Baoguo Zhang

The impacts of cypermethrin pesticide application on the non-target microbial community of the pepper plant phyllosphere.

Although pesticides have been extensively used for controlling insects and disease pathogens of plants, little is known regarding the impacts of applying these pesticides on the microbial community in the plant phyllosphere. Here, we report the effects of cypermethrin pesticide application upon the microbial community of the pepper plant phyllosphere. Assessments were made using culture-independent techniques including phospholipid fatty acid analysis (PLFA) and 16S rRNA gene directed Polymerase Chain Reaction with Denaturing Gradient Gel Electrophoresis (PCR-DGGE). During the 21 day greenhouse study, PLFA results indicated that both total and bacterial biomass increased after application of the pesticide. PLFA profiles also indicated that Gram-negative bacteria became predominant. DGGE analysis confirmed a significant change in bacterial community structure within the phyllosphere following the pesticide application where different dendrogram clusters were observed between control and treated samples. Phylogenetic analysis also suggested a change in bacterial phyla following treatment, where bands sequenced within control cultures were predominantly of the Firmicutes phylum, but those bands sequenced in the treated samples were predominantly members of the Bacteroidetes and gamma-Proteobacteria phyla. In conclusion, this study revealed an increase in bacterial abundance and a shift in community composition within the pepper plant phyllosphere following the pesticide application, and highlighted the effective use of PLFA and PCR-DGGE for studying the effect of pesticides upon indigenous phyllosphere microbes.

Assessing the impact of the biological control agent Bacillus thuringiensis on the indigenous microbial community within the pepper plant phyllosphere.

Although biological control agents (BCAs) have been used extensively for controlling insects and pathogens of plants, little is known regarding the effects of such agents on the indigenous microbial communities within the plant phyllosphere. We assessed the effect of the BCA Bacillus thuringiensis (Bt) on the microbial communities within the pepper plant phyllosphere using culture-independent methodologies. Phospholipid fatty acid (PLFA) analysis suggested that the bacterial and fungal biomass were not significantly affected following Bt application. However, principal component analysis of PLFA data indicated that Bt did change the phyllosphere microbial community structure significantly. 16S rRNA gene-directed PCR with denaturing gradient gel electrophoresis (DGGE) also suggested a significant change in the phyllosphere bacterial community structure following Bt inoculation. Phylogenetic analysis of excised DGGE bands suggested a change in bacterial phyla; bands from untreated samples predominantly belonged to the Firmicutes, while Gammaproteobacteria abounded in the treated samples.

Profiling and Identification of Small rDNA-Derived RNAs and Their Potential Biological Functions

Small non-coding RNAs constitute a large family of regulatory molecules with diverse functions. Notably, some small non-coding RNAs matched to rDNA have been identified as qiRNAs and small guide RNAs involved in various biological processes. However, a large number of small rDNA-derived RNAs (srRNAs) are usually neglected and yet to be investigated. We systematically investigated srRNAs using small RNA datasets generated by high-throughput sequencing, and found srRNAs are mainly mapped to rRNA coding regions in sense direction. The datasets from immunoprecipitation and high-throughput sequencing demonstrate that srRNAs are co-immunoprecipitated with Argonaute (AGO) proteins. Furthermore, the srRNA expression profile in mouse liver is affected by diabetes. Overexpression or inhibition of srRNAs in cultured cells shows that srRNAs are involved in various signaling pathways. This study presents a global view of srRNAs in total small RNA and AGO protein complex from different species, and demonstrates that srRNAs are correlated with diabetes, and involved in some biological processes. These findings provide new insights into srRNAs and their functions in various physiological and pathological processes.

Effect of cypermethrin insecticide on the microbial community in cucumber phyllosphere

Cucumber (Cucumis sativus) is one of the most widely used vegetable in the world, and different pesticides have been extensively used for controlling the insects and disease pathogens of this plant. However, little is known about how the pesticides affect the microbial community in cucumber phyllosphere. This study was the first attempt to assess the impact of pyrethroid insecticide cyperemethrin on the microbial communities of cucumber phyllosphere using biochemical and genetic approaches. Phospholipid fatty acid (PLFA) assay indicated that cyperemethrin insecticide treatment led to a significant increase in both total and bacterial biomass and a decrease in fungal biomass and the ratio of Gram-positive (GP) bacteria to Gram-negative (GN) bacteria within the cucumber phyllosphere. Principal-component analyses (PCA) suggested that the number of unsaturated and cyclopropane PLFAs (16:1 omega 9t, 18:1 omega 7, cy17:0, cy19:0) increased with the insecticide treatment, whereas the saturated PLFA i16:0, i17:0 decreased. The increase of GN bacteria implied that the cypermethrin insecticide might be a nutrient for the growth of these phyllosphere microbes. Terminal restriction fragment length polymorphism (T-RFLP) reinforced the PLFA results. A significant change of bacterial community structure was observed in the separate dendrogram cluster between control and treated samples with the cucumber phyllosphere following cypermethrin insecticide treatment. Moreover, the increased terminal restriction fragments (T-RFs) (58, 62, 89, 99, 119, 195, 239, 311, 340, and 473 bp) indicated that some bacteria might play a significant role in the insecticide degradation within the cucumber phylosphere, whereas the disappeared T-RFs (44, 51, 96, 223, 306, and 338 bp) implied that some other bacteria might potentially serve as microbial indicator of cyperemethrin insecticide exposure.

A novel acidophile, community populating waste ore deposits at an acid mine drainage site

Waste ore samples (pH 3.0) were collected at an acid mine drainage (AMD) site in Anhui, China. The present acidophilic microbial community in the waste ore was studied with 16S rRNA gene clone library and denaturing gradient gel electrophoresis (DGGE). Eighteen different clones were identified and affiliated with Actinobacteria, low G + C Gram-positives, Thermomicrobia, Acidobacteria, Proteobacteria, candidate division TM7, and Planctomycetes. Phylogenetic analysis of 16S rRNA gene sequences revealed a diversity of acidophiles in the samples that were mostly novel. It is unexpected that the moderately thermophilic acidophiles were abundant in the acidic ecosystem and may play a great role in the generation of AMD. The result of DGGE was consistent with that of clone library analysis. These findings help in the better understanding of the generation mechanism of AMD and in developing a more efficient method to control AMD.

Isolation of Photobacterium sp. LuB-1 and its application in rapid assays for chemical toxicants in water

Aims:  To isolate marine bacteria with strong bioluminescence in a wide range of NaCl concentration, especially at low salt conditions.