Biocontrol Activity of Volatile-Producing Bacillus megaterium and Pseudomonas protegens Against Aspergillus and Penicillium spp. Predominant in Stored Rice Grains: Study II원문보기
In our previous studies, Bacillus megaterium KU143, Microbacterium testaceum KU313, and Pseudomonas protegens AS15 have been shown to be antagonistic to Aspergillus flavus in stored rice grains. In this study, the biocontrol activities of these strains were evaluated against Aspergillus candidus, As...
In our previous studies, Bacillus megaterium KU143, Microbacterium testaceum KU313, and Pseudomonas protegens AS15 have been shown to be antagonistic to Aspergillus flavus in stored rice grains. In this study, the biocontrol activities of these strains were evaluated against Aspergillus candidus, Aspergillus fumigatus, Penicillium fellutanum, and Penicillium islandicum, which are predominant in stored rice grains. In vitro and in vivo antifungal activities of the bacterial strains were evaluated against the fungi on media and rice grains, respectively. The antifungal activities of the volatiles produced by the strains against fungal development and population were also tested using I-plates. In in vitro tests, the strains produced secondary metabolites capable of reducing conidial germination, germ-tube elongation, and mycelial growth of all the tested fungi. In in vivo tests, the strains significantly inhibited the fungal growth in rice grains. Additionally, in I-plate tests, strains KU143 and AS15 produced volatiles that significantly inhibited not only mycelial growth, sporulation, and conidial germination of the fungi on media but also fungal populations on rice grains. GC-MS analysis of the volatiles by strains KU143 and AS15 identified 12 and 17 compounds, respectively. Among these, the antifungal compound, 5-methyl-2-phenyl-1H-indole, was produced by strain KU143 and the antimicrobial compounds, 2-butyl 1-octanal, dimethyl disulfide, 2-isopropyl-5-methyl-1-heptanol, and 4-trifluoroacetoxyhexadecane, were produced by strain AS15. These results suggest that the tested strains producing extracellular metabolites and/or volatiles may have a broad spectrum of antifungal activities against the grain fungi. In particular, B. megaterium KU143 and P. protegens AS15 may be potential biocontrol agents against Aspergillus and Penicillium spp. during rice grain storage.
In our previous studies, Bacillus megaterium KU143, Microbacterium testaceum KU313, and Pseudomonas protegens AS15 have been shown to be antagonistic to Aspergillus flavus in stored rice grains. In this study, the biocontrol activities of these strains were evaluated against Aspergillus candidus, Aspergillus fumigatus, Penicillium fellutanum, and Penicillium islandicum, which are predominant in stored rice grains. In vitro and in vivo antifungal activities of the bacterial strains were evaluated against the fungi on media and rice grains, respectively. The antifungal activities of the volatiles produced by the strains against fungal development and population were also tested using I-plates. In in vitro tests, the strains produced secondary metabolites capable of reducing conidial germination, germ-tube elongation, and mycelial growth of all the tested fungi. In in vivo tests, the strains significantly inhibited the fungal growth in rice grains. Additionally, in I-plate tests, strains KU143 and AS15 produced volatiles that significantly inhibited not only mycelial growth, sporulation, and conidial germination of the fungi on media but also fungal populations on rice grains. GC-MS analysis of the volatiles by strains KU143 and AS15 identified 12 and 17 compounds, respectively. Among these, the antifungal compound, 5-methyl-2-phenyl-1H-indole, was produced by strain KU143 and the antimicrobial compounds, 2-butyl 1-octanal, dimethyl disulfide, 2-isopropyl-5-methyl-1-heptanol, and 4-trifluoroacetoxyhexadecane, were produced by strain AS15. These results suggest that the tested strains producing extracellular metabolites and/or volatiles may have a broad spectrum of antifungal activities against the grain fungi. In particular, B. megaterium KU143 and P. protegens AS15 may be potential biocontrol agents against Aspergillus and Penicillium spp. during rice grain storage.
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문제 정의
islandicum are needed to enable these strains to use as biocontrol agents during grain storage. Therefore, the objectives of this study were (1) to evaluate the antifungal activities of the bacterial strains against Aspergillus and Penicillium spp. predominant in stored rice grains, (2) to investigate the antifungal activities of the volatiles produced by the strains against the tested fungi, and (3) to identify bacterial volatile compounds using gas chromatography-mass spectrometry (GC-MS) analysis.
