This study was conducted to examine changes in the fungal community on fallen leaves of pear by treatment with lime sulfur. Although the lime sulfur could reduce the primary inoculum of several pathogens on spring season, the effect of lime sulfur has not been well determined scientifically. Fallen leaves infected by pear diseases in pear orchards in Naju were collected and treated with lime sulfur or water as a control. To determine the fungal diversity from each treatment, rDNA internal transcribed spacer (ITS) regions were analyzed after extraction of fungal genomic DNA from lime sulfur-treated or water-treated fallen leaves, respectively. The most common fungal species were Ascomycota and Basidiomycota in both treated leaves. However, the population dynamics of several fungal species including Alternari sp., Cladosporium sp., and Phomopsis sp., which are known as pear pathogens for skin sooty dapple disease, were quite different from each treated leaves. These results indicated that lime sulfur treatment led to changes of fungal communities on pear fallen leaves and could be applicable as a dormant spray.

Pear scab caused by the fungus Venturia nashicola is the most devastating disease on Asian pear ( Pyrus pyrifolia ). However, mycological and biological traits of V. nashicola were rarely known because the artificial cultivation of the fungus is difficult using typical fungal growth medium in the laboratory. The main objective of this study, was to determine the best culture medium composition suitable for mycelial growth of V. nashicola . A new culture medium using pear extracts, various carbon and nitrogen sources were used to examine the best combination for mycelial growth of V. nashicola . The culture medium ammended with pear extracts showed higher mycelial growth than non-ammended PDA medium. Specifically, the culture medium ammended with pear ground young leaf extract showed the highest fungal mycelial growth. The increased mycelial growth of V. nashicola was observed in growth medium ammended with Ca(NO 3 ) 2 but mycelial growth was negatively influenced by supplement with calcium chelating agent, EGTA. Taken together, we invented a new growth medium ammended with pear extract or Ca(NO 3 ) 2 for growth of V. nashicola , and it could be helpful to study on ecological and mycological traits of the pear scab pathogen.

Fluxapyroxad/pyraclostrobin 합제와 살균제를 9회 처리하는 처리체계의 검은별무늬병 방제효과를 조사하였다. 살균제가 처리되지 않은 무처리구의 이병엽률은 58.4%, 그리고 이병과율은 100%로 살균제 효과를 평가하기에 충분하였다. Fluxapyroxad/pyraclostrobin 합제만을 7회 처리한 처리구의 이병엽률은 평균 6.3%로 나타났고 방제가는 89.2%로 조사되었으며, 이병과율은 평균 16.7%, 방제가는 83.3%로 조사되었다. 서로 다른 계통의 살균제를 9회 처리한 검은별무늬병 처리체계의 효과를 조사한 결과, 이병엽률은 5.0%, 이병과율은 13.3%로, 각각의 방제가가 91.4%와 86.7%로 조사되었다. Fluxapyroxad/pyraclostrobin 합제 처리구와 비교해 보았을 때, 살균제를 9회 처리하는 처리체계에서 잎과 과실 모두에서 검은별무늬병의 발병률이 다소 낮게 나타났으나, 통계적인 유의성은 보이지 않았다. 이러한 결과는 수출전업농가에서 사용 가능한 새로운 살균제의 선발을 통한 효율적인 방제가 이루어 진다면 검은별무늬병 방제체계를 확립하는데 도움을 줄 수 있을 것으로 생각한다. The mixture of fluxapyroxad plus pyraclostrobin or the 9 time-spray schedule with various fungicides was evaluated for their efficacy in controlling pear scab in field trials. It showed sufficient condition to evaluate the efficacy of fungicides for the control of pear scab since disease incidence of the untreated control plot was 58.4% or 100% on leaves or fruits, respectively. Pear scab on leaves was occurred by 6.3% at treated plot with the mixture of fluxapyroxad plus pyraclostrobin, resulting in the 89.2% of control value. Incidence of the fruit disease was 16.7% at treated plot with the mixture of fluxapyroxad plus pyraclostrobin, showing 83.3% of control value. When the 9 time-spray schedule was tested, pear scab was occurred by 5% or 13.3% on leaves or fruits, respectively. Their control values were the 91.4% on leaves or 86.7% on fruits. Although the 9 time-spray schedule showed slightly lower disease incidence than the mixture of fluxapyroxad plus pyraclostrobin, but there is no statistically significant difference. These results suggest that the mixture of fluxapyroxad plus pyraclostrobin can control effectively the pear scab.

