Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (16): 3343-3350.doi: 10.3864/j.issn.0578-1752.2011.16.006

• PLANT PROTECTION • Previous Articles     Next Articles

A Nitrogen-Fixing Bacterium Paenibacillus sp. GD812 Antagonistic Against Plant Pathogenic Fungi

CHEN  Qian, GAO  Miao, HU  Hai-Yan, XU  Jing, ZHOU  Yi-Qing, SUN  Jian-Guang   

  1. 中国农业科学院农业资源与农业区划研究所/农业部作物营养与施肥重点实验室
  • Received:2011-01-20 Revised:2011-05-20 Online:2011-08-15 Published:2011-06-03

Abstract: 【Objective】 The objective of this study is to screen nitrogen-fixing bacteria antagonistic against plant pathogenic fungi and research nitrogenase activity, nifH, physiological and biochemical characteristics, identification, stress resistance of the functional strains and inoculation effect on plant and to prepare strains for microbial fertilizer production. 【Method】 Nitrogen-free medium was used to isolate nitrogen-fixers and confrontation method was used to screen antagonistic bacteria against plant pathogenic fungus. Nitrogenase activity was determined with acetylene reduction assay. 16S rDNA and nifH were amplified with PCR. Strain identification was carried out based on the morphology, physiology, and biochemical test and 16S rDNA sequence analysis. Microbial inoculation effect on plant was tested with a pot culture in green house. 【Result】 One strain designated as GD812 was selected, which showed nitrogenase activity 30.661 nmol C2H4/h•mg protein, and strongly antagonistic against plant pathogenic fungi Gibberella zeae and Verticillium dahliae with the inhibition rate of 59.5% and 49.3%, respectively. nifH of GD812 is 300 bp sharing 98% sequence identity with that of Paenibacillus sp. Bs57. GD812 was identified as Paenibacillus sp. based on the results of morphology, physiology and biochemical test and 16S rDNA sequence analysis. Further investigations showed that GD812 could use 20 of the 35 tested carbon sources, could grow under temperature ranging from 4 to 50℃, and could grow in range of pH4 to11. Pot culture result showed that fresh weight of Chinese cabbage inoculated with GD812 increased by 52% compared with the control. 【Conclusion】 The newly selected Paenibacillus sp. GD812 showed high nitrogenase activity, strong antagonism against plant pathogenic fungi, extensive carbon source utilization, stress resistance and good inoculation effect, which is a candidate to be further developed for microbial fertilizer production.

Key words: biological nitrogen fixation, Paenibacillus, strain screening, microbial fertilizer

[1]陈文新. 生物固氮//中国土壤学会. 氮素循环与农业和环境学术研讨会论文集. 厦门: 厦门大学出版社, 2001: 4-5.

Chen W X. Biological nitrogen fixation// Soil Science Society of China (ed). Proceeding of Conference on Nitrogen Cycling and Agriculre Environment. Xiamen: Xiamen University Press, 2001: 4-5. (in Chinese)

[2]Chen W M, de Faria S M, James E K, Elliott G N, Lin K Y, Chou J H, Sheu S Y, Cnockaert M, Sprent J I, Vandamme P. Burkholderia nodosa sp. nov., isolated from root nodules of the woody Brazilian legumes Mimosa bimucronata and Mimosa scabrella. International Journal of Systememic and Evolutional Microbiology, 2007, 57: 1055-1059.

[3]Dutta D, Gachhui R. Nitrogen-fixing and cellulose-producing Gluconacetobacter kombuchae sp. nov., isolated from Kombucha tea. International Journal of Systememic and Evolutional Microbiology, 2007, 57: 353-357.

[4]Addison S L, Foote S M, Reid N M, Lloyd-Jones G. Novosphingobium nitrogenifigens sp. nov., a polyhydroxyalkanoate- accumulating diazotroph isolated from a New Zealand pulp and paper wastewater. International Journal of Systememic and Evolutional Microbiology, 2007, 57: 2467-2471.

[5]Kämpfer P, Thummes K, Chu H I, Tan C C, Arun A B, Chen W M, Lai W A, Shen F T, Rekha P D, Young C C. Pseudacidovorax intermedius gen. nov., sp. nov., a novel nitrogen-fixing betaproteobacterium isolated from soil. International Journal of Systememic and Evolutional Microbiology, 2008, 58: 491-495.

[6]Zhang G X, Peng G X, Wang E T, Yan H, Yuan Q H, Zhang W, Lou X, Wu H, Tan Z Y. Diverse endophytic nitrogen-fixing bacteria isolated from wild rice Oryza rufipogon and description of Phytobacter diazotrophicus gen. nov. sp. nov. Archives of Microbiology, 2008, 189: 431-439.

[7]孙建光, 张燕春, 徐  晶, 胡海燕. 高效固氮芽孢杆菌选育及其生物学特性研究. 中国农业科学,2009, 42(6): 2043-2051.

