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Rice–Xanthomonas oryzae pv. oryzicola (Xoc) is one of the commonly used rice models of host–pathogen interactions. Xoc causes bacterial leaf streak (BLS) and has quarantine status. As a Gram-negative pathogen, Xoc usually employs type III secretion effectors (T3SEs), including transcription activator-like effectors (TALEs) and non-TALEs, to interfere with the innate immunity of rice. However, few major resistance genes corresponding to Xoc are found in rice cultivations; only Rxo1-AvrRxo1 and Xo1-TALEs interactions have been discovered in rice–Xoc. In this review, we focus on the role of the T3S system (T3SS) in Xoc virulence and consider the reported non-TALEs, including AvrRxo1, AvrBs2, XopN, XopC2, XopAP, and XopAK, as well as TALEs including Tal2g/Tal5d, Tal2h, Tal2a, Tal7, Tal10a, TalI, Tal2b, and Tal2c. Interestingly, AvrRxo1, XopC2, and XopAP disturb stomatal opening to promote infection through targeting diverse signaling pathways in rice. Otherwise, Tal2b and Tal2c, respectively, activate two rice salicylic acid (SA) hydroxylation genes to redundantly suppress the SA-mediated basal defense, and TalI, which has unknown targets, suppresses the SA signaling pathway in rice. In addition, other Xoc virulence factors are discussed. In conclusion, several T3SEs from Xoc interfere with similar defense pathways in rice to achieve invasion, providing an outlook for the control of this disease through manipulating the conserved pathways. [ABSTRACT FROM AUTHOR]

This article discusses the coevolution between rice and the pathogen Xanthomonas oryzae pv. oryzicola (Xoc), which causes bacterial leaf streak (BLS) in rice. The study explores natural sources of resistance and genetic susceptibility in the rice-Xoc interaction. The researchers discovered a variation in the effector-binding element (EBE) region of the OsSULTR3;6 gene that confers resistance against Xoc. They also identified four new susceptibility genes in the SULTR family that increase Xoc susceptibility when artificially induced. This research provides valuable insights into rice's genetic susceptibility and its defenses against Xoc. [Extracted from the article]

Abstract Background Two-tiered plant immune responses involve cross-talk among defense-responsive (DR) genes involved in pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI), effector-triggered immunity (ETI) and effector-triggered susceptibility (ETS). Bacterial leaf streak (BLS), caused by Xanthomonas oryzae pv. oryzicola (Xoc) is an important bacterial disease that causes serious threats to rice yield and quality. Transcriptomic profiling provides an effective approach for the comprehensive and large-scale detection of DR genes that participate in the interactions between rice and Xoc. Results In this study, we used RNA-seq to analyze the differentially expressed genes (DEGs) in susceptible rice after inoculation with two naturally pathogenic Xoc strains, a hypervirulent strain, HGA4, and a relatively hypovirulent strain, RS105. First, bacterial growth curve and biomass quantification revealed that differential growth occurred beginning at 1 day post inoculation (dpi) and became more significant at 3 dpi. Additionally, we analyzed the DEGs at 12 h and 3 days post inoculation with two strains, representing the DR genes involved in the PTI and ETI/ETS responses, respectively. Gene Ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed on the common DEGs, which included 4380 upregulated and 4019 downregulated genes and 930 upregulated and 1383 downregulated genes identified for the two strains at 12 h post inoculation (hpi) and 3 dpi, respectively. Compared to those at 12 hpi, at 3 dpi the number of common DEGs decreased, while the degree of differential expression was intensified. In addition, more disease-related GO pathways were enriched, and more transcription activator-like effector (TALE) putative target genes were upregulated in plants inoculated with HGA4 than in those inoculated with RS105 at 3 dpi. Then, four DRs were randomly selected for the BLS resistance assay. We found that CDP3.10, LOC_Os11g03820, and OsDSR2 positively regulated rice resistance to Xoc, while OsSPX3 negatively regulated rice resistance. Conclusions By using an enrichment method for RNA-seq, we identified a group of DEGs related to the two stages of response to the Xoc strain, which included four functionally identified DR genes.

Background: Two-tiered plant immune responses involve cross-talk among defense-responsive (DR) genes involved in pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI), effector-triggered immunity (ETI) and effector-triggered susceptibility (ETS). Bacterial leaf streak (BLS), caused by Xanthomonas oryzae pv. oryzicola (Xoc) is an important bacterial disease that causes serious threats to rice yield and quality. Transcriptomic profiling provides an effective approach for the comprehensive and large-scale detection of DR genes that participate in the interactions between rice and Xoc. Results: In this study, we used RNA-seq to analyze the differentially expressed genes (DEGs) in susceptible rice after inoculation with two naturally pathogenic Xoc strains, a hypervirulent strain, HGA4, and a relatively hypovirulent strain, RS105. First, bacterial growth curve and biomass quantification revealed that differential growth occurred beginning at 1 day post inoculation (dpi) and became more significant at 3 dpi. Additionally, we analyzed the DEGs at 12 h and 3 days post inoculation with two strains, representing the DR genes involved in the PTI and ETI/ETS responses, respectively. Gene Ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed on the common DEGs, which included 4380 upregulated and 4019 downregulated genes and 930 upregulated and 1383 downregulated genes identified for the two strains at 12 h post inoculation (hpi) and 3 dpi, respectively. Compared to those at 12 hpi, at 3 dpi the number of common DEGs decreased, while the degree of differential expression was intensified. In addition, more disease-related GO pathways were enriched, and more transcription activator-like effector (TALE) putative target genes were upregulated in plants inoculated with HGA4 than in those inoculated with RS105 at 3 dpi. Then, four DRs were randomly selected for the BLS resistance assay. We found that CDP3.10, LOC_Os11g03820, and OsDSR2 positively regulated rice resistance to Xoc, while OsSPX3 negatively regulated rice resistance. Conclusions: By using an enrichment method for RNA-seq, we identified a group of DEGs related to the two stages of response to the Xoc strain, which included four functionally identified DR genes. [ABSTRACT FROM AUTHOR]

Phospholipids are important components of biological membranes, participating in various biological processes, including plant development and responses to biotic and abiotic stresses. A previous study showed that mutation of the rice OsCDS5 (CDP‐DAG Synthase) gene alters lipid metabolism, causing enhanced abiotic stress responses, yellowing of leaves at the seedling stage and delayed plant development. Here, we observed that the Oscds5 mutant shows enhanced resistance to rice blast, bacterial blight and bacterial leaf streak. Mutation of OsCDS5 promotes production of reactive oxygen species and increases the expression level of multiple defence‐related genes. Transcriptomic analyses indicate that genes involved in responses to stress, biotic/abiotic stimuli and metabolic processes are highly upregulated and enriched in mutant Oscds5. Metabolomic analyses showed that differential metabolites were enriched in the lipid metabolic and tryptophan metabolic pathways. The decreased level of phosphatidylinositol and increased level of serotonin probably contribute to enhanced disease resistance of the Oscds5 mutant. Taken together, mutation of OsCDS5 enhances abiotic and biotic stress responses, and OsCDS5 may be a promising target for genetic engineering to enhance the resilience of rice to abiotic and biotic stresses simultaneously. [ABSTRACT FROM AUTHOR]

Bacterial leaf streak (BLS), caused by Xanthomonas oryzae pv. oryzicola (Xoc), is a bacterial disease affecting rice production in Asia and Africa, whose severity is expected to increase with climate change. Identification of new quantitative-trait loci (QTL) or resistance genes for BLS resistance is essential for developing resistant rice. A genome-wide association study to identify QTL associated with BLS resistance was conducted using phenotypic and genotypic data from 429 rice accessions. Of 47 QTL identified, 45 were novel and two co-localized with previously reported QTL or genes conferring BLS resistance. qBLS6.2 on chromosome 6 explained the greatest phenotypic variation. Combined analysis of differential expression and annotations of predicted genes near qBLS6.2 based on haplotype and disease phenotype identified OsBLS6.2 (LOC_Os06g02960) as a candidate gene for qBLS6.2. OsBLS6.2 knockout plants showed higher resistance to Xoc than wild-type plants. Many other candidate genes for resistance to Xoc were identified. [ABSTRACT FROM AUTHOR]

ABSTRACT Metal oxide nanoparticles (NPs) possess antibacterial activities and can be used for the control of phytopathogenic bacteria. The objective of this work was to evaluate the antibacterial activity of pure and doped NPs against Dickeya zeae, Pantoea ananatis, and Xanthomonas vasicola pv. vasculorum in vitro, and to assess the efficacy of preventive and curative application of zinc oxide (ZnO), nickel oxide (NiO), and silver-doped zinc oxide (ZnO:Ag) NPs for the control of white spot (WS) and bacterial leaf streak (BLS) in vivo. Bacterial growth inhibition was first evaluated by measuring the diameter of the inhibition zone formed in Petri dishes. Subsequently, the severity of WS and BLS diseases was evaluated in a greenhouse calculating the area under the disease progress curve. The in vitro antibacterial activity was not influenced by the increase in the concentration of doping elements for most NPs. ZnO NPs doped with Ag, K, and Mo; ZnOCl doped with Ag, and pure NPs (Ag2O, CuO, and NiO) showed antibacterial activity against D. zeae, P. ananatis, and X. vasicola pv. vasculorum with relatively similar inhibition zones at different concentrations. Commercial copper showed antibacterial activity only against D. zeae. NiO NPs in preventive and curative applications reduced WS and BLS severities, whereas commercial copper application increased WS severity and reduced BLS severity. The use of NPs has promising applications and further evaluation of their formulation, application form, and timing is necessary for new strategies to control the activity of phytopathogenic bacteria.

