A new virus-like disease of tomato showing chlorotic spots, malformation and necrosis on leaves, and chlorotic blotching, rings, and necrosis on fruits was observed around Sacheon, Gyeongsangnam-do, Korea in 2004. Host range analysis could not differentiate 4 field isolates collected from tomatoes showing different symptoms but identified them as Tomato bushy stunt virus (TBSV). TBSV-tsf2 isolate induced symptoms in indicator plants similar to those caused by the TBSV-C, -S and -Nf. As the isolate could not systemically infect Chenopodium quinoa, the isolate might belong to the previously described TBSV-S isolate. TBSV-tsf2 isolate caused similar cytological alterations that were similar to that generally caused by previously reported TBSV isolates. TBSV-tsf2 isolate, however, could be considered to belong to new strain of TBSV because masses of small electron-dense patches that were not observed from the previously described TBSV. The complete nucleotide sequences of the genomic RNA of 4739 nt excluding non-translated sequences at both termini have been determined and compared to sequences of other TBSV strains. The complete nucleotide sequence identity among TBSV isolates was 98.9% to 99.7%, and to the other tombusviruses ranged from 80.8% to 94.9%. Comparison of the amino acid sequences all five ORFs with those of other TBSV strains shows a similar genomic organization, and high percentage of amino acid sequence homology with TBSV-Nf than TBSV-S isolate. Since the TBSV symptoms were only observed in Sacheon fields where imported seeds from Japan were planted, the TBSV incidence probably caused by the planting contaminated tomato seeds and thus require more through quarantine procedure to prevent settlement of TBSV in Korea. Altogether, these results support that the Korean isolate of TBSV infecting tomato might be new strain.

Phytoplasma was identified from leaf lettuce (Lactuca sativa) cultivated in commercial green-house in Korea. Diseased leaf lettuce revealed proliferation of shoots, and yellowing and shrinking of leaves (lettuce proliferation-K). Polymerase chain reaction (PCR) with universal primer pair P1/P6, and aster yellows (AY) specific primer pair R16F1/R1 amplified 1.5kb and 1.1kb length of DNA fragments, respectively. Nucleotide sequences of 16S rRNA gene were determined (Gen Bank accession no EF489024). Phylogenetic analysis of 16S rDNA showed the closest relationship with AY phytoplasma (GenBank accession no. AY389822 and AY389826), indicating that lettuce proliferation-K is a member of AY. Phytoplasma bodies were detected in phloem sieve tubes of diseased lettuce by transmission electron microscopy. The structures had round or pleomorphic shapes with a diameter of 130-300nm. Phylogenetic analysis of 16S rRNA gene, microscopic observation of phytoplasma bodies and symptomatology indicated that lettuce proliferation-K is caused by phytoplasma in the AY group. This is the first report of phytoplasma disease in lettuce in Korea.

In order to develop chili pepper bacterial spot resistant cultivars and near-isogenic lines (NILs) to prompt the molecular mapping of the resistance gene, we have run backcross breeding program since 1994. Two resistance genes against Xanthomonas axonopodis pv. vesicatoria Bs2 from Fla. XVR 3-25 and Bs3 from our breeding line 25-11-3-2, were introduced into a land race, Chilseongcho (abbreviated to Chilseong hereafter) with good fruit guality. We report here the testing of $BC_4F_3\;to\;BC_4F_5$. We found that $BC_4F_5$ lines of the crosses were homozygous with respect to the respective genes of introduction. The lines, in which Bs2 gene was introduced, were hypersensitively resistant to both race 1 and race 3 of X. axonopodis pv. vesicatoria, whereas, those in which Bs3 was introduced were resistant to race 1.

Cultivation of a biocontrol agent on a certain medium often results in reduced biocontrol efficacy and alters physiological state. In our previous study, Burkholderia gladioli strain B543 with long-term subculture on tryptic soy agar resulted in significantly reduced biocontrol ability against cucumber damping-off caused by P. ultimum. Therefore, we investigated the influence of laboratory culturing media on biocontrol activity and physiological state of Burkholderia gladioli strain B543 by using long-term repeated culture on a certain medium. When isolate B543 were successionally cultured on King's B agar (KBA), tryptic soy agar, nutrient agar (NA), or soil extract agar more than 20 times, the isolate cultured on KBA or NA showed a significantly enhanced biocontrol efficacy and higher population density in the rhizosphere of cucumber compared to that of the others. However, the isolates cultured on KBA more than 20 times showed the lowest production of protease, siderophore, or antifungal substance(s), measured by skim milk agar, Chrome-Azurol-S agar, and potato dextrose agar amended with 10% of the culture filtrate, respectively. Our results suggest that adaptation to proper culturing medium can alter biocontrol ability and physiological state, and we must consider laboratory media in optimizing the use of biocontrol agents.

