Rice Bacterial Blight Disease Research Articles

Strategies for management of bacterial blight disease of rice caused by Xanthomonas oryzae pv. Oryzae (small oryzae): an overview.

26Jul 2016 Strategies for management of bacterial blight disease of rice caused by Xanthomonas oryzae pv. Oryzae (small oryzae): an overview. Ashwarya L. Tandon and S. D. Chaliganjewar. Department of Plant Pathology, College of Agriculture & Research Station, Janjgir-Champa (C.G.)- 495668. India. Department of Plant Pathology, Indira Gandhi Krishi Vishwavidyalaya, Raipur (C.G.)- 492006. India.

Genotypic and Pathotypic Diversity of Xanthomonas oryzae pv. oryzae Strains in Taiwan

AbstractRice bacterial leaf blight, caused by Xanthomonas oryzae pv. oryzae [(Ishiyama) Swings et al. 1990] (Xoo), is a major rice disease of the second crop season in Taiwan. A total of 88 Xoo strains collected from 10 major rice cultivating areas in Taiwan from 1986, 1997, 2000, 2004, and 2011 were characterized by repetitive‐element PCR (REP‐PCR) fingerprinting and virulence analyses. Among the five genetic clusters identified by the pJEL1/pJEL2 (IS1112‐based) and REP1R‐Dt/REP2‐D [repetitive extragenic palindromic (REP)‐based] primer sets, clusters A, C and D contained Xoo strains from geographically distant regions, which suggests a high frequency of Xoo dispersal in Taiwan. The 88 Xoo strains were evaluated by inoculations on IRBB near‐isogenic lines and five Taiwan rice cultivars. A subset of 45 moderately or highly virulent strains were classified into 15 pathotypes by their compatible or incompatible reactions on IR24 and 12 IRBB near‐isogenic lines, each containing a single resistance gene. Analysis of molecular haplotypes and pathotypes revealed a partial relationship. IRBB5, IRBB21 and IRBB4 were incompatible with 96%, 96% and 73% of the strains, so xa5, Xa21 and Xa4 can recognize most of the Xoo strains in Taiwan and elicit resistance. In contrast, IRBB3 (Xa3), IRBB8 (xa8), IRBB10 (Xa10), IRBB11 (Xa11), IRBB13 (xa13) and IRBB14 (Xa14) were susceptible to almost all of the 45 Xoo strains. Inoculation trials revealed significant differences in the susceptibility of five Taiwan cultivars to Xoo (from high to low susceptibility: Taichung Sen 10 > IR24, Taichung Native 1 > Taichung 192, Taikeng 9, Tainan 11). This study provides useful information for resistance breeding and the development of disease management strategies against bacterial blight disease of rice.

Suppressing activity of staurosporine from Streptomyces sp. MJM4426 against rice bacterial blight disease.

To identify the active compounds from the Streptomyces sp. MJM4426 that can protect rice from bacterial blight disease (BB), and to evaluate the potential of this Streptomyces strains and the compound for biocontrol of rice bacterial blight disease. The ethyl acetate extract of Streptomyces sp. MJM4426 can significantly protect rice leaf explants from the infection of Xanthomonas oryzae pv. oryzaeKACC 10331 (Xoo), the pathogen which cause BB. To identify the active compounds, the ethyl acetate extract of Streptomyces sp. MJM4426 was fractionated through a Sephadex LH-20 column chromatography, and further purified by preparative HPLC guided by the inhibitory activity against BB in rice leaf explants. UPLC-Q-TOF/MS analysis showed the active compound displayed its m/z values at [M+H](+) 467·2086 and [M+FA-H](-) 511·1963, and the molecular formula was estimated as C28 H26 N4 O3 which is identical to commercial standard staurosporine. In this study, the isolated staurosporine dramatically suppressed bacterial blight in rice leaf explants with the lowest concentration at 12·5 μmol l(-1) , however, it exhibited low inhibitory activity against Xoo with the MIC value at 256 μg ml(-1) . In addition, greenhouse study showed both crude extract and purified staurosporine can suppress the bacterial blight at the concentration of 5000 and 200 μg ml(-1) respectively. Streptomyces sp. MJM4426 can protect rice leaf explants from the infection of Xoo by producing staurosporine, but not by direct inhibitory activity against Xoo. This is the first report that staurosporine can protect rice leaf against bacterial blight disease and showed the potential of Streptomyces sp. MJM4426 as an alternative to chemical bactericide for bacterial blight disease in rice.

