[Isolation of endophytic fungi from medicinal plant Brucea javanica and their microbial inhibition activity].

OBJECTIVE: To isolate and identify endophytic fungi from Brucea javanica, and to detect the antimicrobial activity of these strains. METHODS: Endophytic fungi were isolated by tissue inoculation culture and identified by conventional morphological characteristic method. Seven kinds of pathogenic fungi and three kinds of bacteria were used as targeting microbes to test microbial inhibition activities by agar plate antagonistic action and modified agar gel diffusion methods, respectively. RESULTS: A total of 83 endophytic fungi strains were isolated from the root, stem, leaf and fruit of Brucea javanica. 34 strains were obtained from the stem, 32 strains were obtained from the leaf, 15 strains were isolated from the root and 2 strains came from the fruit. These 73 strains which had been identified attribute to 5 orders, 6 families and 12 genera. For the isolated strains, 14 strains had antifungal activities against at least one pathogenic fungi, 9 strains showed antibacterial activities against one or more bacteria. Especially, the strain YJ-17 which belonged to Phomopsis genus showed the best inhibitory effect on the targeting microbes. CONCLUSION: The endophytic fungi from Brucea javanica show diversity and microbial inhibition activity, and are worthy for further study on plant disease controlling.

Enhancement of cypermethrin degradation by a coculture of Bacillus cereus ZH-3 and Streptomyces aureus HP-S-01.

Degradation of cypermethrin was significantly enhanced in a coculture of Bacillus cereus ZH-3 and Streptomyces aureus HP-S-01. In the pure culture, longer half-lives (t(1/2)=32.6-43.0h) of cypermethrin were observed, as compared to the mixed cocultures (t(1/2)=13.0h). The optimal degradation conditions were determined to be 28.2°C and pH 7.5 based on response surface methodology (RSM). Under these conditions, the mixed cultures completely metabolized cypermethrin (50mgL(-1)) within 72h. Analysis of degradation products of cypermethrin indicated that the microbial consortium converted cypermethrin to α-hydroxy-3-phenoxy-benzeneacetonitrile, 3-phenoxybenzaldehyde and 4-phenoxyphenyl-2,2-dimethyl-propiophenone, and subsequently transformed these compounds with a maximum specific degradation rate (q(max)), half-saturation constant (K(s)) and inhibition constant (K(i)) of 0.1051h(-1), 31.2289mgL(-1) and 220.5752mgL(-1), respectively. This is the first report of a proposed pathway of degradation of cypermethrin by hydrolysis of ester linkage and oxidization of 3-phenoxybenzyl in a coculture. Finally, this coculture is the first described mixed microbial consortium capable of metabolizing cypermethrin.

Combination of Kluyveromyces marxianus and sodium bicarbonate for controlling green mold of citrus fruit.

Biocontrol efficacy of an antagonistic yeast Kluyveromyces marxianus was evaluated individually or in combination with sodium bicarbonate (SBC) against green mold of citrus fruit caused by Penicillium digitatum. Their effects on postharvest quality of citrus fruit were also investigated. The results indicated that the antagonistic activity of K. marxianus at 1×108 CFU/mL on green mold of citrus fruit was enhanced by 2% SBC treatment. In artificial inoculation trials, disease control after 3 and 6 days, respectively, with the mixture of K. marxianus and 2% SBC (18.33%, 58.33%) was significantly improved over that obtained with K. marxianus (41.67%, 70.00%) or SBC (43.33%, 81.67%) alone. The combination of K. marxianus with SBC was as effective as the imazalil treatment in natural infection trials, which gave about 90% control of green mold. Addition of 2% SBC significantly stimulated the growth of K. marxianus in citrus fruit wounds after 72 h. Moreover, K. marxianus, SBC and their combination did not impair quality parameters including weight loss, fruit firmness, total soluble solids, titratable acidity and ascorbic acid at 4 °C for 30 days followed by 20 °C for 15 days. These results suggested that the use of SBC is a useful approach to improve the efficacy of K. marxianus for the postharvest green mold of citrus fruit.

Isolation and characterization of a fungus able to degrade pyrethroids and 3-phenoxybenzaldehyde.

Fungal strain HU, isolated from activated sludge and identified as a member of the genus Cladosporium based on morphology and sequencing of 28S rRNA, was shown to degrade 90% of fenvalerate, fenpropathrin, ß-cypermethrin, deltamethrin, bifenthrin, and permethrin (100 mgL(-1)) within 5 days. Fenvalerate was utilized as sole carbon and energy source and co-metabolized in the presence of sucrose. Degradation of fenvalerate occurred at pH 5-10 at 18-38°C. The fungus first hydrolyzed the carboxylester linkage to produce α-hydroxy-3-phenoxy-benzeneacetonitrile and 3-phenoxybenzaldehyde, and subsequently degraded these two compounds with a q(max), K(s) and K(i) of 1.73 d(-1), 99.20 mgL(-1) and 449.75 mgL(-1), respectively. Degradation followed first-order kinetics. These results show that the fungal strain may possess potential to be used in bioremediation of pyrethroid-contaminated environments.

Biodegradation of deltamethrin and its hydrolysis product 3-phenoxybenzaldehyde by a newly isolated Streptomyces aureus strain HP-S-01.

A newly isolated actinomycete strain HP-S-01 from activated sludge could effectively degrade deltamethrin and its major hydrolysis product 3-phenoxybenzaldehyde. Based on the morphological, cultural, physio-biochemical characteristics, and 16S rDNA sequence analysis, strain HP-S-01 was identified as Streptomyces aureus. Strain HP-S-01 was also found highly efficient in degrading cyfluthrin, bifenthrin, fenvalerate, fenpropathrin, permethrin, and cypermethrin. Strain HP-S-01 rapidly degraded deltamethrin without a lag phase over a wide range of temperature (18~38°C) and pH (5~10), and metabolized to produce α-hydroxy-3-phenoxy-benzeneacetonitrile and 3-phenoxybenzaldehyde by hydrolysis of the carboxylester linkage. The 3-phenoxybenzaldehyde was further oxidized to form 2-hydroxy-4-methoxy benzophenone resulting in its detoxification. No persistent accumulative product was detected by gas chromatography-mass spectrometry (GC/MS) analysis. Response surface methodology was used to optimize degradation conditions. Strain HP-S-01 completely removed 50~300 mg L(-1) deltamethrin within 7 days under the optimal degradation conditions. Furthermore, the biodegradation kinetics corresponded with the first-order model. Therefore, strain HP-S-01 is suitable for the efficient and rapid bioremediation of pyrethroid-contaminated environment.