31．
[48] D. N. Patel, B.N. Patel. Evaluation of plant extracts and Trichoderma harzianum rifai against Phytophthora parasitica var. nicotianae [J]. Tobacco Research, 1999, 25(1): 4～8.
[49] 王革，郑小波，陆家云等．云南省烟草黑胫病菌对甲霜灵抗性的检测[J]．南京农业大学学报，1997，20(4)：105～107．
[50] 袁宗胜、张广民，刘延荣等．烟草黑胫病菌对甲霜灵的敏感性测定[J]．中国烟草科学，2001(4)：9～12．
[51] 杨梅莲，孙晓璐，宋未．植物内生细菌的研究[J]．微生物学通报，1998，25(4)：224-227．
[52] 刘云霞．植物内生细菌的研究与应用[J]．植物保护，1994，20(5)：30—32．
[53] 周嘉平、周俭民、梁思信．烟草抗黑胫病突变体的细胞筛选[J]．遗传学报，1990，17(3)：180～188．
[54] 方中达．植病研究方法[M]．北京：中国农业出版社．第三版，1998．
[55] 中国科学院南京土壤研究所微生物室．土壤微生物研究法[M]，北京：科学出版社，1985．
[56] 卢振祖．细菌分类学[M]．武汉：武汉大学出版社．第一版，1994．
[57] 梁元存、刘延荣、王玉军等．烟草黑胫病菌致病性分化和烟草品种的抗病性差异[J]．植物保护学报，2003，30(2)：143～147．
[58] 郑小波．疫霉菌及其研究技术[M]．北京：中国农业出版社，1997．
[59] 刘延荣、谢成颂、王智发等．烟草黑胫病菌孢子囊形成和游动孢子释放条件的研究[J]．山东农业科学，1987(1)：28～30．
[60] Kloepper J W ,Rodriguez-Ubana R, Zehnder G W, et al. Plant root-bacterial interactions in biologiacal control of soil borne diseases and protential extension to systemic and foliar diseases[J]. Australssian Plant Pathology, 1999,28:21-26.
[61] McInroy J A, Kloeper J W. Survey of indig enous bacterial endophytes from cotton and sweet corn[J].Plant and Soil ,1995,173: 337-342.
[2] Mcinory J A. Kloepper J W. Plant and soil, 1995,173:337-342.
[3] Gardner J M. Chandler J A. Feldman A W. Plant soil, 1982,86:333-345.
[4] Kluepfel D A. Annu Rev Phytopathol, 1993, 31:441-472.
[5] Lynch J M., Fletcher M, Latham M J. In "Mycrolial Elology A Conceptual approach". Lynch J M, Poole N J (eds) Black well Scientific Publications, 1979,171-189.
[6] Chanway C P. Endophytes: they're not just fungi[J]. Can. J. Bot.,1996,74:321-322.
[7] Chen C E. Bauske M, Musson G, et al. Biological control of Fusarium wilt on contton by use of endophytic bacteria[J]. Biological Control ,1995,5:83-91.
[8] Andrews J H , Ann Rev Phytopathol, 1992,30:603-635.
[9] Chen C, Banske E M, Myssen G, Kloepper J W. In "Improving plant productivity with rhizosphere bacteria". Ryder M H, Stephens P M, Bbowen G D(eds). CSIRO, Australia, 1994, 191-193.
[10] Sayonara M P Assis, Rosa L R Mariano, Sami J Michereff et al. In Advances in Biological Control of P:lant Diseases Tang Wenhua, R James Cook., Albest Rovira (Eds). China Agricultural University Pressing, 1996,347-353.
[11] 崔林、孙振、孔福在等．马铃薯内生细菌的分离及环腐病拮抗菌的筛选鉴定[J]．植物病理学报，2003，33(4)：353～358．
[12] 何红、蔡学清、洪永聪等．辣椒内生细菌的分离及拮抗菌的筛选[J]．中国生物防治，2002，18(4)：171～175．
[13] 黎起秦、罗宽、林纬等．番茄青枯病内生拮抗细菌的筛选[J]．植物病理学报，2003，33(4)：364～367．
[14] Brdder U S, Reis V D. In Dolsinelli M Marerrassi R, Vincenzini M (Eds). Mtrogen Fixationkluwer Academic Pubishers Dordsecht, 1991:105-111.
