Ke-Qin Zhang

Yunnan University, Yün-nan, Yunnan, China

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Publications (112)315.79 Total impact

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    ABSTRACT: The ascomycete fungus, Fusarium graminearum (teleomorph Gibberella zeae), is the most common causal agent of Fusarium head blight (FHB), a devastating disease for cereal crops worldwide. F. graminearum produces ascospores (sexual spores) and conidia (asexual spores), which can serve as disease inocula of FHB. Meanwhile, Fusarium-infected grains are often contaminated with mycotoxins such as trichothecenes (TRIs), fumonisins, and zearalenones, among which TRIs are related to the pathogenicity of F. graminearum, and these toxins are hazardous to humans and livestock. In recent years, with the complete genome sequencing of F. graminearum, an increasing number of functional genes involved in the production of secondary metabolites, hyphal differentiation, sexual and asexual reproduction, virulence and pathogenicity have been identified from F. graminearum. In this review, the secondary metabolite synthesis, hyphal development and pathogenicity related genes in F. graminearum were thoroughly summarized, and the genes associated with secondary metabolites, sexual reproduction, energy metabolism, and pathogenicity were highlighted.
    Biotechnology advances 01/2014; · 8.25 Impact Factor
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    ABSTRACT: Malate synthase (Mls), a key enzyme in the glyoxylate cycle, is required for virulence in microbial pathogens. In this study, we identified the AoMls gene from the nematode-trapping fungus Arthobotrys oligospora. The gene contains 4 introns and encodes a polypeptide of 540 amino acids. To characterize the function of AoMls in A. oligospora, we disrupted it by homologous recombination, and the ΔAoMls mutants were confirmed by PCR and Southern blot analyses. The growth rate and colony morphology of the ΔAoMls mutants showed no obvious difference from the wild-type strains on potato dextrose agar (PDA) plate. However, the disruption of gene AoMls led to a significant reduction in conidiation, failure to utilize fatty acids and sodium acetate for growth, and its conidia were unable to germinate on minimal medium supplemented with sodium oleate. In addition, the trap formation was retarded in the ΔAoMls mutants, which only produced immature traps containing one or two rings. Moreover, the nematicidal activity of the ΔAoMls mutants was significantly decreased. Our results suggest that the gene AoMls plays an important role in conidiation, trap formation and pathogenicity of A. oligospora.
    Applied Microbiology and Biotechnology 12/2013; · 3.69 Impact Factor
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    ABSTRACT: The biotransformation of the major saponins in Panax notoginseng, including the ginsenosides Rg1, Rh1, Rb1, and Re, by endophytes isolated from P. notoginseng was studied. One hundred and thirty-six endophytes were isolated and screened for their biotransformational abilities. The results showed that five of the tested endophytes were able to transform these saponins. These five strains were identified based on their ITS or 16S rDNA sequences, which revealed that they belonged to the genera Fusarium, Nodulisporium, Brevundimonas, and Bacillus genera. Ten transformed products were isolated and identified, including a new compound 6-O-[α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl]-20-O-β-D-glucopyranosyldammarane-3,6,12,20,24,25-hexaol (3), and nine known compounds, compound K (1), ginsenoside F2 (2), vinaginsenoside R13 (4), vinaginsenoside R22 (5), pseudo-ginsenoside RT4 (6), (20S)-protopanaxatriol (7), ginsenoside Rg1 (8), vinaginsenoside R15 (9), and (20S)-3-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosylprotopanaxatriol (10). This is the first study on the biotransformation of chemical components in P. notoginseng by endophytes isolated from the same plant.
    Chemistry & Biodiversity 11/2013; 10(11):2021-31. · 1.81 Impact Factor
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    ABSTRACT: The Caenorhabditis elegans DAF-16 transcription factor is critical for diverse biological processes, particularly longevity and stress resistance. Disruption of the DAF-2 signaling cascade promotes DAF-16 activation, and confers resistance to killing by pathogenic bacteria, such as Pseudomonas aeruginosa, Staphylococcus aureus, and Enterococcus faecalis. However, daf-16 mutants exhibit similar sensitivity to these bacteria as wild-type animals, suggesting that DAF-16 is not normally activated by these bacterial pathogens. In this report, we demonstrate that DAF-16 can be directly activated by fungal infection and wounding in wild-type animals, which is independent of the DAF-2 pathway. Fungal infection and wounding initiate the Gαq signaling cascade, leading to Ca(2+) release. Ca(2+) mediates the activation of BLI-3, a dual-oxidase, resulting in the production of reactive oxygen species (ROS). ROS then activate DAF-16 through a Ste20-like kinase-1/CST-1. Our results indicate that DAF-16 in the epidermis is required for survival after fungal infection and wounding. Thus, the EGL-30-Ca(2+)-BLI-3-CST-1-DAF-16 signaling represents a previously unknown pathway to regulate epidermal damage response.