OsBAK1 is involved in rice resistance to Xanthomonas oryzae pv. oryzae PXO99

OsBAK1 gene belongs to a receptor like kinase gene family in rice and shares a highly conserved gene structure and sequence homology with Arabidopsis thaliana BAK1 gene. To investigate the role of OsBAK1 in rice immunity, the full-length cDNA of OsBAK1 was isolated by RT-PCR and the transgenic rice lines (over expression and RNA-interference lines) were generated using Agrobacterium-mediated transformation. The expression level of OsBAK1 was determined by q-PCR in overexpression and RNAi transgenic rice lines. Based on quantitative polymerase chain reaction (q-PCR) results, two overexpression lines and two RNAi lines were evaluated in bioassays for resistance to Xanthomonas oryzae pv. oryzae PXO99, the causal agent of rice bacterial blight disease. Pathogenicity bioassays showed overexpression OsBAK1 lines exhibited resistance to blight disease whereas OsBAK1 RNAi lines promoted susceptibility. Besides, OsBAK1 can complement the function of AtBAK1 in Arabidopsis bak1 protoplast, activating FRK1 expression, a marker gene in PTI signaling pathway, after treatment by flg22. Furthermore, the transcriptional level of OsBAK1 was induced significantly in rice by defense signaling molecules such as salicylic acid, jasmonic acid, and PXO99 inoculation. Our results illustrated OsBAK1 positively regulates plant defense against rice bacterium pathogen Xanthomonas oryzae pv. oryzae PXO99.

Hydrogen peroxide is indispensable to Xanthomonas oryzae pv. oryzae-induced hypersensitive response and nonhost resistance in Nicotiana benthamiana

Nonhost resistance is a durable and broad-spectrum resistance in plants that are outside of the given pathogens’ host range. Previously, we reported that the important pathogen Xanthomonas oryzae pv. oryzae (Xoo), which causes bacterial blight disease in rice, rapidly triggers the hypersensitive response (HR) and nonhost resistance in its nonhost plant Nicotiana benthamiana. In this study, we optimized the conditions, including the inoculum concentration, the leaf positions for inoculation and the plant post-inoculation growth temperature, to induce Xoo-triggered HR and nonhost resistance. We demonstrated that inoculating with a Xoo concentration of 1 × 108 cfu mL−1 in fully expanded leaves of N. benthamiana plants resulted in the strongest induction of HR and nonhost resistance. Furthermore, we probed the role of hydrogen peroxide (H2O2) in Xoo-induced HR and nonhost resistance. An exogenous supply of H2O2 accelerated Xoo-induced HR, while the elimination of H2O2 by the application of a catalase blocked the HR-mediated suppression of bacterial growth in N. benthamiana. Moreover, ∆hrpD6 and ∆hrcU Xoo mutants, which are deficient in H2O2 production, did not show an induction of HR, while another Xoo mutant, ∆hpa1, caused a strong H2O2 accumulation prior to its delayed HR induction. Collectively, our results reveal that H2O2 is indispensable to Xoo-induced HR and nonhost resistance in N. benthamiana.