In a trial to select suitable fungicides for developing a spray program that can control apple white rot effectively, after-infection activities in some protective fungicides were detected. Six fungicides, mancozeb, propineb, benomyl, folpet, azoxystrobin and iminoctadine-triacetate, which had been extensively used in apple orchards, were sprayed on 12-year-old apple trees (cv. Fuji) at 15-day intervals from late May to late July. Disease incidences and infection frequencies of the fruit bagged just before and soon after each spray were examined. When the infection frequency or disease incidence of the fruit bagged after each spraying of fungicide was significantly lower than those of the fruit bagged before spraying, the fungicides appeared to confer after-infection activity. The six fungicides showed diverse activities on white rot: folpet showed after-infection activity on disease development, iminoctadine-triacetate showed after-infection activity on infection, azoxystrobin showed after-infection activity on disease development and infection, and mancozeb, propineb and benomyl showed no distinct activity. The activity of a fungicide became much higher when it was sprayed alternately with other fungicide rather than successive spraying of the same fungicide. Analysis of the properties of these protective fungicides could lead to the development of a highly effective spray program against white rot.

Seedlot disinfection techniques to control mung bean sprout rot caused by Colletoricum acutatum and C. gloeosporioides were evaluated for commercial production scheme. Soaking seedlots in propolis (100 X) and ethanol (20% for 30 min) appeared promising with control values of 85.5 and 80.8 respectively, but still resulted in up to 20% rot incidence. None of the C. acutatum conidia survived through hot water immersion treatment (HWT) for 10 min at temperatures of 55, 60 and $65^{\circ}C$, whereas the effective range of the dry heat treatment (DHT) was $60-65^{\circ}C$. Tolerance of mung bean seedlot, as estimated by hypocotyl elongation and root growth, was lower for HWT than for DHT. Germination and growth of sprouts were excellent over the range of $55-65^{\circ}C\;at\;5^{\circ}C$ intervals, except for HWT at $65^{\circ}C$ for 5 min. At this marginal condition, heat damage appeared so that approximately 2% of seeds failed to sprout to normal germling and retarded sprouts were less than 5% with coarse wrinkled hypocotyls. These results suggested that DHT would be more feasible to disinfect mung bean seedlots for commercial sprout production. Heat treatment at above ranges was highly effective in eliminating the epiphytic bacterial strains associated with marketed sprout rot samples. HWT of seedlot at 55 and $60^{\circ}C$ for 5 min resulted in successful control of mung bean sprout rot incidence with marketable sprout quality. DHT at 60 and $65^{\circ}C$ for 30 min also gave good results through the small-scale sprouting system. Therefore, we optimized DHT scheme at 60 and $65^{\circ}C$ for 30 min, considering the practical value of seedlot disinfection with high precision and accuracy. This was further proved to be a feasible and reliable method against anthracnose incidence and those bacterial strains associated with marketed sprout rot samples as well, through factory scale mung bean sprout production system.

The biocontrol agent, Serratia plymuthica A21-4, has been developed for controlling pepper Phytophthora blight. Serratia plymuthica A21-4 strongly inhibits the mycelial growth, zoospore formation, and cyst germination of Phytophthora capsici in vitro. The application of a cell suspension of strain A21-4 to pepper plants in pot experiments and in greenhouse successfully controlled the disease. The bacteria produced a potent antifungal substance which was a key factor in the suppression of Phytophthora capsici. The most active chemical com-pound was isolated and purified by antifungal activity-guided fractionation. The chemical structure was identified as a chlorinated macrolide $(C_{23}H_{31}O_8Cl)$ by spectroscopic (UV, IR, MS, and NMR) data, and was named macrocyclic lactone A21-4. The active compound significantly inhibited the formation of zoosporangia and zoospore and germination of cyst of P. capsici at concentrations lower than $0.0625{\mu}g/ml$. The effective concentrations of the macrocyclic lactone A21-4 for $ED_{50}$ of mycelial growth inhibition were $0.25{\mu}g/ml,\;0.25{\mu}g/ml,\;0.30{\mu}g/ml \;and\;0.75{\mu}g/ml$ against P. capsici, Pythium ultimum, Sclerotinia sclerotiorum and Botrytis cinerea, respectively.