제안 방법
In the tests of bacterial volatile effects on conidial germination, volatiles from B. megaterium KU143 and P. protegens AS15 significantly (p < .05) inhibited the conidial germination of the tested fungi (except the inhibition of A. candidus by strain AS15) compared with M. testaceum KU313, and S. aquatilis KU408, and MgSO4 treatments (Figure 2 and Table 2).
islandicum KU101, in a dual-culture assay. The assay was based on the inhibition of the fungal mycelia on PDA plates (90 mm in diameter), streaked in the center with each tested bacterial strain, Sphingomonas aquatilis KU408 (bacterial negative control), or SDW (untreated control). Streaked plates were inoculated with a conidial suspension (2 ml) of each tested fungus, prepared as described above, to the opposite edges of the plates.
The experiments were designed to be completely randomized designs and were conducted twice with three replicates per treatment. Data from repeated experiments were combined after confirming the homogeneity of variances using Levene’s test [38].
대상 데이터
The tested fungi, A. candidus AC317, A. fumigatus AF8, P. fellutanum KU53, and P. islandicum KU101, isolated from stored rice grains in Korea [8–10], were stored on potato dextrose agar (PDA) at 4 ℃ until used.
데이터처리
Analysis of variance (ANOVA) was conducted using the general linear model (GLM) procedures, and the means were separated using the least significant difference (LSD) test at p < .05.
이론/모형
Data from repeated experiments were combined after confirming the homogeneity of variances using Levene’s test [38].
성능/효과
Among the tested strains, P. protegens AS15 caused the most significant (p < .05) reduction in mycelial growth in all the tested fungi, followed by B. megaterium KU143 (Table 1).
protegens AS15 could degrade aflatoxin B1 and utilize the toxin for bacterial growth in nutrient-deficient conditions. In this study, the selected antagonistic strains were examined if they could show a broad spectrum of antifungal or biocontrol activity against A. candidus, A. fumigatus, P. fellutanum, and P. islandicum, which are deleterious grain fungi predominant in stored rice. Consequently, we found that these strains inhibited the growth and development of all the tested fungi on media and rice grains.
후속연구
flavus in peanuts [49]. Thus, further investigation is needed for purification and characterization of the antifungal compounds produced by the bacterial strains tested in this study.
참고문헌 (59)
1 Magan N , Hope R , Cairns V , et al Post-harvest fungal ecology: impact of fungal growth and mycotoxin accumulation in stored grain . Eur J Plant Pathol . 2003 ; 109 : 723 – 730 .
2 Oh JY , Jee SN , Nam Y , et al Populations of fungi and bacteria associated with samples of stored rice in Korea . Mycobiology . 2007 ; 35 : 36 – 38 . 24015066
3 Mew TW , Gonzales P. A handbook of rice seedborne fungi . Los Baños : International Rice Research Institute ; 2002 .
4 Mannaa M , Kim KD. Influence of temperature and water activity on deleterious fungi and mycotoxin production during grain storage . Mycobiology . 2017 ; 45 : 240 – 254 . 29371792
5 Park JW , Choi SY , Hwang HJ , et al Fungal mycoflora and mycotoxins in Korean polished rice destined for humans . Int J Food Microbiol . 2005 ; 103 : 305 – 314 . 16099315
6 Mannaa M , Kim KD. Microbe-mediated control of mycotoxigenic grain fungi in stored rice with focus on aflatoxin biodegradation and biosynthesis inhibition . Mycobiology . 2016 ; 44 : 67 – 78 . 27433116
8 Oh JY , Kim EN , Ryoo MI , et al Morphological and molecular identification of Penicillium islandicum isolate KU101 from stored rice . Plant Pathol J . 2008 ; 24 : 469 – 473 .
9 Oh JY , Sang MK , Lee HJ , et al First detection of Penicillium fellutanum from stored rice in Korea . Res Plant Dis . 2011 ; 17 : 216 – 221 .
10 Oh JY , Sang MK , Oh JE , et al Microbial population, aflatoxin contamination and predominant Aspergillus species in Korean stored rice . Plant Pathol J . 2010 ; 26 : 121 – 129 .
11 Boller RA , Schroeder HW. Influence of Aspergillus candidus on production of aflatoxin in rice by Aspergillus parasiticus . Phytopathology . 1974 ; 64 : 121 – 123 .