Rice–Xanthomonas oryzae pv. oryzicola (Xoc) is one of the commonly used rice models of host–pathogen interactions. Xoc causes bacterial leaf streak (BLS) and has quarantine status. As a Gram-negative pathogen, Xoc usually employs type III secretion effectors (T3SEs), including transcription activator-like effectors (TALEs) and non-TALEs, to interfere with the innate immunity of rice. However, few major resistance genes corresponding to Xoc are found in rice cultivations; only Rxo1-AvrRxo1 and Xo1-TALEs interactions have been discovered in rice–Xoc. In this review, we focus on the role of the T3S system (T3SS) in Xoc virulence and consider the reported non-TALEs, including AvrRxo1, AvrBs2, XopN, XopC2, XopAP, and XopAK, as well as TALEs including Tal2g/Tal5d, Tal2h, Tal2a, Tal7, Tal10a, TalI, Tal2b, and Tal2c. Interestingly, AvrRxo1, XopC2, and XopAP disturb stomatal opening to promote infection through targeting diverse signaling pathways in rice. Otherwise, Tal2b and Tal2c, respectively, activate two rice salicylic acid (SA) hydroxylation genes to redundantly suppress the SA-mediated basal defense, and TalI, which has unknown targets, suppresses the SA signaling pathway in rice. In addition, other Xoc virulence factors are discussed. In conclusion, several T3SEs from Xoc interfere with similar defense pathways in rice to achieve invasion, providing an outlook for the control of this disease through manipulating the conserved pathways.

Abstract Background Rice is the second-largest food crop in the world and vulnerable to bacterial leaf streak disease. A thorough comprehension of the genetic foundation of agronomic traits was essential for effective implementation of molecular marker-assisted selection. Results Our study aimed to evaluate the vulnerability of rice to bacterial leaf streak disease (BLS) induced by the gram-negative bacterium Xanthomonas oryzae pv. oryzicola (Xoc). In order to accomplish this, we first analyzed the population structure of 747 accessions and subsequently assessed their phenotypes 20 days after inoculation with a strain of Xoc, GX01. We conducted genome-wide association studies (GWAS) on a population of 747 rice accessions, consisting of both indica and japonica subpopulations, utilizing phenotypic data on resistance to bacterial leaf streak (RBLS) and sequence data. We identified a total of 20 QTLs associated with RBLS in our analysis. Through the integration of linkage mapping, sequence analysis, haplotype analysis, and transcriptome analysis, we were able to identify five potential candidate genes (OsRBLS1—OsRBLS5) that possess the potential to regulate RBLS in rice. In order to gain a more comprehensive understanding of the genetic mechanism behind resistance to bacterial leaf streak, we conducted tests on these genes in both the indica and japonica subpopulations, ultimately identifying superior haplotypes that suggest the potential utilization of these genes in breeding disease-resistant rice varieties. Conclusions The findings of our study broaden our comprehension of the genetic mechanisms underlying RBLS in rice and offer significant insights that can be applied towards genetic improvement and breeding of disease-resistant rice in rapidly evolving environmental conditions.

Bacterial leaf streak (BLS) of maize is an emerging foliar disease of maize in the Americas. It is caused by the gram-negative nonvascular bacterium Xanthomonas vasicola pv. vasculorum. There are no chemical controls available for BLS, and thus, host resistance is crucial for managing X. vasicola pv. vasculorum. The objective of this study was to examine the genetic determinants of resistance to X. vasicola pv. vasculorum in maize, as well as the relationship between other defense-related traits and BLS resistance. Specifically, we examined the correlations among BLS severity, severity for three fungal diseases, flg-22 response, hypersensitive response, and auricle color. We conducted quantitative trait locus (QTL) mapping for X. vasicola pv. vasculorum resistance using the maize recombinant inbred line population Z003 (B73 × CML228). We detected three QTLs for BLS resistance. In addition to the disease resistance QTL, we detected a single QTL for auricle color. We observed significant, yet weak, correlations among BLS severity, levels of pattern-triggered immunity response and leaf flecking. These results will be useful for understanding resistance to X. vasicola pv. vasculorum and mitigating the impact of BLS on maize yields. [ABSTRACT FROM AUTHOR]

Bacterial leaf streak (BLS), caused chiefly by the pathogen Xanthomonas translucens pv. translucens, is becoming an increasingly important foliar disease of barley in the Upper Midwest. The deployment of resistant cultivars is the most economical and practical method of control. To identify sources of BLS resistance, we evaluated two panels of breeding lines from the University of Minnesota (UMN) and Anheuser-Busch InBev (ABI) barley improvement programs for reaction to strain CIX95 in the field at St. Paul and Crookston, MN, in 2020 and 2021. The percentage of resistant lines in the UMN and ABI panels with mid-season maturity was 1.8% (6 of 333 lines) and 5.2% (13 of 251 lines), respectively. Both panels were genotyped with the barley 50K iSelect SNP array, and then a genome-wide association study was performed. A single, highly significant association was identified for BLS resistance on chromosome 6H in the UMN panel. This association was also identified in the ABI panel. Seven other significant associations were detected in the ABI panel: two each on chromosomes 1H, 2H, and 3H and one on chromosome 5H. Of the eight associations identified in the panels, five were novel. The discovery of resistance in elite breeding lines will hasten the time needed to develop and release a BLS-resistant cultivar. [ABSTRACT FROM AUTHOR]

Xanthomonas translucens pv. cerealis causes bacterial leaf streak disease on small grain cereals. Type II and III secretion systems (T2SS and T3SS) play a pivotal role in the pathogenicity of the bacterium, while no data are available on the transcriptomic profile of wheat cultivars infected with either wild type (WT) or mutants of the pathogen. In this study, WT, TAL-effector mutants, and T2SS/T3SS mutants of X. translucens pv. cerealis strain NXtc01 were evaluated for their effect on the transcriptomic profile of two wheat cultivars, 'Chinese Spring' and 'Yangmai-158', using Illumina RNA-sequencing technology. RNA-Seq data showed that the number of differentially expressed genes (DEGs) was higher in Yangmai-158 than in Chinese Spring, suggesting higher susceptibility of Yangmai-158 to the pathogen. In T2SS, most suppressed DEGs were related to transferase, synthase, oxidase, WRKY, and bHLH transcription factors. The gspD mutants showed significantly decreased disease development in wheat, suggesting an active contribution of T2SS in virulence. Moreover, the gspD mutant restored full virulence and its multiplication in planta by addition of gspD in trans. In the T3SS-deficient strain, downregulated DEGs were associated with cytochrome, peroxidases, kinases, phosphatases, WRKY, and ethylene-responsive transcription factors. In contrast, upregulated DEGs were trypsin inhibitors, cell number regulators, and calcium transporter. Transcriptomic analyses coupled with quantitative real-time-PCR indicated that some genes are upregulated in Δtal1/Δtal2 compared with the tal-free strain, but no direct interaction was observed. These results provide novel insight into wheat transcriptomes in response to X. translucens infection and pave the way for understanding host-pathogen interactions. [ABSTRACT FROM AUTHOR]

Bacterial leaf streak (BLS) is a disease of monocot plants caused by Xanthonionas translucens on small grains, X vasic·ola on maize and sorghum, and X. oryzae on rice. These three pathogens cause remarkably similar symptomology in their host plants. Despite causing similar symptoms. BLS pathogens are dispersed throughout the larger Xanthomonas phylogeny. Each aforementioned species includes strain groups that do not cause BLS and instead cause vascular disease. In this commentary, we hypothesize that strains of X. trans! ucens, X. vasic<, ta. and X. ory:ae convergently evolved to cause BLS due to shared evolutionary pressures. We examined the diversity of secreted effectors, which may be important virulence factors for BLS pathogens and their evolution. We discuss evidence that differences in gene regulation and abilities to manipulate plant hormones may also separate BLS pathogens from other Xanthoniona.r species or pathovars. BLS is becoming an increasing issue across the three pathosystems. Overall, we hope that a better understanding of conserved mechanisms used by BLS pathogens will enable researchers to translate findings across production systems and guide approaches to control this (re)emerging threat. [ABSTRACT FROM AUTHOR]

