The Auto-Regulation of ATL2 E3 Ubiquitin Ligase Plays an Important Role in the Immune Response against Alternaria brassicicola in Arabidopsis thaliana.

The ubiquitin/26S proteasome system is a crucial regulatory mechanism that governs various cellular processes in plants, including signal transduction, transcriptional regulation, and responses to biotic and abiotic stressors. Our study shows that the RING-H2-type E3 ubiquitin ligase, Arabidopsis Tóxicos en Levadura 2 (ATL2), is involved in response to fungal pathogen infection. Under normal growth conditions, the expression of the ATL2 gene is low, but it is rapidly and significantly induced by exogenous chitin. Additionally, ATL2 protein stability is markedly increased via chitin treatment, and its degradation is prolonged when 26S proteasomal function is inhibited. We found that an atl2 null mutant exhibited higher susceptibility to Alternaria brassicicola, while plants overexpressing ATL2 displayed increased resistance. We also observed that the hyphae of A. brassicicola were strongly stained with trypan blue staining, and the expression of A. brassicicola Cutinase A (AbCutA) was dramatically increased in atl2. In contrast, the hyphae were weakly stained, and AbCutA expression was significantly reduced in ATL2-overexpressing plants. Using bioinformatics, live-cell confocal imaging, and cell fractionation analysis, we revealed that ATL2 is localized to the plasma membrane. Further, it is demonstrated that the ATL2 protein possesses E3 ubiquitin ligase activity and found that cysteine 138 residue is critical for its function. Moreover, ATL2 is necessary to successfully defend against the A. brassicicola fungal pathogen. Altogether, our data suggest that ATL2 is a plasma membrane-integrated protein with RING-H2-type E3 ubiquitin ligase activity and is essential for the defense response against fungal pathogens in Arabidopsis.

Antifungal Potential of Melaleuca alternifolia against Fungal Pathogen Fusarium oxysporum f. sp. cubense Tropical Race 4.

Fusarium wilt of bananas caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4) poses the most serious threat to banana production globally. The disease has been managed using chemical fungicides, yet the control levels are still unsatisfactory. This study investigated the antifungal activities of tea tree (Melaleuca alternifolia) essential oil (TTO) and hydrosol (TTH) against Foc TR4 and their bioactive components. The potential of TTO and TTH in inhibiting the growth of Foc TR4 was evaluated in vitro using agar well diffusion and spore germination assays. Compared to the chemical fungicide, TTO effectively suppressed the mycelial growth of Foc TR4 at 69%. Both the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of TTO and TTH were established at 0.2 µg/µL and 50% v/v, respectively, suggesting the fungicidal nature of the plant extracts. The disease control efficacies were also demonstrated by a (p ≤ 0.05) delayed Fusarium wilt symptom development in the susceptible banana plants with reduced LSI dan RDI scores from 70% to around 20-30%. A GC/MS analysis of TTO identified terpinen-4-ol, eucalyptol, and α-terpineol as the major components. In contrast, an LC/MS analysis of TTH identified different compounds, including dihydro-jasmonic acid and methyl ester. Our findings indicate the potential of tea tree extracts as natural alternatives to chemical fungicides to control Foc TR4.

Functional roles of the pepper leucine-rich repeat protein and its interactions with pathogenesis-related and hypersensitive-induced proteins in plant cell death and immunity.

