• Journal of Microbiology and Biotechnology
• /
• v.19 no.2
• /
• pp.194-203
• /
• 2009

$HpaG_{Xooc}$, from rice pathogenic bacterium Xanthomonas oryzae pv. oryzicola, is a member of the harpin group of proteins, eliciting hypersensitive cell death in non-host plants, inducing disease and insect resistance in plants, and enhancing plant growth. To express and secret the $HpaG_{Xooc}$ protein in Bacillus subtilis, we constructed a recombinant expression vector pM43HF with stronger promoter P43 and signal peptide element nprB. The SDS-PAGE and Western blot analysis demonstrated the expression of the protein $HpaG_{Xooc}$ in B. subtilis. The ELISA analysis determined the optimum condition for $HpaG_{Xooc}$ expression in B. subtilis WBHF. The biological function analysis indicated that the protein $HpaG_{Xooc}$ from B. subtilis WBHF elicits hypersensitive response(HR) and enhances the growth of tobacco. The results of RT-PCR analysis revealed that $HpaG_{Xooc}$ induces expression of the pathogenesis-related genes PR-1a and PR-1b in plant defense response.

• Molecules and Cells
• /
• v.27 no.5
• /
• pp.563-570
• /
• 2009

We previously isolated the OsCBT gene, which encodes a calmodulin (CaM)-binding protein, from a rice expression library constructed from fungal elicitor-treated rice suspension cells. In order to understand the function of OsCBT in rice, we isolated and characterized a T-DNA insertion mutant allele named oscbt-1. The oscbt-1 mutant exhibits reduced levels of OsCBT transcripts and no significant morphological changes compared to wild-type plant although the growth of the mutant is stunted. However, oscbt-1 mutants showed significant resistance to two major rice pathogens. The growth of the rice blast fungus Magnaporthe grisea, as well as the bacterial pathogen Xanthomonas oryzae pv. oryzae was significantly suppressed in oscbt-1 plants. Histochemical analysis indicated that the hypersensitive-response was induced in the oscbt-1 mutant in response to compatible strains of fungal pathogens. OsCBT expression was induced upon challenge with fungal elicitor. We also observed significant increase in the level of pathogenesis-related genes in the oscbt-1 mutant even under pathogen-free condition. Taken together, the results support an idea that OsCBT might act as a negative regulator on plant defense.

• The Plant Pathology Journal
• /
• v.31 no.2
• /
• pp.152-164
• /
• 2015

Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.56 no.2
• /
• pp.124-133
• /
• 2011

Bacterial blight (BB), cuased by the vascular pathogen Xanthomonas oryzae pv. oryzae, is one of the major threats in rice fields worldwide. In Korea, two resistance genes against BB, Xa1 and Xa3 had been intensively used for developing high quality japonica rice cultivars. Those traditional resistance sources have being rapidly ified by the adopting of BB pathogen through mutations of the corresponding avr-genes, such as K3a exhibiting high compatibility to both Xa1 and Xa3. To expanding genetic resource against BB in Korea, the Suweon506, an introgression line between a Korean japonica cultivar, Hwaseong and a wild relative, Oryza minuta, was be subjected for genetic analysis owing to the BB resistance. Through association analyses between the pathotyping and genotyping results for each $F_2$ progenies, derived from a cross between Suweon506 and a Tongil type cultivar, Milyang23, a major resistant dominant gene is localized on the subterminal region of rice chromosome 4, where at least three BB resistancde genes, Xa1, Xa2, and Xa22, were reported previously.

• Journal of Life Science
• /
• v.25 no.10
• /
• pp.1091-1097
• /
• 2015

We have isolated and characterized an ascorbate peroxidase (APx) gene, OsAPx1 from rice. Northern and Western blot analyses indicated that at young seedling stage, OsAPx1 mRNA was expressed highly in root, shoot apical meristem (SAM) and leaf sheath than leaf. In mature plant, OsAPx1 gene expressed highly in root, stem and flower but weakly in leaf. OsAPx1 gene and protein expression level was induced in leaves inoculated with Magnaporthe oryzae (M. oryzae) and Xanthomonas oryzae pv. oryzae (Xoo). Phytohormones treatment showed that OsAPx1 was up-regulated by jasmonic acid (JA), but was down regulated by ABA and SA co-treatments with JA, resulting that they have antagonistic effect on pathogen responsive OsAPx1 expression. Phylogenetic analysis illustrated that Arabidopsis AtAPx1 has a close relationship with OsAPx1. In AtAPx1 knock out lines, the accumulation of O₂^- and H₂O₂ are all highly detected than wild type, revealing that the high concentration of exogenous H₂O₂ cause the intercellular superoxide anion and hydrogen peroxide accumulation in AtAPx1 knockout plant. These results suggested that OsAPx1 gene may be associated with the pathogen defense cascades as the mediator for balancing redox state by acting ROS scavenger and is associated with response to the pathogen defense via Jasmonic acid signaling pathway.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.65 no.4
• /
• pp.314-326
• /
• 2020

This study was carried out to develop a resistant variety against the K3a race of bacterial blight, Xanthomonas oryzae pv. oryzae, through expansion and pyramiding of resistance genes. To develop an elite bacterial blight-resistant cultivar, the breeding process and bacterial blight resistance reactions in advanced backcross lines (ABLs) were analyzed. ABLs21 which contain Xa3 and Xa21, were developed by double backcrossing japonica cultivar Hwanggeumnuri, which has bacterial blight resistant Xa3 gene, and indica variety IRBB21, which havs Xa21 gene, followed by disease resistance bioassay and marker-assisted selection. The resistance genes of ABLs21 were amplified by PCR with the molecular markers 9643.T4 (Xa3) and U1/I1 (Xa21). Hwanggeumnuri and IRBB3 showed resistance reactions against K1, K2, and K3 races, and a susceptible reaction against K3a, K4, and K5 races. IRBB21 showed resistance reactions against K2, K3, K3a, K4 and K5 races, and a susceptible reaction against K1 race. Hwanggeumnuri showed susceptible reactions at the seedling, tillering and adult stages (all stages), whereas ABL21-1 showed moderate resistance at the tillering stage. ABL21-1 showed stable resistance against 18 isolates of K3a race, and the lesion length was shorter than that of the donor parents. In cluster analysis, the HB4032 isolate showed the highest pathogenicity among the 18 isolates. The molecular marker polymorphisms and average substituted chromosome segment lengths of ABLs21 were 63.2 % and 86.1 cM, respectively. Insertion of the donor chromosomal segments occurred in the predicted region of the Xa21 gene of ABLs21.