Journal of Microbiology

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Biocontrol activity of volatile organic compounds from Streptomyces alboflavus TD-1 against Aspergillus flavus growth and aflatoxin production: Mingguan Yang , Laifeng Lu , Jing Pang , Yiling Hu , Qingbin Guo , Zhenjing Li , Shufen Wu , Huanhuan Liu , Changlu Wang; J. Microbiol. 2019;57(5):396-404. Published online May 6, 2019; DOI: https://doi.org/10.1007/s12275-019-8517-9

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Aspergillus flavus is a saprophytic fungus that contaminates crops with carcinogenic aflatoxin. In the present work, the antifungal effects of volatile organic compounds (VOCs) from Streptomyces alboflavus TD-1 against A. flavus were investigated. VOCs from 8-day-old wheat bran culture of S. alboflavus TD-1 displayed strong inhibitory effects against mycelial growth, sporulation, and conidial germination of A. flavus. Severely misshapen conidia and hyphae of A. flavus were observed by scanning electron microscopy after exposure to VOCs for 6 and 12 h, respectively. Rhodamine 123 staining of mitochondria indicated that mitochondria may be a legitimate antifungal target of the VOCs from S. alboflavus TD-1. Furthermore, the VOCs effectively inhibited aflatoxin B1 production by downregulating genes involved in aflatoxin biosynthesis. Dimethyl trisulfide and benzenamine may play important roles in the suppression of A. flavus growth and production of aflatoxin. The results indicate that VOCs from S. alboflavus TD-1 have tremendous potential to be developed as a useful bio-pesticide for controlling A. flavus.

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Isolation and characterization of Aspergillus flavus strains in China: Firew Tafesse Mamo , Bo Shang , Jonathan Nimal Selvaraj , Yan Wang , Yang Liu; J. Microbiol. 2018;56(2):119-127. Published online February 2, 2018; DOI: https://doi.org/10.1007/s12275-018-7144-1

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Abstract

Important staple foods (peanuts, maize and rice) are susceptible to contamination by aflatoxin (AF)-producing fungi such as Aspergillus flavus. The objective of this study was to explore non-aflatoxin-producing (atoxigenic) A. flavus strains as biocontrol agents for the control of AFs. In the current study, a total of 724 A. flavus strains were isolated from different regions of China. Polyphasic approaches were utilized for species identification. Non-aflatoxin and non-cyclopiazonic acid (CPA)-producing strains were further screened for aflatoxin B1 (AFB1) biosynthesis pathway gene clusters using a PCR assay. Strains lacking an amplicon for the regulatory gene aflR were then analyzed for the presence of the other 28 biosynthetic genes. Only 229 (32%) of the A. flavus strains were found to be atoxigenic. Smaller (S) sclerotial phenotypes were dominant (51%) compared to large (L, 34%) and non-sclerotial (NS, 15%) phenotypes. Among the atoxigenic strains, 24 strains were PCR-negative for the fas-1 and aflJ genes. Sixteen (67%) atoxigenic A. flavus strains were PCRnegative for 10 or more of the biosynthetic genes. Altogether, 18 new PCR product patterns were observed, indicating great diversity in the AFB1 biosynthesis pathway. The current study demonstrates that many atoxigenic A. flavus strains can be isolated from different regions of China. In the future laboratory as well as field based studies are recommended to test these atoxigenic strains as biocontrol agents for aflatoxin contamination.

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The biodiversity of Aspergillus flavus in stored rice grain leads to a decrease in the overall aflatoxin B1 production in these species
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Characterization of siderophore produced by Pseudomonas syringae BAF.1 and its inhibitory effects on spore germination and mycelium morphology of Fusarium oxysporum: Sumei Yu , Chunying Teng , Jinsong Liang , Tao Song , Liying Dong , Xin Bai , Yu Jin , Juanjuan Qu; J. Microbiol. 2017;55(11):877-884. Published online October 27, 2017; DOI: https://doi.org/10.1007/s12275-017-7191-z