Protein ubiquitination is one of the major regulatory processes used by eukaryotic cells. The ubiquitin E3 ligase acts as a main determinant of substrate specificity. However, the precise roles of E3 ligase in plants to drought stress are poorly understood. In this study, a gourd family (Lagenaria siceraria) ortholog of Arabidopsis thaliana RING Zinc Finger 1 (AtRZF1) gene, designated LsRZF1, was identified and characterized. LsRZF1 was reduced by abscisic acid (ABA), osmotic stress, and drought conditions. Compared to wild type, transgenic Arabidopsis plants ectopic expressing LsRZF1 were hypersensitive to ABA and osmotic stress during early seedling development, indicating that LsRZF1 negatively regulates drought-mediated control of early seedling development. Moreover, the ectopic expression of the LsRZF1 gene was very influential in drought sensitive parameters including proline content, water loss, and the expression of dehydration stress-related genes. Furthermore, ubiquitin E3 ligase activity and genetic data indicate that AtRZF1 and LsRZF1 function in similar pathway to control proline metabolism in Arabidopsis under drought condition. Together, these results suggest that the E3 ligase LsRZF1 is an important regulator of water deficit stress during early seedling development.

Polyamines in plants are involved in various physiological and developmental processes including abiotic and biotic stress responses. We investigated the expression of ADCs, which are key enzymes in putrescine (Put) biosynthesis, and roles of Put involving defense response in Arabidopsis. The increased expression of ADC1 and ADC2, and the induction of Put were detected in GVG-NtMEK2(DD) transgenic Arabidopsis, whereas, their performance was partially compromised in GVG-NtMEK2(DD)/mpk3 and GVG-NtMEK2(DD)/mpk6 mutant following DEX treatment. The expression of ADC2 was highly induced by Pst DC3000 inoculation, while the transcript levels of ADC1 were slightly up-regulated. Compared to the WT plant, Put content in the adc2 knock-out mutant was reduced after Pst DC3000 inoculation, and showed enhanced susceptibility to pathogen infection. The adc2 mutant exhibited reduced expression of PR-1 after bacterial infection and the growth of the pathogen was about 4-fold more than that in the WT plant. Furthermore, the disease susceptibility of the adc2 mutant was recovered by the addition of exogenous Put. Taken together, these results suggest that Arabidopsis MPK3 and MPK6 play a positive role in the regulation of Put biosynthesis, and that Put contributes to bacterial pathogen defense in Arabidopsis.

catheter-related blood stream infection rates (CR-BSIs) are a substantial problem in the intensive care unit.

A pathogen-inducible WRKY cDNA was cloned from the leaves of watermelon (Citrullus lanatus) seedlings 24 h after inoculation with Cladosporium cucumerinum. The deduced protein of the gene, designated as ClWRKY70, was classified as a group III WRKY protein based on its single WRKY domain containing a Cys2HisCys zinc-finger motif. Its Arabidopsis thaliana sequence homologue (AtWRKY70) has been described as playing an important role in the plant defense response. ClWRKY70 gene transcripts were highly accumulated in watermelon by salicylic acid treatment, but not by jasmonic acid. By evaluating target gene expression in transgenic Arabidopsis overexpressing the ClWRKY70 gene, it is suggested that the watermelon WRKY gene may play a positive regulatory role in plant resistance against pathogen attack.