Dynamic and Coordinated Expression Changes of Rice Small RNAs in Response to Xanthomonas oryzae pv. oryzae

Endogenous small RNAs are newly identified players in plant immune responses, yet their roles in rice (Oryza sativa) responding to pathogens are still less understood, especially for pathogens that can cause severe yield losses. We examined the small RNA expression profiles of rice leaves at 2, 6, 12, and 24 hours post infection of Xanthomonas oryzae pv. oryzae (Xoo) virulent strain PXO99, the causal agent of rice bacterial blight disease. Dynamic expression changes of some miRNAs and trans-acting siRNAs were identified, together with a few novel miRNA targets, including an RLK gene targeted by osa-miR159a.1. Coordinated expression changes were observed among some small RNAs in response to Xoo infection, with small RNAs exhibiting the same expression pattern tended to regulate genes in the same or related signaling pathways, including auxin and GA signaling pathways, nutrition and defense-related pathways. These findings reveal the dynamic and complex roles of small RNAs in rice-Xoo interactions, and identify new targets for regulating plant responses to Xoo.

Identification and fine-mapping of a new resistance gene, Xa40, conferring resistance to bacterial blight races in rice (Oryza sativa L.).

A new bacterial blight resistance gene has been identified through fine-mapping, which confers high levels of resistance to all Korean Xanthomonas oryzae pv. oryzae (Xoo) races, including the new Xoo race K3a. Rice bacterial leaf blight (BB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) is a serious constraint to rice production in Asia and Africa. The japonica advanced backcross breeding lines derived from the indica line IR65482-7-216-1-2 in the background of cultivar Junam are resistant to all Korean BB races, including K3a. To identify the gene(s) involved in resistance to Korean Xoo races, the association of genotypic and phenotypic variations was examined in two F2 populations derived from the crosses between 11325 (IR83261-3-7-23-6-2-1-1-2-1-2)/Anmi and 11325/Ilpum. The segregation ratios of F2 individuals from the crosses of 11325/Anmi and 11325/Ilpum were 578 resistant:209 susceptible and 555 resistant:241 susceptible, respectively, which is consistent with the expected allelic frequency of a 3:1 ratio. Genetic analysis using graphical mapping indicated that resistance (R) was controlled by a new resistance gene linked with the flanking markers RM27320 and ID55.WA18-5 within an approximately 80-kb region between 28.14 and 28.22 Mbp on chromosome 11. The eight candidate genes functionally predicted were included in the target region. Examination of the candidate genes by RT-PCR analysis only corroborated with the significant difference in transcript levels of the WAK3 gene in the presence or absence of pathogen infection. Allelism tests performed with other known BB R-genes revealed that the allele was distinct from others having a similar chromosomal location.

Comparative proteomics reveals differential induction of both biotic and abiotic stress response associated proteins in rice during Xanthomonas oryzae pv. oryzae infection

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight disease in rice and brutally affects the yield up to 50% of total production. Here, we report a comparative proteomics analysis of total foliar protein isolated from infected rice leaves of susceptible Pusa Basmati 1 (PB1) and resistant Oryza longistaminata genotypes. Two-dimensional gel electrophoresis (2-DE) coupled with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) approaches identified 29 protein spots encoding unique proteins from both the genotypes. Identified proteins belonged to a large number of biological and molecular functions related to biotic and abiotic stress proteins which are potentially involved during Xoo infection. Biotic and abiotic stress-related proteins were induced during Xoo infection, indicating the activation of common stress pathway during bacterial blight infection. Candidate genes conferring tolerance against bacterial blight, which include germin-like protein, putative r40c1, cyclin-dependent kinase C, Ent-isokaur-15-ene synthase and glutathione-dependent dehydroascorbate reductase 1 (GSH-DHAR1), were also induced, with germin-like proteins induced only in the resistant rice genotype O. longistaminata. Energy, metabolism and hypothetical proteins were common among both the genotypes. Further, host defence/stress-related proteins were mostly expressed in resistant genotype O. longistaminata, indicating possible co-evolution of the pathogen and the wild rice, O. longistaminata.

