Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.

The objectives of this study were to identify the aecial host ranges of four Korean Gymnosporangium species, G. asiaticum, G. cornutum, Gjaponicum and Gyamadae, and to verify the morphological characteristics of telia as diagnostic keys to the species. Thirteen Korean Rosaceous woody species were artificially inoculated with teliospores obtained from Juniperus species. There was high specificity between telial and aecial hosts and the fungal species, providing the first experimental proof on host alternation of these rust fungi in Korea. Telia on the witches' broom and on the small galls were identified as new telial characteristics in G asiaticum and in G. yamadae, respectively. Aecial hosts of G. asiaticum and G. yamadae showed varying responses in their susceptibility and in the days required for formation and duration of spermogonia and aecia after inoculation. Four telial host species in Juniperus were confirmed for the first time in Korea, which include J. chinensis var. kaizuka, J. chinensis var. horizontalis and J. chinensis var. globosa for G. asiaticum; and J. chinensis var. kaizuka for G. yamadae.

Since 2000 severe rots on aerial and underground parts of paprika (Capsicum annum L.) has occurred in most surveyed glasshouses throughout the country. A total of 56 isolates of a fungus were consistently isolated from various plant parts such as fruit, stem, branch, and root collected from 16 farms in five provinces. Anamorph stage of the fungus was identified as Fusarium solani based on its morphological characteristics. However, the fungus readily produced a sexual structure of perithecia on infected plant tissues and on agar media. Since the fungus formed abundant perithecium by a single isolate, it was considered as a homothallic strain of Nectria haematococca, the teleomorph of F. solani. Irregularly globose perithecia with orange to red color formed sparsely to gregariously on dead tissues of fruits and basal stems at the late infection stage, which is a diagnostic sign for the disease. Perithecia ranged from 125 to 220 ${\mu}m$ in diameter varied among isolates. Asci enveloping eight ascospores were cylindrical and measured 60-80x8-12 ${\mu}m$. Ellipsoid to obovate ascospores are two-celled and measured 11-18x4-7 ${\mu}m$. Ascospores were hyaline, slightly constricted at the central septum, and revealed longitudinal striations that is characteristic of the species. This fungus that has never been reported in Korea has previously become a threat to paprika cultivation because of its strong pathogenicity and nationwide distribution.

Regression models for determining infection periods of apple white rot were developed based on conidial germination and appressorium formation of Botryosphaeria dothidea. A total of 120 apple fruits were inoculated with the fungal conidial suspension and subjected to 6 temperatures and 10 wetness periods. Conidia germinated and produced appressoria, exhibiting swollen tips of germ tubes on the fruit surface. Conidial germination (G) increased with temperature (T) and wetness period (W), and was described as $G=-89.273+7.649T+7.056W-0.109T^{2}-0.085W^{2}-0.066TW(R^{2}=0.75)$. Less than 2 hr of wetness period were enough for conidia to germinate at 25 to $30^{\circ}C$. Effects of temperature and wetness period on appressorium formation (A) could be explained as $A=-1.540-2.375W+0.045W^{2}+0.213TW(R^{2}=0.77)$. The relationship between conidial germination and appressorium formation ($A_g$) was described as$A_g=0.381-0.227G+0.005G^{2}(R^{2}=0.67)$, suggesting that conidial germination may have to reach approximately $43.7\%$ to initiate appressorium formation. Using the regression equation for conidial germination and the criterion of $43.7\%$ conidial germination, an infection model was developed to determine infection periods based on temperature and wetness period. The infection model with the criterion of $43.7\%$ conidial germination was apparently more conservative than the appressorium formation model in determining possibility of apple infection. The infection model seemed sensitive to variable weather conditions, suggesting possible use of the model for timing fungicide sprays to control white rot of apples in practice.

Paenibacillus elgii SD17 (KCTC $10016BP^T$=NBRC $100335^T$) was recently reported as a new species. Based on its inhibitory activity to Thanatephorus cucumeris AG1-1, strain SD17 was further evaluated for its potential as a biocontrol agent against soil-borne diseases of turf grasses in Korea. P. elgii SD17 showed a broad spectrum of antimicrobial activity in vitro test and suppressed development of turf grass diseases; Pythium blight caused by Pythium aphanidermatum and brown patch caused by T. cucumeris AG1-1 on creeping bentgrass (Agrostis palustris) in the growth chamber tests. Under a condition for massive culture in a 5,000 L fermenter, P. elgii SD17 reached $6.4{\times}10^8$ spores/ml that resulted in approximately $1.0{\times}10^7$ cfu/g when formulated into a granule formulation (GR) using the whole culture broth instead of water. Using the GR formulation, biocontrol activity of P. elgii SD17 was confirmed. In the growth chamber tests, the GR formulation was effective against brown patch and Pythium blight with similar level of disease severity compared to each of the standard fungicides at the application rates of 10 g/$m^2$ or above. In the field tests, compared to each untreated control, the GR formulation also effectively controlled Pythium blight, brown patch and large patch at all the application rates of 5, 10 and 20 g/$m^2$, respectively, without significant response by the application rates. However its performance was inferior to each of the standard chemical fungicides. Based on these results, we consider this GR formulation of P. elgii SD17 as an effective biocontol agent to suppress Pythium blight, brown patch and large patch of turf grasses in Korea.