    PLoS Pathogens 10/2013; 9(10):e1003660. · 8.14 Impact Factor
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    ABSTRACT: Advances in genetic transformation techniques have made important contributions to molecular genetics. Various molecular tools and strategies have been developed for functional genomic analysis of filamentous fungi since the first DNA transformation was successfully achieved in Neurospora crassa in 1973. Increasing amounts of genomic data regarding filamentous fungi are continuously reported and large-scale functional studies have become common in a wide range of fungal species. In this review, various molecular tools used in filamentous fungi are compared and discussed, including methods for genetic transformation (e.g., protoplast transformation, electroporation, and microinjection), the construction of random mutant libraries (e.g., restriction enzyme mediated integration, transposon arrayed gene knockout, and Agrobacterium tumefaciens mediated transformation), and the analysis of gene function (e.g., RNA interference and transcription activator-like effector nucleases). We also focused on practical strategies that could enhance the efficiency of genetic manipulation in filamentous fungi, such as choosing a proper screening system and marker genes, assembling target-cassettes or vectors effectively, and transforming into strains that are deficient in the nonhomologous end joining pathway. In summary, we present an up-to-date review on the different molecular tools and latest strategies that have been successfully used in functional genomics in filamentous fungi.
    Biotechnology advances 08/2013; · 8.25 Impact Factor
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    ABSTRACT: Proteins that serve as regulator of G protein signaling (RGS) primarily function as GTPase accelerators that promote GTP hydrolysis by the Gα subunits, thereby inactivating the G protein and rapidly switching off G protein-coupled signaling pathways. Since the first RGS protein was identified from the budding yeast Saccharomyces cerevisiae, more than 30 RGS and RGS-like proteins have been characterized from several model fungi, such as Aspergillus nidulans, Beauveria bassiana, Candida albicans, Fusarium verticillioides, Magnaporthe oryzae, and Metarhizium anisopliae. In this review, the partial biochemical properties and functional domains of RGS and RGS-like proteins were predicted and compared, and the roles of RGS and RGS-like proteins in different fungi were summarized. Moreover, the phylogenetic relationship among RGS and RGS-like proteins from various fungi was analyzed and discussed.
    Applied Microbiology and Biotechnology 08/2013; · 3.69 Impact Factor
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    ABSTRACT: Nematode-trapping fungi live mainly as saprobes in soil environments. When encountering nematodes, these fungi become 'carnivorous' and develop specialized trapping devices to attack their hosts for extracting nutrients, especially nitrogen source. Thus, nematode-trapping fungi are model organisms for understanding the molecular mechanism of the switch between saprobic and parasitic phases of pathogen life cycles. Arthrobotrys oligospora, one of the best-studied nematode-trapping fungi, mainly lives as a saprobe. In the presence of nematodes, A. oligospora enters the parasitic stage by forming adhesive reticulate traps to capture nematodes. In filamentous fungi, autophagy has been shown to be involved in morphogenesis and morphology. In this study, we demonstrate that autophagy is induced by nematodes during the early stage of trap formation in A. oligospora. Disruption of atg8 gene not only abolishes the nematode-induced autophagy, but also suppresses trap formation and reduces pathogenicity for nematodes. During the early stage of trap formation, the expression of genes involved in amino acid biosynthesis is upregulated and the transcriptional activity of GCN4 is induced in A. oligospora, suggesting that nematodes induce autophagy probably by triggering intracellular amino acid starvation. Autophagy is thus crucial for trap formation in A. oligospora during infection of nematodes.