Downy Mildew Resistance 6-like (DMR6-like) genes are identified as salicylic acid (SA) hydroxylases and negative regulators of plant immunity. Previously, we identified two rice DMR6-like genes, OsF3H03g, and OsF3H04g, that act as susceptible targets of transcription activator-like effectors (TALEs) from Xanthomonas oryzae pv. oryzicola (Xoc), which causes bacterial leaf streak (BLS) in rice. Furthermore, all four homologs of rice DMR6-like proteins were identified to predominantly carry the enzyme activity of SA 5-hydroxylase (S5H), negatively regulate rice broad-spectrum resistance, and cause the loss of function of these OsDMR6s, leading to increased resistance to rice blast and bacterial blight (BB). Here, we curiously found that an OsF3H04g knock-out mutant created by T-DNA insertion, osf3h04g, was remarkedly susceptible to BLS and BB and showed an extreme reduction in SA content. OsF3H04g knock-out rice lines produced by gene-editing were mildly susceptible to BLS and reduced content of SA. To explore the susceptibility mechanism in OsF3H04g loss-of-function rice lines, transcriptome sequencing revealed that another homolog, OsS3H, had induced expression in the loss-of-function OsF3H04g rice lines. Furthermore, we confirmed that a great induction of OsS3H downstream and genomically adjacent to OsF3H04g in osf3h04g was primarily related to the inserted T-DNA carrying quadruple enhancer elements of 35S, while a slight induction was caused by an unknown mechanism in gene-editing lines. Then, we found that the overexpression of OsS3H increased rice susceptibility to BLS, while gene-editing mediated the loss-of-function OsS3H enhanced rice resistance to BLS. However, the knock-out of both OsF3H04g and OsS3H by gene-editing only neutralized rice resistance to BLS. Thus, we concluded that the knock-out of OsF3H04g activated the expression of the OsS3H, partially participating in the susceptibility to BLS in rice. [ABSTRACT FROM AUTHOR]

Background: Rice is the second-largest food crop in the world and vulnerable to bacterial leaf streak disease. A thorough comprehension of the genetic foundation of agronomic traits was essential for effective implementation of molecular marker-assisted selection. Results: Our study aimed to evaluate the vulnerability of rice to bacterial leaf streak disease (BLS) induced by the gram-negative bacterium Xanthomonas oryzae pv. oryzicola (Xoc). In order to accomplish this, we first analyzed the population structure of 747 accessions and subsequently assessed their phenotypes 20 days after inoculation with a strain of Xoc, GX01. We conducted genome-wide association studies (GWAS) on a population of 747 rice accessions, consisting of both indica and japonica subpopulations, utilizing phenotypic data on resistance to bacterial leaf streak (RBLS) and sequence data. We identified a total of 20 QTLs associated with RBLS in our analysis. Through the integration of linkage mapping, sequence analysis, haplotype analysis, and transcriptome analysis, we were able to identify five potential candidate genes (OsRBLS1—OsRBLS5) that possess the potential to regulate RBLS in rice. In order to gain a more comprehensive understanding of the genetic mechanism behind resistance to bacterial leaf streak, we conducted tests on these genes in both the indica and japonica subpopulations, ultimately identifying superior haplotypes that suggest the potential utilization of these genes in breeding disease-resistant rice varieties. Conclusions: The findings of our study broaden our comprehension of the genetic mechanisms underlying RBLS in rice and offer significant insights that can be applied towards genetic improvement and breeding of disease-resistant rice in rapidly evolving environmental conditions. [ABSTRACT FROM AUTHOR]

Bacterial panicle blight, bacterial leaf streak, and bacterial brown stripe are common bacterial diseases in rice that represent global threats to stable rice yields. In this study, we used the rice variety HZ, Nekken and their 120 RIL population as experimental materials. Phenotypes of the parents and RILs were quantitatively analyzed after inoculation with Burkholderia glumae, Xanthomonas oryzae pv. oryzicola, and Acidovorax avenae subsp. avenae. Genetic SNP maps were also constructed and used for QTL mapping of the quantitative traits. We located 40 QTL loci on 12 chromosomes. The analysis of disease resistance-related candidate genes in the QTL regions with high LOD value on chromosomes 1, 3, 4, and 12 revealed differential expression before and after treatment, suggesting that the identified genes mediated the variable disease resistance profiles of Huazhan and Nekken2. These results provide an important foundation for cloning bacterial-resistant QTLs of panicle blight, leaf streak, and brown stripe in rice. [ABSTRACT FROM AUTHOR]

Rice (Oryza sativa L.) is a staple food in many countries around the world, particularly in China. The production of rice is seriously affected by the bacterial leaf streak and rice blast, which can reduce rice yield or even cause it to fail to be harvested. In this study, susceptible material 58B was edited by CRISPR/Cas9, targeting a target of the Pi21 gene and a target of the effector-binding element (EBE) of the OsSULTR3;6 gene, and the mutants 58b were obtained by Agrobacterium-mediated method. The editing efficiency of the two targets in the T0 generation was higher than 90.09%, the homozygous mutants were successfully selected in the T0 generation, and the homozygous mutation rate of each target was higher than 26.67%. The expression of the edited pi21 and EBE of Ossultr3;6 was significantly reduced, and the expression of defense responsive genes was significantly upregulated after infected with rice blast. The lesion areas of rice blast and bacterial leaf streak were significantly reduced in 58b, and the resistance of both was effectively improved. Furthermore, the gene editing events did not affect the agronomic traits of rice. In this study, the resistance of 58b to rice blast and bacterial leaf streak was improved simultaneously. This study provides a reference for using Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 (CRISPR/Cas9) to accelerate the improvement of rice varieties and the development of new materials for rice breeding. [ABSTRACT FROM AUTHOR]

The genus Xanthomonas comprises phytopathogenic bacteria which infect about 400 host species, including a wide variety of economically important plants. Xanthomonas oryzae pv. oryzicola (Fang et al., 1957) Swings et al., 1990 is the causal agent of bacterial leaf streak (BLS) being one of the most destructive bacterial diseases of rice. BLS symptoms are very similar to those of bacterial blight caused by closely related Xanthomonas oryzae pv. oryzae. X. o. pv. oryzae and X. o. pv. oryzicola and often occur in rice f ields simultaneously, so separate leaves may show symptoms of both diseases. The quarantine status and high severity of the pathogen require a highly eff icient, fast and precise diagnostic method. We have developed an assay for Xanthomonas oryzae pv. oryzicola detection using real-time polymerase chain reaction (qPCR) and PCR amplicon sequencing. The DNA samples of X. o. pv. oryzae and X. o. pv. oryzicola were obtained from the collection of CIRM-CFBR (France). To evaluate the analytical sensitivity of the assay, a vector construct based on the pAL2-T plasmid was created through the insertion of X. o. pv. oryzicola target fragment (290 bp). Primers and a probe for qPCR were selected for the hpa1 gene site. They allowed identifying all the strains the sequences of which had been loaded in the GenBank NCBI Nucleotide database before November 11, 2021. The SeqX.o.all sequencing primers were selected for the hrp gene cluster sequence, namely for the nucleotide sequence encoding the Hpa1 protein, the sequencing of which allows for eff icient differentiation of X. oryzae species. The analytical specif icity of the system was tested using the DNAs of 53 closely related and accompanying microorganisms and comprised 100 % with no false-positive or false-negative results registered. The system’s analytical sensitivity was not less than 25 copies per PCR reaction. Its eff icacy has been conf irmed using f ive different qPCR detection systems from different manufacturers, so it can be recommended for diagnostic and screening studies.

The severity of the bacterial leaf streak disease (BLS) depends on various factors such as environmental conditions and genetic resistance. However, the effect of diversity and dynamics of seed microbiota on the activity of this bacterium in wheat is unknown. Curtobacterium flaccumfaciens strains with different pathogenic and beneficial traits can colonize seeds in several plant species. This study intends to screen C. flaccumfaciens pv. flaccumfaciens (Cff) isolates, collected from wheat seeds infected by Xanthomonas translucens pv. cerealis (Xtc) to determine their effect on the bacterial leaf streak disease severity. The selected isolates were identified based on morphological, biochemical and molecular characteristics, and also their synergistic or antagonistic effects on Xtc were investigated under in vivo and in vitro conditions. Isolates of Curtobacterium flaccumfaciens, in particular Cff were detected as epiphyte and endophyte in more than 40% of the samples taken from fields in Fars, Markazi, Lorestan and West Azerbaijan provinces of Iran which were rotated with bean cultivation. Antagonistic properties of selected isolates, including the inhibition zone, production of extracellular compounds, enzymes and IAA, phosphate solubility, and autoaggregation, coaggregation and biofilm synthesis, were carried out. All Cff isolates showed high inhibitory potency, and were more successful in biofilm and autoaggregation formation in competition with the Xtc. Seed treatment by Cff isolates successfully reduced the area under the disease progression curve (AUDPC) by 1.7 to 6.2 times. The results provides new information on the Cff transmission through wheat seeds. The Cff can persist in wheat seeds and develop systemically and endophytically in wheat as non-host plant. The overall impact of planting Cff contaminated wheat seeds in wheat-bean rotation system, will be increasing the outbreak of bean bacterial wilt disease in the upcoming years if suitable environmental conditions are provided. Our findings suggest that care needs to be taken about the transmission of this pathogen by wheat seeds. [ABSTRACT FROM AUTHOR]