MAIN CONCLUSION: Pepper leucine-rich repeat protein (CaLRR1) interacts with defense response proteins to regulate plant cell death and immunity. This review highlights the current understanding of the molecular functions of CaLRR1 and its interactor proteins. Plant cell death and immune responses to microbial pathogens are controlled by complex and tightly regulated molecular signaling networks. Xanthomonas campestris pv. vesicatoria (Xcv)-inducible pepper (Capsicum annuum) leucine-rich repeat protein 1 (CaLRR1) serves as a molecular marker for plant cell death and immunity signaling. In this review, we discuss recent advances in elucidating the functional roles of CaLRR1 and its interacting plant proteins, and understanding how they are involved in the cell death and defense responses. CaLRR1 physically interacts with pepper pathogenesis-related proteins (CaPR10 and CaPR4b) and hypersensitive-induced reaction protein (CaHIR1) to regulate plant cell death and defense responses. CaLRR1 is produced in the cytoplasm and trafficked to the extracellular matrix. CaLRR1 binds to CaPR10 in the cytoplasm and CaPR4b and CaHIR1 at the plasma membrane. CaLRR1 synergistically accelerates CaPR10-triggered hypersensitive cell death, but negatively regulates CaPR4b- and CaHIR1-triggered cell death. CaHIR1 interacts with Xcv filamentous hemagglutinin (Fha1) to trigger disease-associated cell death. The subcellular localization and cellular function of these CaLRR1 interactors during plant cell death and defense responses were elucidated by Agrobacterium-mediated transient expression, virus-induced gene silencing, and transgenic overexpression studies. CaPR10, CaPR4b, and CaHIR1 positively regulate defense signaling mediated by salicylic acid and reactive oxygen species, thereby activating hypersensitive cell death and disease resistance. A comprehensive understanding of the molecular functions of CaLRR1 and its interacting protein partners in cell death and defense responses will provide valuable information for the molecular genetics of plant disease resistance, which could be exploited as a sustainable disease management strategy.

Light- and Relative Humidity-Regulated Hypersensitive Cell Death and Plant Immunity in Chinese Cabbage Leaves by a Non-adapted Bacteria Xanthomonas campestris pv. vesicatoria.

Inoculation of Chinese cabbage leaves with high titer (107 cfu/ml) of the non-adapted bacteria Xanthomonas campestris pv. vesicatoria (Xcv) strain Bv5-4a.1 triggered rapid leaf tissue collapses and hypersensitive cell death (HCD) at 24 h. Electrolyte leakage and lipid peroxidation markedly increased in the Xcv-inoculated leaves. Defence-related gene expressions (BrPR1, BrPR4, BrChi1, BrGST1 and BrAPX1) were preferentially activated in the Xcv-inoculated leaves. The Xcv-triggered HCD was attenuated by continuous light but accelerated by a dark environment, and the prolonged high relative humidity also alleviated the HCD. Constant dark and increased relative humidity provided favorable conditions for the Xcv bacterial growth in the leaves. Pretreated fluridone (biosynthetic inhibitor of endogenous abscisic acid [ABA]) increased the HCD in the Xcv-inoculated leaves, but exogenous ABA attenuated the HCD. The pretreated ABA also reduced the Xcv bacterial growth in the leaves. These results highlight that the onset of HCD in Chinese cabbage leaves initiated by non-adapted pathogen Xcv Bv5-4a.1 and in planta bacterial growth was differently modulated by internal and external conditional changes.

Pepper osmotin-like protein 1 (CaOSM1) is an essential component for defense response, cell death, and oxidative burst in plants.

Osmotin or osmotin-like protein, a PR-5 family member, is differentially induced in plants by abiotic and biotic stresses. Here, we demonstrate that the pepper (Capsicum annuum) osmotin-like protein 1 gene, CaOSM1, was required for the defense and hypersensitive cell death response and oxidative burst signaling during Xanthomonas campestris pv. vesicatoria (Xcv) infection. CaOSM1 protein was localized to the plasma membrane in leaf cells of Nicotiana benthamiana. Agrobacterium-mediated transient expression of CaOSM1 in pepper distinctly induced the hypersensitive cell death response and H2O2 accumulation. Knock-down of CaOSM1 in pepper by virus-induced gene silencing increased the susceptibility to Xcv infection, which was accompanied by attenuation of the cell death response and decreased accumulation of H2O2. CaOSM1 overexpression in transgenic Arabidopsis conferred reduced susceptibility and accelerated cell death response and H2O2 accumulation to infection by Pseudomonas syringe pv. tomato and Hyaloperonospora arabidopsidis. Together, these results suggest that CaOSM1 is involved in cell death and oxidative burst responses during plant defense against microbial pathogens.

Phospholipase A(2) is required for PIN-FORMED protein trafficking to the plasma membrane in the Arabidopsis root.

