|本期目录/Table of Contents|

[1]李亚男,蒋芬,张杰,等.高原土壤中稻瘟病拮抗细菌的抑菌效果及抗菌机理[J].应用与环境生物学报,2017,23(01):33-40.[doi:10.3724/SP.J.1145.2016.05021]
 LI Yanan,JIANG Fen,ZHANG Jie,et al.Inhibition effects and antibacterial mechanism against rice blast of antagonistic bacteria in plateau soil[J].Chinese Journal of Applied & Environmental Biology,2017,23(01):33-40.[doi:10.3724/SP.J.1145.2016.05021]
点击复制

高原土壤中稻瘟病拮抗细菌的抑菌效果及抗菌机理()
分享到:

《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
23卷
期数:
2017年01期
页码:
33-40
栏目:
研究论文
出版日期:
2017-02-25

文章信息/Info

Title:
Inhibition effects and antibacterial mechanism against rice blast of antagonistic bacteria in plateau soil
作者:
李亚男蒋芬张杰范永义陈敬彭友林胡运高
西南科技大学水稻所 绵阳 621000
Author(s):
LI Ya’nan JIANG Fen ZHANG Jie FAN Yongyi CHEN Jing PENG Youlin & HU Yungao**
Rice Research Institute, Southwest University of Science and Technology, Mianyang 621000, China
关键词:
萎缩芽孢杆菌稻瘟病培养条件孢子抑制菌丝抑制高原土壤代谢产物
Keywords:
Bacillus atrophaeus rice blast culture condition spore inhibition mycelia inhibition plateau soil metabolite
分类号:
S476.19
DOI:
10.3724/SP.J.1145.2016.05021
摘要:
利用平板稀释法从西藏7个不同地点的土壤中分离筛选细菌,并通过平板对峙法对所分离菌株的稻瘟病菌抑制效果进行初筛,获得一株拮抗细菌YN01,对其进行生理生化和分子生物学鉴定、最适生长和发酵条件筛选,分泌IAA和铁载体能力检测及稻瘟病菌孢子抑制实验. 经形态、生理生化特征和分子生物学鉴定,确定菌株YN01为萎缩芽孢杆菌(Bacillus atrophaeus). YN01对稻瘟病菌丝生长的抑制率超过90%,最适生长碳源、氮源分别为蔗糖和酵母粉,在pH值低于6时不能正常生长,最适生长温度为30-35 ℃;碳源及氮源分别为乳糖和谷氨酸、pH值为8以及培养温度为30-35 ℃时发酵液抑菌活性最强;YN01能够分泌IAA,浓度达到4.287 mg/L,同时能够产生铁载体,可溶性指数为1.72;与阴性对照相比,菌悬液和发酵液对稻瘟病菌的孢子萌发起到显著的抑制效果. 拮抗菌菌体通过与病原菌丝进行营养和空间位点竞争以及产生有抑菌效应的代谢产物,从而导致稻瘟病菌的孢子萌发缓慢,菌丝发育停滞,扰乱其生理活动,达到抑菌目的. 得上,拮抗细菌YNO1具有较强的抑菌活性,是潜在的植物病原真菌生防菌资源,具有一定的开发与应用价值. (图10 表3 参33)
Abstract:
The aim of this study was to isolate antagonistic bacteria from the soil of seven different locations of Tibet. We tested with plate confrontation method the inhibitory activities of these bacteria against rice blast (Magnaporthe grisea) and obtained a strain of antagonistic bacterium named YN01. We also tested the physio-biochemical characteristics and molecular identification, and screened the best condition of growth and fermentation. We determined the IAA secretion ability with Salkowski colorimentric, and qualitatively detected the siderophores and spore inhibition effect with chromazurol S plate. Result based on physio-biochemical characteristics and molecular identification showed YN01 as Bacillus atrophaeus. The inhibition rate of rice blast mycelia exceeded 90%. Its best carbon source and nitrogen source were sucrose and yeast powder. The isolate could not grow normally when pH < 6. The best temperature was 30-35 °C. The best condition of inhibition effect included sucrose, glutamate, pH = 8, and the best temperature region of 30-35 °C. YN01 had secretion abilities of IAA and siderophores. Contrasted with negative control, bacterial suspension and fermentation broth inhibited spore germination significantly. Through nutrition and spatial competition and produced metabolites, YN01 led to slacken spore of rice blast fungal germination, aberrant morphology in mycelia, and disrupted mycelia physiological activity. In summary, for its powerful bacteriostatic activity YN01 is a potential antagonistic bacteria source in the future.