Quantitative polymerase chain reaction (qPCR) is a commonly used technique for plant disease diagnosis, which generally requires DNA extraction from the plant samples. The efficiency of DNA extraction has a significant influence on the accuracy of qPCR. Thus, including a DNA extraction efficiency control is important in qPCR analyses. To develop and evaluate a DNA extraction efficiency control, a Stagonospora nodorum (syn. Phaeosphaeria nodorum , synonym and correct taxonomic name: Parastagonospora nodorum) strain named HUA-22 was created by transferring a green florescent protein (GFP) gene into the genome of the S. nodorum strain Sn15. HUA-22 showed a similar pathogenicity as Sn15 but carried a strong GFP activity. A qPCR primers/probe set named P-GFP, targeting the GFP sequence, was designed. Using P-GFP, qPCR analysis was conducted on DNA extracted from replicated samples of HUA-22 conidia, and confirmed that HUA-22 could be used to characterize the variation in replicated DNA extractions. Conidia of HUA-22 were used to spike soil samples inoculated with the canola clubroot pathogen Plasmodiophora brassicae , canola stem samples infected with the blackleg pathogens Leptosphaeria biglobosa and/or L. maculans and wheat/barley samples infected with Xanthomonas translucens pv. translucens (Xtt) or X. translucens pv. undulosa (Xtu). Duplex qPCR using P-GFP and a primers/probe set specific to P. brassicae , triplex qPCR using P-GFP and primers/probe sets specific to L. biglobosa and L. maculans , and triplex qPCR using P-GFP and primers/probe sets specific to Xtt and Xtu were conducted. The results indicated that HUA-22 could be used as a standard for DNA extraction efficiency in qPCR-based plant disease diagnosis. Adding HUA-22 conidia to plant or soil samples prior to DNA extraction, and subsequent use of the P-GFP detection control, provided an added control that could distinguish truly negative from false-negative qPCR results. • A GFP -labeled S. nodorum strain named HUA-22 was created by transformation. • GFP in HUA-22 can be used as a DNA control in qPCR diagnosis of plant diseases. • HUA-22 was tested in the diagnosis of plant diseases caused by five pathogens. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to illustrate the potential utilization of the gene of resistance to bacterial blight and bacterial leaf streak, so as to provide reference for enriching genetic base of rice breeding.【Method】A total of 102 rice cultivars were used to screen by molecular marker for the gene xa5, which mediated the resistance for both for resistance to bacterial blight and bacterial leaf streak. The parents carrying xa5 gene and having good resistance to bacterial blight and bacterial leaf streak were used as resistant sources to cross with susceptible cultivated rice. Through pedigree method and molecular marker-assisted selection（MAS）, elite hybrid rice restorer lines were cultivated. The excellent restorer lines and sterile lines were combined with strong heterosis combinations to analyze the resistance and comprehensive agronomic traits of strong heterosis combinations.【Result】A total of 17 cultivars were screened, which carried xa5 gene from 102 rice germplasm resources（materials）. Among these cultivars, DP3, IRBB5, N36, N776, N777, N779 and P89 were well resistant to both bacterial blight and bacterial leaf streak. An elite line Y240-1 was selected from the cross between DP3 and 9311. Moreover, 998 R was crossed with elite individual Y240-1. With molecular MAS, an elite restorer line Yexiangzhan with xa5 was developed from the cross of 998 R and Y240-1, after multiple generations of selfing. Yexiangzhan was combined with sterile line Pingfeng A to produce an elite hybrid cultivar Pingfengyouxiangzhan, which showed higher resistance to bacterial blight and bacterial leaf streak in comparison with Pingfeng A. Pingfengyouxiangzhan was resistant to rice blast at moderate level and meet the 2nd class standard of fine quality issued by Ministry of Agriculture of China.【Conclusion】xa5 gene with resistance to bacterial blight and bacterial leaf streak of rice has partial dominant effect, which can be used for breeding improvement of hybrid rice; It is helpful to enrich and broaden the genetic basis of rice breeding by cultivating the superior hybrid combinations of rice with moderate resistance to bacterial blight and stripe disease with wild rice blood. [ABSTRACT FROM AUTHOR]

Plant diseases pose significant threats to agricultural yields and are responsible for nearly 20 % of losses in total food production. Therefore, the rapid detection of plant pathogens is critically important for preventing the rapid development of plant diseases and minimizing crop damage. Raman spectroscopy (RS) has been shown to be effective for detecting living biological samples. Compared with traditional detection methods, RS is fast, sensitive, and non-destructive; it also does not require sample labeling. In this study, we used Laser tweezers Raman spectroscopy combined with convolutional neural networks to detect two closely related strains of bacteria, Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xoc), exuded from bacteria-infected rice leaves. The accuracy of this technique was 97.5 %. For the application of RS in the field, we used the portable Raman spectrometer to detect mock-inoculated as well as Xoo- and Xoc-infected rice leaves at different disease courses. The identification accuracy via this technique was 87.02 % in the early stage, in which no obvious symptoms were apparent. This method also revealed spectral differences in rice leaves caused by the two bacteria, which could be leveraged for subsequent analysis of the molecular mechanism of infection. Our results indicate that RS is a promising approach for the early detection of bacterial diseases in rice in the field, as well as for in-depth single-cell analysis in laboratory settings., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Yu Vincent Fu reports financial support was provided by Beijing Natural Science Foundation grant. Ning Xu reports financial support was provided by National Natural Science Foundation of China Grant. Weilai Lu reports financial support was provided by National Natural Science Foundation of China Grant. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Bacterial leaf streak (BLS) of corn caused by Xanthomonas vasicola pv. vasculorum was first reported in the United States in 2017. The biology and management of BLS are poorly understood. The objective of this work was to determine the effects of hybrids, foliar treatments, and infection conditions (timing, temperature, and inoculation site) on BLS of sweet corn. Field studies were conducted to determine if hybrid or foliar disease management treatments influenced BLS development and yield. Corn leaves were inoculated in plots with X. vasicola pv. vasculorum , and noninoculated plots were used for comparison. The leaf incidence and severity of BLS differed significantly among sweet corn hybrids, suggesting different levels of susceptibility to BLS. Grain yield was significantly reduced (14.7%) by BLS for one hybrid. The corn growth stage at the time of infection influenced BLS, with incidence and severity significantly greater after inoculation at stage V6 than at V9. Foliar application of Kocide, LifeGard, and Kocide and LifeGard significantly reduced leaf severity compared with nontreated controls in field studies. Kocide significantly reduced leaf incidence, but no treatments significantly increased yield versus controls. In comparisons of inoculation methods in a growth chamber, lesion length on leaves was significantly greater on stalk-inoculated than leaf-inoculated plants. Lesions developed on leaf-inoculated plants only at inoculation sites, whereas lesions developed on stalk-inoculated plants on multiple leaves. In controlled environments, lesion length on leaves was significantly greater at 21°C than at 27 and 32°C. This study expands our understanding of factors that influence the development and management of BLS of sweet corn., Competing Interests: The author(s) declare no conflict of interest.

Downy Mildew Resistance 6-like (DMR6-like) genes are identified as salicylic acid (SA) hydroxylases and negative regulators of plant immunity. Previously, we identified two rice DMR6-like genes, OsF3H03g, and OsF3H04g, that act as susceptible targets of transcription activator-like effectors (TALEs) from Xanthomonas oryzae pv. oryzicola (Xoc), which causes bacterial leaf streak (BLS) in rice. Furthermore, all four homologs of rice DMR6-like proteins were identified to predominantly carry the enzyme activity of SA 5-hydroxylase (S5H), negatively regulate rice broad-spectrum resistance, and cause the loss of function of these OsDMR6s, leading to increased resistance to rice blast and bacterial blight (BB). Here, we curiously found that an OsF3H04g knock-out mutant created by T-DNA insertion, osf3h04g, was remarkedly susceptible to BLS and BB and showed an extreme reduction in SA content. OsF3H04g knock-out rice lines produced by gene-editing were mildly susceptible to BLS and reduced content of SA. To explore the susceptibility mechanism in OsF3H04g loss-of-function rice lines, transcriptome sequencing revealed that another homolog, OsS3H, had induced expression in the loss-of-function OsF3H04g rice lines. Furthermore, we confirmed that a great induction of OsS3H downstream and genomically adjacent to OsF3H04g in osf3h04g was primarily related to the inserted T-DNA carrying quadruple enhancer elements of 35S, while a slight induction was caused by an unknown mechanism in gene-editing lines. Then, we found that the overexpression of OsS3H increased rice susceptibility to BLS, while gene-editing mediated the loss-of-function OsS3H enhanced rice resistance to BLS. However, the knock-out of both OsF3H04g and OsS3H by gene-editing only neutralized rice resistance to BLS. Thus, we concluded that the knock-out of OsF3H04g activated the expression of the OsS3H, partially participating in the susceptibility to BLS in rice.

Bacterial panicle blight, bacterial leaf streak, and bacterial brown stripe are common bacterial diseases in rice that represent global threats to stable rice yields. In this study, we used the rice variety HZ, Nekken and their 120 RIL population as experimental materials. Phenotypes of the parents and RILs were quantitatively analyzed after inoculation with Burkholderia glumae, Xanthomonas oryzae pv. oryzicola, and Acidovorax avenae subsp. avenae. Genetic SNP maps were also constructed and used for QTL mapping of the quantitative traits. We located 40 QTL loci on 12 chromosomes. The analysis of disease resistance-related candidate genes in the QTL regions with high LOD value on chromosomes 1, 3, 4, and 12 revealed differential expression before and after treatment, suggesting that the identified genes mediated the variable disease resistance profiles of Huazhan and Nekken2. These results provide an important foundation for cloning bacterial-resistant QTLs of panicle blight, leaf streak, and brown stripe in rice.