Sun J G, Zhang Y C, Xu J, Hu H Y. Isolation and biological characteristic investigation on efficient nitrogen-fixing bacilli. Scientia Agricultura Sinica, 2009, 42(6): 2043-2051. (in Chinese)

[8]张燕春, 孙建光, 徐  晶, 胡海燕. 固氮芽孢杆菌GD272的筛选鉴定及其固氮性能研究. 植物营养与肥料学报, 2009, 15(5): 1196-1201.

Zhang Y C, Sun J G, Xu J, Hu H Y. Isolation and identification and evaluation of nitrogen-fixing bacillus strain GD272. Plant Nutrition and Fertilizer Science, 2009, 15(5): 1196-1201. (in Chinese)

[9]孙建光, 徐  晶, 胡海燕, 张燕春, 刘  君, 王文博, 孙燕华. 中国十三省市土壤中非共生固氮微生物菌种资源研究. 植物营养与肥料学报, 2009, 15(6): 1450-1465.

Sun J G, Xu J, Hu H Y, Zhang Y C, Liu J, Wang W B, Sun Y H. Collection and investigation on asymbiotic nitrogen-fixing microbial resources from 13 provinces over China. Plant Nutrition and Fertilizer Science, 2009, 15(6): 1450-1465. (in Chinese)

[10]刘晓光, 高克祥, 谷建才, 杜建玲, 唐秀光. 毛白杨内生菌优势种毛壳ND35室内拮抗作用的研究. 林业科学, 1999, 35(5): 57-62.

Liu X G, Gao K X, Gu J C, Du J L, Tang X G. Testing on the antagonism of the dominant of endophytic fungi from Populus tomentosa, Chaetomium ND35 in the laboratory. Scientia Silvae Sinicae, 1999, 35 (5): 57-62. (in Chinese)

[11]东秀珠, 蔡妙英. 常见细菌系统鉴定手册. 北京: 科学出版社, 2001.

Dong X Z, Cai M Y. Manual of Systermatic Determinative Bacteriology. Beijing: Science Press, 2001. (in Chinese)

[12]沈  萍, 范秀容, 李广武. 微生物学实验(第三版). 北京: 高等教育出版社, 1999.

Shen P, Fan X R, Li G W. Microbiology Experiment (3rd Edition). Beijing: High Education Press, 1999. (in Chinese)

[13]Ding Y, Wang J, Liu Y, Chen S. Isolation and identification of nitrogen-fixing bacilli from plant rhizospheres in Beijing region. Journal of Applied Microbiology, 2005, 99: 1271-1281.

[14]赵  斌, 何绍江. 微生物学实验. 北京: 科学出版社, 2002.

Zhao B, He S J. Microbiology Experiment. Beijing: Science Press, 2002. (in Chinese)

[15]Kim J F, Jeong H, Park S Y, Kim S B, Park Y K, Choi S K, Ryu C M, Hur C G, Ghim S Y, Oh T K, Kim J J, Park C S, Park S H. Genome sequence of the polymyxin-producing plant-probiotic rhizobacterium Paenibacillus polymyxa E681. Jouranl of Bacteriology, 2010, 192(22): 6103-6104.

[16]Suominen I, Spröer C, Kämpfer P, Rainey F A, Lounatmaa K, Salkinoja-Salonen M. Paenibacillus stellifer sp. nov., a cyclodextrin- producing species isolated from paperboards. International Journal of Systememic and Evolutional Microbiology, 2003, 53: 1369-1374.

[17]Aguilera M, Monteoliva-Sánchez M, Suárez A, Guerra V, Lizama C, Bennasar A, Ramos-Cormenzana A. Paenibacillus jamilae sp. nov., an exopolysaccharide-producing bacterium able to grow in olive-mill wastewater. International Journal of Systememic and Evolutional Microbiology, 2001, 51(5): 1687-1692.

[18]Holt J G. Bergey's Manual of Systematic Bacteriology (1st Edition).  Baltimore: Williams & Wilkins, 1984-1989.

[19]陈文新. 土壤和环境微生物学. 北京: 北京农业大学出版社, 1990: 152-156.

Chen W X. Soil and Enviromental Microbiology. Beijing: Beijing Agricultural University Press, 1990: 152-156. (in Chinese)

[20]Jeong-Hwa C, Im W T, Yoo J S, Lee S M, Moon D S, Kim H J, Rhee S K, Roh D H. Paenibacillus donghaensis sp. nov., a xylan-degrading and nitrogen-fixing bacterium isolated from East Sea sediment. Journal of Microbiology Biotechnology, 2008, 18: 189-193.

[21]Ma Y C, Chen S F. Paenibacillus forsythiae sp. nov., a nitrogen-fixing species isolated from rhizosphere soil of Forsythia mira. International Journal of Systememic and Evolutional Microbiology, 2008, 58: 319-323.

[22]Ma Y C, Xia Z Q, Liu X M, Chen S F. Paenibacillus sabinae sp. nov., a nitrogen-fixing species isolated from the rhizosphere soils of shrubs. International Journal of Systememic and Evolutional Microbiology, 2007, 57: 6-11.