Phospholipase A(2) (PLA(2)), which hydrolyzes a fatty acyl chain of membrane phospholipids, has been implicated in several biological processes in plants. However, its role in intracellular trafficking in plants has yet to be studied. Here, using pharmacological and genetic approaches, the root hair bioassay system, and PIN-FORMED (PIN) auxin efflux transporters as molecular markers, we demonstrate that plant PLA(2)s are required for PIN protein trafficking to the plasma membrane (PM) in the Arabidopsis thaliana root. PLA(2)alpha, a PLA(2) isoform, colocalized with the Golgi marker. Impairments of PLA(2) function by PLA(2)alpha mutation, PLA(2)-RNA interference (RNAi), or PLA(2) inhibitor treatments significantly disrupted the PM localization of PINs, causing internal PIN compartments to form. Conversely, supplementation with lysophosphatidylethanolamine (the PLA(2) hydrolytic product) restored the PM localization of PINs in the pla(2)alpha mutant and the ONO-RS-082-treated seedling. Suppression of PLA(2) activity by the inhibitor promoted accumulation of trans-Golgi network vesicles. Root hair-specific PIN overexpression (PINox) lines grew very short root hairs, most likely due to reduced auxin levels in root hair cells, but PLA(2) inhibitor treatments, PLA(2)alpha mutation, or PLA(2)-RNAi restored the root hair growth of PINox lines by disrupting the PM localization of PINs, thus reducing auxin efflux. These results suggest that PLA(2), likely acting in Golgi-related compartments, modulates the trafficking of PIN proteins.

Vapours from plant essential oils to manage tomato grey mould caused by Botrytiscinerea.

Tomato grey mould has been a great concern during tomato production. The in vitro antifungal activity of vapours emitted from four plant essential oils (EOs) (cinnamon oil, fennel oil, origanum oil, and thyme oil) were evaluated during in vitro conidial germination and mycelial growth of Botrytis cinerea, the causal agent of grey mould. Cinnamon oil vapour was the most effective in suppressing conidial germination, whereas the four EOs showed similar activities regarding inhibiting mycelial growth in dose-dependent manners. The in planta protection effect of the four EO vapours was also investigated by measuring necrotic lesions on tomato leaves inoculated by B. cinerea. Grey mould lesions on the inoculated leaves were reduced by the vapours from cinnamon oil, origanum oil and thyme oil at different levels, but fennel oil did not limit the spread of the necrotic lesions. Decreases in cuticle defect, lipid peroxidation, and hydrogen peroxide production in the B. cinerea-inoculated leaves were correlated with reduced lesions by the cinnamon oil vapours. The reduced lesions by the cinnamon oil vapour were well matched with arrested fungal proliferation on the inoculated leaves. The cinnamon oil vapour regulated tomato defence-related gene expression in the leaves with or without fungal inoculation. These results suggest that the plant essential oil vapours, notably cinnamon oil vapour, can provide eco-friendly alternatives to manage grey mould during tomato production.

Co-treatment with Origanum Oil and Thyme Oil Vapours Synergistically Limits the Growth of Soil-borne Pathogens Causing Strawberry Diseases.

Vapours from origanum oil (O) and thyme oil (T) were applied to the four soil-borne strawberry pathogens Fusarium oxysporum f. sp. fragariae, Colletotrichum fructicola, Lasiodiplodia theobromae, and Phytophthora cactorum, causing Fusarium wilt, anthracnose, dieback, and Phytophthora rot, respectively. Increasing T vapour doses in the presence of O vapour strongly inhibited mycelial growths of the four pathogens and vice versa. When mycelia of F. oxysporum f. sp. fragariae and P. cactorum exposed to the combined O + T vapours were transferred to the fresh media, mycelial growth was restored, indicating fungistasis by vapours. However, the mycelial growth of C. fructicola and L. theobromae exposed to the combined O + T vapours have been slightly retarded in the fresh media. Prolonged exposure of strawberry pathogens to O + T vapours in soil environments may be suggested as an alternative method for eco-friendly disease management.

Population Structure of Stagonosporopsis Species Associated with Cucurbit Gummy Stem Blight in Korea.