Two-component systems (TCSs) (cognate sensor histidine kinase/response regulator pair, HK/RR) play a crucial role in bacterial adaptation, survival, and productive colonization. An atypical orphan single-domain RR VemR was characterized by the non-vascular pathogen Xanthomonas oryzae pv. oryzicola (Xoc) is known to cause bacterial leaf streak (BLS) disease in rice. Xoc growth and pathogenicity in rice, motility, biosynthesis of extracellular polysaccharide (EPS), and the ability to trigger HR in non-host tobacco were severely compromised in the deletion mutant strain RΔ vemR as compared to the wild-type strain RS105. Site-directed mutagenesis and phosphotransfer experiments revealed that the conserved aspartate (D56) residue within the stand-alone phosphoacceptor receiver (REC) domain is essential for phosphorelay and the regulatory activity of Xoc VemR. Yeast two-hybrid (Y2H) and co-immunoprecipitation (co-IP) data identified CheA as the HK co-opting the RR VemR for phosphorylation. Affinity proteomics identified several downstream VemR-interacting proteins, such as 2-oxoglutarate dehydrogenase (OGDH), DNA-binding RR SirA, flagellar basal body P-ring formation protein FlgA, Type 4a pilus retraction ATPase PilT, stress-inducible sensor HK BaeS, septum site-determining protein MinD, cytoskeletal protein CcmA, and Type III and VI secretion system proteins HrpG and Hcp, respectively. Y2H and deletion mutant analyses corroborated that VemR interacted with OGDH, SirA, FlgA, and HrpG; thus, implicating multi-layered control of diverse cellular processes including carbon metabolism, motility, and pathogenicity in the rice. Physical interaction between VemR and HrpG suggested cross-talk interaction between CheA/VemR- and HpaS/HrpG-mediated signal transduction events orchestrating the hrp gene expression. [ABSTRACT FROM AUTHOR]

Two-component systems (TCSs) (cognate sensor histidine kinase/response regulator pair, HK/RR) play a crucial role in bacterial adaptation, survival, and productive colonization. An atypical orphan single-domain RR VemR was characterized by the non-vascular pathogen Xanthomonas oryzae pv. oryzicola (Xoc) is known to cause bacterial leaf streak (BLS) disease in rice. Xoc growth and pathogenicity in rice, motility, biosynthesis of extracellular polysaccharide (EPS), and the ability to trigger HR in non-host tobacco were severely compromised in the deletion mutant strain RΔvemR as compared to the wild-type strain RS105. Site-directed mutagenesis and phosphotransfer experiments revealed that the conserved aspartate (D56) residue within the stand-alone phosphoacceptor receiver (REC) domain is essential for phosphorelay and the regulatory activity of Xoc VemR. Yeast two-hybrid (Y2H) and co-immunoprecipitation (co-IP) data identified CheA as the HK co-opting the RR VemR for phosphorylation. Affinity proteomics identified several downstream VemR-interacting proteins, such as 2-oxoglutarate dehydrogenase (OGDH), DNA-binding RR SirA, flagellar basal body P-ring formation protein FlgA, Type 4a pilus retraction ATPase PilT, stress-inducible sensor HK BaeS, septum site-determining protein MinD, cytoskeletal protein CcmA, and Type III and VI secretion system proteins HrpG and Hcp, respectively. Y2H and deletion mutant analyses corroborated that VemR interacted with OGDH, SirA, FlgA, and HrpG; thus, implicating multi-layered control of diverse cellular processes including carbon metabolism, motility, and pathogenicity in the rice. Physical interaction between VemR and HrpG suggested cross-talk interaction between CheA/VemR- and HpaS/HrpG-mediated signal transduction events orchestrating the hrp gene expression.

Bacterial leaf streak (BLS) caused by bacterium Xanthomonas oryzae pv. oryzicola (Xoc) is one of the most prominent rice diseases. BLS causes a significant reduction in paddy yields. However, there are limited studies and a lack of information regarding the mechanisms and cells affected on leaf tissues severed from this disease. Therefore, in this study, sensitive paddy variety IR24 was inoculated against BLS, and the pathogen colonised mesophyll cells and some bundle sheath cells. The infection spreads rapidly towards the base and apex of the leaf, but rather slowly to the left and right sides of the leaf veins. Another experiment was performed to unravel anatomical characteristics in sensitive paddy varieties (TN1, IR24, IR5) and resistant paddy varieties (IR26, Dular, IR36) against BLS. Susceptible paddy varieties have less thick midrib and leaf lamina, a high number of bundle sheath cells at primary vascular tissue (midrib), one layer of sclerenchyma cells at the secondary vein, and more than two metaxylems at the primary vein. Resistant paddy varieties, on the other hand, consist of a relatively thickened midrib and leaf lamina, fewer bundle sheath cells at the primary vascular tissue (midrib), more than one sclerenchyma layers at the secondary vein, and two metaxylems at the primary vein. This study contributes new knowledge in identifying the level of infection in paddy fields, and helps breeders in producing resistant paddies to this disease. [ABSTRACT FROM AUTHOR]

Bacterial diseases affecting wheat production in Uruguay are an issue of growing concern yet remain largely uninvestigated in the region. Surveys of 61 wheat fields carried out from 2017 to 2019 yielded a regional collection of 63 strains identified by 16S rRNA gene analysis as Xanthomonas spp. A real-time PCR protocol with species-specific primers previously reported allowed the identification of 44 strains as X. translucens, the causal agent of bacterial leaf streak (BLS) in wheat and other cereal crops. Multilocus sequence analysis of four housekeeping genes (dnaK, fyuA, gyrB, and rpoD) revealed that these strains were most closely related to X. translucens pv. undulosa, the pathovar that is most commonly associated with BLS of wheat. Multilocus sequence typing was applied to examine the genetic diversity of X. translucens strains. Strains were assigned to four different sequence types, three of which were previously reported globally. Additionally, 17 Xanthomonas strains not belonging to X. translucens were obtained from diseased wheat leaves. Phylogenetic analysis showed that these strains are closely related to X. prunicola and clustered together with previously uncharacterized Xanthomonas strains isolated from wheat in Minnesota. In planta pathogenicity assays carried out on a BLS-susceptible wheat cultivar showed that X. translucens pv. undulosa strains caused brown necrosis symptoms typical of BLS, whereas non-translucens Xanthomonas sp. strains elicited an atypical symptom of dry necrosis. These findings suggest that local wheat fields are affected by X. translucens pv. undulosa and by a new wheat pathogen within the Xanthomonas genus. [ABSTRACT FROM AUTHOR]

Summary: Transcription activator‐like (TAL) effectors are major virulence factors secreted by the type III secretion systems of Xanthomonas oryzae pv. oryzicola (Xoc) and X. oryzae pv. oryzae (Xoo), causing bacterial leaf streak and bacterial blight, respectively, in rice. However, the knowledge of Xoc TAL effector function in promoting bacterial virulence remains limited.Here, we isolated the highly virulent Xoc strain HGA4 from the outbreak region of Huanggang (Hubei, China), which contains four TAL effectors not found in the Chinese model strain RS105. Among these, Tal2b was selected for introduction into RS105, which resulted in a longer lesion length than that in the control.Tal2b directly binds to the promoter region of the gene and activates the expression of OsF3H03g, which encodes 2‐oxoglutarate‐dependent dioxygenase in rice. OsF3H03g negatively regulates salicylic acid (SA)‐related defense by directly reducing SA, and it plays a positive role in susceptibility to both Xoc and Xoo in rice.OsF3H03g interacts with a uridine diphosphate‐glycosyltransferase protein (OsUGT74H4), which positively regulates bacterial leaf streak susceptibility and may inactivate SA via glycosylation modification. [ABSTRACT FROM AUTHOR]

Abstract Rice (Oryza sativa L.) is a staple food crop, feeding more than 50% of the world’s population. Diseases caused by bacterial, fungal, and viral pathogens constantly threaten the rice production and lead to enormous yield losses. Bacterial blight (BB) and bacterial leaf streak (BLS), caused respectively by gram-negative bacteria Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xoc), are two important diseases affecting rice production worldwide. Due to the economic importance, extensive genetic and genomic studies have been conducted to elucidate the molecular mechanism of rice response to Xoo and Xoc in the last two decades. A series of resistance (R) genes and their cognate avirulence and virulence effector genes have been characterized. Here, we summarize the recent advances in studies on interactions between rice and the two pathogens through these R genes or their products and effectors. Breeding strategies to develop varieties with durable and broad-spectrum resistance to Xanthomonas oryzae based on the published studies are also discussed.

