A half-sib family of two 4-year-old seedlings of Pinus $\times$ rigitaeda was inoculated with each of 20 Pinus thunbergii isolates of Fusarium circinatum (syn. Fusarium subglutinans f. sp. pini) from two pitch canker damaged sites in Jeju Island, South Korea. Initial symptoms of needle damages were visible on most of the seedlings at 18 days after inoculation. The 20 tested isolates were not significantly different in virulence, based on lesion lengths at the site of inoculation (P = 0.217). The most virulent isolate FT-7 showed the longest lesion length. Some seedlings began to die 46 days after inoculation. All seedlings were dead by 68 days after inoculation except two seedlings inoculated with each of isolates FS-2 and FS-13, respectively. Using the FT-7, 38-year-old 11 P. $\times$ rigitaeda trees, which were survived from a seed orchard severely damaged by pitch canker, were inoculated on branches in the seed orchard in Jeju Island to assess differences in susceptibility to pitch canker. The 11 trees differed significantly (P < 0.001) in susceptibility to F. circinatum based on average lesion lengths measured 56 days after inoculation. It is possible that induced resistance contributed to their capacity to limit lesion development. The susceptibility of natural selection P. $\times$ rigitaeda trees are more likely affected by interaction with F. circinatum rather than environmental conditions.

The pathotypes of Cercospora beticola, causal agent of sugar beet leaf spot disease, were identified by application of pathogenicity test using 100 isolates obtained from the provinces with intensive sugar beet cultivation. For the identification of pathotypes, five sugar beet cultivars were used each with different resistance factors. Cultivar reactions were determined by inoculation of cultivars with the isolates under controlled conditions and measuring disease severity on the $15^{th}$ day according to the 1-9 KWS Scale. Based on the reactions of the five cultivars, a total of 15 pathotypes were detected. All employed sugar beet cultivars were resistant to Pathotype no:1 comprising most of the isolates. Genetic diversity of the causal agent was characterized by AFLP reaction. The products acquired at the end of AFLP reaction were detected by means of Beckman CEQ 8800 DNA Capillary Series Analysis and the results obtained were evaluated according to the similarity index UPGMA. For the genetic analysis of C. beticola isolates, 9874 polymorphic fragments of sizes between 100 and 500 bp were analysed which were generated by nine primers. The dendrogram derived from AFLP analysis depicted the existence of five different subgroups. The polymorphism rate among isolates was 91.13% and the dendrogram distribution of the pathotypes obtained by pathogenicity indicated that pathotypes were not discriminated and did not compose any groups.

Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.

We performed diagnostic PCR assays and a phylogenetic analysis using partial sequences of TEF1 (translation elongation factor-1) to determine the trichothecene chemotypes and genetic diversity of F. graminearum isolates from maize and rice samples collected in 2009 in Korea. PCR using a species-specific primer set revealed a total of 324 isolates belonging to the putative F. graminearum species complex. PCR with trichothecene chemotypespecific primers revealed that the nivalenol (NIV) chemotype was predominant among the fungal isolates from rice (95%) in all provinces examined. In contrast, the predominant chemotype among the corn isolates varied according to region. The deoxynivalenol (DON) chemotype was found more frequently (66%) than the NIV chemotype in Gangwon Province, whereas the NIV chemotype (70%) was predominant in Chungbuk Province. Phylogenetic analysis showed that all DON isolates examined were clustered into lineage 7, while the NIV isolates resided within lineage 6 (F. asiaticum). Compared with previous studies, the lineage 6 isolates in rice have been predominantly maintained in southern provinces, while the dominance of lineage 7 in maize has been evident in Gangwon at a slightly reduced level.