[15] Shishido M, Loeb B M, Chanway C P. External and internal root colonization of lodgepole pine seedings by two growth-promoting Bacillus strains originated from different root microsites[J]. Can.
J. Microbiol.,1989, 35:456-463.
[16] Van Peer R. Schippers B. Plant growth responses to bacterization with selected Pseudomonas spp. strains and rhizosphere microbial development in hydroponic culture[J]. Can. J. Microbiol., 1989,35:456-463.
[17] Brooks D S, Gonzalez, C F, Appel D N. (abstract) Phytopathol, 1988,78:1626.
[18] 何红，蔡学清，关雄等．辣椒内生枯草芽孢杆菌(Bacillus subtilis)BS-2和BS-1防治辣椒炭疽病研究[J]．植物病理学报，2003，33(2)：170～173．
[19] Chanway C P. Endophytes: they're not just fungil[J].Can. J. Bot, 1996,74:321-322.
[20] Buchenaver H. Biological control of soil-borne disease by rhizobacteris[J]. J. Plant Disease and Protection, 1995,44:40-50.
[21] Velazhahan R., Samiyappan R. Relationship between antagonistic activties of Pseudomonas fluarescens isolates against Rhizochonic solani and their production of lyric enzymes[J]. J. Plant Disease and Protection, 1999,106(3):244-250.
[22] 孔庆科，丁爱云．内生细菌作为生防因子的研究进展[J]．山东农业大学学报(自然科学版)，2001，32(2)：256～260．
[23] Lemanceau P, Lantour X. Effects of the plant on the soil-borne microflofta; application to the fluorescent pseudomonas[R], the 7th international Cogress of Plant Pathology(ICPP),2.7.4a.
[24] 金玲、巴峰、计平生等．小麦内生有害细菌的发现和作用研究[J]．植物病理学报，1996，26(2)：123～126．
[25] 中国农业科学院植物研究所．中国农作物病虫害(下)[M]．1996：575～580．
[26] Broadbent P. Bactetia and actinomyctes antagonistic to fungal root pathogens in Australian siol[J].Aust. J. Biol. Sci.,1971,24:925～944.
[27] English J. T., Mitchell D. J. relationship between the development of root systems of tobacco and infection by Phytophthora parasitica var. nicotianae[J]. Phytopathology, 1988,78:1478-1483.
[28] 沈其益．棉花病害基础研究与防治[M]．北京：科学出版社，1995．69—187．
[29] 王远山，王平，胡正嘉．绿针假单胞菌PL9菌株对烟草黑胫病菌的拮抗作用研究[J]．华中农业大学学报，2002，21(3)：248～251．
[30] 方敦煌，顾金刚，李江涛等．烟草黑胫病菌的拮抗根际细菌的筛选[J]．云南农业大学学报，2001，16(2)：93～97．
[31] 王万能，肖崇刚．烟草内生细菌118防治黑胫病的机理研究[J]．西南农业大学学报，2003，25(1)：28—31．
[32] English J. T., Mitchell D. J. Influence of an introduced composite of microorganism on infection of tobacco by Phytophthora parasitica var. nicotianae[J]. Phytopathology, 1988,78:1484-1490.
[33] Cartwright D.K., Spurr H.W. Biological control of Phytophthora parasitica var. nicotianae on Tobacco Seedlings with Non-pathogenic Binucleate Rhizoctonia fung[J]. Soil Biology & Biochemistry, 1998, 30(14):1879-1884.
[34] Patel D.N., Patel B.N. Evaluation of plant extracts and Trichoderma harzianum rifai against Phytophthora parasitica var. nicotianae[J]. Tobacco Research, 1999, 25(1):4-8.
[35] Sreeramulu K.R., Onkarappa T., Swamy H.N. Biocontrol of damping-off and blanck shank disease in tobacco nursey[J]. Tobacco Research, 1998,24(1):1-4.