Abstract Polygalacturonase-inhibiting proteins (PGIPs) have been shown to recognize fungal polygalacturonases (PGs), which initiate innate immunity in various plant species. Notably, the connection between rice OsPGIPs and PGs in Xanthomonas oryzae pv. oryzicola (Xoc), which causes bacterial leaf streak (BLS), remains unclear. Here, we show that OsPGIP1 was strongly induced after inoculating rice with the Xoc strain RS105. Furthermore, OsPGIP1-overexpressing (OV) and RNA interference (RNAi) rice lines increased and decreased, respectively, the resistance of rice to RS105, indicating that OsPGIP1 contributes to BLS resistance. Subsequently, we generated the unique PG mutant RS105Δpg, the virulence of which is attenuated compared to that of RS105. Surprisingly, the lesion lengths caused by RS105Δpg were similar to those caused by RS105 in the OV lines compared with wild-type ZH11 with reduced Xoc susceptibility. However, the lesion lengths caused by RS105Δpg were still significantly shorter in the OV lines than in ZH11, implying that OsPGIP1-mediated BLS resistance could respond to other virulence factors in addition to PGs. To explore the OsPGIP1-mediated resistance, RNA-seq analysis were performed and showed that many plant cell wall-associated genes and several MYB transcription factor genes were specifically expressed or more highly induced in the OV lines compared to ZH11 postinoculation with RS105. Consistent with the expression of the differentially expressed genes, the OV plants accumulated a higher content of jasmonic acid (JA) than ZH11 postinoculation with RS105, suggesting that the OsPGIP1-mediated resistance to BLS is mainly dependent on the plant cell wall-associated immunity and the JA signaling pathway.

Wheat (Triticum aestivum) loses 21.5% yield to pests and diseases annually (Savary et al. 2019). Among the wheat diseases, bacterial leaf streak (BLS) is a growing problem, costing $78.5 million in losses (https://cropprotectionnetwork.org/). In July 2022, we sampled winter wheat leaf samples at Volga (44.30, -96.92), South Dakota, USA with an estimated disease incidence of 40% (n=100). The typical symptoms were water-soaking with large necrotic and chlorotic streaks extending the length of the leaves and were strikingly similar to BLS. To isolate the pathogen, leaves were cut lengthwise into 1 cm pieces and surface-sterilized using a 10% NaOCl solution for 3 min, followed by 70% ethanol for 3 min, and then rinsed with sterile distilled water and placed in 500 ul of sterile distilled water for 5 min and using a sterile loop the water was streaked over a plate of Nutrient Agar (NA). Following Duveiller et al. (1997), the streaked plate was incubated in the dark at 28℃ for 48 h. Observed single colonies were sub-cultured thrice onto fresh NA plates to obtain a pure culture. We named the culture SD101. Bacteria were found to be gram-negative with a colony morphology initially raised, smooth, and white that later turned yellow. DNA was extracted using the Wizard HMW DNA Extraction Kit (Promega, Madison, WI) following the manufacturer's protocol, and sequenced using Nanopore MinION R9.4 (Oxford Nanopore Technology). We used the Rapid Annotation Using Subsystems Technology approach (Aziz eal. 2008) to extract the 16S rDNA, DNA gyrase subunit B (gyrB), and translation initiation factor IF-2 (infB) gene sequences that were deposited in GenBank under accession numbers PP329908.1 for 16S rDNA, PP496481 for infB, and PP328920.1 for gyrB. Homology analysis using CLC Genomics Workbench 22.0.2 (QIAGEN) and BLASTn against the GenBank nucleotide database resulted in a 99.74% match (1543/ 1547 bp) of the 16S sequence, 99.59% match (2674/ 2685 bp) of the infB sequence, and 99.42% match (2396/ 2410 bp) of the gyrB sequence with Pantoea ananatis strain AJ13355 (AP012032). To test pathogenicity, seeds of spring wheat breeding line SD4892 were planted in 30 cm × 30 cm pots in a greenhouse under a 16 h light photoperiod. The inoculum was prepared from 48-h-old NA plates of SD101 rinsed with 1X Phosphate Buffer Saline (PBS buffer), adjusted to an OD600 = 1.0, and amended with two drops of Tween 20 (polyoxyethylene sorbitol ester, Millipore Sigma). PBS with Tween 20 was used as a negative control. The inoculum was sprayed on 15 replicates of 15-day-old seedlings, kept at 95% relative humidity for 48 h, then moved to the greenhouse at 23 to 25°C. The symptoms appeared as water soaking that later turned to necrotic streaks with surrounding chlorosis on all 15 inoculated plants while control plants remained healthy. The pathogen was re-isolated from the leaves as described above. The 16S rDNA, infB, and gyrB sequences amplified and sequenced were identical to the gene sequences from the whole genome sequencing. The experiment was repeated with the same results, completing Koch's postulates. Although P. ananatis is pathogenic on corn, rice, and other plant species in the United States (Coutinho et al. 2009), and was reported pathogenic on wheat in Poland (Krawczyk et al. 2020), this is the first report of its pathogenicity on wheat in the United States. The prevalence, and incidence of BLS disease caused by P. ananatis in wheat is needed to estimate its threat to wheat production and to develop management strategies.

Bacterial leaf streak caused by Xanthomonas oryzae pv. oryzicola (Xoc) is a continuous threat to rice cultivation, leading to substantial yield losses with socioeconomic implications. Iron ions are essential mineral nutrients for plant growth, but little information is available on how they influence mechanisms of rice immunity against Xoc. Here, we investigated the role of the myeloblastosis-related (MYB) transcriptional repressor OsMYBxoc1 in modulation of rice resistance through control of iron ion transport. Overexpression of OsMYBxoc1 significantly increased rice resistance, whereas OsMYBxoc1 RNA-interference lines and knockout mutants showed the opposite result. Suppression of OsMYBxoc1 expression dampened the immune response induced by pathogen-associated molecular patterns. We demonstrated that OsMYBxoc1 binds specifically to the OsNRAMP5 promoter and represses transcription of OsNRAMP5. OsNRAMP5, a negative regulator of rice resistance to bacterial leaf streak, possesses metal ion transport activity, and inhibition of OsMYBxoc1 expression increased the iron ion content in rice. Activity of the ion-dependent H 2 O 2 scavenging enzyme catalase was increased in plants with suppressed expression of OsMYBxoc1 or overexpression of OsNRAMP5. We found that iron ions promoted Xoc infection and interfered with the production of reactive oxygen species induced by Xoc. The type III effector XopAK directly inhibited OsMYBxoc1 transcription, indicating that the pathogen may promote its own proliferation by relieving restriction of iron ion transport in plants. In addition, iron complemented the pathogenicity defects of the RS105&#95;ΔXopAK mutant strain, further confirming that iron utilization by Xoc may be dependent upon XopAK. In conclusion, our study reveals a novel mechanism by which OsMYBxoc1 modulates rice resistance by regulating iron accumulation and demonstrates that Xoc can accumulate iron ions by secreting the effector XopAK to promote its own infection., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola (Xoc), impacts the production of rice. However, several rice cultivars displayed resistance to Xoc in the field, but scarce information is available about the role of endophytic microbiota in disease resistance. In the present study, the endophytic bacterial communities of resistant and susceptible rice cultivars "CG2" and "IR24", respectively, were analyzed using high throughput 16S rRNA gene amplified sequencing and culture dependent method was further used for bacterial isolation. A total of 452,716 high-quality sequences representing 132 distinct OTUs (Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes) and 46 isolates of 16 genera were explored from rice leaves infected with Xoc. Community diversity of endophytic bacteria were higher in the leaves of the resistant cultivars compared to susceptible cultivars upon Xoc infection. Strikingly, this diversity might contribute to natural defense of the resistant cultivar against pathogen. Pantoea, which is pathogen antagonist, was frequently detected in two cultivars and higher abundance were recorded in resistant cultivars. Different abundance genus includes endophytic isolates with marked antagonistic activity to Xoc. The increased proportions of antagonistic bacteria, may contribute to resistance of rice cultivar against Xoc and the Pantoea genus was recruited by Xoc infection play a key role in suppressing the development of BLS disease in rice. Taken together, this work reveals the association between endophytic bacteria and BLS resistance in rice and identification of antagonism-Xoc bacterial communities in rice. [ABSTRACT FROM AUTHOR]

Summary: Bacterial leaf streak (BLS) is a major bacterial disease of rice. Utilization of host genetic resistance has become one of the most important strategies for controlling BLS. However, only a few resistance genes have been characterized. Previously, a recessive BLS resistance gene bls1 was roughly mapped on chromosome 6. Here, we further delineated bls1 to a 21 kb region spanning four genes. Genetic analysis confirmed that the gene encoding a mitogen‐activated protein kinase (OsMAPK6) is the target of the allelic genes BLS1 and bls1. Overexpression of BLS1 weakened resistance to the specific Xanthomonas oryzae pv. oryzicola (Xoc) strain JZ‐8, while low expression of bls1 increased resistance. However, both overexpression of BLS1 and low expression of bls1 could increase no‐race‐specific broad‐spectrum resistance. These results indicate that BLS1 and bls1 negatively regulate race‐specific resistance to Xoc strain JZ‐8 but positively and negatively control broad‐spectrum resistance, respectively. Subcellular localization demonstrated that OsMAPK6 was localized in the nucleus. RGA4, which is known to mediate resistance to Xoc, is the potential target of OsMAPK6. Overexpression of BLS1 and low expression of bls1 showed increase in salicylic acid and induced expression of defense‐related genes, simultaneously increasing broad‐spectrum resistance. Moreover, low expression of bls1 showed increase an in jasmonic acid and abscisic acid, in company with an increase in resistance to Xoc strain JZ‐8. Collectively, our study provides new insights into the understanding of BLS resistance and facilitates the development of rice host‐resistant cultivars. Significance Statement: A recessive bacterial leaf streak resistance gene bls1, which encodes a mitogen‐activated protein kinase (OsMAPK6), was cloned from common wild rice. The gene bls1 and its dominant allele BLS1 negatively regulate race‐specific resistance to Xanthomonas oryzae pv. oryzicola strain JZ‐8, but negatively and positively control broad‐spectrum resistance, respectively. [ABSTRACT FROM AUTHOR]