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Abstract

In this study, an antagonistic bacterium against Fusarium oxysporum was identified and designated as Pseudomonas syringae strain BAF.1 on the basis of 16S rDNA sequence analysis and physiological-biochemical characteristics. It produced catechol-species siderophore at a molecular weight of 488.59 Da and a maximum amount of 55.27 μg/ml with glucose as a carbon source and asparagine as a nitrogen source at a C/N ratio of 10:1, 30°C and pH 7. The siderophore exhibited prominent antagonistic activity against Fusarium oxysporum with a maximum inhibition rate of 95.24% and had also suppressive effects on other kinds of 11 phytopathogenic fungi in the absence of FeCl3·6H2O. Spore germination was completely inhibited by 50 μl of the siderophorecontaining solution, and the ultrastructures of mycelia and spores were also considerably suppressed by siderophore treatment as established by electron microscopy observation. These results indicate that the siderophore produced by Pseudomonas syringae BAF.1 could be potentially used for biocontrol of pathogenic Fusarium oxysporum.

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Poly-γ-glutamic acid productivity of Bacillus subtilis BsE1 has positive function in motility and biocontrol against Fusarium graminearum: Luyao Wang , Ning Wang , Dandan Mi , Yuming Luo , Jianhua Guo; J. Microbiol. 2017;55(7):554-560. Published online June 30, 2017; DOI: https://doi.org/10.1007/s12275-017-6589-y

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Abstract

In this study, we investigate the relationship between γ-PGA productivity and biocontrol capacity of Bacillus subtilis BsE1; one bacterial isolate displayed 62.14% biocontrol efficacy against Fusarium root rot. The γ-PGA yield assay, motility assay, wheat root colonization assay, and biological control assay were analysed in different γ-PGA yield mutants of BsE1. The pgsB (PGA-synthase-CapB gene) deleted mutant of BsE1 reduced γ-PGA yield and exhibited apparent decline of in vitro motile ability. Deletion of pgsB impaired colonizing capacity of BsE1 on wheat root in 30 days, also lowered biocontrol efficacies from 62.08% (wild type BsE1) to 14.22% in greenhouse experiment against Fusarium root rot. The knockout of pgdS and ggt (genes relate to two γ-PGA degrading enzymes) on BsE1, leads to a considerable improvement in polymer yield and biocontrol efficacy, which attains higher level compared with wild type BsE1. Compared with ΔpgsB mutant, defense genes related to reactive oxygen species (ROS) and phytoalexin expressed changes by notable levels on wheat roots treated with BsE1, demonstrating the functional role γ-PGA plays in biocontrol against Fusarium root rot. γ-PGA is not only important to the motile and plant root colonization ability of BsE1, but also essential to the biological control performed by BsE1 against Fusarium root rot. Our goal in this study is to reveals a new perspective of BCAs screening on bacterial isolates, without good performance during pre-assays of antagonism ability.

Citations

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Double-Wing Motif Protein is a Novel Biofilm Regulatory Factor of the Plant Disease Biocontrol Agent, Bacillus subtilis
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Bo Zhang, Xiaoying Hu, Donglin Zhao, Yuping Wang, Jianhua Qu, Yue Tao, Zhonghui Kang, Hongqi Yu, Jingyi Zhang, Ying Zhang
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Research Support, Non-U.S. Gov't

The Use of Pseudomonas fluorescens P13 to Control Sclerotinia Stem Rot (Sclerotinia sclerotiorum) of Oilseed Rape: Hui Li , Huaibo Li , Yan Bai , Jing Wang , Ming Nie , Bo Li , Ming Xiao; J. Microbiol. 2011;49(6):884-889. Published online December 28, 2011; DOI: https://doi.org/10.1007/s12275-011-1261-4

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Abstract: Sclerotinia stem rot (SSR) caused by the fungus Sclerotinia sclerotiorum has been an increasing threat to oilseed rape (Brassica napus L.) cultivation. Efficient and environment‐friendly treatments are much needed. Here we focus on microbial control. The Pseudomonas fluorescens P13 that was isolated from oilseed rape cultivation soil, proved to be a useful biocontrol strain for application. Morphology, physiological and biochemical tests and 16S rDNA analysis demonstrated that it was P. fluorescens P13 and that it had a broad antagonistic spectrum, significantly lessening the mycelial growth of S. sclerotiorum by 84.4% and suppressing sclerotial formation by 95‐100%. Scanning electron microscopy studies attested that P13 deformed S. sclerotiorum mycelia when they were cultured together. P13 did not produce chitinase but did produce hydrogen cyanide (HCN) which was likely one of the antagonistic mechanisms. The density of P13 remained at a high level (≥106 CFU/ml) during 5 weeks in the rhizosphere soil and roots. P13 reduced SSR severity at least by 59% in field studies and also promoted seedling growth (p<0.05) at the seedling stage. From these data, our work provided evidence that P13 could be a good alternative biological resource for biocontrol of S. sclerotiorum.