We carried out functional analysis of a putative novel Arabidopsis plasma membrane glucose-responsive regulator, designated AtPGR, which contains seven predicted transmembrane domains. Several evidences showed that AtPGR is a glucose-related protein, but its biological functions have yet to be reported in any plant. Analyses of the AtPGR promoter-β-glucuronidase (GUS) construct and RNA in situ hybridization revealed substantial gene expression in the vasculature of various tissues, especially in the phloem region. Glucose treatment induced the highest levels of GUS activity, reaching a peak at 3 h and declining thereafter, consistent with glucose-mediated regulation of the AtPGR promoter. We generated an atpgr RNAi knockdown mutant and found that this plant grew and developed normally. Ectopic expression of AtPGR gene modulated the induction of glucose and 2-deoxyglucose insensitivity under stress conditions. By way of contrast, cotyledon greening of atpgr RNAi knockdown mutant seeds enhanced sensitivity to glucose and 2-deoxyglucose. Taken together, these results suggest that AtPGR functions as a potential glucose-responsive regulator in carbohydrate metabolism.

We previously demonstrated that root colonization of the rhizobacterium, Pseudomonas chlororaphis O6, induced expression of a galactinol synthase gene (CsGolS1), and resulting galactinol conferred induced systemic resistance (ISR) against fungal and bacterial pathogens in cucumber leaves. To examine the role of galactinol on ISR, drought or high salt stress, we obtained T-DNA insertion Arabidopsis mutants at the AtGolS1 gene, an ortholog of the CsGolS1 gene. The T-DNA insertion mutant compromised resistance induced by the O6 colonization against Erwinia carotovora. Pharmaceutical application of 0.5 - 5 mM galactinol on roots was sufficient to elicit ISR in wild-type Arabidopsis against infection with E. carotovora. The involvement of jasmonic acid (JA) signaling on the ISR was validated to detect increased expression of the indicator gene PDF1.2. The T-DNA insertion mutant also compromised tolerance by increasing galactinol content in the O6-colonized plant against drought or high salt stresses. Taken together, our results indicate that primed expression of the galactinol synthase gene AtGolS1in the O6-colonized plants can play a critical role in the ISR against infection with E. carotovora, and in the tolerance to drought or high salt stresses.

Previously, root colonization by the rhizobacterium, Pseudomonas chlororaphis O6, was shown to induce expression of galactinol synthase conferring systemic resistance against a fungal pathogen in cucumber leaves. Here, the Arabidopsis – Botrytis cinerea system is introduced to better understand signal transduction of galactinol and (or) raffinose family oligosaccharides (RFO) during O6-mediated induced systemic resistance (ISR). Among the 10 Arabidopsis galactinol synthase genes, only AtGolS1 was specifically induced upon infection with the fungal pathogen B. cinerea. AtGolS1 was primed by O6 colonization against the pathogen in Arabidopsis leaves. Arabidopsis T-DNA insertion mutants at the AtGolS1 gene site compromised O6-mediated ISR against the pathogen, thereby suggesting that AtGolS1 plays an important role in ISR. O6 colonization increased AtGolS1 transcription as well as ISR in several Arabidopsis signaling mutants, but not in the jar1-1 and coi1 mutant lines. Exogenous jasmonate treatment induced transcription of AtGolS1 in wild-type Col-0 plants, but salicylic acid and 1-aminocyclopropane-1-carboxylate did not. These studies on signaling mutants and target gene expression indicate that expression of AtGolS1 in response to O6 colonization is mediated through the jasmonate-dependent pathway, stimulating ISR in Arabidopsis against B. cinerea infection.

Root colonization of plants with certain rhizobacteria, such as Pseudomonas chlororaphis O6, induces tolerance to biotic and abiotic stresses. Tolerance to drought was correlated with reduced water loss in P. chlororaphis O6-colonized plants and with stomatal closure, indicated by size of stomatal aperture and percentage of closed stomata. Stomatal closure and drought resistance were mediated by production of 2R,3R-butanediol, a volatile metabolite of P. chlororaphis O6. Root colonization with bacteria deficient in 2R,3R-butanediol production showed no induction of drought tolerance. Studies with Arabidopsis mutant lines indicated that induced drought tolerance required the salicylic acid (SA)-, ethylene-, and jasmonic acid-signaling pathways. Both induced drought tolerance and stomatal closure were dependent on Aba-1 and OST-1 kinase. Increases in free SA after drought stress of P. chlororaphis O6-colonized plants and after 2R,3R-butanediol treatment suggested a primary role for SA signaling in induced drought tolerance. We conclude that the bacterial volatile 2R,3R-butanediol was a major determinant in inducing resistance to drought in Arabidopsis through an SA-dependent mechanism.