Genetic engineering of the Xa10 promoter for broad-spectrum and durable resistance to Xanthomonas oryzae pv. oryzae.

Many pathovars of plant pathogenic bacteria Xanthomonas species inject transcription activator-like (TAL) effectors into plant host cells to promote disease susceptibility or trigger disease resistance. The rice TAL effector-dependent disease resistance gene Xa10 confers narrow-spectrum race-specific resistance to Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of bacterial blight disease in rice. To generate broad-spectrum and durable resistance to Xoo, we developed a modified Xa10 gene, designated as Xa10(E5) . Xa10(E5) has an EBE-amended promoter containing 5 tandemly arranged EBEs each responding specifically to a corresponding virulent or avirulent TAL effector and a stable transgenic rice line containing Xa10(E5) was generated in the cultivar Nipponbare. The Xa10(E5) gene was specifically induced by Xoo strains that harbour the corresponding TAL effectors and conferred TAL effector-dependent resistance to the pathogens at all developmental stages of rice. Further disease evaluation demonstrated that the Xa10(E5) gene in either Nipponbare or 9311 genetic backgrounds provided broad-spectrum disease resistance to 27 of the 28 Xoo strains collected from 11 countries. The development of Xa10(E5) and transgenic rice lines provides new genetic materials for molecular breeding of rice for broad-spectrum and durable disease resistance to bacterial blight.

Proteomic and functional analyses of a novel porin-like protein in Xanthomonas oryzae pv. oryzae.

Proteomic analysis is a useful technique for postulating and elucidating protein functions. In the present work, a shotgun proteomic analysis was used to identify functions of the PXO_03968 gene (previously known as the ax21) from Xanthomonas oryzae pv. oryzae (Xoo), a causal agent for bacterial blight disease in rice. Structural prediction performed on the protein sequence encoded by PXO_03968 reveals that it encodes a putative porin-like protein, possessing a β-barrel domain with 10 β-strands and a signal peptide at the N-terminus. We renamed the gene as an omp1X (outer membrane protein 1 in Xoo), generated its knock out mutant (XooΔomp1X), and compared the protein expression level in the mutant to that in the wild type. A total of 106 proteins displayed more than 1.5-fold difference in expression between the mutant and the wild type strains. COG analysis revealed that these proteins are involved in cell motility as well as signal transduction. In addition, phenotypic analysis demonstrated that motility and biofilm formation in XooΔomp1X are lower than the wild type. These results provide new insights into the functions of outer membrane proteins in Gram-negative bacteria.

Development of Defense Signaling Pathways Against Bacterial Blight Disease in Rice Using Genome-Wide Transcriptome Data

Bacterial blight (BB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) drives severe yield and quality losses in rice (Oryza sativa Xa1, Xa3/Xa26, xa5, xa13, Xa21, and Xa27). Here we employ a transcriptomics approach to elucidate the Xa21-, NH1- (NPR1 homolog 1) (NH1)-, and NRR- (negative regulator of disease resistance) mediated defense response to Xoo. Among the candidate genes, we focused on 288 genes showing significant change in at least two of the above comparisons to support the association with an enhanced defense response. Gene Ontology enrichment analysis for this gene list revealed that response to biotic stimulus was 25.0-fold more enriched compared to the control, well qualifying the candidate genes for enhanced resistant response. The biotic stress overview installed in the MapMan toolkit was used to identify diverse components consisting of defense signaling pathways such as genes involved in disease resistance, redox, signaling, regulation of transcription, pathogenesis-related functions, secondary metabolism, and protein degradation. Of these, we validated the expression patterns of genes related to regulation of transcription and pathogenesis-related functions and suggest a functional network model for WRKY transcription factors mediating defense signaling pathways against Xoo. We expect that our analysis will contribute to increasing the depth of knowledge on the molecular mechanism for enhanced disease resistance against bacterial blight disease in rice.