    Environmental Microbiology Reports 08/2013; 5(4):511-7. · 2.71 Impact Factor
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    ABSTRACT: Microorganisms can attack and kill nematodes by diverse processes such as capturing, parasitizing, and producing toxins and enzymes. Extracellular enzymes, including serine proteases, chitinases, and collagenases are shown to be important virulence factors that can degrade the main chemical constituents of the nematode cuticle and eggshell. Here, we review the structure, function, regulation, and evolution of these nematicidal enzymes and provide insights into the mechanisms of microbial infections against nematodes. We discuss the practical applications of these nematicidal enzymes in agriculture and other areas.
    Applied Microbiology and Biotechnology 07/2013; · 3.69 Impact Factor
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    ABSTRACT: Nematode-trapping fungi can secrete many extracellular hydrolytic enzymes such as serine proteases and chitinases to digest and penetrate nematode/egg-cuticles. However, little is known about the structure and function of chitinases in these fungi. In this study, 16 ORFs encoding putative chitinases, which all belong to glycoside hydrolase (GH) family 18, were identified from the Arthrobotrys oligospora genome. Bioinformatics analyses showed that these 16 putative chitinases differ in their functional domains, molecular weights and pI. Phylogenetic analysis grouped these A. oligospora chitinases into four clades: clades I, II, III and IV, respectively, including an A. oligospora-specific subclade (Clade IV-B) that contained high-molecular weight chitinases (≥100 kDa). Transcriptional analysis of A. oligospora chitinases suggested that the expression of most chitinases was repressed by carbon starvation, and all chitinases were up-regulated under nitrogen starvation. However, chitinase AO-190 was up-regulated under carbon and/or nitrogen starvation. Moreover, several chitinases (such as AO-59, AO-190 and AO-801) were up-regulated in the presence of chitinous substrates or a plant pathogenic fungus, indicating that they could play a role in biocontrol applications of A. oligospora. Our results provided a basis for further understanding the functions, diversities and evolutionary relationships between chitinase genes in nematode-trapping fungi.
    Archives of Microbiology 05/2013; · 1.91 Impact Factor
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    ABSTRACT: The carnivorous fungus Arthrobotrys entomopaga (Drechsler) could develop adhesive knobs to capture nematodes. Chemical study on the culture medium of adhesive knobs producing A. entomopaga led to isolation of six trace amounts of metabolites, including two new metabolites paganins A-B (1-2), blumenol A (3), talathermophilins A and B (4-5), and cyclo(glycyltryptophyl) (6). Compounds 3-6 were reported for the first time from carnivorous fungi. Compounds 1-2 promoted the formation of the predatory adhesive knobs with an increasing rate up to 118% at a concentration of 50 uM, while showed moderate inhibitory activity at a concentration of 5 uM. Moreover, compounds 1-2 displayed strong inhibitory activities towards the formation of A. entomopaga conidiophores with inhibitory rates of 40-75%. Growth experiments suggested that compounds 1-2 could be involved in the regulating the fungal predatory and reproductive abilities. Nematode chemotaxis bioassay indicated that compounds 1 and 3 displayed strong nematode-attracting abilities. These findings provided a new type of regulatory metabolites and support for the hypothesis that predators often evolve to respond to their metazoan prey.
    Journal of Agricultural and Food Chemistry 04/2013; · 2.91 Impact Factor
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    ABSTRACT: Proteins are dynamic entities in cellular solution with functions governed essentially by their dynamic personalities. We review several dynamics studies on serine protease proteinase K and HIV-1 gp120 envelope glycoprotein to demonstrate the importance of investigating the dynamic behaviors and molecular motions for a complete understanding of their structure-function relationships. Using computer simulations and essential dynamic (ED) analysis approaches, the dynamics data obtained revealed that: (i) proteinase K has highly flexible substrate-binding site, thus supporting the induced-fit or conformational selection mechanism of substrate binding; (ii) Ca(2+) removal from proteinase K increases the global conformational flexibility, decreases the local flexibility of substrate-binding region, and does not influence the thermal motion of catalytic triad, thus explaining the experimentally determined decreased thermal stability, reduced substrate affinity, and almost unchanged catalytic activity upon Ca(2+) removal; (iii) substrate binding affects the large concerted motions of proteinase K, and the resulting dynamic pocket can be connected to substrate binding, orientation, and product release; (iv) amino acid mutations 375 S/W and 423 I/P of HIV-1 gp120 have distinct effects on molecular motions of gp120, facilitating 375 S/W mutant to assume the CD4-bound conformation, while 423 I/P mutant to prefer for CD4-unliganded state. The mechanisms underlying protein dynamics and protein-ligand binding, including the concept of the free energy landscape (FEL) of the protein-solvent system, how the ruggedness and variability of FEL determine protein's dynamics, and how the three ligand-binding models, the lock-and-key, induced-fit, and conformational selection are rationalized based on the FEL theory are discussed in depth.