SIPK와 WIPK의 상위 단계 인산화 효소로 알려진 NtMEK2가 DEX 유도성 시스템에 의해 밝혀졌다. 이 NtMEK2 유전자가 지속적으로 활성화된 돌연변이체인 $NtMEK2^{DD}$의 발현은 SIPK와 WIPK를 활성화 시켜 주므로 과민감 반응과 같은 세포 괴사를 야기하는 것으로 나타나 NtMEK2-SIPK/WIPK 체계가 담배에서 방어 반응을 조절하고 있음을 알 수 있었다. 그러나 NtMEK2-SIPK/WIPK 체계에 의해서 조절 되는 하위 기질이나 방어관련 유전자들에 대한 연구는 아직 미비한 상태이다. 그래서 본 연구는 NtMEK2-SIPK/WIPK 체계에 매개되는 하위 유전자들을 분리하기 위하여 $NtMEK2^{DD}$ 형질전환 식물체를 이용해 ACP에 기초한 DDRT-PCR을 수행하였다. 그 결과 본 연구를 통해 처음으로 pI2-4, MTS2, SINA, CDM1, HRGP 및 DEG45를 포함해 여섯 개의 DEG들을 선발하였다. 이 유전자들의 발현은 $NtMEK2^{DD}$ 형질전환에서 다시 확인하였으며 특히 pI2-4, CDM1, HRGP의 유전자 발현은 다른 유전자들과 비교해 볼 때 살리실산과 담배모자이크바이러스에 강하게 반응하여 증폭됨을 알 수 있었다. 이러한 결과를 볼 때 NtMEK2-SIPK/WIPK 체계에 의해 조절되는 세 개의 유전자는 병저항성에 관여하고 있음을 제시한다 하겠다. NtMEK2, which is the tobacco MAPK kinase that is upstream of SIPK and WIPK, was identified using the dexamethasone (DEX)-inducible gain-of-function transgenic system. Expression of $NtNEK2^{DD}$, a constitutively active mutant of NtNEK2, leads to HR-like cell death, which indicates that the NtMEK2-SIPK/WIPK cascade controls defense responses in tobacco. However, little is known about the downstream target substrates or defense-related genes that are regulated by the NtMEK2-SIPK/ WIPK cascade. In this study, ACP-based differential display RT-PCR was used to isolate the downstream effectors mediated by the NtMEK2-SIPK/WIPK cascade in $NtNEK2^{DD}$ transgenic plants. The results identified 6 novel differentially expressed genes (DEGs). These included pathogen induced protein 2-4 (pI2-4), monoterpene synthase 2 (MTS2), seven in absentia protein (SINA), cell death marker protein 1 (CDM1), hydroxyproline-rich glycoprotein (HRGP) and unknown genes (DEG45). The induction of these genes was confirmed by RT-PCR of samples obtained from $NtNEK2^{DD}$ plants. Additionally, when compared with other isolated DEGs, the pI2-4, CDM1 and HRGP genes were significantly up-regulated in response to treatment with salicylic acid and tobacco mosaic virus. Taken together, these results suggest that three novel DEGs were regulated by the NtMEK2-SIPK/WIPK cascade involved in disease resistance in tobacco.

Root colonization by Pseudomonas chlororaphis O6 in cucumber elicited an induced systemic resistance (ISR) against Corynespora cassiicola. In order to gain insight into O6-mediated ISR, a suppressive subtractive hybridization technique was applied and resulted in the isolation of a cucumber galactinol synthase (CsGolS1) gene. The transcriptional level of CsGolS1 and the resultant galactinol content showed an increase several hours earlier under O6 treatment than in the water control plants following C. cassiicola challenge, whereas no difference was detected in the plants without a pathogen challenge. The CsGolS1-overexpressing transgenic tobacco plants demonstrated constitutive resistance against the pathogens Botrytis cinerea and Erwinia carotovora, and they also showed an increased accumulation in galactinol content. Pharmaceutical application of galactinol enhanced the resistance against pathogen infection and stimulated the accumulation of defense-related gene transcripts such as PR1a, PR1b, and NtACS1 in wild-type tobacco plants. Both the CsGolS1-overexpressing transgenic plants and the galactinol-treated wild-type tobacco plants also demonstrated an increased tolerance to drought and high salinity stresses.