The prototype Chlorella virus PBCV-l encodes 11 tRNA genes and over 350 protein-encoding genes in its 330 kbp genome. Initial attempts to overexpress the recombinant A189/192R protein, a putative virus attachment protein, in E. coli strain BL21(DE3) SI were unsuccessful, and multiple protein bands were detected on Western blots. However, the full-length A189/192R recombinant protein or fragments derived from it were detected when they were expressed in E. coli BL21 CodonPlus (DE3) RIL, which contains extra tRNAs. Codon usage analysis of the a189/192r gene showed highly biased usage of the AGA and AVA codons compared to genes encoded by E. coli and Chlorella. In addition, there were biases of XXA/U($56\%$) and XXG/ C($44\%$) in the codons recognized by the viral tRNAs, which correspond to the codon usage bias in the PBCV-1 genome of XXA/U ($63\%$) over those ending in XXC/G ($37\%$). Analysis of the codon usage in the major capsid protein and DNA polymerase showed preferential usage of codons that can be recognized by the viral tRNAs. The Asn (AAC) and Lys (AAG) codons whose corresponding tRNA genes are duplicated in the tRNA gene cluster were the most abundant (i.e., preferred) codons in these two proteins. The tRNA genes encoded in the PBCV-l genome seem to play a very important role during the synthesis of viral proteins through supplementing the tRNAs that are frequently used in viral proteins, but are rare in the host cells. In addition, these tRNAs would help the virus to adapt to a wide range of hosts by providing tRNAs that are rare in the host cells.

The partial nucleotide sequences of the genomic dsRNA mycoviruses infecting Pleurotus ostreatus (isolates ASI2596, ASI2597, and Bupyungbokhoe) and Agaricus blazei Murrill were determined and compared with those of the other dsRNA mycoviruses. Partial nucleotide sequences of the purified dsRNA from ASI2596 and ASI2597 revealed RNA-dependent RNA polymerase sequences that are closely related to Oyster mushroom isometric virus 2, while nucleotide sequences and the deduced amino acid sequence from dsRNA mycovirus infecting Agaricus blazei did not show any significant homology to the other dsRNA mycoviruses. Specific primers were designed for RT-PCR detection of these dsRNA viruses and were found to specifically detect each dsRNA virus. Northern blot analysis confirmed the homogeneity of RT-PCR products to each purified dsRNA. Altogether, our results suggest that these virus-specific primer sets can be employed for the specific detection of each dsRNA mycovirus in infected mushrooms.

A virus isolate collected from infected paprika (Capsicum annuum var. grossum) was characterized as Potato virus Y (PVY) based on biological, serological, cytopathological, and molecular properties. In host range studies, the paprika isolate produced the mosaic symptom on some tobacco, tomato and pepper (Capsicum annuum). A new paprika isolate also infected potato cultivars which is different biological characteristic compared to the other popular potyvirus infecting paprika, Pepper mottle virus (PepMoV). Previously reported PVY strains, $PVY^o$ and $PVY^N$ did not infect pepper and typical PepMoV isolates did not infect potato. Distinctive inclusion patterns of the scroll, pinwheel, long laminated inclusions, and helper components in the cytoplasm of infected cells were also different to those observed by the typical PVY isolate infections. However, the paprika isolate reacted to the monoclonal antibody of $PVY^N$ strain with high absorbance readings. RT-PCR amplification, cloning, and sequencing of the 3' untranslated region and a part of coat protein gene also added additional evidence of the paprika isolate as the $PVY^N$-related isolate. Multiple alignments as well as cluster dendrograms of PVY-paprika isolate revealed close phylogenetic relationship to the $PVY^N$ subgroup. Altogether, these results suggest that a new PVY isolate infecting paprika contained distinct characteristics compared to the other previously described PVY strains with closer relationship to the $PVY^N$ strain.