    Journal of biomolecular structure & dynamics 03/2013; · 4.99 Impact Factor
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    ABSTRACT: Nematophagous fungi can trap and capture nematodes and other small invertebrates. This unique ability has made them ideal organisms from which to develop biological control agents against plant- and animal-parasitic nematodes. However, effective application of biocontrol agents in the field requires a comprehensive understanding about the ecology and population genetics of the nematophagous fungi in natural environments. Here, we genotyped 228 strains of the nematode-trapping fungus using 12 single nucleotide polymorphic markers located on eight random DNA fragments. The strains were from different ecological niches and geographical regions from China. Our analyses identified that ecological niche separations contributed significantly, whereas geographic separation contributed relatively little to the overall genetic variation in our samples of . Interestingly, populations from stressful environments seemed to be more variable and showed more evidence for recombination than those from benign environments at the same geographic areas. We discussed the implications of our results to the conservation and biocontrol application of in agriculture and forestry.
    Ecology and Evolution 02/2013; · 1.18 Impact Factor
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    ABSTRACT: A novel gene (designated as cen219) encoding endoglucanase was isolated from a Bursaphelenchus xylophilus metagenomic library by functional screening. Sequence analysis revealed that cen219 encoded a protein of 367 amino acids. SDS-PAGE analysis of purified endoglucanase suggested that Cen219 was a monomeric enzyme with a molecular mass of 40 kDa. The optimum temperature and pH for endoglucanase activity of Cen219 was separately 50°C and 6.0. It was stable from 30 to 50°C, and from pH 4.0 to 7.0. The activity was significantly enhanced by Mn(2+) and dramatically reduced by detergent SDS and metals Fe(3+), Cu(2+) or Hg(2+). The enzyme hydrolyzed a wide range of β-1, 3-, and β-1, 4-linked polysaccharides, with varying activities. Activities towards microcrystalline cellulose and filter paper were relatively high, while the highest activity was towards oat gum. The Km and Vmax of Cen219 towards CMC was 17.37 mg/ml and 333.33 U/mg, respectively. The findings have an insight into understanding the molecular basis of host-parasite interactions in B. xylophilus species. The properties also make Cen219 an interesting enzyme for biotechnological application.
    PLoS ONE 01/2013; 8(12):e82437. · 3.73 Impact Factor
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    ABSTRACT: Macrocyclic PKS-NRPS hybrid metabolites represent a unique family of natural products mainly from bacteria with broad and outstanding biological activities. However, their distribu-tion in fungi has rarely been reported, and few work has been reported regarding to their nematocidal activity. Here we describe an unprecedented class of PKS-NRPS hybrid metabolites possessing a 13-membered lactam-bearing macrolactone, thermolides A-F (1-6) from a thermophilic fungus Talaromyces thermophilus. We showed that compounds 1 and 2 displayed potent inhibitory activity against three notorious nematodes with LC50 values of 0.5-1 ug/mL, as active as commercial avermectins. This work provided a new class of promising lead compounds for nematocide dis-covery.
    Journal of the American Chemical Society 12/2012; · 10.68 Impact Factor
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    ABSTRACT: The basidiomycete genus Stereum fam Stereaceae is a rich source of sesquiterpenoids. This phytochemical investigation of the basidiomycete Stereum sp. CCTCC AF 2012007 led to isolation of six cadinane-type sesquiterpenes named stereumins K-P. Stereumin O was very likely an artifact formed from stereumin N during isolation. Their structures were elucidated mainly by 1D and 2D NMR spectroscopy. Structures of stereumins K, L, M and P were further confirmed by single-crystal X-ray diffraction, and the absolute configuration of stereumin K was determined.