Biological control of Xanthomonas Oryzae pv. Oryzae causing rice bacterial blight disease by Streptomyces toxytricini VN08-A-12, isolated from soil and leaf-litter samples in Vietnam.

A total of 2690 actinomycete strains were screened as potential biological control agents in controlling rice bacterial blight (BB) in Vietnam. From these microorganisms, seventeen actinomycete strains were found to be capable of inhibiting all 10 major Xoo races isolated from Xoo-infected rice leaves. One strain, namely VN08-A-12, contained effective characteristics in selectively inhibiting all 10 races in vitro, but did not inhibit most of the other tested microorganisms. Therefore, VN08-A-12 was subsequently selected for rice field trials for two seasons on two rice cultivars SS1 and KD18. Results showed VN08-A-12 was not only able to reduce Xoo lesion lengths in the two rice cultivars (lesion length reduction of up to 38.3%), but it also significantly reduced Xoo-related yield loss in infected rice cultivars from the field (yield loss reduction of up to 43.2%). Interestingly, the culture of this strain also increased the rice yield in healthy rice cultivars (from 2.66% to 16.98% for SS1 and from 3.11% to 5.94% for KD18 cultivar). The strain VN08-A-12 was shown to be identical to Streptomyces toxytricini. To our knowledge, this is the first study reporting S. toxytricini as a beneficial biological agent for the control of BB in rice.

Prediction and characterization of protein–protein interaction network in Xanthomonas oryzae pv. oryzae PXO99A

Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of bacterial blight disease in rice, is one of the most serious plant pathogens worldwide. In the current analysis, we constructed a protein–protein interaction network of Xoo strain PXO99A with two computational approaches (interolog method and domain combination method), and verified by K-Nearest Neighbors classification method. The predicted PPI network of Xoo PXO99A contains 36,886 interactions among 1988 proteins. KNN verification and GO annotation confirm the reliability of the network. Detailed analysis of flagellar synthesis and chemotaxis system shows that σ factors (especially σ28, σ54) in Xoo PXO99A are very important for flagellar synthesis and motility, and transcription factors RpoA, RpoB and RpoC are hubs to connect most σ factors. Furthermore, Xoo PXO99A may have both cAMP and c-di-GMP signal transduction system, and the latter is especially important for this plant pathogen. This study therefore provides valuable clues to explore the pathogenicity and metabolic regulation of Xoo PXO99A.

Molecular Phylogeny, Homology Modeling, and Molecular Dynamics Simulation of Race-Specific Bacterial Blight Disease Resistance Protein (xa5) of Rice: A Comparative Agriproteomics Approach

Rice (Oryza sativa L.), a model plant belonging to the family Poaceae, is a staple food for a majority of the people worldwide. Grown in the tropical and subtropical regions of the world, this important cereal crop is under constant and serious threat from both biotic and abiotic stresses. Among the biotic threats, Xanthomonas oryzae pv. oryzae, causing the damaging bacterial blight disease in rice, is a prominent pathogen. The xa5 gene in the host plant rice confers race-specific resistance to this pathogen. This recessive gene belongs to the Xa gene family of rice and encodes a gamma subunit of transcription factor IIA (TFIIAγ). In view of the importance of this gene in conferring resistance to the devastating disease, we reconstructed the phylogenetic relationship of this gene, developed a three-dimensional protein model, followed by long-term molecular dynamics simulation studies to gain a better understanding of the evolution, structure, and function of xa5. The modeled structure was found to fit well with the small subunit of TFIIA from human, suggesting that it may also act as a small subunit of TFIIA in rice. The model had a stable conformation in response to the atomic flexibility and interaction, when subjected to MD simulation at 20 nano second in aqueous solution. Further structural analysis of xa5 indicated that the protein retained its basic transcription factor function, suggesting that it might govern a novel pathway responsible for bacterial blight resistance. Future molecular docking studies of xa5 underway with its corresponding avirulence gene is expected to shed more direct light into plant-pathogen interactions at the molecular level and thus pave the way for richer agriproteomic insights.

