Search Results (23)

Bacterial leaf streak (BLS) is one of the internationally significant quarantine diseases in rice. Effectively utilizing BLS resistance genes from wild rice (Oryza rufipogon Griff.) to breed new varieties offers a fundamental solution for BLS control. This study focused on the fine mapping of the BLS resistance gene bls2 and the development of closely linked molecular markers for breeding BLS-resistant lines. Using a Guangxi common wild rice accession DY19 (carrying bls2) as the donor parent and the highly BLS-susceptible indica rice variety 9311 as the recipient parent, BLS-resistant rice lines were developed through multiple generations of backcrossing and selfing, incorporating molecular marker-assisted selection (MAS), single nucleotide polymorphism(SNP) chip genotyping, pathogen inoculation assays, and agronomic trait evaluation. The results showed that bls2 was delimited to a 113 kb interval between the molecular markers ID2 and ID5 on chromosome 2, with both markers exhibiting over 98% accuracy in detecting bls2. Four stable new lines carrying the bls2 segment were obtained in the BC₅F₄ generation. These four lines showed highly significant differences in BLS resistance compared with 9311, demonstrating moderate resistance or higher with average lesion lengths ranging from 0.69 to 1.26 cm. Importantly, no significant differences were observed between these resistant lines and 9311 in key agronomic traits, including plant height, number of effective panicles, panicle length, seed setting rate, grain length, grain width, length-to-width ratio, and 1000-grain weight. Collectively, two molecular markers closely linked to bls2 were developed, which can be effectively applied in MAS, and four new lines with significantly enhanced resistance to BLS and excellent agronomic traits were obtained. These findings provide technical support and core germplasm resources for BLS resistance breeding. Full article

Rice bacterial blight (BB) and bacterial leaf streak (BLS), caused by Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xoc), respectively, are among the most devastating bacterial diseases threatening global rice production. The interactions between rice and Xanthomonas oryzae are complex and dynamic, involving recognition, attack, defense, and adaptation mechanisms enacted by both the rice host and the pathogens. This review summarizes recent advances in understanding rice–Xanthomonas oryzae interactions, focusing on infection models, pathogenic mechanisms, and immune responses elicited by Xanthomonas oryzae. Special attention is devoted to the roles of transcription activator-like effectors (TALEs) and non-TALE effectors in pathogenicity, the functions of resistance (R) genes in defense, and the interconnected molecular networks of interactions derived from multi-omics approaches. Understanding these interactions is essential for developing effective disease-resistance strategies and creating elite disease-resistant rice varieties. Full article

Rice (Oryza sativa L.) bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola (Xoc) seriously threatens rice production. This disease can be controlled by cultivating lines with appropriate resistance genes. To discover new resistance loci, a natural population of 246 sequenced backbone varieties of indica rice was inoculated with Xoc in the field, and their disease level was evaluated. The population structure, distribution of resistance phenotypes, and effect of combinations of resistance genes were analyzed in a genome-wide association study. On the basis of single nucleotide polymorphism data and disease phenotypes at the seedling stage, seven resistance loci (qBls1.1, qBls4.1, qBls5.1, qBls7.1, qBls8.1, qBls9.1, and qBls12.1) were identified to determine the superior haplotype of the corresponding loci, which showed stronger BLS resistance. The effects of combining loci were analyzed. As the number of superior haplotypes increased, BLS resistance tended to increase, indicating that BLS resistance can be significantly improved by pyramiding multiple resistance loci in ideal germplasm. This study provides new resistance loci and genetic resources for breeding BLS-resistant rice varieties. These genetic resources will be useful for the fine mapping of resistance loci, gene cloning, and the breeding of BLS-resistant varieties in follow-up research. Full article