参考文献/References:

1 肖军, 杨镇, 陈珣, 王红, 肇莹, 刘国丽, 杨涛. 内生菌醇提取物对水稻抗稻瘟病的诱导作用[J]. 中国生物防治学报, 2015, 31 (3): 433-438 [Xiao J, Yang Z, Chen X, Wang H, Zhao Y, Liu GL, Yang T. Induced resistance against blast disease of rice by ethanol extract from endophytesto [J]. Chin J Biol Contr, 2015, 31 (3): 433-438]
2 张传清, 周明国, 朱国念. 稻瘟病化学防治药剂的历史沿革与研究现状[J]. 农药学学报, 2009, 11 (1): 72-80 [Zhang CQ, Zhou MG, Zhu GN. History and research advance of chemicals for controlling rice blast disease [J]. Chin J Pesticide Sci, 2009, 11 (1): 72-80]
3 刘小红, 张福生, 王国平, 陈锋, 章初龙, 林福呈. ZJUF0986活性代谢产物对稻瘟病菌致病性的影响[J]. 植物保护学报, 2008, 35 (6): 481-485 [Liu XH, Zhang FS, Wang GP, Chen F, Zhang CL, Lin FC. The effect of ZJUF0986 bioactivem etabolite on the pathogenicity of Magnaporthe oryzae [J]. Acta Phytoph Sin, 2008, 35 (6): 481-485]
4 李永刚, 宋兴舜, 马凤鸣, 文景芝, 赵雪莹. 水稻稻瘟病拮抗菌L1鉴定及抑菌特性的初步研究[J]. 微生物学通报, 2008, 35 (6): 898-902 [Li YG, Song XS, Ma FM, Wen JZ, Zhao XY. Identification and characterization of an antagonistic strain L1 against Phyricularia grisea [J]. Microbiology, 2008, 35 (6): 898-902]
5 姜海波, 徐文静, 杜茜, 许修宏, 李启云. BPS28发酵液抗菌谱及抗菌物质稳定性测试[J]. 吉林农业科学, 2009, 34 (1): HB, Xu WJ, Du Q, Xu XY, Li QY. The Inhibition spectrum and stability determination of BPS28 fermentation production [J]. J Jilin Agric Sci, 2009, 34 (1): 14-16]
6 Kawamata H, Narisawa K, HashiSa T. Suppression of riceblast by phylloplane fungi isolated from rice plants [J]. J Gen Plant Pathol, 2004, 70 (2): 131-138
7 穆常青, 刘雪, 陆庆光, 蒋细良, 朱昌雄. 枯草芽孢杆菌B-332菌株对稻瘟病的防治效果及定殖作用[J]. 植物保护学报, 2007, 34 (2): 123-128 [Mu CQ, Liu X, Lu QG, Jiang XL, Zhu CX. Biological control of rice blast by Bacillus subtilis B-332 strain [J]. Acta Phytophy Sin, 2007, 34 (2): 123-128]
8 Foldes T, Banhegyi I, Hepai Z. Isolation of Bacillus strains from the rhizosphere of cereals and in vitro screening for antagonism against phytopathogenic,foodborne pathogenic and spoilage microorganisms [J]. J Appl Microbiol, 2000, 89 (5): 840-846
9 王梦颖. 香蕉内生菌多样性分析及广谱拮抗菌筛选[D]. 海口: 海南大学, 2014 [Wang MY. Diversity research of endophytes and broad-spectrum antagonistic bacterias screening from banana [D]. Haikou: Hainan University, 2014]
10 刘起丽, 张建新, 徐瑞富, 段长勇. 柑橘皮内生细菌分离及柑橘青霉病菌拮抗菌筛选研究[J]. 中国农学通报, 2011, 27 (28): 235-239 [Liu QL, Zhang JX, Xu RF, Duan CY. Studies on Isolation of endophytic bacteria from citrus skins and screening of antagonistic strains against Penicillium italicum [J]. Chin Agric Sci Bulln, 2011, 27 (28): 235-239]
11 冯云利, 奚家勤, 马莉, 莫明和, 方敦煌, 夏振远, 雷丽萍, 杨发祥, 周峰. 烤烟品种NC297内生细菌中拮抗烟草黑胫病的生防菌筛选及种群组成分析[J]. 云南大学学报(自然科学版), 2011, 33 (4): 488-496 [Feng YL, Xi JQ, Ma L, Mo MH, Fang DH, Xia ZY, Lei LP, Yang FX, Zhou F. Isolation and phylogenetic analysis of the antagonistic endophytic bacteria against Phytophthora parasitica var. nicotianae from tobacco variety NC297 [J]. J Yunnan Univ, 2011, 33 (4): 488-496]
