In this study, 200 isolates of fluorescent pseudomonads were isolated from different fields of East and West Azarbaijan and Ardebil provinces of Iran. These bacterial isolates were screened on the basis of a dual culture assay, the presence of known antibiotic genes, and their ability to successfully colonize roots and to promote plant growth. Twelve isolates exhibited 30% or more inhibition of mycelia growth of $P.$ $drechsleri$. Genes encoding production of the antibiotics 2,4-diacetylphloroglucinol, phenazine-1-carboxylic acid, and pyoluteorin were detected in some strains but none of the strains possessed the coding gene for production of antibiotic pyrrolnitrin. In an $in$ $vitro$ test for root colonization, the population density on roots of plants treated with most of the above strains was more than 6 $\log_{10}$ CFU $g^{-1}$ roots, with a maximum of 7.99 $\log_{10}$ CFU $g^{-1}$ roots for strain 58A. Most of the strains promoted significant plant growth in comparison to non-treated controls. In green house studies, the percentage of healthy plants in pots treated with strains 58A and 8B was 90.8% and 88.7%, respectively. The difference between these treatments and treatment with the fungicide metalaxyl was not significant.

This study investigated the distribution of endophytic fungi obtained from the surface sterilized tissues of leaves, stems and roots of chili pepper ($Capsicum$ $annuum$ L.) plants in seedling, flowering and fruiting stages in Korea and their antifungal activity. A total of 481 isolates were recovered and were identified using molecular techniques. Based on rDNA ITS gene sequence and phylogenetic analysis, 21 fungal genera were characterized, belonging to 16 Ascomycota and 5 Basidiomycota. $Penicillium$ in seedling stage, $Fusarium$ in flowering stage, $Colletotrichum$ followed by $Fusarium$, $Alternaria$ and $Xylaria$ in fruiting stage was predominant and $Alternaria$, $Cladosporium$ and $Fusarium$ were common in all growth stages. Among 481 endophytes 90 phenotypes were evaluated for the antimicrobial activity against three major pathogens ($Phytophthora$ $capsici$, $Colletotrichum$ $acutatum$ and $Fusarium$ $oxysporum$) of chili pepper. Among them 16 isolates inhibited the growth of at least one test microorganisms. Three strains showed a broad spectrum antifungal activity and displayed strong inhibition against chili pepper pathogenic fungi.

Endophytic bacterial communities of tidal flat plants antagonistic to oomycete plant pathogens were studied by the isolation of 256 root colonizing endophytic bacteria from surface-disinfected root tissues of six plants ($Rosa$ $rugosa$, $Suaeda$ $maritima$, $Vitex$ $rotundifolia$, $Carex$ $scabrifolia$, $Glehnia$ $littoralis$ and $Elymus$ $mollis$) growing in a tidal flat area of Namhae Island, Korea. To understand the antagonistic potential, an $in$ $vitro$ antagonistic assay was performed to characterize and identify strains that were antagonistic to the oomycete plant pathogens $Phytophthora$ $capsici$ and $Pythium$ $ultimum$ from the total population. Nine percent of the total number of isolated bacteria exhibited in vitro inhibitory activity against target plant pathogenic oomycetes. Taxonomic and phylogenetic placement of the antagonistic bacteria was investigated by analysis of the 16S rRNA gene sequences. The sequence analysis classified the antagonistic strains into four major classes of the domain bacteria ($Firmicutes$, ${\alpha}-Proteobacteria$, ${\gamma}-Proteobacteria$ and $Actinomycetes$) and 10 different genera. Further production of secondary metabolites, hydrolytic enzymes and plant growth promoting traits were determined for the putative new species of antagonistic endophytic bacteria. These new strains could not be identified as known species of ${\alpha}-Proteobacteria$, and so may represent novel bacterial taxa. The unexpected high antagonistic bacterial diversity associated with the tidal flat plants may be indicative of their importance in tidal flat plants as a promising source of novel antimicrobial compounds and biocontrol agents.

