Root rot caused by Cylindrocarpon destructans is one of the most important diseases of ginseng (Panax ginseng C. A. Meyer). Two types of symptoms found in ginseng root rot are black root rot and rusty root (rusty spots), in which disease severities are high and low, respectively. Symptom development and related histopathological changes were examined in an inoculation test on 2-year-old ginseng roots using virulent (Cy9801) and avirulent (Cy0001) isolates of C. destructans under different temperature conditions (13, 18, 23, and $28^{\circ}C$). Black root rot was only induced by Cy9801 in the lower temperature range (13, 18, and $23^{\circ}C$) and not at the higher temperature ($28^{\circ}C$). No black root rot, but only rusty spot symptoms, were induced by Cy0001 at all temperatures tested except $13^{\circ}C$, at which no symptoms occurred on over half of inoculation sites, suggesting disease development was influenced by pathogen virulence and temperature. Wound periderms were formed in all root tissues with rust spot symptoms at $28^{\circ}C$ caused by Cy9801 and at 18, 23, and $28^{\circ}C$ temperatures caused by Cy0001. No wound periderm was formed at $13^{\circ}C$ by either Cy9801 or Cy0001. Light microscopy revealed that the wound periderm was formed by initial cell divisions in cell wall formation and/or additional cell wall layering in parenchyma cells without obvious nuclear division, followed by layering of the divided cells adjacent to the inoculation sites, blocking the spread of the rot. These results suggest that disease development declined at lower temperatures and by the formation of a wound periderm at higher temperatures, and that ginseng rusty root may develop under conditions unfavorable for further disease development of C. destructans.

Snow molds are the most important winter diseases of turfgrass in the United States and Canada. Eight isolates of three snow mold fungal species (three isolates of Typhula ishikariensis, three of T. incarnata, and two of Microdochium nivale) were collected from infected turfgrasses on golf courses. The isolates were evaluated for their relative aggressiveness on three cultivars (L-93, Penncross, and Providence) of creeping bentgrass (Agrostis palustris) under the same controlled conditions. Four plant ages (15, 19, 23 and 27 week-old plants from germination to inoculation) were evaluated for their susceptibility to the three pathogens and for the recovery of the plants. Regardless of age or cultivar of the host plant, M. nivale was found to be more aggressive and faster to infect and colonize than Typhula species. After three weeks recovery, M. nivale-inoculated plants showed higher disease severity than plants inoculated with the two Typhula species. Plants infected by Typhula species displayed no significant difference in disease severity. As creeping bentgrass plants get older, the severity of disease caused by three snow molds gradually decreases. This effect was observed in all cultivars tested, suggesting expression of age-related resistance as the bentgrass plants matured.

Ninety isolates of Colletotrichum species from new persimmon tree twigs and 50 isolates from pepper plant fruits were isolated via single-spore isolation. Of the 140 isolates, 26 were examined for mycelial growth, carbendazim sensitivity, and ITS sequence. Four of the isolates from the persimmon trees, which were cultivated exclusively in an orchard, showed fast mycelial growth and sensitivity to carbendazim, while five of the pepper isolates showed slower mycelial growth and were resistant to the fungicide. However, 17 isolates from persimmon trees cultivated with pepper plants in the same orchard showed slow mycelial growth like the pepper isolates and they were sensitive to carbendazim like the persimmon isolates. ITS sequence analysis of these 27 isolates led to the identification of the 22 persimmon isolates as C. gloeosporioides and the five pepper isolates as C. acutatum. PCR with species-specific primers confirmed that the 90 isolates from persimmon were C. gloeosporioides whereas the 50 isolates from pepper were C. acutatum. The 90 persimmon isolates of C. gloeosporioides and 50 pepper isolates of C. acutatum were compared by a wound inoculation test to determine their capacity for host cross-infection. All of the C. acutatum isolates from pepper caused typical symptoms of anthracnose on the fruits of pepper plants and twigs of persimmon; they differed from the C. gloeosporioides isolates from persimmon, more than 90% of which were able to infect only persimmon. Amplified fragment length polymorphism analysis revealed the existence of two groups (C. gloeosporioides and C. acutatum isolates group). At 80% genetic similarity, the C. gloeosporioides group was defined within four clusters, while the C. acutatum group was within three clusters. However, these clusterings were unrelated with the virulence of Colletotrichum species against pepper fruits.