12 杨斌, 薛泉宏, 陈占全, 郭志英, 张晓鹿, 周永强, 许英俊, 孙德福. 青海沙珠玉人工植被系统土壤放线菌生态分布及拮抗性[J]. 应用生态学报, 2008, 19 (8): 1694-1701 [Yang B, Xu HQ, Chen ZQ, Guo ZY, Zhang XL, Zhou YQ, Xu YJ, Sun DF. Ecological distribution and antimicrobial effects of soil actinomycetes in artificial vegetation Systems in Shazhuyu of Qinghai [J]. Chin J Appl Ecol, 2008, 19 (8): 1694-1701
13 关统伟, 张利莉, 赵震宇. 新疆干旱区土壤放线菌拮抗植物病原真菌的研究[J]. 甘肃农业大学学报, 2007, 42 (2): 71-74 [Guan TW, Zhang LL, Zhao ZY, Antimicrobial activity of soil actinomycetes in the arid area of Xinjiang to plant pathogenic fungi [J]. J Gansu Agric Univ, 2007, 42 (2): 71-74]
14 何建清, 吴云锋, 袁耀锋. 西藏色季拉山土壤拮抗放线茵的筛选[J]. 西北农林科技大学学报(自然科学版), 2005, 33: 187-189 [He JQ, Wu YF, Yuan YF. Screening of antagonistica actinomycetes from Shegyla Mountain in Tibet [J]. J Northwest Sci-Tech Univ Agric For, 2005, 33: 187-189]
15 王建林, 钟志明, 王忠红, 陈宝雄, 余成群, 胡兴祥, 沈振西, 大次卓嘎, 张宪洲. 青藏高原高寒草原生态系统土壤碳氮比的分布特征[J]. 生态学报, 2014, 34 (22): 6678-6691 [Wang JL, Zhong ZM, Wang ZH, Chen BX, Yu CQ, Hu XX, Shen ZX, Da CZG, Zhang XZ. Soil C /N distribution characteristics of alpine steppe ecosystem in Qinhai-Tibetan Plateau [J]. Acta Ecol Sin, 2014, 34 (22): 6678-6691]
16 迟莉. 水稻稻瘟病拮抗菌筛选及施用后对水稻植株的影响[D]. 北京: 中国农业科学院, 2014 [Chi L. Effect on rice plants antimicrobial rice blast screening and application of antagonistic [D]. Beijing: Chinese Academy of Agriculture Science, 2014]
17 靳西彪. 海洋中稻瘟病菌的分离、发酵优化及其活性成分分析[D]. 雅安: 四川农业大学, 2012 [Jin XB. The isolation and ferment optimization of antifungal bacterium from marine samples and analysis of its active compounds [D]. Ya′an: Sichuan Agricultural University, 2012]
18 Glickmann E, Dessaux Y. A critical examination of the specificity of the Salkowski reagent for indolic compounds produced by phytopathogenic bacteria [J] . Appl Environ Microbiol, 1995, 619 (2): 793-796
19 赵翔, 谢志翔, 陈绍兴, 沈萍. 适合高产铁载体细菌筛选、检测体系的改进与探析[J]. 微生物学通报, 2006, 33 (6): 13-18 [Zhao X, Xie ZX, Chen SX, Shen P. Improvement and analysis in over-siderophores production bacteria filtrating and detecting [J].Microbiol China, 2006, 33 (6): 13-18
20 Neilands JB. Siderophore: Structure and function of microbial iron transport compounds [J]. J Biol Chem, 1995, 270 (45): 26723-26726
21 任争光, 张志勇, 魏艳敏. 芽胞杆菌防治园艺植物病害的研究进展[J]. 中国生物防治, 2006, 22 (S1): 194-198 [Ren ZG, Zhang ZY, Wei YM. Research progress on biological control of horticultural plant diseases by Bacillus spp. [J]. Chin J Biol Contr, 2006, 22 (S1): 194-198]
22 张淑梅, 沙长青, 赵晓宇, 王玉霞, 张先成, 李晶, 孟利强, 于德水. 一株抗真菌内生多粘芽孢杆菌的分离鉴定及对水稻恶苗病菌的抑制作用[J]. 中国生物工程杂志, 2010, 30 (2): 84-88 [Zhang SM, Sha CQ, Zhao XY, Wang YX, Zhang XC, Li J, Meng LQ, Yu DS. Identification of an endophytic Paenibacillus polymyxa strain producing antifungal protein and the inhibition to Fusarium moniliforme causing rice bakanae disease [J]. Chin Biotechnol, 2010, 30 (2): 84-88]