Members of the XB3 Family from Diverse Plant Species Induce Programmed Cell Death in Nicotiana benthamiana

Programmed cell death has been associated with plant immunity and senescence. The receptor kinase XA21 confers resistance to bacterial blight disease of rice (Oryza sativa) caused by Xanthomonas oryzae pv. oryzae (Xoo). Here we show that the XA21 binding protein 3 (XB3) is capable of inducing cell death when overexpressed in Nicotiana benthamiana. XB3 is a RING finger-containing E3 ubiquitin ligase that has been positively implicated in XA21-mediated resistance. Mutation abolishing the XB3 E3 activity also eliminates its ability to induce cell death. Phylogenetic analysis of XB3-related sequences suggests a family of proteins (XB3 family) with members from diverse plant species. We further demonstrate that members of the XB3 family from rice, Arabidopsis and citrus all trigger a similar cell death response in Nicotiana benthamiana, suggesting an evolutionarily conserved role for these proteins in regulating programmed cell death in the plant kingdom.

Analysis of nucleotide diversity among alleles of the major bacterial blight resistance gene Xa27 in cultivars of rice (Oryza sativa) and its wild relatives

Xa27 is one of the important R-genes, effective against bacterial blight disease of rice caused by Xanthomonas oryzae pv. oryzae (Xoo). Using natural population of Oryza, we analyzed the sequence variation in the functionally important domains of Xa27 across the Oryza species. DNA sequences of Xa27 alleles from 27 rice accessions revealed higher nucleotide diversity among the reported R-genes of rice. Sequence polymorphism analysis revealed synonymous and non-synonymous mutations in addition to a number of InDels in non-coding regions of the gene. High sequence variation was observed in the promoter region including the 5'UTR with 'π' value 0.00916 and 'θ w '=0.01785. Comparative analysis of the identified Xa27 alleles with that of IRBB27 and IR24 indicated the operation of both positive selection (Ka/Ks>1) and neutral selection (Ka/Ks≈0). The genetic distances of alleles of the gene from Oryza nivara were nearer to IRBB27 as compared to IR24. We also found the presence of conserved and null UPT (upregulated by transcriptional activator) box in the isolated alleles. Considerable amino acid polymorphism was localized in the trans-membrane domain for which the functional significance is yet to be elucidated. However, the absence of functional UPT box in all the alleles except IRBB27 suggests the maintenance of single resistant allele throughout the natural population.

Secretome analysis of the rice bacteriumXanthomonas oryzae(Xoo) using in vitro and in planta systems

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight disease in rice, and that severely affects yield loss (upto 50%) of total rice production. Here, we report a proteomics investigation of Xoo (compatible race K3)-secreted proteins, isolated from its in vitro culture and in planta infected rice leaves. 2DE coupled with MALDI-TOF-MS and/or nLC-ESI-MS/MS approaches identified 139 protein spots (out of 153 differential spots), encoding 109 unique proteins. Identified proteins belonged to multiple biological and molecular functions. Metabolic and nutrient uptake proteins were common up to both in vitro and in planta secretomes. However, pathogenicity, protease/peptidase, and host defense-related proteins were highly or specifically expressed during in planta infection. A good correlation was observed between protein and transcript abundances for nine proteins secreted in planta as per semiquantitative RT-PCR analysis. Transgenic rice leaf sheath (carrying PBZ1 promoter::GFP cell death reporter), when used to express a few of the identified secretory proteins, showed a direct activation of cell death signaling, suggesting their involvement in pathogenicity related with secretion effectors. This work furthers our understanding of rice bacterial blight disease, and serves as a resource for possible translation in generating disease resistant rice plants for improved seed yield.