[23]Lee F L, Kuo H P, Tai C J, Yokota A, Lo C C. Paenibacillus taiwanensis sp. nov., isolated from soil in Taiwan. International Journal of Systememic and Evolutional Microbiology, 2007, 57: 1351-1354.

[24]Ma Y C, Zhang J, Chen S F. Paenibacillus zanthoxyli sp. nov., a novel nitrogen-fixing species isolated from the rhizosphere of Zanthoxylum

 

simulans. International Journal of Systememic and Evolutional Microbiology, 2007, 57: 873-877.

[25]Beneduzi A, Costa P B, Parma M, Melo I S, Bodanese-Zanettini M H, Passaglia L M. Paenibacillus riograndensis sp. nov., a nitrogen-fixing species isolated from the rhizosphere of Triticum aestivum. International Journal of Systememic and Evolutional Microbiology, 2010, 60: 128-133.

[26]Jin H J, Lv J, Chen S F. Paenibacillus sophorae sp. nov., a nitrogen-fixing species isolated from the rhizosphere of Sophora japonica. International Journal of Systememic and Evolutional Microbiology, 2011, 61: 767-771.

[27]Jin H J, Zhou Y G, Liu H C, Chen S F. Paenibacillus jilunlii sp. nov., a novel nitrogen-fixing species isolated from the rhizosphere of Begonia semperflorens. International Journal of Systememic and Evolutional Microbiology, 2011, 61: 1350-1355.

[28]Hong Y Y, Ma Y C, Zhou Y G, Gao F, Liu H C, Chen S F. Paenibacillus sonchi sp. nov., a nitrogen-fixing species isolated from the rhizosphere of Sonchus oleraceus. International Journal of Systememic and Evolutional Microbiology, 2009, 59: 2656-2661.
[1] LI Qiang, HUANG YingXin, ZHONG RongZhen, SUN HaiXia, ZHOU DaoWei. Influence of Medicago sativa Proportion on Its Individual Nitrogen Fixation Efficiency and Underlying Physiological Mechanism in Legume-Grass Mixture Grassland [J]. Scientia Agricultura Sinica, 2020, 53(13): 2647-2656.
[2] GONG AnDong, ZHU ZiYu, LU YaNan, WAN HaiYan, WU NanNan, Cheelo Dimuna, GONG ShuangJun, WEN ShuTing, HOU Xiao. Functional Analysis of Burkholderia pyrrocinia WY6-5 on Phosphate Solubilizing, Antifungal and Growth-Promoting Activity of Maize [J]. Scientia Agricultura Sinica, 2019, 52(9): 1574-1586.
[3] LIU Peng,TIAN YingZhe,ZHONG YongJia,LIAO Hong. Isolation and Application of Effective Rhizobium Strains in Peanut on Acidic Soils [J]. Scientia Agricultura Sinica, 2019, 52(19): 3393-3403.
[4] ZHANG Bing-huo, LI Han-quan, LUO Juan-yan, YANG Jian-yuan, SHI Hong-qiu, SUN Feng-zhen. Influences of Actinomycete Strain JXJ-0136 on the Growth of Brassica chinensis and Vigna unguiculata and Its Phosphate Solubilization [J]. Scientia Agricultura Sinica, 2016, 49(16): 3152-3161.
[5] MA Ming-chao, LIU Li, JIANG Xin, GUAN Da-wei, LI Jun. Evaluation of the Effect of Co-Inoculant of Paenibacillus mucilaginosus and Bradyrhizobium japonicum in Application [J]. Scientia Agricultura Sinica, 2015, 48(18): 3600-3611.
[6] XIE Kai-yun, ZHANG Ying-jun, LI Xiang-lin, HE Feng, WAN Li-qiang, WANG Dong, QIN Yan. Competition and Coexistence of Alfalfa (Medicago sativa L.) and Smooth Brome (Bromus inermis Layss.) in Mixture [J]. Scientia Agricultura Sinica, 2015, 48(18): 3767-3778.
[7] WEN Feng-yun,LIAO Fu-pin,LIN Jian-rong,ZHONG Yang-sheng
.

Cloning, Expression and Application of β-1,3-1,4-glucanase Gene from Paenibaccillus polymyxa CP7

[J]. Scientia Agricultura Sinica, 2010, 43(22): 4614-4623 .
[8] CHEN Hai-ying,LIAO Fu-ping,LIN Jian-rong. Isolation and Structure Analysis of the Components of Anti-Gram- Negative Bacteria by the Strain CP7 of Paenibacillus polymyxa
[J]. Scientia Agricultura Sinica, 2009, 42(6): 2105-2110 .
[9] SUN Jian-guang,ZHANG Yan-chun,XU Jing,HU Hai-yan
. Isolation and Biological Characteristic Investigation on Efficient Nitrogen-Fixing Bacilli
[J]. Scientia Agricultura Sinica, 2009, 42(6): 2043-2051 .
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!