Bacterial leaf streak (BLS), caused by Xanthomonas translucens pv. undulosa , is a devastating cereal disease that is becoming more common in Iran. A cost-effective and realistic strategy to cope with BLS is planting of BLS-resistant cultivars. To discover BLS-resistant genotypes, 360 hexaploid wheat genotypes were selected and inoculated with a local strain of Xanthomonas translucens pv. undulosa in the field and greenhouse in two years (2019 & 2020). In the field experiments, disease reaction of wheat plants was assessed based on both percentage of disease incidence (PDI) and the percentage of disease severity (PDS). In the greenhouse experiment, disease reaction was evaluated as the disease severity (DS). The genotypes with PDS, PDI, and DS values ≤ 20% were considered BLS-resistant or semi-resistant. Genotypes with zero or very low PDS, PDI and DS during the first and second years were considered highly resistant to the disease. Despite resistant genotypes being more prevalent than susceptible genotypes in each year, genotype responses to BLS varied over the two years, but in both years, most genotypes showed a resistance response. Forty-six genotypes (12.8%) were consistently BLS-resistant across years in the field and greenhouse trials. In 2020, field BLS intensity ratings (PDI and PDS) were positively correlated with flag leaf width and days to maturity. In contrast, PDI and PDS were negatively correlated with plant height, peduncle length, and spike length. In 2019, no significant correlations were found between PDI, PDS, and phenotypic traits. Cluster heatmaps and principal component analysis (PCA) separated resistant and susceptible genotypes. In BLS-resistant genotypes, plant height, peduncle length, and spike length were significantly longer, but flag leaf width and days to maturity were shorter. This study identified new and diverse sources of resistance to BLS that could be used in wheat breeding efforts. • Three hundred and sixty wheat genotypes were screened for bacterial leaf streak response. • Field-disease and phenotypic values varied greatly among wheat genotypes. • Some disease-resistant wheat genotypes remained stable in greenhouse and fields. • A negative or positive correlation was found between disease and phenotypic traits. • Heat maps separated wheat genotypes by disease response and phenotypic traits. [ABSTRACT FROM AUTHOR]

β-Glucosidase belongs to the glycoside hydrolase I family, which is widely present in multiple species and responds to various biotic and abiotic stresses. In rice, whether β-glucosidase is involved in the interaction between plants and microorganisms is not clear. In this study, we found that the expression of several genes encoding β-glucosidases, including OsBGLU19 and OsBGLU23, were upregulated after inoculation with Xanthomonas oryzae pv. oryzicola (Xoc) and downregulated after inoculation with X. oryzae pv. oryzae (Xoo). The respective insertion mutants of OsBGLU19 and OsBGLU23, bglu19 and bglu23, were more susceptible to Xoc infection. The expression of OsAOS2, a key gene in the jasmonic acid signal pathway, was dramatically downregulated after inoculation with Xoc in the bglu19 and bglu23 mutants. Simultaneously, the expression of downstream disease resistance-related genes, such as OsPR1a, OsPR5 and a key transcription factors OsWRKY72 were obviously downregulated. The resistance mediated by OsBGLU19 and OsBGLU23 to bacterial leaf streak is related to disease resistance-related genes above mentioned.

Abstract Background The function of Arabidopsis enhanced disease susceptibility 1 (AtEDS1) and its sequence homologs in other dicots have been extensively studied. However, it is unknown whether rice EDS1 homolog (OsEDS1) plays a role in regulating the rice-pathogen interaction. Results In this study, a OsEDS1-knouckout mutant (oseds1) was characterized and shown to have increased susceptibility to Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xoc), suggesting the positive role of OsEDS1 in regulating rice disease resistance. However, the following evidence suggests that OsEDS1 shares some differences with AtEDS1 in its way to regulate the host-pathogen interactions. Firstly, OsEDS1 modulates the rice-bacteria interactions involving in jasmonic acid (JA) signaling pathway, while AtEDS1 regulates Arabidopsis disease resistance against biotrophic pathogens depending on salicylic acid (SA) signaling pathway. Secondly, introducing AtEDS1 could reduce oseds1 mutant susceptibility to Xoo rather than to Xoc. Thirdly, exogenous application of JA and SA cannot complement the susceptible phenotype of the oseds1 mutant, while exogenous application of SA is capable of complementing the susceptible phenotype of the ateds1 mutant. Finally, OsEDS1 is not required for R gene mediated resistance, while AtEDS1 is required for disease resistance mediated by TIR-NB-LRR class of R proteins. Conclusion OsEDS1 is a positive regulator in rice-pathogen interactions, and shares both similarities and differences with AtEDS1 in its way to regulate plant-pathogen interactions.

The Gram-negative bacterium Xanthomonas translucens infects a wide range of gramineous plants with a notable impact on small grain cereals. However, genomics-informed intra-species population structure and virulence repertories of the pathogen have rarely been investigated. In this study, the complete genome sequences of seven X. translucens strains representing an entire set of genetic diversity of two pathovars X. translucens pv. undulosa and X. translucens pv. translucens is provided and compared with those of seven publicly available complete genomes of the pathogen. Organization of the 25 type III secretion system genes in all the 14 X. translucens strains was exactly the same, while TAL effector genes localized singly or in clusters across four loci in X. translucens pv. translucens and five to six loci in X. translucens pv. undulosa. Beside two previously unreported endogenous plasmids in X. translucens pv. undulosa, and variations in repeat variable diresidue (RVD) of the 14 strains, tal1a of X. translucens pv. translucens strain XtKm8 encode the new RVDs HE and YI which have not previously been reported in xanthomonads. Further, a number of truncated tal genes were predicted among the 14 genomes lacking conserved BamHI site at N-terminus and SphI site at C-terminus. Our data have doubled the number of complete genomes of X. translucens clarifying the population structure and genomics of the pathogen to pave the way in the small grain cereals industry for disease resistance breeding in the 21st century’s agriculture.

The Gram-negative bacterium Xanthomonas translucens infects a wide range of gramineous plants with a notable impact on small grain cereals. However, genomics-informed intra-species population structure and virulence repertories of the pathogen have rarely been investigated. In this study, the complete genome sequences of seven X. translucens strains representing an entire set of genetic diversity of two pathovars X. translucens pv. undulosa and X. translucens pv. translucens is provided and compared with those of seven publicly available complete genomes of the pathogen. Organization of the 25 type III secretion system genes in all the 14 X. translucens strains was exactly the same, while TAL effector genes localized singly or in clusters across four loci in X. translucens pv. translucens and five to six loci in X. translucens pv. undulosa. Beside two previously unreported endogenous plasmids in X. translucens pv. undulosa , and variations in repeat variable diresidue (RVD) of the 14 strains, tal1a of X. translucens pv. translucens strain XtKm8 encode the new RVDs HE and YI which have not previously been reported in xanthomonads. Further, a number of truncated tal genes were predicted among the 14 genomes lacking conserved Bam HI site at N-terminus and Sph I site at C-terminus. Our data have doubled the number of complete genomes of X. translucens clarifying the population structure and genomics of the pathogen to pave the way in the small grain cereals industry for disease resistance breeding in the 21st century's agriculture. [ABSTRACT FROM AUTHOR]

Bacterial leaf streak (BLS) caused by bacterium Xanthomonas oryzae pv. oryzicola (Xoc) is one of the most prominent rice diseases. BLS causes a significant reduction in paddy yields. However, there are limited studies and a lack of information regarding the mechanisms and cells affected on leaf tissues severed from this disease. Therefore, in this study, sensitive paddy variety IR24 was inoculated against BLS, and the pathogen colonised mesophyll cells and some bundle sheath cells. The infection spreads rapidly towards the base and apex of the leaf, but rather slowly to the left and right sides of the leaf veins. Another experiment was performed to unravel anatomical characteristics in sensitive paddy varieties (TN1, IR24, IR5) and resistant paddy varieties (IR26, Dular, IR36) against BLS. Susceptible paddy varieties have less thick midrib and leaf lamina, a high number of bundle sheath cells at primary vascular tissue (midrib), one layer of sclerenchyma cells at the secondary vein, and more than two metaxylems at the primary vein. Resistant paddy varieties, on the other hand, consist of a relatively thickened midrib and leaf lamina, fewer bundle sheath cells at the primary vascular tissue (midrib), more than one sclerenchyma layers at the secondary vein, and two metaxylems at the primary vein. This study contributes new knowledge in identifying the level of infection in paddy fields, and helps breeders in producing resistant paddies to this disease.