Gummy stem blight (GSB), a common and serious disease in cucurbits worldwide, is caused by three genetically distinct species: Stagonosporopsis cucurbitacearum (syn. Didymella bryoniae), S. citrulli, and S. caricae. In Korea, however, the three species of Stagonosporopsis have been barely characterized. In this study, 21 Stagonosporopsis isolates were recovered from watermelon (Citrullus lanatus) and muskmelon (Cucumis melo) leaves and stem showing blight symptoms collected from 43 fields in Korea. Sequence analysis performed with an internal transcribed spacer region was not competent to differentiate the Stagonosporopsis isolates. On the contrary, analysis of ß-tubulin (TUB) genes and three microsatellite markers, Db01, Db05, and Db06, successfully differentiated Stagonosporopsis isolates. Further sequence analysis identified two Stagonosporopsis species, S. citrulli and S. caricae, and one previously unknown species of Stagonosporopsis. Representative isolates from three species caused dark water-soaked lesions on the detached watermelon and muskmelon leaves with no significant differences in the aggressiveness. Our results indicate that the S. citrulli, S. caricae, and unknown Stagonosporopsis sp. are all causal agents of GSB for both watermelon and muskmelon. This is the first report of a new species and the population structure of Stagonosporopsis species causing GSB in Korea.

Heavy metal toxicity in plants and the potential NO-releasing novel techniques as the impending mitigation alternatives.

Environmental pollutants like heavy metals are toxic, persistent, and bioaccumulative in nature. Contamination of agricultural fields with heavy metals not only hampers the quality and yield of crops but also poses a serious threat to human health by entering the food chain. Plants generally cope with heavy metal stress by regulating their redox machinery. In this context, nitric oxide (NO) plays a potent role in combating heavy metal toxicity in plants. Studies have shown that the exogenous application of NO donors protects plants against the deleterious effects of heavy metals by enhancing their antioxidative defense system. Most of the studies have used sodium nitroprusside (SNP) as a NO donor for combating heavy metal stress despite the associated concerns related to cyanide release. Recently, NO-releasing nanoparticles have been tested for their efficacy in a few plants and other biomedical research applications suggesting their use as an alternative to chemical NO donors with the advantage of safe, slow and prolonged release of NO. This suggests that they may also serve as potential candidates in mitigating heavy metal stress in plants. Therefore, this review presents the role of NO, the application of chemical NO donors, potential advantages of NO-releasing nanoparticles, and other NO-release strategies in biomedical research that may be useful in mitigating heavy metal stress in plants.

Genetic diversity of tomato-infecting Tomato yellow leaf curl virus (TYLCV) isolates in Korea.

Epidemic outbreaks of Tomato yellow leaf curl virus (TYLCV) diseases occurred in greenhouse grown tomato (Solanum lycopersicum) plants of Busan (TYLCV-Bus), Boseong (TYLCV-Bos), Hwaseong (TYLCV-Hwas), Jeju Island (TYLCV-Jeju), and Nonsan (TYLCV-Nons) in Korea during 2008-2009. Tomato disease by TYLCV has never occurred in Korea before. We synthesized the full-length genomes of each TYLCV isolate from the tomato plants collected at each area and determined their nucleotides (nt) sequences and deduced the amino acids of six open reading frames in the genomes. TYLCV-Bus and -Bos genomes shared higher nt identities with four Japanese isolates -Ng, -Omu, -Mis, and -Miy. On the other hand, TYLCV-Hwas, -Jeju, and -Nons genomes shared higher nt identities with five Chinese isolates TYLCV-AH1, -ZJ3, -ZJHZ12, -SH2, -Sh10, and two Japanese isolates -Han and -Tosa. On the basis of a neighbor-joining tree, five Korean TYLCV isolates were separated into three clades. TYLCV-Bus and -Bos formed the first clade, clustering with four Japanese isolates TYLCV-Mis, -Omu, -Ng, and -Miy. TYLCV-Jeju and -Nons formed the second clade, clustering with two Chinese isolates -ZJHZ212 and -Sh10. TYLCV-Hwas was clustered with two Japanese isolates -Han and -Tosa and three Chinese isolates -AH1, -ZJ3, and -SH2. Two fragments that had a potentially recombinant origin were identified using the RDP, GENECONV, BootScan, MaxChi, Chimaera, SiScan, and 3Seq methods implemented in RDP3.41. On the basis of RDP analysis, all TYLCV isolates could originated from the interspecies recombination between TYLCV-Mld[PT] isolated from Portugal as a major parent and TYLCTHV-MM isolated from Myanmar as a minor parent.

Chemical Fungicides and Bacillus siamensis H30-3 against Fungal and Oomycete Pathogens Causing Soil-Borne Strawberry Diseases.

