Shining Zhou

Inhibition of Vibrio biofilm formation by a marine actinomycete strain A66

China remains by far the largest aquaculture producer in the world. However, biofilms formed by pathogenic Vibrio strains pose serious problems to marine aquaculture. To provide a strategy for biofilm prevention, control, and eradication, extracts from 88 marine actinomycetes were screened. Thirty-five inhibited the biofilm formation of Vibrio harveyi, Vibrio vulnificus, and Vibrio anguillarum at a concentration of 2.5% (v/v). Thirty-three of the actinomycete extracts dispersed the mature biofilm. Six extracts inhibited the quorum-sensing system of V. harveyi by attenuating the signal molecules N-acylated homoserine lactones’ activity. Strain A66, which was identified as Steptomyces albus, both attenuated the biofilms and inhibited their quorum-sensing system. It is suggested that strain A66 is a promising candidate to be used in future marine aquaculture.

Isolation and characterization of endophytic Streptomyces strains from surface-sterilized tomato (Lycopersicon esculentum) roots

Aims:  To isolate endophytic Streptomyces strains from tomato and examine their antimicrobial activity.

Isolation of Endophytic Actinomycetes From Roots and Leaves of Banana (Musa Acuminata) Plants and Their Activities Against Fusarium oxysporumf. sp. cubense

Two hundred and forty-two actinomycete strains were isolated from the interior of leaves and roots of healthy and wilting banana plants. Most of them were streptomycetes, Streptomyces griseorubiginosus-like strains were the most frequently isolated strains. Community analysis demonstrated increased actinomycete diversity in wilting leaves compared to that in healthy leaves, similar actinomycete communities were found in wilting and healthy roots. Screening of the isolates for antagonistic activity against Fusarium oxysporumf. sp. cubenserevealed that the proportion of antagonistic streptomycetes in healthy roots was higher than that in wilting roots (P < 0.01), but no difference was found between antagonistic strains isolated from healthy and wilting leaves. The potential biological control of Panama disease of banana by endophytic streptomycetes, especially Streptomyces griseorubiginosus-like strains was discussed.

Diversity of cultivated and uncultivated actinobacterial endophytes in the stems and roots of rice

A dual approach consisting of cultivation and molecular retrieval of actinobacterial 16S rRNA genes was used to characterize the diversity of actinobacterial community inhabiting interior of rice stems and roots. Streptomyces is the most frequently isolated genus from rice stems and roots. Forty-five clones chosen randomly among 250 clones in the 16S rRNA gene clone library from roots were affiliated with nine genera of actinobacteria and uncultured actinobacteria (Mycobacterium, Streptomyces, Micromonospora, Actinoplanes, Frankia, Dactylosporangium, Amycolatopsis, Corynebacterium, Rhodococcus, and uncultured actinobacterium). However, 33 clones from stems were affiliated with four genera and uncultured actinobacteria (Streptomyces, Mycobacterium, Nocardiodies, Janibacter, uncultured earthworm cast bacterium, uncultured earthworm intestine bacterium, and uncultured actinobacterium). Species similar to S. cyaneus were isolated from surface-sterilized roots and stems of rice and detected inside rice roots by culture-independent methods. Species similar to S. caviscabies, S. scabies, and S. turgidiscabies were simultaneously detected from the interior of rice stems by the culture-dependent and culture-independent methods. S. galilaeus was detected from the interior of rice stems and roots. These results indicated that some actinobacterial populations in rice stems were correlated with those in roots.

Screening for novel quorum-sensing inhibitors to interfere with the formation of Pseudomonas aeruginosa biofilm

The objective of this study was to screen for novel quorum-sensing inhibitors (QSIs) from traditional Chinese medicines (TCMs) that inhibit bacterial biofilm formation. Six of 46 active components found in TCMs were identified as putative QSIs based on molecular docking studies. Of these, three compounds inhibited biofilm formation by Pseudomonas aeruginosa and Stenotrophomonas maltophilia at a concentration of 200 µM. A fourth compound (emodin) significantly inhibited biofilm formation at 20 µM and induced proteolysis of the quorum-sensing signal receptor TraR in Escherichia coli at a concentration of 3-30 mM. Emodin also increased the activity of ampicillin against P. aeruginosa. Therefore, emodin might be suitable for development into an antivirulence and antibacterial agent.

Characterization of a new keratinolytic Trichoderma atroviride strain F6 that completely degrades native chicken feather

Aims: To isolate novel nonpathogenic fungus that completely degrades native chicken feather and characterize its keratinases.