Sulfur, an important component of plants, is regulated by a variety of stresses in sulfate assimilation and metabolism. Increase has been observed in the expression of O-acetylserine(thiol)lyase (OASTL) through two-dimensional electrophoresis with the shoot tips of Arabidopsis infected by beet severe curly top geminivirus (BSCTV). With the three- to six-fold increases in the transcript expression of OASTL, serine acetyltransferase (SAT) and $\gamma$-glutmylcysteine synthetase (GSH) were induced over the mock-inoculated organization in each organization through real-time RT-PCR analysis. The expression of those genes might affect the accumulation of anthocyanin in symptomatic tissues and the induction of abnormal callus-like structures formed by additional cell divisions as typical disease symptoms of BSCTV-infected Arabidopsis. This is the first report to describe the collaborative induction of OASTL, SAT, and GSH in virus-infected plants. The changed expressions of OASTL, SAT, and GSH in Arabidopsis infected with BSCTV raises new aspects regarding the biological function of symptomatic tissues related to sulfate metabolism.

A Korean isolate of Pepper mild mottle virus (PMMoV-Kr) was isolated from a diseased hot pepper plant and its biological and molecular properties were compared to that of PMMoV-J and PMMo V -So The genomic RNA of PMMoV-Kr consists of 6,356 nucleotides. The nucleotide and amino acid sequences identities of four viral proteins and two noncoding regions among PMMoV-Kr, PMMoV-S and PMMoV-J were $96.9\%\;to\;100.0\%\;and\;97.5\%\;to\;98.6\%$, respectively. Full-length cDNA amplicon of PMMoV-Kr was directly amplified by RT-PCR with a set of 5'-end primer anchoring T7 RNA promoter sequence and 3'-end virus-specific primer. Capped transcript RNAs from the full-length cDNA clone were highly infectious and caused characteristic symptoms of wild type PMMoV when mechanically inoculated to systemic host plants such as Nicotiana benthamiana and pepper plants.

Turnip mosaic virus (TuMV) is an infectious viral pathogen on the cruciferous crops, predominantly Chinese cabbage (Brassica campestris subsp. pekinensis) and radish (Raphanus sativus). On the basis of the symptom development in selective differential hosts from indicator host species, Chinese cabbage and Korean radish inbred lines, the representative eight isolates of TuMV were divided into two major groups/or six types. Group I includes Th 1, Ca-ad7, and Cj-ca2-1 isolates, while group II includes the other isolates (rg-pfl, r 9-10, Rhcql-2, Stock and Mustard). According to the molecular phylogenetic analysis, these isolates, however, divided into two groups and two independent isolates. Phylogenetic analysis indicated that four isolates (Tu 1, r9-10, Stock and Rh-cql-2) formed a distinct phylogenetic group, and the other two isolates (Ca-ad7 and Cj-ca2-1) also formed another group. Mustard and rg-pfl isolates did not seem to have any relationship with these two groups. Taken together, these results indicated that virulence differentiation on host plants, molecular phylogenetic analysis of the nucleotide and the deduced amino acid of TuMV coat proteins did not show any relationship. The multi-resistant lines, Wonyae 20026 and BP058 in Chinese cabbage represent valuable genetic materials that can be used for crucifer breeding programs on TuMV resistance, but not in Korean radish.

Infection rate of Chrysanthemum stunt viroid (CSVd) in 64 commercial chrysanthemum cultivars cultivated in Korea ranged from 9.7 to 66.8$\%$. Symptoms on leaves of CSVd-infected chrysanthemum included yellow spots, chlorosis, vein clearing, vein bending and crumpling. CSVd induced flower malformation in 'Scot', color change in 'Sharotte', and color breaking in 'Sharon'. CSVd caused reduction of plant height, leaf size, flower size and the flowers number by $32-50\%,\;26-35\%$, $14-36\%\;and\;14-75\%$, respectively. In conclusion, CSVd affected plant height, leaf size and flower quality in chrysanthemum plants.

In 2003 typical phytoplasma symptoms of witches' broom and flower malformation were observed on statice (Limonium sinuatum) plants grown at commercial greenhouses in Busan, South Korea. The DNA extracted from the infected leaves was amplified using universal primer pair of Pl/P6 derived from conserved 16S rRNA gene of Mollicutes giving the expected Polymerase chain reaction (PCR) product of 1.5 kb. In the nested PCR assays, the expected DNA fragment of 1.1 kb was amplified with the specific primer pair 16Fl/Rl that was designed on the basis of aster yellows (AY) phytoplasma 16S rDNA sequences. The 1.1 kb PCR products were cloned and nucleotide sequences were determined. The sequences were identical to that of Onion yellows OY phytoplasma (GenBank accession no. D12569) isolated from Onion in Japan. Electron microscopy of thin sections of leaf veins showed phytoplasma bodies in the phloem. Statice witches' broom symptom occurred on statice in commercial greenhouses in Korea was confirmed as infection of AY phytoplasma by transmission electron microscopy observation, and by determination of 16S rRNA gene sequences of phytoplasma.