Jujube trees infected with phytoplasma exhibit symptoms of typical witches' broom, such as yellowing, abnormally small leaves, short internodes and proliferation of shoots. A 1.2 kb fragment of the 16S rDNA from jujube phytoplasma was generated by R16F2n/R16R2 primer pair from earlier amplified P1/P7 PCR products of cloned jujube witches' broom phytoplasmas. Enzymatic restriction fragment length polymorphism (RFLP) and sequence analysis of 16S rDNA revealed that the jujube tree was infected with 16S rDNA I and V groups of phytoplasmas. Extensive comparative analyses of restriction enzyme profiles from Alu I, Hha I, Msp I, and Rsa I clearly classified the two into different phytoplasma groups. The phylogenie analyses based on 16S rDNA showed that the similarity of the two different clones was 87.5%. This is the first report of a mixed phytoplasmal infection in a single jujube tree.

Tomato spotted wilt virus (TSWV) was identified from seven plants at two areas, Anyang and Dangjin, in Korea. The isolates of TSWV were seven as TSWV-KATm from tomato, TSWV-KAPo from potato, TSWV-KABal from balsam, TSWV-KACTm from cherry tomato and TSWV-KAIxe from Ixeris dentata at Anyang area, and TSWV-KDSe from sesame and TSWV-KDRP from red pepper at Dangjin area. Pathogenicity of seven TSWV isolates was various on the assay plants, and could not be grouped definitely. Three isolates of TSWV-KAIxe, TSWV-KACTm and TSWV-KABal had relatively narrower host ranges among the seven isolates. Percentage of nucleotide substitution in nucleotide sequences encoding nucleocapsid protein (NCP) was 1.2-1.7% among seven TSWV isolates and TSWV-KP. Korean TSWV isolates were divided into three groups by nucleotide homology or amino acid compositions. From the analysis of nucleotide sequences of Korean TSWV isolates compared with those of TSWV reported from other 5 countries including Japan, the Korean seven isolates of TSWV was grouped with German TSWV (D13926). No Korean TSWV isolates were grouped with those from The Netherlands, Brazil and USA.

Lichen-forming fungi (LEF) were isolated from 67 Hungarian lichen species from ascospores or thallus fragments. LFF were successfully isolated from 26 species with isolation rate of 38.8%. Of the total number of isolation from ascospores (27 species) and thallus fragments (40 species), 48% and 32.5% of the species were successfully isolated, respectively. Comparison of rDNA sequences of ITS regions between the isolated LFF and the original thallus confirmed that all the isolates originated from the thallus fragments were LEF. The following 14 species of LEF were newly isolated in this study; Acarospora cervina, Bacidia rubella, Cladonia pyxidata, Lasallia pustulata, Lecania hyaline, Lecanora argentata, Parmelina tiliacea, Parmotrema chinense, Physconia distorta, Protoparmeliopsis muralis, Ramalina pollinaria, Sarcogyne regularis, Umbilicaria hirsuta, Xanthoparmelia conspersa and X. stenophylla. Antifungal activity of the Hungarian LFF was evaluated against plant pathogenic fungi of Colletotrichum acutatum, C. coccodes and C. gloeosporioides, causal agent of anthracnose on hot pepper. Among the 26 isolates, 11 LFF showed more than 50% of inhibition rates of mycelial growth of at least one target pathogen. Especially, LFF of Evernia prunastri, Lecania hyalina and Lecanora argentata were remarkably effective in inhibition of mycelial growth of all the tested pathogens with antibiotic mode of action. On the other hands, five isolates of Cladonia furcata, Hypogymnia physodes, Lasallia pustulata, Ramalina fastigiata and Ramalina pollinaria exhibited fungal lytic activity against all the three pathogens. Among the tested fungal pathogens, C. coccodes seemed to be most sensitive to the LFF. The Hungarian LFF firstly isolated in this study can be served as novel bioresources to develop new biofungicides alternative to current fungicides to control hot pepper anthracnose pathogenic fungi.