Endophytic fungi from Musa acuminata leaves and roots in South China

One hundred and sixty-three endophytic fungal cultures were isolated from 200 leaf samples of Musa acuminata trees, which were soaked in 36% formaldehyde solution for surface sterilization. They belonged to the genera of Gloeosporium musae (45%), Myxosporium spp. (11%), Deightoniella torulosa (8.5%), Alternaria tenuis (7.9%), Sphaceloma spp. (7.4%), Aureobasidium spp. (4.3%), Melida spp. (1.8%), Uncinula spp. (1.8%), Penicillium spp. (1.8%), Aspergillus spp. (1.2%), Sarcinella spp. (1.2%), Cladosporium sp. (0.6%), Cephalosporium sp. (0.6%) and sterile mycelium (6.7%). Sixty-eight endophytic fungal cultures were isolated from 100 root samples. They respectively belonged to the genera of Aspergillus spp. (31%), Paecilomyces spp. (16%), Penicillium spp. (15%), Fusarium spp. (10%), Gloeosporium musae (6%), yeast (3%), Deightoniella torulosa (3%), Spicaria sp. (1.4%), Cephalosporrium sp. (1.4%), Meliola sp. (1.4%) and sterile mycelium (10%). Water agar (containing 50 μg chloramphenicol ml−1 and 50 μg streptomycin ml−1) seemed to be a better medium for isolation of endophytic fungi than potato-dextrose agar (PDA, containing 50 μg chloramphenicol ml−1 and 50 μg streptomycin ml−1).

Enhanced phytoremediation of toxic metals by inoculating endophytic Enterobacter sp. CBSB1 expressing bifunctional glutathione synthase.

To engineer plant-bacteria symbionts for remediating complex sites contaminated with multiple metals, the bifunctional glutathione (GSH) synthase gene gcsgs was introduced into endophytic Enterobacter sp. CBSB1 to improve phytoremediation efficiency of host plant Brassica juncea. The GSH contents of shoots inoculated with CBSB1 is 0.4μMg(-1) fresh weight. However, the GSH concentration of shoots with engineered CBSB1-GCSGS increased to 0.7μMg(-1) fresh weight. The shoot length, fresh weight and dry weight of seedlings inoculated with CBSB1-GCSGS increased 67%, 123%, and 160%, compared with seedlings without inoculation, respectively. The Cd and Pb concentration in shoots with CBSB1-GCSGS increased 48% and 59% compared with seedlings without inoculation, respectively. The inoculation of CBSB1 and CBSB1-GCSGS could increase the Cd and Pb extraction amounts of seedlings significantly compared with those without inoculation (P<0.05), the seedlings inoculated with CBSB1-GCSGS showed the highest Cd and Pb extraction amounts. It was concluded that the gcsgs gene introduced into Enterobacter sp. CBSB1 upgraded the phytoremediation efficacy of B. juncea. So the engineered Enterobacter sp. CBSB1-GCSGS showed potentials in remediation sites contaminated with complex contaminants by inoculating into remediating plants.

Temporal and spatial diversity of bacterial communities in coastal waters of the South china sea.

Bacteria are recognized as important drivers of biogeochemical processes in all aquatic ecosystems. Temporal and geographical patterns in ocean bacterial communities have been observed in many studies, but the temporal and spatial patterns in the bacterial communities from the South China Sea remained unexplored. To determine the spatiotemporal patterns, we generated 16S rRNA datasets for 15 samples collected from the five regularly distributed sites of the South China Sea in three seasons (spring, summer, winter). A total of 491 representative sequences were analyzed by MOTHUR, yielding 282 operational taxonomic units (OTUs) grouped at 97% stringency. Significant temporal variations of bacterial diversity were observed. Richness and diversity indices indicated that summer samples were the most diverse. The main bacterial group in spring and summer samples was Alphaproteobacteria, followed by Cyanobacteria and Gammaproteobacteria, whereas Cyanobacteria dominated the winter samples. Spatial patterns in the samples were observed that samples collected from the coastal (D151, D221) waters and offshore (D157, D1512, D224) waters clustered separately, the coastal samples harbored more diverse bacterial communities. However, the temporal pattern of the coastal site D151 was contrary to that of the coastal site D221. The LIBSHUFF statistics revealed noticeable differences among the spring, summer and winter libraries collected at five sites. The UPGMA tree showed there were temporal and spatial heterogeneity of bacterial community composition in coastal waters of the South China Sea. The water salinity (P=0.001) contributed significantly to the bacteria-environment relationship. Our results revealed that bacterial community structures were influenced by environmental factors and community-level changes in 16S-based diversity were better explained by spatial patterns than by temporal patterns.

Seasonal and spatial diversity of microbial communities in marine sediments of the South China Sea

This study was conducted to characterize the diversity of microbial communities in marine sediments of the South China Sea by means of 16S rRNA gene clone libraries. The results revealed that the sediment samples collected in summer harboured a more diverse microbial community than that collected in winter, Deltaproteobacteria dominated 16S rRNA gene clone libraries from both seasons, followed by Gammaproteobacteria, Acidobacteria, Nitrospirae, Planctomycetes, Firmicutes. Archaea phylotypes were also found. The majority of clone sequences shared greatest similarity to uncultured organisms, mainly from hydrothermal sediments and cold seep sediments. In addition, the sedimentary microbial communities in the coastal sea appears to be much more diverse than that of the open sea. A spatial pattern in the sediment samples was observed that the sediment samples collected from the coastal sea and the open sea clustered separately, a novel microbial community dominated the open sea. The data indicate that changes in environmental conditions are accompanied by significant variations in diversity of microbial communities at the South China Sea.