The anthracnose disease on pepper fruits in Korea was caused by Colletotrichum acutatum as well as C. gloeosporioides. Since C. acutatum showed less sensitivity to benomyl, it was analyzed whether the less sensitivity was given by the same mechanism for the fungicide resistance of C. gloeosporioides. The isolates of C. acutatum were less sensitive to the three benzimidazole fungicides tested, benomyl, carbendazim, and thiophanate-methyl. However, the of C. acutatum isolates were different from the resistant isolates of C. gloeosporioides in their response to diethofencarb, one of N-phenyl-carbamates; the former was still less sensitive to diethofencarb than the latter. The differences in the resistance mechanisms in two species were conspicuous in sequence analysis of the tub2 genes. The genes from C. acutatum did not show any non-synonymous base substitutions at the regions known to be correlated with the benzimidazole-resistance. All of these data may indicate that the less sensitivity of C. acutatum to benomyl is based on different mechanism(s) from that of C. gloeosporioides.

Typical susceptible symptoms of the bacterial pustule disease caused by Xanthomonas axonopodis pv. glycines are pustule and chlorotic haloes that usually occur in leaves of Glycine max plants. The soybean genotype 'PI96188' showed an atypical response to all tested races X. axonopodis pv. glycines, accompanied with necrosis without chlorotic haloes on the underside of the necrotic symptoms. X. axonopodis pv. glycines 8ra grew to levels from 10 to 100 fold lower on PI96188 than on susceptible cultivar Jinjoo1, but 10-fold higher than on the resistant cultivar CNS. The chlorophyll content in PI96188 leaves remained unchanged until 12 days after bacterial infection. Ultrastructural observation showed that the infected leaf cells of PI96188 had intact normal chloroplasts compared to those of the susceptible cultivar Jinjoo1. Chloroplast degradation or the absence of chloroplasts in cells of the infected tissues was observed in Jinjoo1. Senescence-related ACS7 gene was significantly induced in PI96188 compared to those in Jinjoo1 at 2 days after inoculation. While photosynthesis-related rbcS gene showed the dramatic change in Jinjoo1, this gene was constitutively expressed in PI96188. However, expression of the defense-related genes, such as peroxidase and isoflavone synthase in the infected PI96188 leaves was similar to that in Jinjoo1. Together, these results suggest that the novel necrotic symptom in PI96188 is a kind of resistant response different from a typical hypersensitive response in the resistant genotypes.

Infection structures were observed using a fluorescence microscope at the penetration sites on the leaves of potato plants pre-inoculated with the bacterial strains Pseudomonas putida TRL2-3, Micrococcus luteus TRK2-2, and Flexibacteraceae bacterium MRL412, which mediated an induced systemic resistance on potato plants against late blight disease caused by Phytophthora infestans. In order to compare the infection structures on the leaves expressing systemic acquired resistance, the leaves of potato plants pre-treated with DL-3-amino butyric acid (BABA) were also observed after challenge inoculation with the same pathogen. The infection structures were investigated. The total number of germination and appressorium formation of P. infestans were counted. Furthermore, the frequencies of fluorescent epidermal cells at the penetration sites, which indicate a defense response of plant cell, were estimated. There were no differences on the germination rates of the fungal cysts among the untreated control, BABA pre-treated, and bacterial strains pre-inoculated plants. However, appressorium formation was slightly decreased on the leaves of BABA pre-treated plants compared to those of untreated as well as bacterial strains pre-inoculated plants. Furthermore, the frequencies of fluorescent cells of BABA pre-treated and bacterial strains pre-inoculated were higher than that of untreated plants, indicating an active defense reaction of the host cells against the fungal attack. On the other hand, the pre-treatment with BABA caused a stronger fluorescent of epidermal cells at the penetration sites compared to the pre-inoculation with the bacterial strains. Interestingly, the frequency of fluorescent cells by BABA, however, was lower than that by the bacterial strains. Based on the results it is suggested that the infection structures showing resistance reaction on the leaves of potato plants were different between by pre-inoculation with bacterial strains and by pre-treatment with BABA against the late blight pathogen.