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. Full article

LuxR-type regulators play pivotal roles in regulating numerous bacterial processes, including bacterial motility and virulence, thereby exerting a significant influence on bacterial behavior and pathogenicity. Xanthomonas oryzae pv. oryzicola, a rice pathogen, causes bacterial leaf streak. Our research has identified VmsR, which is a response regulator of the two-component system (TCS) that belongs to the LuxR family. These findings of the experiment reveal that VmsR plays a crucial role in regulating pathogenicity, motility, biofilm formation, and the production of extracellular polysaccharides (EPSs) in Xoc GX01. Notably, our study shows that the vmsR mutant exhibits a reduced swimming motility but an enhanced swarming motility. Furthermore, this mutant displays decreased virulence while significantly increasing EPS production and biofilm formation. We have uncovered that VmsR directly interacts with the promoter regions of fliC and fliS, promoting their expression. In contrast, VmsR specifically binds to the promoter of gumB, resulting in its downregulation. These findings indicate that the knockout of vmsR has profound effects on virulence, motility, biofilm formation, and EPS production in Xoc GX01, providing insights into the intricate regulatory network of Xoc. Full article

Calcium (Ca²⁺) is a versatile intracellular second messenger that regulates several signaling pathways involved in growth, development, stress tolerance, and immune response in plants. Autoinhibited Ca²⁺-ATPases (ACAs) play an important role in the regulation of cellular Ca²⁺ homeostasis. Here, we systematically analyzed the putative OsACA family members in rice, and according to the phylogenetic tree of OsACAs, OsACA9 was clustered into a separated branch in which its homologous gene in Arabidopsis thaliana was reported to be involved in defense response. When the OsACA9 gene was knocked out by CRISPR/Cas9, significant accumulation of reactive oxygen species (ROS) was detected in the mutant lines. Meanwhile, the OsACA9 knock out lines showed enhanced disease resistance to both rice bacterial blight (BB) and bacterial leaf streak (BLS). In addition, compared to the wild-type (WT), the mutant lines displayed an early leaf senescence phenotype, and the agronomy traits of their plant height, panicle length, and grain yield were significantly decreased. Transcriptome analysis by RNA-Seq showed that the differentially expressed genes (DEGs) between WT and the Osaca9 mutant were mainly enriched in basal immune pathways and antibacterial metabolite synthesis pathways. Among them, multiple genes related to rice disease resistance, receptor-like cytoplasmic kinases (RLCKs) and cell wall-associated kinases (WAKs) genes were upregulated. Our results suggest that the Ca²⁺-ATPase OsACA9 may trigger oxidative burst in response to various pathogens and synergically regulate disease resistance and leaf senescence in rice. Full article

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. Full article

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. Full article

Bacterial leaf streak (BLS) of maize (Zea mays), caused by the bacterium Xanthomonas vasicola pv. vasculorum (Xvv), was first reported in Brazil in 2018. In this study, we evaluated 52 species of cultivated plants, cover crops, forage, and grasses that are used in succession or crop rotation with maize, and weeds with natural occurrence in maize-producing regions, to determine their potentials as alternative hosts for Xvv. We investigated (i) the pathogenicity of Xvv based on symptom development, (ii) epiphytic colonization of the bacterium in asymptomatic plants, and (iii) bacterial colonization in plant tissues using scanning electron microscopy (SEM) in symptomatic and asymptomatic species. Ten species, all belonging to the Poaceae family, presented symptoms after Xvv infection, including Avena sativa (cvs. IPR Afrodite and IPR Esmeralda), A. strigosa (cv. IPR 161), Hordeum vulgare (cv. BRS Cauê), Oryza sativa (cv. IPR 117), Brachiaria brizantha (Brizantha and cv. Marandu), Digitaria horizontalis, D. insularis, Echinochloa colonum, Eleusine indica, and Sorghum arundinaceum. Furthermore, epiphytic colonization by Xvv was observed in 23 asymptomatic species. Scanning micrographs revealed that Xvv cells and their aggregates were distributed throughout the leaf surface. In addition, bacterial colonization in the intercellular tissues of the substomatal chambers of white oat, black oat, and maize was observed across the tissue fractures. Despite showing typical symptoms of Xvv infection, SEM examination revealed evidence of Xvv colonization only on the leaf surface of rice. In asymptomatic species, such as rye, sorghum, and millet, a low number of bacterial cells were found on the leaf surface. However, no evidence of internal tissue colonization was observed in millet fractures, suggesting that Xvv survives only epiphytically in this species. Full article