Rice (Oryza sativa L.) is a crucial staple crop worldwide, and bacterial diseases are among the primary factors affecting rice yield. In late October 2022, bacterial leaf streak disease was observed on the leaves of the rice variety Meixiangzhan 2 across multiple fields (approximately 130 hm2) in Leizhou City, Guangdong Province, China. The incidence rate was up to 30% in each field. Infected rice leaves exhibited distinctive symptoms at the boundary between diseased and healthy tissue, featuring dark green to yellow-brown streaks, while most of the leaf margin exhibited symptoms of either leaf edge or sheath rot. Disease progression from the leaf tip inwards revealed gray-white or dehydrated lesions with a bluish-gray color. Some leaves exhibited wrinkling at the edges, and severe symptoms at the leaf tip resembled those of bacterial leaf blight in rice. Ten leaves were collected from 10 infected rice plants in three distinct fields, and leaf pieces at the border of diseased and healthy areas were surface disinfected with 75% anhydrous ethanol for 60 seconds, rinsed three times with sterile water, and then soaked in sterile water for 8 hours. The obtained bacterial suspension was diluted at a ratio of 1: 106, and 100 μL of the diluted samples were plated on Potato Dextrose Agar (PDA) plates. After incubation at 28°C for 48 hours, the yellow bacterial colonies that appeared, were purified on PDA plates. To confirm the bacterial species, the amplification of genes gyrB, leuS, rpoB, and 16S rDNA was performed on six randomly selected isolates from the three different fields using the primers 27F/1492R, gyrB-F/R, leuS-F/R and rpoB-F/R, as reported by Yu et al (2022), respectively. PCR products were sequenced. All six isolates had identical sequences for all genes sequenced.The gene sequences of 16S rDNA (960 bp), gyrB (953 bp), leuS (733 bp), and rpoB (877 bp) for LZ1, were deposited in the NCBI database under accession numbers PP048830 , PP068625 , PP068626, and PP068627, respectively. These sequences were subsequently compared using BLASTn tool against the NCBI nr/nt database. The 16S rDNA, gyrB, leuS, and rpoB of LZ1 showed similarities of 99.90%, 99.16%, 99.73%, and 99.89%, with the corresponding sequences of P. ananatis TZ39 (GenBank accession numbers MZ800600.1 for 16S rDNA, and CP081342.1 for gyrB, leuS and rpoB ). MLSA analysis using concatenated sequences of gyrB, leuS, and rpoB genes indicated that the isolated strain LZ1 belongs to P. ananatis. In the tillering stage of rice varieties Meixiangzhan 2 and Huahangyuzhan, P. ananatis LZ1 was inoculated at a concentration of 108 CFU/mL using the leaf-cutting method, with sterile water used as a control (Toh et al., 2019). After 14 days of bacterial inoculation, the inoculated leaves gradually became necrotic, changing from light green to brown showing identical symptoms as those in the field, while the control plants remained symptom-free. Subsequent 16S rDNA, gyrB, leuS and rpoB gene sequencing results further confirmed the identity of the pathogen as P. ananatis, thereby fulfilling Koch's postulates. Previous reports have already identified P. ananatis as the pathogen causing rice bacterial leaf streak (Kini et al., 2017; Arayaskul et al., 2019; Yu et al., 2022; Lu et al., 2022; Luna et al., 2023; Yuan et al., 2023). This is the first report of rice bacterial leaf streak caused by P. ananatis in Guangdong Province, China, laying the foundation for future research to establish strategies for the prevention and control of this disease.

Bacterial leaf streak and black chaff are important bacterial diseases of wheat, which have been reported to be caused by Xanthomonas translucens. In 2016, symptoms of bacterial leaf streak and black chaff were observed in Algeria, at experimental wheat breeding stations and in farmers’ fields under sprinkler irrigation on two wheat cultivars, ‘Hiddab’ and ‘Simeto’. Yellow Xanthomonas-like bacterial colonies were isolated from plant material, including leaves, spikes and post-harvest crop residues. Initial characterisation using biochemical, physiological and pathogenicity tests identified the bacteria as Xanthomonas. Diagnostic PCR targeting the 16S-23S rRNA intergenic region indicated that the strains were X. translucens, a clade-1 xanthomonad. However, partial DNA sequences of the housekeeping genes gyrB and rpoD revealed that the strains belong to clade 1, but likely represent a new Xanthomonas species that has not been previously described on wheat or other Gramineae. The most closely related strain, NCPPB 2654, was isolated from a bean plant in the United Kingdom in 1974. Further characterization is required to clarify the taxonomic status of the Algerian Xanthomonas isolates from wheat, and to determine their host ranges and impacts on plant cultivation.

Xanthomonas oryzae delivers transcription activator-like effectors (TALEs) into plant cells to facilitate infection. Following economic principles, the redundant TALEs are rarely identified in Xanthomonas. Previously, we identified the Tal2b, which activates the expression of the rice 2-oxoglutarate-dependent dioxygenase gene OsF3H03g to promote infection in the highly virulent strain of X. oryzae pv. oryzicola HGA4. Here, we reveal that another clustered TALE, Tal2c, also functioned as a virulence factor to target rice OsF3H04g, a homologue of OsF3H03g. Transferring Tal2c into RS105 induced expression of OsF3H04g to coincide with increased susceptibility in rice. Overexpressing OsF3H04g caused higher susceptibility and less salicylic acid (SA) production compared to wild-type plants. Moreover, CRISPR–Cas9 system-mediated editing of the effector-binding element in the promoters of OsF3H03g or OsF3H04g was found to specifically enhance resistance to Tal2b- or Tal2c-transferring strains, but had no effect on resistance to either RS105 or HGA4. Furthermore, transcriptome analysis revealed that several reported SA-related and defense-related genes commonly altered expression in OsF3H04g overexpression line compared with those identified in OsF3H03g overexpression line. Overall, our results reveal a functional redundancy mechanism of pathogenic virulence in Xoc in which tandem Tal2b and Tal2c specifically target homologues of host genes to interfere with rice immunity by reducing SA.

Bacterial leaf streak (BLS) is now the fourth-most devastating disease in rice. Dular and H359 are two indica rice varieties with contrasting responses to BLS. Dular displays high resistance, while H359 is susceptible. In this study, RNA-seq was used to examine the early molecular processes deployed during the resistance response of Dular and H359 at different times after inoculation. Differentially expressed gene (DEG) analysis identified 3031 genes in Dular and 7161 in H359 that were modulated in response to infection after 12 and 24 h. There were significantly more DEGs in H359 than in Dular, and there were significantly more downregulated genes than upregulated genes. Gene ontology (GO) and KEGG enrichment analyses revealed a similar set of GO terms and KEGG pathways enriched in both varieties. However, KEGG analysis of upregulated DEGs revealed that some phenylpropane metabolism-related pathways were specially enriched in Dular. Further comparison and analysis showed that the numbers of resistance-related DEGs in the two varieties were significantly reduced at 24 h compared with 12 h after BLS infection and genes critically involved in conferring resistance during the early stage mainly included WRKY transcription factors, receptor kinases and disease, exocyst, MAPK signalling pathway and hormones related genes. Our study suggests that resistance-related genes may play an important role at an early stage of infection and phenylpropane metabolism related genes may partly response for BLS resistance of Dular, thus providing valuable information for future studies on the molecular mechanisms of BLS resistance in rice. [ABSTRACT FROM AUTHOR]

This paper reports original evidence regarding the potential role of seed transmission of Xanthomonas vasicola pv. vasculorum in the epidemiology of bacterial leaf streak (BLS) in maize. We evaluated the occurrence of the pathogen on seeds from diseased fields and its subsequent transmission to seedlings. In 2016 and 2017, X. vasicola pv. vasculorum was detected by TaqMan PCR from 22 of 41 maize seed lots harvested from naturally infected fields in Colorado, Nebraska, and Iowa. However, many of the PCR-positive samples did not yield culturable X. vasicola pv. vasculorum colonies. The highest levels of seed contamination were detected in dent maize and popcorn from NE and CO. Seed transmission was evaluated in greenhouse grow-outs from eight seed lots, totaling more than 14,000 plants. Putative seed transmission events from naturally contaminated seed lots, estimated from PCR results, occurred at a frequency between 0.1 and 0.5% in 10-seedling pooled samples and at a frequency of 2.7% from individual plant assays. However, no seedling symptoms were observed during these assays and live X. vasicola pv. vasculorum colonies were not recovered from PCR-positive seedlings. In contrast, seed transmission was readily demonstrated from artificially contaminated seed lots, including typical symptoms and recovery of live bacteria. Seed transmission consistently occurred from seeds soaked in bacterial suspensions with concentrations of 106 CFU/ml, suggesting that a threshold population of the bacterium is necessary for the development of BLS symptoms and recovery of live bacteria. The low bacterial populations on naturally contaminated seeds apparently were not sufficient to result in diseased seedlings. [ABSTRACT FROM AUTHOR]