Sequence analysis of the Chlorella virus SS-2 revealed one putative nuclease gene that is 807 bp long and encodes a 31kDa protein. Multiple sequence alignment analysis reveals the presence of highly conserved PD-(D/E)XK residues in the encoded protein. The gene cloned into an expression vector was expressed as a His-tagged fusion protein in chaperone containing pKJE7 cells. The recombinant protein was purified using a His-Trap chelating HP column and used for functional analysis. Exonuclease activity of the SS-2 nuclease was detected when the DNA substrates, such as linear ssDNA, PCR amplicon, linear dsDNA with 5'-overhang ends, 3'-overhang ends, or blunt ends were used. Covalently closed circular DNA was also degraded by the SS-2 recombinant protein, suggesting that the SS-2 nuclease has an endonuclease activity. Stable activity of SS-2 nuclease was observed between $10^{\circ}C$ and $50^{\circ}C$. The optimum pH concentrations for the SS-2 nuclease were pH 6.0-8.5. Divalent ions inhibited the SS-2 nuclease activity.

An infectious full-length clone of Soybean mosaic virus (SMV) strain G5H was constructed under the control of the cauliflower mosaic virus 35S promoter. The cloned SMV G5H established infections upon simple rub-inoculation of soybean leaves with intact plasmid DNA. We demonstrated that this SMV G5H infectious DNA clone caused typical characteristic symptoms and virulence of SMV strain G5H in twelve tested soybean cultivars. Soybean cultivars Lee74, Somyungkong and Sowonkong developed systemic mosaic symptom while Kwanggyo, Taekwangkong, Hwangkeumkong and Geumjeongkong-l showed systemic necrosis. In contrast, Geumjeongkong-2, Jinpumkong-2, L29, V94-5152 and Ogden showed resistant response against SMV-G5H infection. We also determined full-length sequence of cloned SMV-G5H. The phyogenetic analyses reveal that SMV-G5H is most closely related to SMV-G5, and support that SMV-G5H might be derived from SMV-G5 by recombination rather than mutation.

In this study, we characterized the bacterial strain KJ2C12 in relation with its biocontrol activity against Phytophthora capsici on pepper, and identified this strain using morphological, physiological, biochemical, fatty acid methyl ester, and 16S rRNA gene sequence analyses. Strain KJ2C12 significantly (P=0.05) reduced both final disease severity and areas under the disease progress curves of 5-week-old pepper plants inoculated with P. capsici compared to buffer-treated controls. As for the production of antibiotics, biofilms, biosurfactant, extracellular enzyme, HCN, and swarming activity, strain KJ2C12 produced an extracellular enzyme with protease activity, but no other productions or swarming activity. However, Escherichia coli produced weak biofilm only. Strain KJ2C12 could colonize pepper roots more effectively in a gnotobiotic system using sterile quartz sand compared to E. coli over 4 weeks after treatments. However, no bacterial populations were detected in 10 mM $MgSO_4$ buffer-treated controls. Strain KJ2C12 produced significantly higher microbial activity than the $MgSO_4$-treated control or E. coli over 4 weeks after treatments. Bacterial strain KJ2C12 was identified as Bacillus luciferensis based on morphological, physiological, and biochemical characteristics as well as FAME and 16S rRNA gene sequence analyses. In addition, these results suggested that B. luciferensis strain KJ2C12 could reduce Phytophthora blight of pepper by protecting infection courts through enhanced effective root colonization with protease production and an increase of soil microbial activity.