A tobamovirus, Ribgrass mosaic virus (RMV), was identified newly from chinese cabbage (Brassica campestris L. pekinensis) in Korea. Virus disease incidence of RMV on chinese cabbage was 37.9% in alpine area on August in 1993. RMV induced the symptoms of necrotic ring spots, necrotic streak on midrib and malformation. RMV, Ca1 and Ca3 isolate, could infect 35 species out of 45 plants including Chenopodium amaranticolor. Physical properties of RMV Ca1 isolate were very stable as 10.8 over for dilution end point, $95^{\circ}C$ for temperature inactivation point and 18 weeks for longevity in vitro. RMV had the soil transmission rate of 75.0% for the chinese cabbages, 'Chunhawang' and 'Seoul' cultivars. The purified virions of RMV had the typical ultraviolet absorption spectrum of maximum at 260 nm and minimum at 247 nm. RMV of Ca1 isolate was related serologically with antisera of Tobacco mosaic virus (TMV)-Cym, TMV-O and Pepper mottle virus, but not related with antiserum of Odontoglossum ring spot virus. coat protein gene of RMV-Ca1, sized 473 nucleotides, encoded 158 amino acid residues. Nucleotide identity of RMV-Ca1 CP gene was 96.4% with RMV-Shanghai (GenBank accession No. of AF185272) from China and 96.0% with RMV-Impatiens (GenBank accession No. of AM040974) from Germany. Identity of amino acids between RMV-Ca1 and the two RMV isolates was 96.8%. Specific three primers were selected for rapid and easy genetic detection of RMV using Virion Captured (VC)/RT-PCR method.

Two similar microbial fungicides (termed as MA and MB) developed in a Korean biopesticide company were analyzed and compared each other in their biocontrol activities against the phytophthora blight of chili pepper caused by Phytophthora capsici. MA and MB contained the microbe Paenibacillus polymyxa and Bacillus subtilis, respectively, with concentrations over those posted on the microbial products. In comparison of the isolated microbes (termed as MAP from MA and MBB from MB) in the antagonistic activities against P. capsici was effective, prominently against zoospore germination, while MBB only significantly inhibited the mycelia growth of the pathogen. Some effectiveness of MAP and MBB was noted in the inhibition of zoosporangium formation and zoospore release from zoosporangia; however, no such large difference between MAP and MBB was noted. In a pot experiment, MA reduced the severity of the phytophthora blight more than MB, suggesting that the disease control efficacy would be more attributable to the inhibition of zoospore germination than mycelia growth of P. capsici. These results also suggest that the similar microbes MA and MB targeting different points in the life cycle of the pathogen differ in the disease control efficacies. Therefore, to develop microbial fungicides it is required to examine the targeting points in the pathogen's life cycle as well as the action mode of antagonistic microorganisms.

This study was conducted in Bahar and Lalehjin, Hamadan, Iran to assess the biocontrol efficacy of Coniothyrium minitans Campbell against potato white mold caused by Sclerotinia sclerotiorum (Lib.) de Bary under field and greenhouse conditions. In addition, the resistance of common potato cultivars against S. sclerotiorum was determined in a greenhouse experiment. After straw inoculation of six potato cultivars (Pashandi, Istambouli, Agria, Marfauna, Alpha and Spartaan) with S. sclerotiorum, the least disease severity was observed in Spartaan and Marfauna. Agria showed the most susceptibility to S. sclerotiorum. Compared with the healthy control, different concentrations of C. minitans conidia ($10^7$, $10^8$ and $10^9$ conidia/mL) reduced disease severity under greenhouse condition, and a concentration $10^9$ was the most effective treatment. During 2008 and 2009, four field trials were conducted to evaluate the efficacy of C. minitans in different soil and aerial applications on disease incidence of potato white mold. In 2008, soil application of $Contans^{(R)}$ WG (a commercial product of C. minitans) showed the greatest biocontrol capacity whereas soil application of solid-substrate C. minitans was found inferior when compared with other treatments in both Bahar and Lalehjin field sites. In 2009, benomyl application was the most effective treatment in reducing disease incidence in both tested field sites.

