• Journal of Microbiology and Biotechnology
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• v.21 no.7
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• pp.679-685
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• 2011

Xanthomonas oryzae pv. oryzae (Xoo) produces a putative effector, XoAvrBs2. We expressed XoAvrBs2 homologously in Xoo with a TAP-tag at the C-terminus to enable quantitative analysis of protein expression and secretion. Addition of rice leaf extracts from both Xoo-sensitive and Xoo-resistant rice cultivars to the Xoo cells induced expression of the XoAvrBs2 gene at the transcriptional and translational levels, and also stimulated a remarkable amount of XoAvrBs2 secretion into the medium. In a T3SS-defective Xoo mutant strain, secretion of the TAPtagged XoAvrBs2 was blocked. Thus, we elucidated the transcriptional and translational expressions of the XoAvrBs2 gene in Xoo was induced in vitro by the interaction with rice and the induced secretion of XoAvrBs2 was T3SSdependent. It is the first report to measure the homologous expression and secretion of XoAvrBs2 in vitro by rice leaf extract. Our system for the quantitative analysis of effector protein expression and secretion could be generally used for the study of host-pathogen interactions.

• The Plant Pathology Journal
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• v.36 no.3
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• pp.289-296
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• 2020

Type VI secretion system (T6SS) is a contact-dependent secretion system, employed by most gram-negative bacteria for translocating effector proteins to target cells. The present study was conducted to investigate T6SS in Xanthomonas oryzae pv. oryzae (Xoo), which causes bacterial blight in rice, and to unveil its functions. Two T6SS clusters were found in the genome of Xoo PXO99^A. The deletion mutants, Δhcp1, Δhcp2, and Δhcp12, targeting the hcp gene in each cluster, and a double-deletion mutant targeting both genes were constructed and tested for growth rate, pathogenicity to rice, and inter-bacterial competition ability. The results indicated that hcp in T6SS-2, but not T6SS-1, was involved in bacterial virulence to rice plants. However, neither T6SS-1 nor T6SS-2 had any effect on the ability to compete with Escherichia coli or other bacterial cells. In conclusion, T6SS gene clusters in Xoo have been characterized, and its role in virulence to rice was confirmed.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.562-569
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• 2024

Xanthomonas oryzae pv. oryzae (Xoo) causes a devastating bacterial leaf blight in rice. Here, the antimicrobial effects of ᴰ-limonene, ᴸ-limonene, and its oxidative derivative carveol against Xoo were investigated. We revealed that carveol treatment at ≥ 0.1 mM in liquid culture resulted in significant decrease in Xoo growth rate (> 40%) in a concentration-dependent manner, and over 1 mM, no growth was observed. The treatment with ᴰ-limonene and ᴸ-limonene also inhibited the Xoo growth but to a lesser extent compared to carveol. These results were further elaborated with the assays of motility, biofilm formation and xanthomonadin production. The carveol treatment over 1 mM caused no motilities, basal level of biofilm formation (< 10%), and significantly reduced xanthomonadin production. The biofilm formation after the treatment with two limonene isomers was decreased in a concentration-dependent manner, but the degree of the effect was not comparable to carveol. In addition, there was negligible effect on the xanthomonadin production mediated by the treatment of two limonene isomers. Field emission-scanning electron microscope (FE-SEM) unveiled that all three compounds used in this study cause severe ultrastructural morphological changes in Xoo cells, showing shrinking, shriveling, and holes on their surface. Moreover, quantitative real-time PCR revealed that carveol and ᴰ-limonene treatment significantly down-regulated the expression levels of genes involved in virulence and biofilm formation of Xoo, but not with ᴸ-limonene. Together, we suggest that limonenes and carveol will be the candidates of interest in the development of biological pesticides.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.655-659
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• 2000

A 11.5-kb DNA fragment containing an endo-inulinase gene was cloned from Xanthomonas oryzae #5. It contained a single open reading frame of 3,999bp, encoding a polypeptide composed of signal peptide of 32 amino acids and mature protein of 1,301 amino acids. From the comparison of amino acids sequences with fructan hydrolases, inulinase, levanase and CFTase, the sequence of the endo-inulinase had highly homology of 72% with CFTase of B. circulans, and six highly conserved regions including the ${\beta}-fructosidase$ motif were found.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.740-747
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• 2014

Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a very serious disease in rice-growing regions of the world. In spite of their economic importance, there are no effective ways of protecting rice plants from this disease. Bacteriophages infecting Xoo affect the population dynamics of the pathogen and consequently the occurrence of the disease. In this study, we investigated the diversity, host range, and infectivity of Xoo phages, and their use as a bicontrol agent on BLB was tested. Among the 34 phages that were isolated from floodwater in paddy fields, 29 belonged to the Myoviridae family, which suggests that the dominant phage in the ecosystem was Myoviridae. The isolated phages were classified into two groups based on plaque size produced on the lawn of Xoo. In general, there was a negative relationship between plaque size and host range, and interestingly the phages having a narrow host range had low efficiency of infectivity. The deduced protein sequence analysis of htf genes indicated that the gene was not a determinant of host specificity. Although the difference in host range and infectivity depending on morphotype needs to be addressed, the results revealed deeper understanding of the interaction between the phages and Xoo strains in floodwater and damp soil environments. The phage mixtures reduced the occurrence of BLB when they were treated with skim milk. The results indicate that the Xoo phages could be used as an alternative control method to increase the control efficacy and reduce the use of agrochemicals.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.494-500
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• 2010

Bacterial blight, an important and potentially destructive bacterial disease in rice, is caused by Xanthomonas oryzae. Recently, this organism has developed resistance to available antibiotics, prompting scientists to find a suitable alternative. This study focused on secondary metabolites of Phomopsis longicolla to target X. oryzae. Five bioactive compounds were isolated by activity-guided fractionation from ethyl acetate extracts of mycelia and were identified by LC/MS and NMR spectroscopy as dicerandrol A, dicerandrol B, dicerandrol C, deacetylphomoxanthone B, and fusaristatin A. This is the first time fusaristatin A has been isolated from Phomopsis sp. Deacetylphomoxanthone B showed a higher antibacterial effect against X. oryzae KACC 10331 than the positive control (2,4-diacetyphloroglucinol). Dicerandrol A also showed high antimicrobial activity against Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis) and yeast (Candida albicans). In addition, high production yields of these compounds were obtained at the stationary and death phases.

• Microbiology and Biotechnology Letters
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• v.46 no.4
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• pp.346-359
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• 2018

Xanthomonas oryzae, a bacterial pathogen causing leaf blight disease (BLB) in rice, can cause widespread disease and has caused epidemics globally, resulting in severe crop losses of 50% in Asia. The pathogen is seed-borne and is transmitted through seeds. Thus, control of BLB requires the elimination of the pathogen from seeds. Concern about environment-friendly organic production has spurred improvements in a variety of biological disease control methods, including the use of bacteriophages, against bacterial plant pathogens. The present study explored the potential of bacteriophages isolated from diseased plant leaves and soil samples in killing the bacterial pathogen in rice seeds. Eight different phages were isolated and evaluated for their bacteriolytic activity against different pathogenic X. oryzae strains. Of these, a phage designated ${\varphi}XOF4$ killed all the pathogenic X. oryzae strains and showed the broadest host range. Transmission electron microscopy of ${\varphi}XOF4$ revealed it to be a tailed phage with an icosahedral head. The virus was assigned to the family Siphoviridae, order Caudovirales. Seedlings raised from the seeds treated with $1{\times}10^8pfu/ml$ of ${\varphi}XOF4$ phage displayed reduced incidence of BLB disease and complete bacterial growth inhibition. The findings indicate the potential of the ${\varphi}XOF4$ phage as a potential biological control agent against BLB disease in rice.

• Research in Plant Disease
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• v.17 no.1
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• pp.90-94
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• 2011

Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a very serious disease in rice growing regions of the world. There are no effective ways of protecting rice from the disease. In this study, the bacteriophage (phage) mixtures infecting Xoo were investigated as biological control agent on BLB. The effects of pH, heat and ultraviolet on the stability of phages were investigated to check and increase the possibility of practical use in the field. Phages were rather stable between pH 5 and pH 10. The infectivity dropped sharply when the phages were incubated at $50^{\circ}C$ and more than 90% of the phages were inactivated after two minutes of ultraviolet treatment. The phages were stable for 7 days at the rice plant leaves, and the phages survived 10 times more than other treatments when mixed with skim milk. Although the skim milk increased the stability of the phages, the control efficacy was not effective. However, the phage mixtures reduced the occurrence of BLB when they were treated with Tecloftalam WP or Acibenzolar-S-methyl simultaneously. The results indicated that the Xoo phages could be used as an alternative control method to increase the control efficacy and reduce the use of agrochemicals.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1995.06b
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• pp.21-25
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• 1995

;Bacterial blight caused by Xanthomonas oryzae pv. Olyzae is one of the most important diseases of rice. Host resistance, which relies on single, dominant resistance genes, is the only reliable method to control the disease at present. Pathogenic variation of the bacteria has been shown to follow the deployment of resistance genes in commercial cultivars. Information on the factors and the mechanisms for genetic variation of this pathogen is limited. Further, we have no clear evidence of whether population variability is due to sexual recombination or to variation introduced by mutations or intragenic recombination in a clonally maintained population.(omitted)itted)

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2005.04a
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• pp.49.1-49.1
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• 2005