Journal Article

Cyclic Lipopeptide Profile of Three Bacillus subtilis Strains; Antagonists of Fusarium Head Blight: Christopher A. Dunlap , David A. Schisler , Neil P. Price , Steven F. Vaughn; J. Microbiol. 2011;49(4):603-609. Published online September 2, 2011; DOI: https://doi.org/10.1007/s12275-011-1044-y

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Abstract

The objective of the study was to identify the lipopetides associated with three Bacillus subtilis strains. The strains are antagonists of Gibberella zeae, and have been shown to be effective in reducing Fusarium head blight in wheat. The lipopeptide profile of three B. subtilis strains (AS43.3, AS43.4, and OH131.1) was determined using mass spectroscopy. Strains AS43.3 and AS43.4 produced the anti-fungal lipopeptides from the iturin and fengycin family during the stationary growth phase. All three strains produced the lipopeptide surfactin at different growth times. Strain OH131.1 only produced surfactin under these conditions. The antifungal activity of the culture supernatant and individual lipopeptides was determined by the inhibition of G. zeae. Cell-free supernatant from strains AS43.3 and AS43.4 demonstrated strong antibiosis of G. zeae, while strain OH131.1 had no antibiosis activity. These results suggest a different mechanism of antagonism for strain OH131.1, relative to AS43.3 and AS43.4.

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Phytopathology®.2013; 103(2): 146. CrossRef
Biological control of winter wheat pathogens with the use of antagonisticSphingomonasbacteria under greenhouse conditions
Urszula Wachowska, Witold Irzykowski, Małgorzata Jędryczka, Anna D. Stasiulewicz-Paluch, Katarzyna Głowacka
Biocontrol Science and Technology.2013; 23(10): 1110. CrossRef
Production of schizophyllan from distiller’s dried grains with solubles by diverse strains of Schizophyllum commune
Nongnuch Sutivisedsak, Timothy D Leathers, Neil PJ Price
SpringerPlus.2013;[Epub] CrossRef
Utilization of agricultural biomass in the production of the biopolymer schizophyllan
Nongnuch Sutivisedsak, Timothy D Leathers, Melinda S Nunnally, Neil P J Price, Girma Biresaw
Journal of Industrial Microbiology and Biotechnology.2013; 40(1): 105. CrossRef
Analysis of free amino acids during fermentation by Bacillus subtilis using capillary electrophoresis
Yanli Ren, Jinyan Zhou, Xiaoyong Zhang, Zhidong Li, Juan Zhong, Jie Yang, Tan Xu, Hong Tan
Biotechnology and Bioprocess Engineering.2012; 17(6): 1244. CrossRef

Research Support, Non-U.S. Gov'ts

Application of Terminal Restriction Fragment Length Polymorphism (T-RFLP) Analysis to Monitor Effect of Biocontrol Agents on Rhizosphere Microbial Community of Hot Pepper (Capsicum annuum L.): Young Tae Kim , Myoungho Cho , Je Yong Jeong , Hyang Burm Lee , Seung Bum Kim; J. Microbiol. 2010;48(5):566-572. Published online November 3, 2010; DOI: https://doi.org/10.1007/s12275-010-0126-6

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Abstract: Microbial communities in hot pepper (Capsicum annuum L.) cultivation fields under different cultivation methods were investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis. Rhizosphere soil and leaf samples were collected from control, conventional and nature-friendly cultivation fields between May and July, 2009. Two Bacillus subtilis strains were applied to nature-friendly cultivation fields as biocontrol agents during the sampling period. Relative abundances of bacteria and plant pathogenic fungi related T-RFs were also measured to monitor the effect of biocontrol agents on potential plant pathogenic fungi. In the principal component analysis (PCA) based on T-RFLP profiles, the microbial communities from rhizosphere soil samples in July, including bacteria and fungi, showed distinct difference between nature-friendly cultivation fields and other cultivation fields. However, there was no correlation between cultivation methods and leaf microbial communities at any sampling period. Changes in the abundance of bacteria related T-RF in the rhizosphere of nature-friendly cultivation fields were observed clearly two months after application of biocontrol agent, while the abundance of plant pathogenic fungi related T-RFs significantly decreased.