In order to control postharvest spoilage of satsuma mandarin fruits, rhizobacteria were isolated from soil samples. The Pantoea agglomerans strain 59-4 (Pa 59-4) which suppresses the decay of mandarin fruit by green and blue mold, was tested for the control efficacy and its mode of action was investigated. Pa 59-4 inhibited infection by green and blue mold on wounded mandarins, which were artificially inoculated with a spore suspension of Penicillium digitatum and P. italicum with control efficacies of 85-90% and 75-80%, respectively. The biocontrol efficacy was increased by raising the concentration of cells to between $10^8$ and $10^9\;cfu/ml$, and pretreatment with the antagonist prevented subsequent infection by green mold. The population of Pa 59-4 was increased more than 10 fold during the 24 hr incubation at $20^{\circ}C$, indicating that colonization of the wound site might prevent the infection by green mold. Despite poor antifungal activity, the Pa 59-4 isolate completely inhibited the germination and growth of P. digitatum spores at $1{\times}10^8\;cfu/ml$. We argue that the control efficacy was mediated by nutrient competition. Overall, the effective rhizobacterium, Pa 59-4, was shown to be a promising biocontrol agent for the postharvest spoilage of mandarin fruits by green and blue mold.

In the course of a searhing environmental friendly antifungal compounds, we found that mixture of methyl esters of fatty acids obtained from soybean oil had potent control efficacy against barley powdery mildew (Blumeria graminis f. sp. hordei). In this study, ten alkyl fatty acid esters (AFAEs) were tested for in vivo antifungal activity against five plant diseases such as rice blast, rice sheath blight, tomato gray mold, tomato late blight and barley powdery mildew. Some AFAEs showed the most control efficacy against barley powdery mildew among the tested plant diseases. By 5-hr protective and 1-day curative applications, six AFAEs ($3,000\;{\mu}g/ml$), including methyl and ethyl palmitates, methyl and ethyl oleates, methyl linoleate, and methyl linolenate demonstrated both curative and protective activities against barley powdery mildew. In contrary, methyl laurate strongly controlled the development of powdery mildew on barley plants by curative treatment at a concentration of $333\;{\mu}g/ml$, but did not show protective activity even at $3,000\;{\mu}g/ml$. Under greenhouse conditions, the seven AFAEs ($1,000\;{\mu}g/ml$) except for methyl and ethyl stearates, and methyl caprylate also effectively controlled cucumber powdery mildew caused by Podosphaera xanthii. Among them, methyl and ethyl palmitates ($333\;{\mu}g/ml$) represented the most control activity of more than 68% against the disease. The results are the first report on the antifungal activity of methyl and ethyl esters of fatty acids against plant pathogenic fungi.

In this study, we evaluated the efficacy of fluopicolide to inhibit Phytophthora capsici in vitro, and to control pepper Phytophthora blight in a greenhouse and pepper fields. Fluopicolide was tested on various developmental stages of P. capsici 06-143 (a sensitive isolate to metalaxyl) and JHAW1-2 (a resistant isolate to metalaxyl). Mycelial growth and zoosporangium germination of both isolates were completely inhibited at $4.0\;{\mu}g/ml$ of the fungicide in vitro. The $EC_{50}$ (effective concentrations reducing 50%) of P. capsici 06-143 against zoospore were $0.219\;{\mu}g/ml$, while those of JHAW1-2 were $3.829\;{\mu}g/ml$. When fluopicolide was applied at 100 and $1,000\;{\mu}g/ml$ 7 days before inoculation with P. capsici 06-143 in the greenhouse test, the disease was controlled completely until 6 days after inoculation. However, the curative effect of fluopicolide was not as much as the protective effect. When fluopicolide was applied by both soil drenching and foliar spraying, the treatments strongly protected pepper against the Phytophthora blight disease. Based on these results, fluopicolide can be a promising candidate for a fungicide to control P. capsici in the pepper fields.