12 Krysińska-Traczyk E , Dutkiewicz J. Aspergillus candidus : a respiratory hazard associated with grain dust . Ann Agric Environ Med . 2000 ; 7 : 101 – 109 . 11153039
13 Takahashi C , Sekita S , Yoshihira K , et al The structures of toxic metabolites of Aspergillus candidus . II. The compound B (xanthoascin), a hepato- and cardio-toxic xanthocillin analog . Chem Pharm Bull . 1976 ; 24 : 2317 – 2321 . 1017077
14 Takahashi C , Yoshihira K , Natori S , et al The structures of toxic metabolites of Aspergillus candidus . I. The compounds A and E, cytotoxic p-terphenyls . Chem Pharm Bull . 1976 ; 24 : 613 – 620 . 949813
15 Abad A , Fernández-Molina JV , Bikandi J , et al What makes Aspergillus fumigatus a successful pathogen? Genes and molecules involved in invasive aspergillosis . Rev Iberoam Micol . 2010 ; 27 : 155 – 182 . 20974273
17 Marumo S. Islanditoxin, a toxic metabolite produced by Penicillium islandicum sopp: part III . Bull Agric Chem Soc Jpn . 1959 ; 23 : 428 – 437 .
18 Kozlovsky AG , Vinokurova NG , Adanin VM , et al New diketopiperazine alkaloids from Penicillium fellutanum . J Nat Prod . 2000 ; 63 : 698 – 700 . 10843594
19 Vinokurova NG , Boichenko LV , Arinbasarov MU. Production of alkaloids by fungi of the genus Penicillium grown on wheat grain . Appl Biochem Microbiol . 2003 ; 39 : 403 – 406 .
20 Mannaa M , Oh JY , Kim KD. Microbe-mediated control of Aspergillus flavus in stored rice grains with a focus on aflatoxin inhibition and biodegradation . Ann Appl Biol . 2017 ; 171 : 376 – 392 .
21 Mannaa M , Oh JY , Kim KD Biocontrol activity of volatile-producing Bacillus megaterium and Pseudomonas protegens against Aspergillus flavus and aflatoxin production on stored rice grains . Mycobiology . 2017 ; 45 : 213 – 219 . 29138628
22 Mannaa M , Kim KD. Control strategies for deleterious grain fungi and mycotoxin production from preharvest to postharvest stages of cereal crops: a review . Life Sci Nat Resour Res . 2017 ; 25 : 13 – 27 .
23 Muir WE , Wallace HAH. Effects of treating damp grain with formaldehyde to prevent storage deterioration . Can J Plant Sci . 1972 ; 52 : 375 – 379 .
25 Sauer DB , Burroughs R. Efficacy of various chemicals as grain mold inhibitors . Trans ASAE . 1974 ; 17 : 557 – 559 .
26 Dunkel F , Lung PZ , Chuan L , et al Insect and fungal response to sorbic acid-treated wheat during storage in south China . J Econ Entomol . 1982 ; 75 : 1083 – 1088 .
27 White DG , Toman J , Burnette DC , et al The effect of postharvest fungicide application on storage fungi of corn during ambient air drying and storage . Plant Dis . 1993 ; 77 : 562 – 568 .
28 White DG , Toman J. Effects of postharvest oil and fungicide application on storage fungi in corn following high-temperature grain drying . Plant Dis . 1994 ; 78 : 38 – 43 .
29 Gerhardson B. Biological substitutes for pesticides . Trends Biotechnol . 2002 ; 20 : 338 – 343 . 12127281
30 Sang MK , Kim JD , Kim BS , et al Root treatment with rhizobacteria antagonistic to Phytophthora blight affects anthracnose occurrence, ripening, and yield of pepper fruit in the plastic house and field . Phytopathology . 2011 ; 101 : 666 – 678 . 21405997
32 Hua SST , Beck JJ , Sarreal SBL , et al The major volatile compound 2-phenylethanol from the biocontrol yeast, Pichia anomala , inhibits growth and expression of aflatoxin biosynthetic genes of Aspergillus flavus . Mycotoxin Res . 2014 ; 30 : 71 – 78 . 24504634
33 Wilson MC , Mori T , Rückert C , et al An environmental bacterial taxon with a large and distinct metabolic repertoire . Nature . 2014 ; 506 : 58 – 62 . 24476823
34 Lee SY , Oh JY , Ryoo MI , et al Biological control of the rice storage fungi Aspergillus and Penicillium species by antagonistic bacteria originated from rice . Plant Pathol J . 2007 ; 23 : 328 .