The Endophyte Curtobacterium flaccumfaciens Reduces Symptoms Caused by Xylella fastidiosa in Catharanthus roseus: Paulo Teixeira Lacava , Wenbin Li , Welington Luiz Araujo , Joao Lucio Azevedo , John Stephen Hartung; J. Microbiol. 2007;45(5):388-393.; DOI: https://doi.org/2599 [pii]

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Abstract: Citrus variegated chlorosis (CVC) is a disease of the sweet orange [Citrus sinensis (L.)], which is caused by Xylella fastidiosa subsp. pauca, a phytopathogenic bacterium that has been shown to infect all sweet orange cultivars. Sweet orange trees have been occasionally observed to be infected by Xylella fastidiosa without evidencing severe disease symptoms, whereas other trees in the same grove may exhibit severe disease symptoms. The principal endophytic bacterial species isolated from such CVC-asymptomatic citrus plants is Curtobacterium flaccumfaciens. The Madagascar periwinkle [Citrus sinensis (L.)] is a model plant which has been used to study X. fastidiosa in greenhouse environments. In order to characterize the interactions of X. fastidiosa and C. flaccumfaciens, periwinkle plants were inoculated separately with C. flaccumfaciens, X. fastidiosa, and both bacteria together. The number of flowers produced by the plants, the heights of the plants, and the exhibited disease symptoms were evaluated. PCR-primers for C. flaccumfaciens were designed in order to verify the presence of this endophytic bacterium in plant tissue, and to complement an existing assay for X. fastidiosa. These primers were capable of detecting C. flaccumfaciens in the periwinkle in the presence of X. fastidiosa. X. fastidiosa induced stunting and reduced the number of flowers produced by the periwinkle. When C. flaccumfaciens was inoculated together with X. fastidiosa, no stunting was observed. The number of flowers produced by our doubly- inoculated plants was an intermediate between the number produced by the plants inoculated with either of the bacteria separately. Our data indicate that C. flaccumfaciens interacted with X. fastidiosa in C. roseus, and reduced the severity of the disease symptoms induced by X. fastidiosa. Periwinkle is considered to be an excellent experimental system by which the interaction of C. flaccumfaciens and other endophytic bacteria with X. fastidiosa can be studied.

Isolation and Characterization of Bacillus Strains for Biological Control: Han-Soo Kim , Jiyong Park , Sung-Won Choi , Kee-Hyun Choi , Gung Pyo Lee , Soo Jung Ban , Chang Hoo Lee , Chung Sun Kim; J. Microbiol. 2003;41(3):196-201.

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Abstract: The object of this study was to characterize Bacillus strains GB-017 and GB-0356, which produce antifungal substances, especially for plant pathogens. In addition, this study was undertaken to characterize the culture conditions required for the production of antifungal substances and to document some of the properties of the antifungal substance produced by these soil-isolated strains. Strains GB-0365 and GB-017 were found to be bacillus-shaped, gram-positive and motile, and to inhibit Botrytis cineria, Fusarium sp., Pythium sp., and Rhizoctonia solani. Antagonistic activity was maintained up to pH 9.0, and the antifungal activity was stable to heat at 80oC for 1 h. Antifungal substances were separated and purified using ion exchange and adsorption columns including WK-10(H^+) (pH 7.0), HP-20 column (pH 3.0) and IPA (pH 3.0). and IPA. Its UV absorption spectrum showed major peaks at 231 and 259 nm, corresponding to polyene and lactone. A fast atom bombardment mass spectrum (FAB MS) showed a highest peak at 441 m/z and major peaks at 192, 205, and 370 m/z.

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