    Phytochemistry 11/2012; · 3.05 Impact Factor
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    ABSTRACT: Background: Soil-dwelling Bacillus nematocida B16 can kill Caenorhabditis elegans via a Trojan horse-like mechanism. However, colonization is a key problem that must be solved during the infection process. Aims: To study the molecular mechanism involved in the colonization of B. nematocida B16 against the host C. elegans. Methods: GFP-expressing strain B16g was constructed and its nematocidal activity was assayed. 'Feeding transfer' experiments were carried out separately using of B16 and B16g strains to explore the colonization mode of the bacteria. Fluorescence microscopy was used to observe the interactions between fluorescent signal and the quantity of bacteria in the intestine. A mariner-based transposon called TnYLB-1 was also applied in the random mutagenesis of B16 to screen the mutants with impaired colonization of nematode worms and identify potential localization-related genes. Results and Conclusion: A small inoculum of the bacteria resulted in its proliferation in the C. elegans intestine. The fluorescence signal was enhanced with increasing bacterial density in the intestine. Several candidate genes with possibly important roles in colonization were found. These results provide a solid foundation for further elucidation of the infection process at the molecular level and enrichment of our knowledge of bacterial pathogenesis.
    Journal of Molecular Microbiology and Biotechnology 10/2012; 22(4):258-67. · 1.95 Impact Factor
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    ABSTRACT: Novel autoregulatory metabolites, arthrosporols A-C (1-3), involved in regulating the morphological switch in fungi, were purified and characterized from the carnivorous fungus Arthrobotrys oligospora. These compounds possess a novel hybrid carbon skeleton consisting of an epoxy-cyclohexenol combined with a rare monocyclic sesquiterpenol substructure. This is the first report of a monocyclic sesquiterpenol of this type of fungal origin. Compounds 1-3 displayed significant inhibitory activities toward the formation of conidiophores, while compounds 1 and 3 showed the opposite effects on the formation of a two-dimensional network with increasing rates of 40-90% and inhibiting rates of 30-90%, respectively.
    Journal of Natural Products 07/2012; 75(7):1419-23. · 3.29 Impact Factor
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    ABSTRACT: Chemical investigation of one entomopathogenic fungus Paecilomyces cateniobliquus YMF1.01799 led to the isolation and identification of six metabolites, which include three new compounds (2-3, and 5) and three known metabolites. Their structures were established by spectroscopic studies such as 1D and 2D NMR and MS analysis. Insect growth experiments suggested that polyketide-derived compound 1 showed significant inhibitory effect on the growth of cotton bollworm Helicoverpa armigera, while terpenoid-derived metabolite 5 promoted the growth of the larvae. The findings revealed that the entomopathogenic fungus P. cateniobliquus could produce different types of metabolites to regulate growth of the insect.
    Journal of Agricultural and Food Chemistry 05/2012; 60(22):5604-8. · 2.91 Impact Factor
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    02/2012; , ISBN: 978-953-51-0037-9
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    ABSTRACT: Malachite green (MG), N-methylated diaminotriphenylmethane, is one of the most common dyes in textile industry and has also been used as an effective antifungal agent. However, due to its negative impact on the environment and carcinogenic effects to mammalian cells, there is a significant interest in developing microbial agents to degrade this type of recalcitrant molecules. Here, an Exiguobacterium sp. MG2 was isolated from a river in Yunnan Province of China as one of the best malachite green degraders. This strain had a high decolorization capability even at the concentration of 2500 mg/l and maintained its stable activity within the pH range from 5.0 to 9.0. High-pressure liquid chromatography, liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry were employed to detect the catabolic pathway of MG. Six intermediate products were identified and a potential biodegradation pathway was proposed. This pathway involves a series of reactions of N-demethylation, reduction, benzene ring-removal, and oxidation, which eventually converted N-methylated diaminotriphenylmethane into N, N-dimethylaniline that is the key precursor to MG. Furthermore, our molecular biology experiments suggested that both triphenylmethane reductase gene tmr and cytochrome P450 participated in MG degradation, consistent with their roles in the proposed pathway. Collectively, our investigation is the first report on a biodegradation pathway of triphenylmethane dye MG in bacteria.
    PLoS ONE 01/2012; 7(12):e51808. · 3.73 Impact Factor

Publication Stats

704 Citations
315.79 Total Impact Points

Institutions

  • 2004–2014
    • Yunnan University
      • • Laboratory for Conservation and Utilization of Bio-resources
      • • The Key Laboratory for Microbial Resources of the Ministry of Education
      Yün-nan, Yunnan, China
  • 2011
    • McMaster University
      • Department of Biology
      Hamilton, Ontario, Canada
  • 2008
    • Kunming Medical College
      Yün-nan, Yunnan, China