Polyphosphate levels are modulated by the actions of polyphosphate kinase, encoded by ppk , and exopolyphosphatase, encoded by ppx . The genes ppk and ppx are adjacent to each other in the genome of the root colonizer, Pseudomonas chlororaphis O6. A ppk -deficient mutant was more sensitive to oxidative stress than the wild-type and the ppx mutant. Transcripts from ppx increased as cultures matured from mid- to late-logarithmic and stationary phases, whereas abundance was greater for ppk in the late-logarithmic phase than in the stationary phase. Transcript accumulation from the rpoS gene, encoding the stationary-phase sigma factor RpoS, was decreased in the mid- and late-logarithmic and stationary phases in the ppk mutant. Thus, ppk regulates rpoS transcript accumulation in P. chlororaphis O6. However, mutations in either the ppk or ppx genes had no effect on induction of systemic resistance in plants colonized by P. chlororaphis O6.

Seed coating by a phenazine-producing bacterium, Pseudomonas chlororaphis O6, induced dose-dependent inhibition of germination in wheat and barley seeds, but did not inhibit germination of rice or cucumber seeds. In wheat seedlings grown from inoculated seeds, phenazine production levels near the seed were higher than in the roots. Deletion of the gacS gene reduced transcription from the genes required for phenazine synthesis, the regulatory phzI gene and the biosynthetic phzA gene. The inhibition of seed germination and the induction of systemic disease resistance against a bacterial soft-rot pathogen, Erwinia carotovora subsp. carotovora, were impaired in the gacS and phzA mutants of P chlororaphis O6. Culture filtrates of the gacS and phzA mutants of P chlororaphis 06 did not inhibit seed germination of wheat, whereas that of the wild-type was inhibitory. Our results showed that the production of phenazines by P chlororaphis O6 was correlated with reduced germination of barley and wheat seeds, and the level of systemic resistance in tobacco against E. carotovora.

Aims: The primary objective of this study was to determine the role of 6-phosphogluconate dehydratase in root colonization and the induction of systemic resistance by the rhizobacterium, Pseudomonas chlororaphis O6.Methods and Results: The edd gene encoding for 6-phosphogluconate dehydratase, which is one of the key enzymes in glucose utilization, was cloned. Transcription of the gene was higher in medium containing sugars than with organic acids. An edd mutant failed to grow on glucose but grew on organic acids. The edd mutant colonized tobacco roots at wild-type levels early after inoculation, but levels were lower by 12 days. The edd mutant failed to induce the systemic resistance in tobacco to a soft-rot pathogen at wild-type level.Conclusions: 6-Phosphogluconate dehydratase in P. chlororaphis O6 contributes to root colonization and induction of systemic resistance presumably as the consequence of its essential role in the Entner-Doudoroff (ED) pathway.Significance and Impact of the study: Metabolism of sugars through the ED pathway in P. chlororaphis O6 may be important because it facilitates the production of inducers of systemic resistance including butanediol.

Root colonization by a rhizobacterium, Pseudomonas chlororaphis O6, elicited induced systemic resistance (ISR) in the leaves of cucumber plants against fungal and bacterial pathogens. To understand the role of unique genes during strain O6-mediated ISR, a suppressive subtractive hybridization method was undertaken and led to isolation of twenty-five distinct genes. The transcriptional levels of all the genes showed an increase much earlier under O6 treatment than in water control plants only after challenge with pathogen, while no difference detected on the plants without pathogen challenge. This suggests that O6-mediated ISR is associated with the priming phenomenon, an enhanced capacity for the rapid and effective activation of cellular defense responses after challenge inoculation.