The rhizobacteria of Panax notoginseng were isolated from six sites in Yanshan, Maguan and Wenshan Counties, Yunnan Province of China, and their antagonistic activity against P. notoginseng root-rot fungal pathogens was determined. Of the 574 rhizobacteria isolated, 5.8% isolates were antagonistic in vitro to at least one of the five pathogens, Cylindrocarpon didynum, Fusarium solani, Phytophthora cactorum, Phoma herbarum, and Rhizoctonia solani. The number of rhizo bacteria and the number that inhibited fungi differed depending on sampling sites and isolation methods. Rhizobacteria isolated from the site in Yanshan and Maguan showed more antagonistic effect than them in Wenshan. Heat treatment of rhizosphere soil at $80^{\circ}C$ for 20 min scaled the antagonists up to 14.0%. Antagonistic bacteria in the roots proportioned 3.9% of the total isolates. The most antagonistic isolates 79-9 and 81-4 are Bacillus subtilis based on their 168 rDNA sequence and biochemical and physiological characteristics. Identification and evaluation of antagonistic bacteria against P. notoginseng root-rot pathogens in the main planting areas improved our understanding of their distribution in rhizosphere soil. Furthermore these results indicated that the interactions between biocontrol agent and soil microbes should be seriously considered for the successful survival and biocontrol efficacy of the agents in soil.

Pseudomonas fluorescens strain Gpf01, isolated from ginseng rhizosphere showed antiviral activity against Cucumber mosaic virus, when tested in a local host of CMV, Chenopodium amaranticolor. Transposon mutant library of Gpf01 was prepared using pGS9::Tn5 and the mutant Gpf01-RS19 was found to loose antiviral production. We developed primers from the flanking region of Tn5 and found a cosmid clone pAV1123, harboring 1.2 kb antiviral compound producing (avcf01) locus. When a sub-clone pPH9, which carried 9.3 kb region of pAV1123, was introduced into antivirus deficient P. fluorescens wild type strain B16, it exhibited antiviral activity. Using Tn3-gus mutagenesis and complementation analysis, it was found that the genes related to antiviral activity production resided in a 9.3 kb HindIII-HindIII fragment of pAV1123, indicating that the plasmid carries an essential genes promoting antiviral activity.

In November 2008, typical powdery mildew symptoms were observed on leaves of Arabidopsis thaliana ecotype Col-0 plants in a growth room under controlled laboratory conditions at Korea University, Seoul. The disease was characterized by the appearance of white powder-like fungal growth on the surface of infected leaves. As the disease progressed, infected leaves exhibited chlorotic or necrotic brown lesions, and leaf distortion and senescence. Conidiophores of the causal fungus were hyaline, unbranched, 3-4 celled, cylindrical, and $80-115{\times}6-9{\mu}m$ in size. Singly produced conidia (pseudoidium type) were hyaline, oblong to cylindrical or oval in shape, and $26-55{\times}15-20{\mu}m$ in size with a length/width ratio of average 3, angular/rectangular wrinkling of outer wall and no distinct fibrosin bodies. Appressoria on the hyphae were multi-lobed. These structures are typical of the powdery mildew Oidium subgenus Pseudoidium, anamorph of the genus Erysiphe. The measurements of the fungal structures coincided with those of Erysiphe cruciferarum. The phylogenetic analysis using ITS rDNA sequences revealed that the causal fungus Erysiphe sp. KUS-F23994 is identical to E. cruciferarum. The isolated fungus incited powdery mildew symptoms on the inoculated Arabidopsis leaves, which proved Koch's postulates. Taken all data together, we first report the occurrence of powdery mildew disease of A. thaliana caused by Erysiphe cruciferarum in Korea.