Chemical and biological agents were evaluated to inhibit Colletotrichum fructicola, Phytophthora cactorum, and Lasiodiplodia theobromae causing strawberry diseases. Mycelial growths of C. fructicola were gradually arrested by increasing concentrations of fungicides pyraclostrobin and iminoctadine tris (albesilate). P. cactorum and L. theobromae were more sensitive to pyraclostrobin compared to C. fructicola, but iminoctadine tris (albesilate) was not or less effective to limit P. cactorum or L. theobromae, respectively. Bacillus siamensis H30-3 was antagonistic against the three pathogens by diffusible as well as volatile molecules, and evidently reduced aerial mycelial formation of P. cactorum. B. siamensis H30-3 growth was declined by at least 0.025 mg/ml of pyraclostrobin. The two fungicides additively inhibited mycelial growths of C. fructicola, but not of P. cactorum and L. theobromae. B. siamensis H30-3 volatiles led to less growth of C. fructicola than one reduced by the fungicides. Taken together, in vitro antimicrobial activities of the two fungicides together with or without B. siamensis H30-3 volatiles may be cautiously incorporated into integrated management of strawberry diseases dependent on causal pathogens.

Menadione Sodium Bisulfite-Protected Tomato Leaves against Grey Mould via Antifungal Activity and Enhanced Plant Immunity.

Tomato grey mould has been one of the destructive fungal diseases during tomato production. Ten mM of menadione sodium bisulfite (MSB) was applied to tomato plants for eco-friendly control of the grey mould. MSB-reduced tomato grey mould in the 3rd true leaves was prolonged at least 7 days prior to the fungal inoculation of two inoculum densities (2 × 104 and 2 × 105 conidia/ml) of Botrytis cinerea. Protection efficacy was significantly higher in the leaves inoculated with the lower disease pressure of conidial suspension compared to the higher one. MSB-pretreatment was not effective to arrest oxalic acid-triggered necrosis on tomato leaves. Plant cell death and hydrogen peroxide accumulation were restricted in necrotic lesions of the B. cinereainoculated leaves by the MSB-pretreatment. Decreased conidia number and germ-tube elongation of B. cinerea were found at 10 h, and mycelial growth was also impeded at 24 h on the MSB-pretreated leaves. MSBmediated disease suppressions were found in cotyledons and different positions (1st to 5th) of true leaves inoculated with the lower conidial suspension, but only 1st to 3rd true leaves showed decreases in lesion sizes by the higher inoculum density. Increasing MSB-pretreatment times more efficiently decreased the lesion size by the higher disease pressure. MSB led to inducible expressions of defence-related genes SlPR1a, SlPR1b, SlPIN2, SlACO1, SlChi3, and SlChi9 in tomato leaves prior to B. cinerea infection. These results suggest that MSB pretreatment can be a promising alternative to chemical fungicides for environment-friendly management of tomato grey mould.

Regulation and function of the pepper pectin methylesterase inhibitor (CaPMEI1) gene promoter in defense and ethylene and methyl jasmonate signaling in plants.

Analysis of the promoters of defense-related genes is valuable for determining stress signaling and transcriptional activation during pathogen infection. Here, we have isolated and functionally characterized the promoter region of the pepper (Capsicum annuum) pectin methylesterase inhibitor 1 (CaPMEI1) gene in transiently transformed tobacco plants and stably transformed Arabidopsis plants. Among four 5' deletion constructs analyzed, the -958-bp CaPMEI1 promoter induced a high level of GUS reporter activity in tobacco leaf tissue, driven by pathogen infection as well as by ethylene and methyl jasmonate (MeJA) treatment. The 204-bp region from -958 bp to -754 bp of the CaPMEI1 promoter is responsible for the stress-responsive expression. In addition, the pepper transcription factor CARAV1 activated the CaPMEI1 promoter in tobacco leaves, whereas the transcription factor CAbZIP1 did not. In the transgenic Arabidopsis plants, the -958 bp CaPMEI1 promoter was functionally regulated by developmental cues, bacterial and oomycete pathogen infections, and treatment with ethylene and MeJA. Histochemical GUS staining analyses of Arabidopsis tissues revealed that the CaPMEI1 promoter was mainly activated in leaf veins in response to various biotic and abiotic stimuli. Together, these results suggest that CaPMEI1 promoter activation may be a critical molecular event for host defense response and ethylene- and MeJA-mediated CaPMEI1 gene expression.