A survey was conducted during February-March 2003 to determine the occurrence and population density of plant-parasitic nematodes in strawberry (Fragaria ananassa) fields, cultivated under plastic houses in major strawberry growing regions of Gyeongbuk and Chonbuk provinces, Korea. The survey revealed presence of eleven species of plant-parasitic nematodes viz., Aphelenchoides fragariae, Criconemoides morgensis, Ditylenchus dipsaci, Helicotylenchus dihystera, Hirschmanniella imamuri, Meloidogyne arenaria, M. incognita, Pratylenchus penetrans, Psilenchus hilarulus, Tylenchorhynchus claytoni and Xiphinema sp. Frequency and density of each species were highly variable from field to field and within the field. Plant growth was not uniform in the surveyed fields under plastic houses; stunted growth, chlorotic leaves, small curled or crinkled leaves, deformed buds and flowers and wilted plants with fewer fruits were observed in patches.

Tomato seedlings (20- days old) were transplanted to infested soil with Orobanche cernua and non-infested soils. All plants were maintained under 0, 25, 50 and 75 mM NaCl soil salinity levels throughout their growing period under greenhouse conditions. Plants grown in O. cernua infested soil infIltrated with 0, 25, and 50 mM NaCl solution for salinity showed significant reduction in their growth as well as their total soluble carbohydrate and protein contents in compared with those grown in non-infested soil. However, under 75 mM NaCI salinity level all plants showed similar growth values whether they were grown in O. cernua infested or non-infested soil. Starting at the fifth and through out the eighth week after transplantation there was a significant increase in plant height in the 0, 25 and 50 mM NaCl irrigated plants over other treatments. Irrigation with 50 mM NaCl significantly reduced the emergence of O. cernua (2/plant) and the number of attachments (4.4 attachments) on roots of tomato. Furthermore, irrigation with 75 mM NaCl resulted in complete elimination of O. cernua emergence.

The dlm (disease lesion mimic) mutant of soybean (Glycine max L. Merr) shows the similar lesion of a soybean disease caused by a fungus, Corynespora cassilcola. The lesion was examined at cellular and molecular level. Trypan blue staining result indicated that cell death was detectable in the entire region of leaves excluding veins when the lesions had already been developed. We found that the mesophyll cells of palisade layer in the dim mutant appeared to be wider apart from each other. The chloroplasts of the dim mutant cells contained bigger starch granules than those in normal plants. We also found that the lesion development of dlm plant was light-dependent and the starch degradation during the dark period of diurnal cycle was impaired in the mutant. Three soybean pathogenesis-related genes, PR-1a, PR-4, and PR-10, were examined for their expression patterns during the development of disease lesion mimic. The expression of all three genes was up-regulated to some extent upon the appearance of the disease lesion mimic. Although the exact function of DLM protein remains elusive, our data would provide some insight into mechanism underling the cell death associated with the dim mutation.

We have previously identified genes for four different protease inhibitors (PIs) that were induced upon rice blast infection in a rice blast resistant mutant SHM-11. Our expression analysis of the PIs indicated that induction of the PIs was the highest 24 hr after rice blast inoculation in the rice mutant SHM-11. Three PIs in the group of serine PIs were highly expressed while a cystein PI was weakly expressed upon rice blast inoculation. Four PIs were weakly induced 48 hr after pathogen inoculation in rice blast susceptible wild type rice plant. The simultaneous expression of three serine PIs was apparent from SHM-11 and two of them were induced in rice blast resistant Taebaegbyeo. One of them was induced in rice blast resistant Hwayeongbyeo while none of them were expressed in rice blast susceptible Nagdongbyeo and rice blast resistant Dongjinbyeo. Our results suggest that the expression of PI gene is rice cultivar specific and may be linked with the rice blast resistance in a specific rice mutant by the simultaneous expression of the PI genes.

Web blight symptoms were frequently observed on soybean plants grown in a farmer's fields located in Jincheon in Korea during a disease survey in August, 2005. Incidence of the disease was $5-20\%$ infected plants in two of four soybean fields investigated. A total of 31 isolates of Rhizoctonia sp. were obtained from leaves, leaf petioles, and pods of diseased soybean plants. The isolates were identified as Rhizoctonia solani AG-l(IA) by anastomosis test and based on the morphological and cultural characteristics. Three isolates of R. solani AG-l(IA) were tested for pathogenicity to five cultivars of soybean by artificial inoculation. All the isolates induced blight symptoms on the leaves of soybean and formed sclerotia on the lesions, which were similar to those observed in the field. The pathogenicity tests revealed that all the soybean cultivars tested were susceptible to the pathogen. There was no difference in the pathogenicity among the isolates. The present study first reveals that R. solani AG-l(IA) causes web blight of soybean in Korea.