Macrophomina phaseolina causing charcoal rot was isolated from sesame seeds (cvs. Giza 32 and Shandawel-3) collected from different localities of Assiut, Sohage and El-Minia Governorates. The fungus was found in the highest frequently in samples collected from Assiut Governorate followed by Sohag and finally EL Minia Governorate. The obtained isolates were different in their virulence on the tested sesame cvs. Also, they differed in their growth nature including colony color and sclerotial production. The color of colonies of the pathogen seem to be correlated with density of sclerotial formation. Aqueous extracts of Majorna, Wild chamomile, Geranium oil and Nees plants were highly toxic to tested isolates of M. phaseolina, in vitro. On the other hand the rest of the tested aqueous extracts had no effect. Under greenhouse conditions in 2005 and 2006 seasons, soaking seeds of sesame before sowing in aqueous extracts of Eucalyptus, Nerium, Ocimum and Roesmary plants decreased the disease incidence. Aqueous extracts of Eucalyptus and Ocimum were the most effective treatment. Dipping sesame seeds in hot water at $60^{\circ}C$ for 5 minutes increased seed germination of Giza 32 and Shandawel-3 cvs. followed by $55^{\circ}C$, $50^{\circ}C$ and $45^{\circ}C$, while $40^{\circ}C$ treatment resulted the lowest seed germination rate. Dipping sesame seeds in hot water at different temperature before planting decreased seed, seedling and charcoal rots. Soaking seeds in hot water at $60^{\circ}C$ increased greatly plant height and decreased seed, seedling rot and charcoal rot followed by $55^{\circ}C$ and $50^{\circ}C$, under greenhouse condition.

Suppression subtractive hybridization was used to isolate wound-induced genes from soybean. One of the wound-induced genes, gmwi143 designated as GmCCR, showed high homology with genes encoding cinnamoyl-CoA reductase (CCR; EC 1.2.1.44). Deduced amino acid sequences encoded by GmCCR showed the highest identity (77%) with those of Acacia CCR. There are 2 CCR genes highly homologous to GmCCR in soybean genome based on Phytozome DB analysis. RNA expression of GmCCR was specifically induced by local and systemic wounding, drought, high salinity or by ultraviolet stress. Our study suggests that GmCCR may be involved in resistance mechanism during abiotic stresses in plants.

In order to study disease resistance mechanisms in rice against the rice blast fungus Magnaporthe grisea, we screened fungal elicitor-responsive genes from rice suspension-cultured cells treated with fungal elicitors employing differential hybridization (DH). By DH screening, 31 distinct rice clones were isolated and a majority of them were full-length cDNAs encoding pathogenesisrelated (PR) genes. Sixteen of the 31 genes were upregulated at 4, 8, and 12 h following fungal elicitor treatment. To elucidate the effect of signal molecules and biotic elicitors on the regulation of rice defense genes, we further characterized the transcriptional expression patterns of representative isolated PR genes; OsGlu1, OsGlu2, OsTLP, OsRLK, and OsPR-10, following treatment with fungal elicitor, phytohormones, cycloheximide, and inhibitors of protein phosphorylation. Jasmonic acid (JA) induced transcriptional expression of OsGlu1, OsTLP, and OsRLK, but not of OsGlu2 and OsPR-10 at any of the tested time points. Salicylic acid (SA) and abscisic acid weakly induced the expression of OsTLP and OsRLK. SA showed an antagonistic effect with fungal elicitor and JA. Cycloheximide suppressed all these genes upon elicitor treatment, except for OsGlu2. Staurosporine only induced the expression of OsRLK. Application of calyculin A strongly induced OsRLK expression, but suppressed the expression of OsGlu2. Our study yielded a number of PR genes that play a role in defense mechanisms against the rice blast fungus, as well as contribute towards the elucidation of crosstalk between phytohormones and other modifications during defense signaling.