Rice diseases are extremely harmful to rice growth, and achieving the identification and rapid classification of rice disease spots is an essential means to promote intelligent rice production. However, due to the large variety of rice diseases and the similar appearance of some rice diseases, the existing deep learning methods are less effective at classification and detection. Aiming at such problems, this paper took the spot images of five common rice diseases as the research object and constructed a rice disease data set containing 2500 images of rice bacterial blight, sheath blight, flax leaf spot, leaf streak and rice blast, including 500 images of each disease. An improved lightweight deep learning network model was proposed to realize the accurate identification of disease types and disease spots. A rice disease image classification network was designed based on the RlpNet (rice leaf plaque net) network model, Which is the underlying network, in addition to the YOLOv3 target detection network model in order to achieve the optimization of the feature extraction link, i.e., upsampling by transposed convolution and downsampling by dilated convolution. The improved YOLOv3 model was compared with traditional convolutional neural network models, including the AlexNet, GoogLeNet, VGG-16 and ResNet-34 models, for disease recognition, and the results showed that the average recall, average precision, average F1-score and overall accuracy of the network model for rice disease classification were 91.84%, 92.14%, 91.87% and 91.84%, respectively, which were all improved compared with the traditional algorithms. The improved YOLOv3 network model was compared with FSSD, Faster-RCNN, YOLOv3 and YOLOv4 for spot detection studies, and the results showed that it could achieve a mean average precision (mAP) of 86.72%, a detection rate (DR) of 93.92%, a frames per second (FPS) rate of 63.4 and a false alarm rate (FAR) of only 5.12%. In summary, the comprehensive performance of the proposed model was better than that of the traditional YOLOv3 algorithm, so this study provides a new method for rice disease identification and disease spot detection. It also had good performance in terms of the common detection and classification of multiple rice diseases, which provides some support for the common differentiation of multiple rice diseases and has some practical application value. Full article

Bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola (Xoc) is one of the most important quarantine diseases in the world. Breeding disease-resistant varieties can solve the problem of prevention and treatment of BLS from the source. The discovery of the molecular mechanism of resistance is an important driving force for breeding resistant varieties. In this study, a BLS-resistant near isogenic line NIL-bls2 was used as the material. Guangxi Xoc strain gx01 (abbreviated as WT) and its mutant strain (abbreviated as MT) with a knockout type III effectors (T3Es) gene were used to infect rice material NIL-bls2. The molecular interaction mechanism of rice resist near isogenic lines in response to infection by different pathogenic strains was analyzed by transcriptome sequencing. The results showed that there were 415, 134 and 150 differentially expressed genes (DEGs) between the WT group and the MT group at 12, 24 and 48 h of post inoculation (hpi). Through GO and KEGG enrichment analysis, it was found that, compared with non-pathogenic strains, the T3Es secreted by pathogenic strains inhibited the signal transduction pathway mediated by ethylene (ET), jasmonic acid (JA) and salicylic acid (SA), and the MAPKK (MAPK kinase) and MAPKKK (MAPK kinase kinase) in the MAPK (mitogen-activated protein kinase) cascade reaction, which prevented plants from sensing extracellular stimuli in time and starting the intracellular immune defense mechanism; and inhibited the synthesis of lignin and diterpenoid phytochemicals to prevent plants from establishing their own physical barriers to resist the invasion of pathogenic bacteria. The inhibitory effect was the strongest at 12 h, and gradually weakened at 24 h and 48 h. To cope with the invasion of pathogenic bacteria, rice NIL-bls2 material can promote wound healing by promoting the synthesis of traumatic acid at 12 h; at 24 h, hydrogen peroxide was degraded by dioxygenase, which reduced and eliminated the attack of reactive oxygen species on plant membrane lipids; and at 48 h, rice NIL-bls2 material can resist the invasion of pathogenic bacteria by promoting the synthesis of lignin, disease-resistant proteins, monoterpene antibacterial substances, indole alkaloids and other substances. Through transcriptome sequencing analysis, the molecular interaction mechanism of rice resistance near isogenic lines in response to infection by different pathogenic strains was expounded, and 5 genes, Os01g0719300, Os02g0513100, Os03g0122300, Os04g0301500, and Os10g0575100 closely related to BLS, were screened. Our work provides new data resources and a theoretical basis for exploring the infection mechanism of Xoc strain gx01 and the resistance mechanism of resistance gene bls2. Full article

