Jong Myong Park

Diversity of Endophytic Fungi Associated with the Roots of Four Aquatic Plants Inhabiting Two Wetlands in Korea.

Abstract A total of 4 aquatic plants, Eleocharis kuroguwai Ohwi, Hydrocharis dubia Backer, Salvinia natans All., and Zizania latifolia Turcz., were sampled from representative two wetlands of South Korea. A total of 38 endophytic fungal strains were isolated from aquatic plants native to the Daepyeong wetland, and 27 strains were isolated from the Jilnal wetland. The internal transcribed spacer regions of fungal isolates were sequenced and a phylogenetic analysis was performed. In addition, endophytic fungal diversity from each wetland and host plant species was deduced. A total of 25 fungal genera were purely isolated, and 16 fungal genera were isolated from each of the two wetlands. Commonly isolated genera from both wetlands were Aspergillus, Cladosporium, Clonostachys, Fusarium, Leptosphaeria, Penicillium, and Talaromyces. This study revealed that fungal diversity varied with environmental conditions and by host plant in representative two wetlands.

Fungal load in Bradysia agrestis, a phytopathogen-transmitting insect vector

Larvae of Bradysia agrestis, a phytopathogen-transmitting insect vector in East Asia, were sampled from geographically (ecologically) segregated regions to identify their intestinal fungal flora. A total of 24 fungal strains were isolated from the insect vectors and selected based on morphological differences. In addition, 38 fungal strains were isolated from the ulcerated parts of invaded host plants by the same method, revealing the impact of vector fungal flora on their host plants. For molecular identification of the fungi, internal transcribed spacer (ITS) regions were amplified and sequenced. Their sequences were compared with sequences of other fungal strains obtained from NCBI GenBank, and their phylogeny was determined. The dominant fungal genera in the insect vector were Penicillium (25%), Aspergillus (21%), and Cladosporium (13%). In plant scar lesions, most fungal isolates belonged to the genera Fusarium (31.6%), Phoma (7.8%), Didymella (7.8%), and Epicoccum (7.8%). Fungal genera in vectors or host plant lesions differed by study site. Furthermore, diversity indices by study site showed clear differences based on Margalef’s richness (2.06, 2.40, 3.04), and Menhinick’s (1.89, 2.12, 2.53), and Simpson’s indices (0.14, 0.07, 0.07). In addition, common fungal strains in insect vectors were found to be closely related to members of the genera Cladosporium, Penicillium, or Aspergillus. Among these strains, those showing the highest homology with Aspergillus terreus, which regarded as beneficial fungal genera could be considered ideal paratransgenesis candidates. Some other fungal strains from vectors or ulcerated plant parts from each study site after B. agrestis invasion may be harmful in terms of plant disease or agrifood safety. This study provides information on the fungal microbiota of B. agrestis, an emerging problem in East Asia, and proposes paratransgenesis candidates to control this insect vector. Furthermore, potential transferable pathogens or commensal fungi were revealed by comparing the fungal biota between the insect gut and the ulcerated parts of the invaded host plants.

Foodborne Infectious Diseases Mediated by Inappropriate Infection Control in Food Service Businesses and Relevant Countermeasures in Korea

Objectives The objective of this review is to propose an appropriate course of action for improving the guidelines followed by food handlers for control of infection. For this purpose, previous epidemiological reports related to acute gastroenteritis in food service businesses mediated by food handlers were intensively analyzed. Methods Relevant studies were identified in international databases. We selected eligible papers reporting foodborne infectious disease outbreaks. Among primary literature collection, the abstract of each article was investigated to find cases that absolutely identified a causative factor to be food handlers’ inappropriate infection control and the taxon of causative microbial agents by epidemiological methodologies. Information about the sites (type of food business) where the outbreaks occurred was investigated. Results A wide variety of causative microbial agents has been investigated, using several epidemiological methods. These agents have shown diverse propagation pathways based on their own molecular pathogenesis, physiology, taxonomy, and etiology. Conclusion Depending on etiology, transmission, propagation, and microbiological traits, we can predict the transmission characteristics of pathogens in food preparation areas. The infected food workers have a somewhat different ecological place in infection epidemiology as compared to the general population. However, the current Korean Food Safety Act cannot propose detailed guidelines. Therefore, different methodologies have to be made available to prevent further infections.

Cutaneous Microflora from Geographically Isolated Groups of Bradysia agrestis, an Insect Vector of Diverse Plant Pathogens

Abstract Larvae of Bradysia agrestis, an insect vector that transports plant pathogens, were sampled from geographically isolated regions in Korea to identify their cutaneous fungal and bacterial flora. Sampled areas were chosen within the distribution range of B. agrestis; each site was more than 91 km apart to ensure geographical segregation. We isolated 76 microbial (fungi and bacteria) strains (site 1, 29; site 2, 29; site 3, 18 strains) that were identified on the basis of morphological differences. Species identification was molecularly confirmed by determination of universal fungal internal transcribed spacer and bacterial 16S rRNA gene sequences in comparison to sequences in the EzTaxon database and the NCBI GenBank database, and their phylogenetic relationships were determined. The fungal isolates belonged to 2 phyla, 5 classes, and 7 genera; bacterial species belonged to 23 genera and 32 species. Microbial diversity differed significantly among the geographical groups with respect to Margalef’s richness (3.9, 3.6, and 4.5), Menhinick’s index (2.65, 2.46, and 3.30), Simpson’s index (0.06, 0.12, and 0.01), and Shannon’s index (2.50, 2.17, and 2.58). Although the microbial genera distribution or diversity values clearly varied among geographical groups, common genera were identified in all groups, including the fungal genus Cladosporium, and the bacterial genera Bacillus and Rhodococcus. According to classic principles of co-evolutionary relationship, these genera might have a closer association with their host insect vector B. agrestis than other genera identified. Some cutaneous bacterial genera (e.g., Pseudomonas) displaying weak interdependency with insect vectors may be hazardous to agricultural environments via mechanical transmission via B. agrestis. This study provides comprehensive information regarding the cutaneous microflora of B. agrestis, which can help in the control of such pests for crop management.