Effect of Bacillus aryabhattai H26-2 and B. siamensis H30-3 on Growth Promotion and Alleviation of Heat and Drought Stresses in Chinese Cabbage.

Plants are exposed to biotic stresses caused by pathogen attack and complex abiotic stresses including heat and drought by dynamic climate changes. To alleviate these stresses, we investigated two bacterial stains, H26-2 and H30-3 in two cultivars ('Ryeokkwang' and 'Buram-3-ho') of Chinese cabbage in plastic pots in a greenhouse. We evaluated effects of bacterial strains on plant growth-promotion and mitigation of heat and drought stresses; the role of exopolysaccharides as one of bacterial determinants on alleviating stresses; biocontrol activity against soft rot caused by Pectobacterium carotovorum subsp. carotovorum PCC21. Strains H26-2 and H30-3 significantly increased fresh weights compared to a MgSO4 solution; reduced leaf wilting and promoted recovery after re-watering under heat and drought stresses. Chinese cabbages treated with H26-2 and H30-3 increased leaf abscisic acid (ABA) content and reduced stomatal opening after stresses treatments, in addition, these strains stably colonized and maintained their populations in rhizosphere during heat and drought stresses. As well as tested bacterial cells, exopolysaccharides (EPS) of H30-3 could be one of bacterial determinants for alleviation of tested stresses in Chinese cabbages, however, the effects were different to cultivars of Chinese cabbages. In addition to bacterial activity to abiotic stresses, H30-3 could suppress incidence (%) of soft rot in 'Buram-3-ho'. The tested strains were identified as Bacillus aryabhattai H26-2 and B. siamensis H30-3 based on 16S rRNA gene sequence analysis. Taken together, H26-2 and H30-3 could be candidates for both plant growth promotion and mitigation of heat and drought stresses in Chinese cabbage.

Bio-Sulfur Pre-Treatment Suppresses Anthracnose on Cucumber Leaves Inoculated with Colletotrichum orbiculare.

Bio-sulfur can be produced in the process of desulfurization from a landfill and collected by some microorganism such as Thiobacillus sp. as a sulfur element. In order to investigate practical use of bio-sulfur as an agent for controlling plant disease, in vitro antifungal activity of bio-sulfur was tested against Colletotrichum orbiculare known to cause cucumber anthracnose. Efficacy of bio-sulfur for suppressing anthracnose disease was also evaluated in vivo using cucumber leaves. Mycelial growth of C. orbiculare on medium containing bio-sulfur was inhibited. Disease severity of cucumber leaves pre-treated with bio-sulfur was significantly decreased compared to that of untreated ones. To illustrate how bio-sulfur could suppress anthracnose disease, structures of cucumber leaves infected with C. orbiculare were observed under a fluorescent microscope and a scanning electron microscope (SEM). Cucumber leaves pre-treated with bio-sulfur showed a low rate of appressorium formation whereas untreated ones showed abundant appressoria. Shrunk fungal hyphae were mostly observed on bio-sulfur-pretreated leaves by SEM. Similar results were observed on leaves pre-treated with a commercial fungicide Benomyl®. These results suggest that inhibition of appressorium formation of C. orbiculare by bio-sulfur may contribute to its suppression of cucumber anthracnose.

Nitric oxide function and signalling in plant disease resistance.

Nitric oxide (NO) is one of only a handful of gaseous signalling molecules. Its discovery as the endothelium-derived relaxing factor (EDRF) by Ignarro revolutionized how NO and cognate reactive nitrogen intermediates, which were previously considered to be toxic molecules, are viewed. NO is now emerging as a key signalling molecule in plants, where it orchestrates a plethora of cellular activities associated with growth, development, and environmental interactions. Prominent among these is its function in plant hypersensitive cell death and disease resistance. While a number of sources for NO biosynthesis have been proposed, robust and biologically relevant routes for NO production largely remain to be defined. To elaborate cell death during an incompatible plant-pathogen interaction NO functions in combination with reactive oxygen intermediates. Furthermore, NO has been shown to regulate the activity of metacaspases, evolutionary conserved proteases that may be intimately associated with pathogen-triggered cell death. NO is also thought to function in multiple modes of plant disease resistance by regulating, through S-nitrosylation, multiple nodes of the salicylic acid (SA) signalling pathway. These findings underscore the key role of NO in plant-pathogen interactions.