Actinomycetes play a vital role as one of the most important natural resources for both pharmaceutical and agricultural applications. The actinomycete strain SPRI-371, isolated from soil collected in Jiangsu province, China, was classified as Streptomyces aureus based on its morphological, physiological, biochemical and molecular biological characteristics. Its bacterial activity metabolites were identified as aureonuclemycin (ANM), belonging to adenosine derivatives with the molecular formula C₁₆H₁₉N₅O₉ for ANM A and C₁₀H₁₃N₅O₃ for ANM B. Simultaneously, the industrial fermentation process of a mutated S. aureus strain SPRI-371 was optimized in a 20 m³ fermentation tank, featuring a rotation speed of 170 rpm, a pressure of 0.05 MPa, an inoculum age of 36–40 h and a dissolved oxygen level maintained at 1–30% within 40–80 h and at >60% in the later period, resulting in an ANM yield of >3700 mg/L. In the industrial separation of fermentation broth, the sulfuric acid solution was selected to adjust pH and 4# resin was used for adsorption. Then, it was resolved with 20% ethanol solution and concentrated in a vacuum (60–65 °C), with excellent results. Antibacterial experiments showed that ANM was less active or inactive against Xanthomonas oryzae pv. oryzae, Xanthomonas citri subsp. citri and Xanthomonas oryzae pv. oryzicola and most bacteria, yeast and fungi in vitro. However, in vivo experiments showed that ANM exhibited extremely significant protective and therapeutic activity against diseases caused by X. oryzae pv. oryzae and X. oryzae pv. oryzicola in rice and X. citri in oranges and lemons. In field trials, ANM A 150 gai/ha + ANM B 75 gai/ha exhibited excellent therapeutic activity against rice bacterial leaf blight, citrus canker and rice bacterial leaf streak. Furthermore, as the dosage and production cost of ANM are lower than those of commercial drugs, it has good application prospects. Full article

Bacterial blight (BB) and bacterial leaf streak (BLS), caused by phytopathogenic bacteria Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xoc), respectively, are the most serious bacterial diseases of rice, while blast, caused by Magnaporthe oryzae (M. oryzae), is the most devastating fungal disease in rice. Generating broad-spectrum resistance to these diseases is one of the key approaches for the sustainable production of rice. Executor (E) genes are a unique type of plant resistance (R) genes, which can specifically trap transcription activator-like effectors (TALEs) of pathogens and trigger an intense defense reaction characterized by a hypersensitive response in the host. This strong resistance is a result of programed cell death induced by the E gene expression that is only activated upon the binding of a TALE to the effector-binding element (EBE) located in the E gene promoter during the pathogen infection. Our previous studies revealed that the E gene Xa23 has the broadest and highest resistance to BB. To investigate whether the Xa23-mediated resistance is efficient against Xanthomonas oryzae pv. oryzicola (Xoc), the causal agent of BLS, we generated a new version of Xa23, designated as Xa23p1.0, to specifically trap the conserved TALEs from multiple Xoc strains. The results showed that the Xa23p1.0 confers broad resistance against both BB and BLS in rice. Moreover, our further experiment on the Xa23p1.0 transgenic plants firstly demonstrated that the E-gene-mediated defensive reaction is also effective against M. oryzae, the causal agent of the most devastating fungal disease in rice. Our current work provides a new strategy to exploit the full potential of the E-gene-mediated disease resistance in rice. Full article

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. Full article

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 OsF3H_03g 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 OsF3H_04g, a homologue of OsF3H_03g. Transferring Tal2c into RS105 induced expression of OsF3H_04g to coincide with increased susceptibility in rice. Overexpressing OsF3H_04g 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 OsF3H_03g or OsF3H_04g 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 OsF3H_04g overexpression line compared with those identified in OsF3H_03g 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. Full article