Efficacy of different control methods was evaluated for disease management of tomato bacterial wilt caused by $Ralstonia$ $solanacearum$. All six chemical pesticides applied to the bacterial suspension showed $in$ $vitro$ bactericidal activities against $R.$ $solanacearum$. Minimal inhibitory concentrations (MICs) of copper hydroxide (CH), copper hydroxide-oxadixyl mixture (CH+O), and copper oxychloride-dithianon mixture (CO+D) were all 200 ${\mu}g/ml$; MIC of copper oxychloride-kasugamycin (CO+K) mixture was 100 ${\mu}g/ml$; MICs of both streptomycin- validamycin (S+V) and oxine copper-polyoxine B mixture (OC+PB) were 10 ${\mu}g/ml$. Among these chemical pesticides, treatment of the detached tomato leaves with the 5 pesticides (1 mg/ml), except for OC+PB delayed early wilting symptom development caused by the bacterial inoculation ($10^6$ and $10^7$ cfu/ml). Four pesticides, CH, CH+O, CO+K and S+V, showed disease protection in pot analyses. Six plant essential oils, such as cinnamon oil, citral, clove oil, eugenol, geraniol and limonene, differentially showed their antibacterial activities $in$ $vitro$ against $R.$ $solanacearum$ demonstrated by paper disc assay. Among those, cinnamon oil and clove oil exert the most effective activity for protection from the wilt disease caused by the bacterial infection ($10^6$ cfu/ml). Treatment with cinnamon oil and clove oil also suppressed bacterial disease by a higher inoculum concentration ($10^7$ cfu/ml). Clove oil could be used for prevention of bacterial wilt disease of tomato plants without any phytotoxicity. Thus, we suggest that copper compounds, antibiotics and essential oils have potency as a controlling agent of tomato bacterial wilt.

$Potato$ $virus$ $X$ (PVX) replication is precisely regulated by regulatory viral sequences and by viral and/or host proteins. In a previous study, we identified a 54-kDa cellular tobacco protein that bound to a region within the first 46 nucleotides (nt) of the 5' non-translated region (NTR) of the viral genome. Optimal binding was dependent upon the presence of an ACCA sequence at nt 10-13. To identify host factors that bind to 5' NTR elements including AC-rich sequences as well as stemloop 1 (SL1), we used northwestern blotting and matrixassisted laser desorption/ionization time-of-flight mass spectrometry for peptide mass fingerprinting. We screened several host factors that might affect PVX replication and selected a candidate protein, $Nicotiana$ $tabacum$ WRKY transcription factor 1 (NtWRKY1). We used a $Tobacco$ $rattle$ $virus$ (TRV)-based virus-induced gene silencing (VIGS) system to investigate the role of NtWRKY1 in PVX replication. Silencing of $WRKY1$ in $Nicotiana$ $benthamiana$ caused lethal apical necrosis and allowed an increase in PVX RNA accumulation. This result could reflect the balancing of PVX accumulation in a systemic $N.$ $benthamiana$ host to maintain PVX survival and still produce a suitable appearance of mosaic and mottle symptoms. Our results suggest that PVX may recruit the WRKY transcription factor, which binds to the 5' NTR of viral genomic RNA and acts as a key regulator of viral infection.

In August 2010, a severe crown rot was observed on chrysanthemum ($Chrysanthemum$ $morifolium$ Ramat., variety Sinro) in several greenhouses located at Damyang and Muan, Jeonnam province, Korea. Three isolates (EML-CHS1, -CHS2, and -CHS3) of $Fusarium$ were isolated from the affected plants and identified based on morphological characteristics and rDNA internal transcribed spacer (ITS) sequence analysis. Sequence analysis by BLAST indicated that EMLCHS1, -CHS2 and CHS3 were closest to a $Fusarium$ species, $F.$ $solani$ with > 99% sequence similarity. Pathogenicity tests were performed on chrysanthemum with spore suspensions containing $3.4{\times}10^6$ spores/ml using the dipping method. Ten days after inoculation, similar symptoms to those observed in the greenhouses were seen on the inoculated plants. The causal fungus was reisolated from the artificially inoculated basal stems, fulfilling Koch's postulates. To our knowledge, this is the first report of crown rot by $Fusarium$ $solani$ on chrysanthemum ($Chrysanthemum$ $morifolium$) in Korea.

Northern stem canker caused by $Diaporthe$ $phaseolorum$ var. $caulivora$ ($Dpc$) has become a serious disease in soybean. The objectives of this study were to survey the existence of $Dpc$ on soybean in Korea, and to examine the potential pathogenicity of $Dpc$ in seed decay. One such isolate, SSLP-4, isolated from a field-grown plant of the Korean soybean cultivar Danbaekkong, was identified as $Dpc$, based on its morphological and molecular characteristics by sequences of internal transcribed spacer (ITS), translation elongation factor (TEF) 1-${\alpha}$ and ${\beta}$-tubulin regions, as well as pathogenic analyses. Moreover, morphological and molecular analyses revealed that isolate SSLP-4 was nearly identical to $Dpc$ strains from the United States. Pathogenicity tests on hypocotyls of soybean seedlings and detached leaves resulted in typical symptoms of soybean northern stem canker and inoculation on plants at R5-R7 stage caused seed decay. All results suggest that the $Dpc$ strain SSLP-4 can cause both stem canker and seed decay on soybean. Thus, the SSLP-4 isolate has the potential to contribute greatly to understanding of host plant resistance mechanisms, both at vegetative and reproductive growth stages in soybean.