Ultrastructures of Colletotrichum orbiculare in Cucumber Leaves Expressing Systemic Acquired Resistance Mediated by Chlorella fusca.

Chlorella, one single-cell green algae organism that lives autotrophically by photosynthesis, can directly suppress some plant diseases. The objective of this study was to determine whether pre-spraying with Chlorella fusca suspension could induce systemic acquired resistance (SAR) in cucumber plants against anthracnose caused by Colletotrichum orbiculare. In order to illustrate SAR induced by algae, infection structures in host cells were observed under a transmission electron microscope (TEM). Cytological changes as defense responses of host mesophyll cells such as accumulation of vesicles, formation of sheath around penetration hyphae, and thickness of cell wells adjoining with intracellular hyphae were demonstrated in cucumber leaves. Similar defense responses were also found in the plant pre-treated with DL-3-aminobutyric acid, another SAR priming agent. Images showed that defense response of host cells was scarcely observed in untreated leaf tissues. These cytological observations suggest that C. fusca could induce SAR against anthracnose in cucumber plants by activating defense responses of host cells.

Reduced Bacterial Wilt in Tomato Plants by Bactericidal Peroxyacetic Acid Mixture Treatment.

Peroxyacetic acid mixture Perosan, composed of peroxyacetic acid, hydrogen peroxide and acetic acid, was evaluated for eco-friendly management of tomato bacterial wilt by Ralstonia pseudosolanacearum. Perosan drastically suppressed in vitro growth of R. pseudosolanacearum in liquid cultures in dose- and incubation time-dependent manners. Higher perosan doses (0.1 and 1%) caused lowered pH and phytotoxicity to detached leaves of two tomato cultivars Cupirang and Benekia 220 in aqueous solution. Treatment with 0.01% of Perosan delayed wilting symptom significantly in the detached leaves of two cultivars inoculated with R. pseudosolanacearum (107 cfu/ml). Soil drenching of 5% Perosan solution in pots caused severe tissue collapse of tomato seedlings at the four-week-old stage of two tomato cultivars. Treatment with 1% Perosan by soil-drenching significantly reduced bacterial wilt in the tomato seedlings of two cultivars. These findings suggest that Perosan treatment can be applied to suppress bacterial wilt during tomato production.

Enhanced Tolerance of Chinese Cabbage Seedlings Mediated by Bacillus aryabhattai H26-2 and B. siamensis H30-3 against High Temperature Stress and Fungal Infections.

Two rhizobacteria Bacillus aryabhattai H26-2 and B. siamensis H30-3 were evaluated whether they are involved in stress tolerance against drought and high temperature as well as fungal infections in Chinese cabbage plants. Chinese cabbage seedlings cv. Ryeokgwang (spring cultivar) has shown better growth compared to cv. Buram-3-ho (autumn cultivar) under high temperature conditions in a greenhouse, whilst there was no difference in drought stress tolerance of the two cultivars. In vitro growth of B. aryabhattai H26-2 and B. siamensis H30-3 were differentially regulated under PEG 6000-induced drought stress at different growing temperatures (30, 40 and 50°C). Pretreatment with B. aryabhattai H26-2 and B. siamensis H30-3 enhanced the tolerance of Chinese cabbage seedlings to high temperature, but not to drought stress. It turns out that only B. siamensis H30-3 showed in vitro antifungal activities and in planta crop protection against two fungal pathogens Alternaria brassicicola and Colletotrichum higginsianum causing black spots and anthracnose on Chinese cabbage plants cv. Ryeokgwang, respectively. B. siamensis H30-3 brings several genes involved in production of cyclic lipopeptides in its genome and secreted hydrolytic enzymes like chitinase, protease and cellulase. B. siamensis H30-3 was found to produce siderophore, a high affinity iron-chelating compound. Expressions of BrChi1 and BrGST1 genes were up-regulated in Chinese cabbage leaves by B. siamensis H30-3. These findings suggest that integration of B. aryabhattai H26-2 and B. siamensis H30-3 in Chinese cabbage production system may increase productivity through improved plant growth under high temperature and crop protection against fungal pathogens.