An unreported disease of garlic was observed in commercial fields in Jiangsu province, China. The symptoms started as water soaked lesions at the base of the leaves. Later, water-soaked areas developed on stems and spread to the internal tissues, followed by yellowing and necrosis along leaf edges and soft rot of the stems. The causal organism isolated from symptomatic plants was identified as Pseudomonas fluorescens based on its biochemical and physiological characteristics and confirmed by the cellular fatty acid composition and Biolog data as well as 168 rRNA gene sequence analysis. The bacterial isolates caused similar symptoms when inoculated onto garlic plants. In addition, leek and shallot were susceptible to the P. fluorescens pathogen. However, the P. fluorescens pathogen failed to cause any symptoms when it was inoculated onto 15 other plants. This is the first report of a bacterial disease of garlic caused by P. fluorescens in China.

The incidence and occurrence of virus infecting paprika (Capsicum annuum var. grossum) in Jeonnam province, the main areas of cultivation in Korea is undetermined. In this study, a total of 1,020 samples with virus-like symptoms were collected in Jeonnam province during summer season for 3 consecutive years (2002-2005) and were tested using enzyme linked immunosorbent assay (ELISA). Results showed that Pepper mottle virus (PepMoV), Broad bean wilt virus (BBWV), and Cucumber mosaic virus (CMV) were found to be the most prevalent viruses with a 3-year average percent incidence of 41.3, 19.8, and 4.4 respectively. Mixed infection with more than two viruses was also found with 3.5%, 17.0%, and 8.3%, respectively. Symptoms of these virus diseases were not evident at the seedling stage but slowly appeared at the transplanting stage and increased to the middle stage (4-5 months after transplanting) during the 3-year cultivation periods. Symptom appearance of infected plants however varied largely with transplanting time. Those plants transplanted from November to January were found to be infected with viruses in June, whereas symptoms appeared with in a month those plants transplanted from June to August. There were differences in the virus incidence from primary factor such as district, type of green house and variety, but these were not statistically significant (data not shown). Recommended control measures of paprika against these viruses is also discussed in this paper.

An antagonistic bacterium of Bacillus subtilis BD0310 against Colletotrichum theae-sinensis was isolated from the phylloplane of tea trees at a tea plantation in Korea. SC (suspension concentrate)-type biofungicide was formulated with the antagonist. Cell viability and antifungal activity of B. subtilis were maintained in the formulation more than 12 months at room temperature. The antagonist was sensitive only to copper sulfate among the chemical pesticides currently registered for tea trees in Korea. Greenhouse application demonstrated that the biofungicide controlled more effectively the disease in a protective mode than in a curative mode. Field trial showed that alternate applications of the biofungicide and chemical fungicide were more effective in controlling tea anthracnose than single application of the biofungicide or chemical fungicide with less use of chemicals. This study suggests that the biofungicide of B. subtilis 8D0310 is an effective method for biological control of anthracnose in tea plantations.

In the course of a searching natural antifungal compounds from plant seeds, we found that the methanol extract of Psoralea corylifolia seeds showed potent control efficacy against tomato late blight caused by Phytophthora infestans and wheat leaf rust Puccinia recondita. Under bioassay-guided purification, we isolated two furanocoumarins, psoralen and isopsoralen, with anti-oomycete activity against P. infestans. By 1-day protective application, both compounds strongly reduced the disease development of P. infestans on tomato seedlings, but hardly controlled development of leaf rust on wheat seedlings. This is the first report on the anti-oomycete activity of P. corylifolia as well as that of psoralen and isopsoralen.

DNA-based studies, including cloning and genotyping, have become routine in fungal research laboratories. However, preparation of high-quality DNA from fungal tissue requires much time and labor and is often a limiting step for high-throughput experiments. We have developed a quick and safe (QS) DNA extraction method for fungi. Time efficiency and safety in the QS method were achieved by using plate-grown mycelia as the starting material, by eliminating phenol-chloroform extraction procedures, and by deploying a simple electric grinder. This QS method is applicable not only to a broad range of microbial eukaryotes, including true fungi and oomycetes, but also to lichens and plants.