For the genetic differentiation of $Pseudomonas$ $syringae$ pathovar $tomato$, a total of 51 $P.$ $syringae$ pv. strains infecting 33 different host plants were analyzed using repetitive element PCR(REP-PCR) and universal rice primer PCR(URP-PCR). The entire DNA fingerprint profiles were analyzed using unweighted pair-group method with arithmetic averages (UPGMA). The 51 $P.$ $syringae$ pv. strains could be divided into five clusters based on 65% similarity by Rep-PCR using BOX, ERIC, and REP primers. $P.$ $syringae$ pv. $tomato$ cluster was well separated from other 31 $P.$ $syringae$ pathovars. $P.$ $syringae$ pv. $tomato$ cluster included only $P.$ $syringae$ pv. $maculicola$ and $P.$ $syringae$ pv. $tomato$. $P.$ $syringae$ pv. $tomato$ strains could be divided into two genetic groups. Meanwhile, the Pseudomonas pv. strains could be divided into four clusters based on 63% similarity by URP-PCR using 2F, 9F, and 17R primers. $P.$ $syringae$ pv. $tomato$ cluster was also well separated from 30 other $P.$ $syringae$ pathovars. In this case, $P.$ $syringae$ pv. $tomato$ cluster included $P.$ $syringae$ pv. $maculicola$, $P.$ $syringae$ pv. $berberidi$, and $P.$ $syringae$ pv. $tomato$. $P.$ $syringae$ pv. $tomato$ strains was also separated into two genetic groups by URP-PCR analysis. Overall, our work revealed that $P.$ $syringae$ pv. $tomato$ can be genetically differentiated from other $P.$ $syringae$ pathovars by the DNA fingerprint profiles of REP-PCR and URP-PCR. We first report that there are two genetically diverged groups in $P.$ $syringae$ pv. $tomato$ strains.

Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$ $vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$ $carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$ $Mosaic$ $Virus$ and the gray mold disease caused by $Botrytis$ $cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$ $manteilii$ B001 and $Bacillus$ $cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.

$Potato$ $virus$ $X$ (PVX) contains $cis$-acting elements including stem-loop 1 (SL1) RNA at the 5' region; SL1 is conserved among all potexviruses. The SL1 at the positive-sense RNA, SL1(+), is required for PVX RNA replication, cell-to-cell movement, and translation. Previous research demonstrated that SL1(+) RNA also serves as the origin of assembly for encapsidation of PVX RNA. To identify the essential sequences and/or regions for capsid protein (CP) subunit recognition within SL1(+) RNA, we used electrophoretic mobility shift assays (EMSA), UV cross-linking, and yeast three-hybrid analyses. The EMSA and UV cross-linking analyses with PVX CP subunits and RNA transcripts corresponding to the SL1(+) RNA showed that the SL1(+) RNA formed complexes with CP subunits. We also conducted EMSA and yeast three-hybrid analyses with RNAs containing various mutations of SL1(+) RNA elements. These analyses indicated that SL1(+) RNA is required for the interaction with PVX CP and that the RNA sequences located at the loop C and tetra loop of the SL1(+) are crucial for CP binding. These results indicate that, in addition to being important for RNA accumulation, the SL1(+) RNA from the 5' region of the PVX genome is also required for specific binding of PVX CP.

We developed a reassortant RNA virus vector derived from $Cucumber$ $mosaic$ $virus$ (CMV), which has advantages of very wide host range and can efficiently induce gene silencing in a few model plants. Certain CMV isolates, however, show limited host ranges presumably because they naturally co-evolved with their own hosts. We used a reassortant comprised of two strains of CMV, Y-CMV and Gn-CMV, to broaden the host range and to develop a virus vector for virus-induced gene silencing (VIGS). Gn-CMV could infect chili pepper and tomato more efficiently than Y-CMV. Gn-CMV RNA1, 3 and Y-CMV RNA2-A1 vector were newly reconstructed, and the transcript mixture of RNA1 and 3 genomes of Gn-CMV and RNA2 genome of Y-CMV RNA2 containing portions of the endogenous phytoene desaturase (PDS) gene (CMV2A1::PDSs) was inoculated onto chili pepper (cv. Chung-yang), tomato (cvs. Bloody butcher, Tigerella, Silvery fir tree, and Czech bush) and $Nicotiana$ $benthamiana$. All the tested plants infected by the reassortant CMV vector showed typical photo-bleaching phenotypes and reduced expression levels of $PDS$ mRNA. These results suggest that the reassortant CMV vector would be a useful tool for the rapid induction of the RNA silencing of endogenous genes in chili pepper and tomato plants.