23 韦露, 陈偿, 龙云映, 蔡奕明, 黄晓纯. 一株短小芽孢杆菌B1的筛选鉴定及其抗菌特性研究[J]. 水产科学, 2015, 34 (3): 161-168 [Wei L, Chen C, Long YY, Cai YM, Huang XC. I, Isolation, identification and antimicrobial activity of Bacillus pumilus B1 [J]. Fish Sci, 2015, 34 (3): 161-168]
24 辛海峰, 孟艳艳, 李建宏, 马鸿翔, 张旭. 一株萎缩芽孢杆菌在小麦中的定植及对赤霉病的防治[J]. 生物学杂志, 2013, 32 (6): 1490-1496 [Xin HF, Meng YY, Li JH, Ma HX, Zhang X. Bacillus atrophaeus strain’s colonization in wheat plant and its inhibition efficiency to Fu-sarium head blight [J]. Chin J Ecol, 2013, 32 (6): 1490-1496
25 刘丁. 萎缩芽孢杆菌(Bacillus atrophaeus)抑制黄曲霉作用研究[D]. 雅安: 四川农业大学, 2013 [Liu D. Study on interact of Bacillus atrophaeus to Aspergillus flabus [D]. Ya′an: Sichaung Agricultural University, 2013
26 郭荣君, 李世东, 张晶, 张旋, 穆光远, 王忠玉. 基于营养竞争原理的大豆根腐病生防芽孢杆菌的筛选及其特性研究[J]. 植物病理学报, 2010, 40 (3): 307-314 [Guo RJ, Li SD, Zhang J, Zhang X, Mu GY, Wang ZY. Characterization of Bacillus strains screened via nutritional competition for biocontrol of soybean root rot disease [J]. Acta Phytopathol Sin, 2010, 40 (3): 307-314]
27 张宝俊, 张家榕, 韩巨才, 刘慧平, 王建明. 内生解淀粉芽胞杆菌LP-5抗菌蛋白的分离纯化及特性[J]. 植物保护学报, 2010, 37 (2): 143-146 [Zhang BJ, Zhang JR, Han JC, Liu HP, Wang JM. Purification and characterization of an antifungal protein from endophytic Bacillus amyloliquefaciens LP-5[J]. Acta Phytophyl Sin, 2010, 37 (2): 143-146]
28 Simon CA, Andrea KR, Francisco RQ. Bacterial iron homeo-stasis [J]. FEMS Microbiol Rev, 2003, 27 (2): 215-237
29 Kloepper JW, John L, Teintze M. Enhanced plant growth bysiderophores produced by plant growth-promoting Rhizobacteria [J]. Nature, 1980, 286: 885-886
30 Petra M, David C, Zed R. Rhizosphere interactions between microorganisms and plants govern iron and phosphorus acquisition along the root axis-model and research methods [J]. Soil Biol Biochem, 2011, 43 (5): 883-894
31 Saravanan VS, Madhaiyan M, Osborne J, Thangaraju M, Sa TM. Ecological occurrence of Gluconacetobacter diazotrophicus and nitrogen-fixing Acetobacteraceae members: their possible role in plant growth promotion [J]. Microb Ecol, 2008, 55 (1): 130-140
32 Saunders DGO, Aves SJ, Nicholas J. Talbot cell cycle–mediated regulation of plant infection by the rice blast fungus [J]. Plant Cell, 2010, 22 (2): 497-507
33 Howard RJ, Valent B. Breaking and entering: host penetration by the fungal rice blast pathogen Magnaporthe grisea [J]. Annu Rev Microbiol, 1996, 50 (2): 491-512

相似文献/References:

[1]李芙蓉,丁涛,白林含.拟茎点霉属Phomopsis sp. S4菌株发酵产物对稻瘟病菌细胞膜的抑制作用[J].应用与环境生物学报,2018,24(02):342.[doi:10.19675/j.cnki.1006-687x.2017.04013]
 LI Furong,DING Tao,BAI Linhan.Inhibition mechanism of fermentation broth extract of Phomopsis sp. strain S4 on cell membranes of Magnaporthe oryzae[J].Chinese Journal of Applied & Environmental Biology,2018,24(01):342.[doi:10.19675/j.cnki.1006-687x.2017.04013]

更新日期/Last Update: 2017-02-25