Plant Growth Promotion and Gibberellin A3 Production by Aspergillus flavus Y2H001

Perilla frutescens var. japonica Hara was collected from farmland in Seongju-gun. Fifteen endophytic fungal strains with different colony morphologies were isolated from roots of P. frutescens. Waito-c rice seedlings were treated with the concentrated culture filtrates (CF) of endophytic fungi for observation of their plant growth-promoting activities. In the results, the CF of Y2H001 fungal strain promoted the growth of the waito-c rice seedlings. The phylogenetic tree of Y2H001 strain was analyzed by the combined sequences of the partial internal transcribed spacer region (ITS) and partial betatubulin gene. Molecular and morphological studies identified the Y2H001 strain as belonging to Aspergillus flavus. In gas chromatography mass spectrometry (GC/MS) analysis of the CF of Y2H001 strain, gibberellic acid (GA) was detected and quantified. Therefore, we describe Y2H001 strain as a new -producing A. flavus based on morphological, molecular characteristics and analysis of secondary metabolite.

Comparative Analysis of Endophytic Fungi Isolated from Dominant Hydrophytes in Junam and Dongpan Wetland

Hydrocharis dubia Backer and Salvinia natans All. were sampled from the Junam and Dongpan reservoirs, representative freshwater wetlands of Korea. A total of 19 endophytic fungal strains were isolated from hydrophytes native to the Junam wetlands and 5 strains were isolated from the Dongpan wetlands. Depending on phylogenetic analysis based on internal transcribed spacer (ITS) region, strains from Junam belonged to 11 genera and from Dongpan belonged to 5 genera. Fusarium, Phoma and Talaromyces were commonly distributed genera from two wetlands. The fungal diversity index showed clear differences between each wetlands or each host hydrophyte. Above all, the highest diversity value was observed from Salvinia natans All., which have been reported as promising biological resources as eutrophication controller in environmental ecology.

Diversity and Phylogenetic Analysis of Culturable Marine Bacteria Isolated from Rhizosphere Soils of Suaeda japonica Makino in Suncheon Bay

Bacterial diversity was studied in the rhizosphere of Suaeda japonica Makino, which is native to Suncheon Bay in South Korea. Soil samples from several sites were diluted serially, and pure isolation was performed by subculture using marine agar and tryptic soy agar media. Genomic DNA was extracted from 29 pure, isolated bacterial strains, after which their 16S rDNA sequences were amplified and analyzed. Phylogenetic analysis was performed to confirm their genetic relationship. The 29 bacterial strains were classified into five groups: phylum Firmicutes (44.8%), Gamma proteobacteria group (27.6%), Alpha proteobacteria group (10.3%), phylum Bacteriodetes (10.3%), and phylum Actinobacteria (6.8%). The most widely distributed genera were Bacillus (phylum Firmicutes), and Marinobacterium, Halomonas, and Vibrio (Gamma proteobacteria group). To confirm the bacterial diversity in rhizospheres of S. japonica, the diversity index was used at the genus level. The results show that bacterial diversity differed at each of the sampling sites. These 29 bacterial strains are thought to play a major role in material cycling at Suncheon Bay, in overcoming the sea/mud flat-specific environmental stress. Furthermore, some strains are assumed to be involved in a positive interaction with the halophyte S. japonica, as rhizospheric flora, with induction of growth promotion and plant defense mechanism.To study the halobacterial diversity at the rhizospheric soil of coastal plant native to Dokdo islands, several host plant were selected and its rhizospheric soil was sampled. Soil sample was diluted serially and pure isolation was done by sub-culture using marine agar media. 26 halophilic strains cultivable at the marine medium containig concentration of 9.0% sodium chloride were selected among total 161 isolates. Their partial 16S rRNA gene sequences extracted from genomic DNA were analyzed and partially identified. Furthermore, to identify their genetic relationship, phylogenetic tree was deduced. Total 26 strains were belongs to Firmicutes (30.8%), Gamma proteobacteria (53.8%), Bacteroidetes (7.7%), Alpha proteobacteria (7.7%), and Actinobacteria (7.7%). These results showed the specific difference from previous researches which has been reported the microbial flora of soil or sea water around the Dokdo islands. Furthermore, 4 among 26 halophilic strains grew at above 12.0% NaCl concentrated marine broth, and 2 strains Idiomarina abyssalis LM4H23 and Halomonas huangheensis AS4H13 grew at 15.0% concentration. These halophilic strains thought to overcoming the severe stress like high salt concentration or variation derived from Dokdo-specific climate and might have unknown, specific relationship with their host coastal plant native to Dokdo islands.