$Tomato$ $spotted$ $wilt$ $virus$ (TSWV)-infected $Capsicum$ $annuum$ plants were collected from open fields during June to July in 2010. The TSWV isolates were designated as Gneung, Ghang-Kjj, Gchang-Njc, Ghae, and Pap. The nucleotide sequence of the nucleocapsid protein (N) and movement protein (NSm) of the five isolates was determined. The pathogenicity of the five isolates was determined on 14 $C.$ $annuum$ cultivars two times by using mechanical inoculation. The five isolates induced different response: Both Gneung and Gchang-Kjj did not infect any of the cultivars in the 2nd trial, while Gchang-Njc, Ghae and Pap infected 11, 6 and 13 of 14 cultivars, respectively. The five isolates also were tested on $Solanum$ $lycopersicum$ breeding line TGC09-71 and three $Nicotiana$ species. $S.$ $lycopersicum$ showed a similar response to the five isolates as did $C.$ $annuum$. Both Gchang-Njc and Ghae infected systemically all three $Nicotiana$ species tested. While both Pap and Gneung did not infect any of the $Nicotiana$ species tested. In conclusion, five TSWV isolates induced different infection spectra in $C.$ $annuum$ cultivars, $Nicotiana$ species and an $S.$ $lycopersicum$ breeding line. Amino acid sequence analysis of the NSm gene could not support or explain the different infection spectra of the five isolates. This study indicated that various isolates must be used as virus inocula for evaluation of $C.$ $annuum$ and $S.$ $lycopersicum$ cultivars in breeding programs for TSWV resistance.

The rhizobacterium NJ134, showing strong $in$ $vitro$ antifungal activity against $Fusarium$ $oxysporum$, was isolated from field grown tomato plants and identified as $Pseudomonas$ sp. based on 16S ribosomal DNA sequence and biochemical analyses. The antifungal compound purified by gas chromatography-mass spectrometry, infrared, and nuclear magnetic resonance analyses from NJ134 cultures was polyketide 2,4-diacetylphloroglucinol (DAPG). Analysis of the sequence of part of one of the genes associated with DAPG synthesis, $phlD$, indicated that the DAPG producer NJ134 was a novel genotype or variant of existing genotype termed O that have been categorized based on isolates from Europe and North America. A greenhouse study indicated that about $10^8$ CFU/g of soil NJ134 culture application was required for effective biocontrol of Fusarium wilt in tomato. These results suggest that a new variant genotype of a DAPG-producing strain of $Pseudomonas$ has the potential to control Fusarium wilt under the low disease pressure conditions.

Trifloxystrobin is a strobilurin fungicide, which possesses broad spectrum control against fungal plant diseases. We demonstrated that pre-treating red pepper plants with trifloxystrobin resulted in increased plant growth and leaf chlorophyll content compared with those in control plants. Relative water content of the leaves and the survival rate of intact plants indicated that plants acquired systemic tolerance to drought stress following trifloxystrobin pre-treatment. The recovery rate by rehydration in the drought treated plant was better in those pre-treated with trifloxystrobin than that in water treated plants. Induced drought tolerance activity by trifloxystrobin was sustained for 25 days after initial application. The trifloxystrobin treated red pepper plants also had induced systemic tolerance to other abiotic stresses, such as frost, cold, and high temperature stresses. These findings suggest that applying the chemical fungicide trifloxystrobin induced systemic tolerance to certain abiotic stresses in red pepper plants.

Trichomes are specialized epidermal structure having the functions of physical and chemical block against biotic and abiotic stresses. Several studies on $Capsicum$ species revealed that virus and herbivore resistance is associated with trichome-formation. However, there is no research on the structural characterization of trichomes developed on the epidermis of $Capsicum$ spp. Thus, this study attempts to charaterize the trichome morphologies in 5 species of $Capsicum$ using a Field Emission Scanning Electron Microscopy (FESEM). Six main trichome types were identified by their morphology under FESEM. Both glandular and non-glandular types of trichomes were developed on the epidermal tissues of $Capsicum$ spp. The glandular trichome were further classified into type I, IV and VII according to their base, stalk length, and stalk. Non-glandular trichomes were also classified into type II, III, and V based on stalk cell number and norphology. Almost all the species in $C.$ $chinense$ and $C.$ $pubescens$ had glandular trichomes. To our knowledge, this is the first study on classification of trichomes in the genus $Capsicum$ and, our results could provide basic informations for understanding the structure and function of trichomes on the epidermal differentiation and association with biotic stress tolerance.