35 Gu Q , Han N , Liu J , et al Expression of Helicobacter pylori urease subunit B gene in transgenic rice . Biotechnol Lett . 2006 ; 28 : 1661 – 1666 . 16912927
36 Hocking AD , Pitt JI. Dichloran-glycerol medium for enumeration of xerophilic fungi from low-moisture foods . Appl Environ Microbiol . 1980 ; 39 : 488 – 492 . 7387151
38 Levene H. Robust tests for equality of variances In: Olkin I , Ghurye SG , Hoeffeling W , Madow WG , Mann HB , editors. Contributions to probability and statistics: essays in honor of Harold Hotelling . Stanford (CA) : Stanford University Press ; 1960 .
39 Sharma RR , Singh D , Singh R. Biological control of postharvest diseases of fruits and vegetables by microbial antagonists: a review . Biol Control . 2009 ; 50 : 205 – 221 .
40 Wisniewski ME , Wilson CL. Biological control of postharvest diseases of fruit and vegetables: recent advances . HortScience . 1992 ; 27 : 94 – 98 .
41 Gourama H , Bullerman LB. Inhibition of growth and aflatoxin production of Aspergillus flavus by Lactobacillus species . J Food Prot . 1995 ; 58 : 1249 – 1256 .
42 Reddy KRN , Reddy CS , Muralidharan K. Potential of botanicals and biocontrol agents on growth and aflatoxin production by Aspergillus flavus infecting rice grains . Food Control . 2009 ; 20 : 173 – 178 .
43 Klich MA , Lax AR , Bland JM. Inhibition of some mycotoxigenic fungi by iturin A, a peptidolipid produced by Bacillus subtilis . Mycopathologia . 1991 ; 116 : 77 – 80 . 1780001
44 Gueldner RC , Reilly CC , Pusey PL , et al Isolation and identification of iturins as antifungal peptides in biological control of peach brown rot with Bacillus subtilis . J Agric Food Chem . 1988 ; 36 : 366 – 370 .
45 Klich MA , Arthur KS , Lax AR , et al Iturin A: a potential new fungicide for stored grains . Mycopathologia . 1994 ; 127 : 123 – 127 . 7984212
47 Bottone EJ , Peluso RW. Production by Bacillus pumilus (MSH) of an antifungal compound that is active against Mucoraceae and Aspergillus species: preliminary report . J Med Microbiol . 2003 ; 52 : 69 – 74 . 12488568
48 Dikin A , Sijam K , Kadir J , et al Mode of action of antimicrobial substances from Burkholderia multivorans and Microbacterium testaceum against Schizophyllum commune Fr . Int J Agric Biol . 2007 ; 9 : 311 – 314 .
49 Zhang T , Shi ZQ , Hu LB , et al Antifungal compounds from Bacillus subtilis B-FS06 inhibiting the growth of Aspergillus flavus . World J Microbiol Biotechnol . 2008 ; 24 : 783 – 788 .
51 Fernando WD , Ramarathnam R , Krishnamoorthy AS , et al Identification and use of potential bacterial organic antifungal volatiles in biocontrol . Soil Biol Biochem . 2005 ; 37 : 955 – 964 .
53 Li Q , Ning P , Zheng L , et al Fumigant activity of volatiles of Streptomyces globisporus JK-1 against Penicillium italicum on Citrus microcarpa . Postharvest Biol Technol . 2010 ; 58 : 57 – 165 .
54 Rao RM , Reddy GN , Sreeramulu J. Synthesis of some new pyrazolo-pyrazole derivatives containing indoles with antimicrobial activity . Der Pharma Chem . 2011 ; 3 : 301 – 309 .
55 Sarada K , Margret RJ , Mohan VR. GC-MS Determination of bioactive components of Naringi crenulata (Roxb) Nicolson . Int J Chem Tech Res . 2011 ; 3 : 1548 – 1555 .
56 Muthulakshmi A , Margret R , Mohan VR. GC-MS analysis of bioactive components of Feronia elephantum Correa ( Rutaceae ) . J Appl Pharm Sci . 2012 ; 2 : 69 – 74 .
57 Raghunath SA , Manjunatha Y , Rayappa K. Synthesis, antimicrobial, and antioxidant activities of some new indole analogues containing pyrimidine and fused pyrimidine systems . Med Chem Res . 2012 ; 21 : 3809 – 3817 .
58 Siddiquee S , Cheong BE , Taslima K , et al Separation and identification of volatile compounds from liquid cultures of Trichoderma harzianum by GC-MS using three different capillary columns . J Chromatogr Sci . 2012 ; 50 : 358 – 367 . 22407347
59 Wang C , Wang Z , Qiao X , et al Antifungal activity of volatile organic compounds from Streptomyces alboflavus TD-1 . FEMS Microbiol Lett . 2013 ; 341 : 45 – 51 . 23351181
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