Screening for actinomyces isolated from soil with the ability to inhibit Xanthomonas oryzae pv. oryzae causing rice bacterial blight disease in Vietnam

Bacterial blight disease caused by Xanthomonas oryzae pv. oryzae ( Xoo ) is one of the major diseases in rice culture of Northen Vietnam, as well as other rice-growing regions of the world. In this study, we isolated and screened for actinomycete strains from Vietnam with the ability to inhibit Xoo isolates from northern Vietnam. From 90 actinomycete strains taken from soil in northern Vietnam in 2010, we screened for their antagonistic activity against 10 races of Xoo causing rice bacterial blight disease. Three actinomycete strains were found to inhibit all 10 Xoo races. Among the three strains, a strain namely VN10-A-44 was shown not to have the ability to produce toxic compounds and was selected for further study. The strain was identified as Streptomyces virginiae by 16S ribosomal RNA gene sequencing. We replaced soybean meal with tofu waste in antibiotic producing medium to improve antagonistic activity of VN10-A-44 against the Xoo pathogen and to make use of tofu waste for large-scale fermentation of VN10-A-44. We found that replacing soybean meal with 20 and 30 g of tofu waste/litter in the antibiotic producing medium gave the largest inhibition zone against the Xoo pathogen. Key words : Xanthomonas oryzae pv. oryzae , rice bacterial blight disease, Streptomyces virginiae , Vietnam.

A CCCH-Type Zinc Finger Nucleic Acid-Binding Protein Quantitatively Confers Resistance against Rice Bacterial Blight Disease

Bacterial blight is a devastating disease of rice (Oryza sativa) caused by Xanthomonas oryzae pv oryzae (Xoo). Zinc finger proteins harboring the motif with three conserved cysteine residues and one histidine residue (CCCH) belong to a large family. Although at least 67 CCCH-type zinc finger protein genes have been identified in the rice genome, their functions are poorly understood. Here, we report that one of the rice CCCH-type zinc finger proteins, C3H12, containing five typical CX(8)-CX(5)-CX(3)-H zinc finger motifs, is involved in the rice-Xoo interaction. Activation of C3H12 partially enhanced resistance to Xoo, accompanied by the accumulation of jasmonic acid (JA) and induced expression of JA signaling genes in rice. In contrast, knockout or suppression of C3H12 resulted in partially increased susceptibility to Xoo, accompanied by decreased levels of JA and expression of JA signaling genes in rice. C3H12 colocalized with a minor disease resistance quantitative trait locus to Xoo, and the enhanced resistance of randomly chosen plants in the quantitative trait locus mapping population correlated with an increased expression level of C3H12. The C3H12 protein localized in the nucleus and possessed nucleic acid-binding activity in vitro. These results suggest that C3H12, as a nucleic acid-binding protein, positively and quantitatively regulates rice resistance to Xoo and that its function is likely associated with the JA-dependent pathway.

Crystallization and preliminary X-ray crystallographic analysis of β-ketoacyl-ACP synthase I (XoFabB) fromXanthomonas oryzaepv.oryzae

The proteins in the fatty-acid synthesis pathway in bacteria have significant potential as targets for the development of antibacterial agents. An essential elongation step in fatty-acid synthesis is performed by β-ketoacyl-acyl carrier protein synthase I (FabB). The organism Xanthomonas oryzae pv. oryzae (Xoo) causes a destructive bacterial blight disease of rice. The XoFabB protein from Xoo was expressed, purified and crystallized for the three-dimensional structure determination that is essential for the development of specific inhibitors of the enzyme. An XoFabB crystal diffracted to 3.0 Å resolution and belonged to the tetragonal space group P4(1), with unit-cell parameters a = b = 82.2, c = 233.2 Å. Assuming that the crystallographic structure contains four molecules in the asymmetric unit, the corresponding V(M) would be 2.18 Å(3) Da(-1) and the solvent content would be 43.5%. The initial structure was determined by the MOLREP program with an R factor of 44.0% and does contain four monomers in the asymmetric unit.