Aspergillus flavus and Aspergillus parasiticus are saprophytic fungi which can infect and contaminate preharvest and postharvest food/feed with production of aflatoxins (B 1 , B 2 , and G). They are also an opportunistic pathogen causing aspergillosis diseases of animals and humans. In this study, the volatile organic compounds (VOCs) from Streptomyces yanglinensis 3-10 were found to be able to inhibit mycelial growth, sporulation, conidial germination, and expression of aflatoxin biosynthesis genes in A. flavus and A. parasiticus in vitro. On peanut kernels, the VOCs can also reduce the disease severity and inhibit the aflatoxins production by A. flavus and A. parasiticus under the storage conditions. Scanning electron microscope (SEM) observation showed that high dosage of the VOCs can inhibit conidial germination and colonization by the two species of Aspergillus on peanut kernels. The VOCs also showed suppression of mycelial growth on 18 other plant pathogenic fungi and one Oomycetes organism. By using SPME-GC-MS, 19 major VOCs were detected, like in other Streptomyces, 2-MIB was found as the main volatile component among the detected VOCs. Three standard chemicals, including methyl 2-methylbutyrate (M2M), 2-phenylethanol (2-PE), and β-caryophyllene (β-CA), showed antifungal activity against A. flavus and A. parasiticus. Among them, M2M showed highest inhibitory effect than other two standard compounds against conidial germination of A. flavus and A. parasiticus. To date, this is the first record about the antifungal activity of M2M against A. flavus and A. parasiticus. The VOCs from S. yanglinensis 3-10 did not affect growth of peanut seedlings. In conclusion, our results indicate that S. yanglinensis 3-10 may has a potential to become a promising biofumigant in for control of A. flavus and A. parasiticus.
The bacterium, Streptomyces yanglinensis 3-10, shows promise in the control of many phytopathogenic fungi. In this study, S. yanglinensis and its antifungal substances, culture filtrate (CF3-10) and crude extracts (CE3-10), were evaluated for their activity in reducing growth and aflatoxin AFB1 production by Aspergillus flavus, both in vitro and in vivo on peanut kernels. The results showed that in dual culture conditions, S. yanglinensis reduced the mycelial growth of A. flavus about 41% as compared to control. The mycelial growth of A. flavus was completely inhibited on potato dextrose agar amended with CF3-10 at 3% (v/v) or CE3-10 at 2.5 μg/ml. In liquid culture experiments, growth inhibition ranged from 32.3 to 91.9% with reduction in AFB1 production ranging from 46.4 to 93.4% using different concentrations of CF3-10 or CE3-10. For in vivo assays, CF3-10 at 0.133 ml/g (v/w) or CE3-10 at 13.3 μg/g (w/w) reduced the postharvest decay of peanut kernels by inhibiting visible growth of A. flavus leading to an 89.4 or 88.1% reduction in AFB1 detected, respectively. Compared with the controls, CF3-10 and CE3-10 in A. flavus shake culture significantly reduced expression levels of two AFB1 biosynthesis genes, aflR and aflS. Furthermore, electron microscopy observation showed that CF3-10 (2%, v/v) caused hyphae growth to be abnormal and shriveled, cell organelles to degenerate and collapse, large vacuoles to appear. These results suggest that S. yanglinensis 3-10 has potential as an alternative to chemical fungicides in protecting peanut kernels and other agricultural commodities against postharvest decay from A. flavus.
This study was conducted to determine the antifungal activity of the metabolites from Streptomyces sp. 3–10, and to purify and identify the metabolites. Meanwhile, the taxonomic status of strain 3–10 was re-evaluated. The cultural filtrates of strain 3–10 in potato dextrose broth were extracted with ethyl acetate. The resulting crude extract at 1 and 5 μg/ml inhibited growth of 22 species in 18 genera of plant pathogenic fungi and Oomycetes, accounting for 92% of the total 24 tested species, suggesting that it has a wide antifungal spectrum. Two compounds were purified from the crude extract and were identified as reveromycins A and B, which demonstrated high antifungal activity against Botrytis cinerea, Mucor hiemails, Rhizopus stolonifer, and Sclerotinia sclerotiorum under acidic pH conditions. Both the crude extract and reveromycin A from strain 3–10 at 10, 50, and 100 μg/ml showed high efficacy in suppression of strawberry fruit rot caused by the above-mentioned four pathogens. The efficacy was comparable to that of corresponding commercial fungicides (pyrimethanil, captan, dimetachlone) used in management of these pathogens. Morphological, physiological, and phylogenetic characterization showed that strain 3–10 is closely related to Streptomyces yanglinensis 1307T, representing a novel phylotype in that species. This study reported a new strain with reveromycins-producing capability. The finding is important for further exploitation of reveromycins for agricultural use.
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