[36] 薛宝娣，李娟，陈永萱．木霉(TR-5)对病原真菌的拮抗机制和防病效果研究[J]．南京农业大学学报，1995，18(1)：31～36．
[37] 徐同，柳良好．木霉几丁质酶及其对植物病原真菌的拮抗作用[J]．植物病理学报，2002，32(2)：97～103．
[38] 王革，李梅云，段玉琪等．木霉菌对烟草黑胫病菌的拮抗机制及其生物防治研究[J]．云南农业大学学报，2001，23(3)：222～226．
[39] 李梅云，李天飞，刘开启等．烟草黑胫病木霉生防菌株的筛选[J]．中国烟草科学，2001(2)：43～46．
[40] Kuc J. Expression of latent mechanisms for resistance to blue mold andother disease in tobacco, In CORESTA Inf. Bull., 9th Tob. Sci. cong., 198,25～43.
[41] Chawla H S. In vitro selection of barley and wheat for resitance against Helminthosporium sativum. Their. Appl. Genet, 1987, 74:841.
[42] 王荔，杨艳琼，杨德等．运用致病毒素筛选抗烟草黑胫病细胞突变体Ⅱ抗毒素愈伤组织的再生植株及其后代的抗病性鉴定[J]．云南农业大学学报，1999，14(3)：273～279．
[43] 王荔，罗文富，杨艳琼等．运用致病毒素筛选抗烟草黑胫病细胞突变体Ⅰ抗毒素愈伤组织的有效筛选[J]．云南农业大学学报，1999，14(1)：16～21．
[44] 黄丽华，陈廷俊，宋军．烟草黑胫病粗毒素高产培养条件的筛选[J]．安徽农业科学，1999，27(4)：368～370．
[45] 赵蕾，梁元存，刘延荣．壳聚糖对烟草抗黑胫病的作用[J]．应用与环境生物学报，2000，6(5)：436～439．
[46] 徐后娟，张军，慕立义等．二氢茉莉酸丙酯诱导烟草抗黑胫病作用研究[J]．农药学报，2003，5(1)：73～76．
[47] 杨献营．烟草病害防治的新途径—诱导抗病性的研究进展[J]．中国烟草，1993(4)：29～31．
[48] D. N. Patel, B.N. Patel. Evaluation of plant extracts and Trichoderma harzianum rifai against Phytophthora parasitica var. nicotianae [J]. Tobacco Research, 1999, 25(1): 4～8.
[49] 王革，郑小波，陆家云等．云南省烟草黑胫病菌对甲霜灵抗性的检测[J]．南京农业大学学报，1997，20(4)：105～107．
[50] 袁宗胜、张广民，刘延荣等．烟草黑胫病菌对甲霜灵的敏感性测定[J]．中国烟草科学，2001(4)：9～12．
[51] 杨梅莲，孙晓璐，宋未．植物内生细菌的研究[J]．微生物学通报，1998，25(4)：224-227．
[52] 刘云霞．植物内生细菌的研究与应用[J]．植物保护，1994，20(5)：30—32．
[53] 周嘉平、周俭民、梁思信．烟草抗黑胫病突变体的细胞筛选[J]．遗传学报，1990，17(3)：180～188．
[54] 方中达．植病研究方法[M]．北京：中国农业出版社．第三版，1998．
[55] 中国科学院南京土壤研究所微生物室．土壤微生物研究法[M]，北京：科学出版社，1985．
[56] 卢振祖．细菌分类学[M]．武汉：武汉大学出版社．第一版，1994．
[57] 梁元存、刘延荣、王玉军等．烟草黑胫病菌致病性分化和烟草品种的抗病性差异[J]．植物保护学报，2003，30(2)：143～147．
[58] 郑小波．疫霉菌及其研究技术[M]．北京：中国农业出版社，1997．
[59] 刘延荣、谢成颂、王智发等．烟草黑胫病菌孢子囊形成和游动孢子释放条件的研究[J]．山东农业科学，1987(1)：28～30．
[60] Kloepper J W ,Rodriguez-Ubana R, Zehnder G W, et al. Plant root-bacterial interactions in biologiacal control of soil borne diseases and protential extension to systemic and foliar diseases[J]. Australssian Plant Pathology, 1999,28:21-26.
[61] McInroy J A, Kloeper J W. Survey of indig enous bacterial endophytes from cotton and sweet corn[J].Plant and Soil ,1995,173: 337-342.