Dactylella pseudoclavata that captures nematodes in adhesive networks was isolated from nematode-infested strawberry plants from Andong, Korea. It produces obclavate, 0-1 septate conidia, $30-40{\mu}m$ long and $8-11{\mu}m$ wide, with round distal ends and bases shaped like bottle-necks. The conidiophores were simple, occasionally branched, $150-300{\mu}m$ long, producing 1-4 conidia at the apex. Chlamydospores were abundant, intercalary or catenulate, yellowish to brown, globate or subglobate, wart on the surface, $30-35{\times}25-30{\mu}m$ in size. This is the first report of Dactylella pseudoclavata in Korea.

In 2005 and 2007, a basal stem rot of Disporum smilacinum caused by Sclerotium rolfsii occurred sporadically in a herb farm at Hamyang, Korea. The symptom initiated with water-soaking lesion and progressed into stem rot and wilt of a whole plant. Severely infected plants were blighted and died eventually. White mycelial mats appeared on the lesion at early stage and a number of sclerotia were formed on the stem near the soil line. The sclerotia were globoid in shape, 1-3 mm in size and white to brown in color. The optimum temperature for the growth and sclerotia formation was 30 on PDA and the hyphal width was measured $3-8{\mu}m$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. On the basis of symptom, mycological characteristics and pathogenicity to the host plant, this fungus was identified as Sclerotium rolfsii Saccardo. This is the first report on the stem rot of D. smilacinum caused by S. rolfsii in Korea.

The occurrence and relative incidence of viruses including Cucumber mosaic virus (CMV), Zucchini yellow mosaic virus (ZYMV), Papaya rings pot virus (PRSV), and Watermelon mosaic virus (WMV), Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), and Melon necrotic spot virus (MNSV) were surveyed from main melon (Cucumis melo L.) production areas in Jeonnam province during 2000-2002. Virus disease incidences of melon cultivating fields were 0% and 11% in spring and fall 2000; 40%, 2.1%, and 8.8% in spring, summer, and fall 2001; and 6.3 % in spring 2002 in main cultivated areas in Jeonnam province, respectively. Field disease incidences of melon virus infections were 0% and 18.8% in spring and fall 2000; 50%, 38.5%, and 82.6% in spring, summer, and fall 2001; and 47.4% in spring 2002, respectively. Total of 101 melon samples showing typical disease symptoms were collected from 2000 to 2002 and tested for virus infection by RT-PCR. Potyvirus-specific DNA fragments for WMV, ZYMV, and PRSV were amplified from 46, 5, and 4 samples, respectively. MNSV specific DNA fragment was amplified from 18 samples. CMV-specific DNA fragment was detected from only 3 samples.

A total of the 261 Phytophthora infestans isolates collected from 2003 to 2005 in Korea were investigated for their physiological race composition. Among the isolates, we detected 18 physiological races and the dominant races were R0.1.3.5.6.10.11 and R0.1.3.5.6.7.10.11 with frequencies of 18.4% and 11.4%, respectively. All of the P. infestans races carried multiple virulence genes and showed virulence to the potato resistance genes R1, R3, R5, R6, R7, R10 and R11, but not to R8 and R9. Therefore, it is likely that the physiological races of P. infestans were diverse in Korea.

Many plant growth-promoting rhizobacteria (PGPRs) have been known for beneficial effects on plants including biological control of soilborne pathogens, induced systemic resistance to plant pathogens, phytohormone production, and improvement of nutrient and water uptake of plants. We developed a simple and rapid method for screening potential PGPR, especially phytohormone producing rhizobacteria, or for analyzing their functions in plant growth using cucumber seedling cuttings. Surface-sterilized cucumber seeds were grown in a plastic pot containing steamed vermiculite. After 7 days of cultivation, the upper part 2 cm in length of cucumber seedling, was cut and used as cucumber cuttings. The base of cutting stem was then dipped in a microcentrifuge tube containing 1.5ml of a bacterial suspension and incubated at $25^{\circ}C$ with a fluorescent light for 10 days. Number and length of developed adventitious roots from cucumber cuttings were examined. The seedling cuttings showed various responses to the isolates tested. Some isolates resulted in withering at the day of examination or in reduced number of roots developed. Several isolates stimulated initial development of adventitious roots showing more adventitious root hair number than that of untreated cuttings, while some isolate had more adventitious root hair number and longer adventitious roots than that of untreated control. Similar results were obtained from the trial with rose cuttings. Our results suggest that this bioassay method may provide a useful way for differentiating PGPR's functions involved in the development of root system.