Resin flow, symptom development, and lignin biosynthesis in response to wounding and fungal inoculation were investigated in Pinus rigida and Pinus densiflora. The two-year-old seedling stems were subjected to three types of treatments: (i) wounding without inoculation, (ii) wound-inoculation with a conidial suspension of Fusarium circinatum, and (iii) pre-wounding woundinoculation with the fungus 20 days after the initial wounding. Resin flow from wounding sites was more evident in P. rigida than P. densiflora in all treatments. The wound-inoculation with the fungus induced almost two-fold higher levels of resin flow than the other treatments in both species. The pre-wounding woundinoculation appeared to result in a decrease in pitch canker development in the two pine species. Some reductions in disease severity were observed in the prewounding wound-inoculated P. rigida, showing a mean disease severity of less than 85%, compared with approximately 100% in the wound-inoculated stems. Disease severity was approximately 50% in the woundinoculated P. densiflora, whereas 10% in the pre-wounding wound-inoculated stems. Higher amounts of lignin were found from bark (ca. 40%) than from xylem (ca. 30%). The wound-inoculated bark and the pre-wounding wound-inoculated bark exhibited higher amounts of lignin among the other treatments. These results suggest that the wound-inoculation apparently prompt the increase in resin flow and lignin biosynthesis from the two pine species, and the prior wounding may be involved in decreased disease severity against the further invasion of F. circinatum.

Plant matrix metalloproteases (MMPs) are a family of apoplastic metalloproteases closely related to human matrilysins. Up-regulation of Nicotiana benthamiana matrix metalloprotease 1 (NMMP1) expression by treatment with pathogens, ethephon and aging indicates that the gene is related to plant defense and the aging process through ethylene signaling. NMMP1 expression was higher than in normal growth leaves following infection with an incompatible pathogen Pseudomonas syringae pv. tomato T1 or a compatible pathogen P. syringae pv. tabaci and in aged leaves. Transient overexpression of NMMP1 in N. benthamiana leaves lowered the growth of P. syringae pv. tabaci. However, NMMP1-silenced leaves showed increased growth of P. syringae pv. tabaci. These data strongly suggest that NMMP1 in N. benthamiana is a defense related gene, which is positively regulated by ethylene.

Fusarium species, a large group of plant pathogens, potentially pose quarantine concerns worldwide. Here, we focus on the development of a method for detecting four Fusarium species in quarantined plants in Korea: F. solani f. sp. cucurbitae, F. stilboides, F. redolens, and F. semitectum var. majus. Species-specific primers were designed from the nucleotide sequences of either the translation elongation factor-1 alpha (TEF1) gene or RNA polymerase II subunit (RPB2) gene. Two different primer sets derived from TEF1, all specific to F. solani f. sp. cucurbitae, were able to differentiate the two races (1 and 2) of this species. A set of nested primers for each race was designed to confirm the PCR results. Similarly, two primer sets derived from RPB2 successfully amplified specific fragments from five F. stilboides isolates grouped within a single phylogenetic clade. A specific TEF1 primer set amplified a DNA fragment from only four of the 12 F. redolens strains examined, which were grouped within a single phylogenetic clade. All of the F. semitectum var. majus isolates could be specifically detected with a single RPB2 primer set. The specificity of the primer sets developed here was confirmed using a total of 130 Fusarium isolates.

The expression of HR-like lesion inducing gene of Oryza sativa (OsHRL) was slightly increased by Xanthomonas oryzae pv. oryzae (Xoo) infection. Transgenic rice plants over-expressing OsHRL gene were challenged with Xoo and the development of disease symptoms were examined to investigate the effect of OsHRL gene expression on plant defense responses. The over-expression of OsHRL increased disease resistance against Xoo compared with wild type plants.

New sources of barley (Hordeum vulgare L.) resistant to spot blotch, caused by Cochliobolus sativus, are needed to provide effective resistance because of the rapid change pathotype patterns of C. sativus in fields. The purposes of our study were to develop a method to screen barley for resistance to spot blotch disease and then use this methodology to screen barley genotypes for resistance to the major virulent pathotype Pt4 in barley populations in Syria. A transparent tape method, in which a conidial suspension of C. sativus was dropped onto transparent tape and placed, treated-side down, on the second leaf surface of barley plants. Disease symptoms of fungus were easily detected on the leaves covered by the transparent tape after 48h of inoculation. The transparent tape method was repeatable and the disease scores obtained were correlated (r = 0.91, P = 0.001) with those obtained by the seedling assay. This method may be beneficial in various plant pathology breeding programs.