Xanthomonas Oryzae Research Articles

Confirmations on gene introgression events and hybridity for BLB resistance and yield in rice

A superior variety of rice with high yield and resistance to diseases is expected to meet the increasing demand for this one of the most important carbohydrate sources worldwide. Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the major biotic stresses limiting rice production. Combining BLB resistance and high-yield traits is possible through gene pyramiding. Introgression of targeted traits in the parental genotypes and hybridity confirmation in the F1 generation are important initial steps in gene pyramiding. This study aimed to confirm the presence of targeted genes in parent genotypes using specific molecular markers and to confirm the hybridity of F1 plants generated from three crosses combinations of Inpari 32 (as the female parent) and three Code-qTSN4 lines (as the male parents) using SSR markers. This study successfully confirmed the introgression of BLB-resistance genes in Inpari 32 (Xa4, Xa7, and Xa21) and in Code-qTSN4 (Xa4, Xa7). The introgression of yield-related QTL, qTSN4, was confirmed in the three Code-qTSN4 lines (A10-1, B12-2, and A16-5) by RM17483 marker. Five SSR markers (RM5, RM55, RM105, RM223, and RM561) successfully confirmed the hybridity of F1 derived from Inpari 32xA10-1 (five individuals), Inpari 32xB12-2 (one individual), Inpari 32xA16-5 (two individuals). These confirmed hybrids can be further evaluated in the F2 generation.
 Keywords: Gene pyramiding; molecular marker; SSR; qTSN4; Xoo.

Selection and Characterization of Antagonistic Bacteria as a High Potential Control Agent for Xanthomonas oryzae pv. oryzae Causing Bacterial Leaf Blight Disease in Purple Rice

Bacterial leaf blight disease is one of the most serious diseases in rice caused by Xanthomonas oryzae pv. oryzae. Purple rice is one of the rice varieties that has been infected by this pathogen. Thus, screening and selecting highly efficient antagonistic bacteria for controlling the pathogen is necessary for disease control to reduce epidemics and crop loss. Forty-eight isolates of antagonistic bacteria were isolated from rice rhizosphere soil. The highly effective antagonistic bacteria were selected by dual culture method. The results showed bacteria strains KY16 and KY17 were the most effective antagonists that were able to inhibit X. oryzae pv. oryzae. The inhibition growth zone of antagonists were approximately 12.70 mm and 11.95 mm, respectively. The morphological and some biochemical characteristics of antagonistic bacteria belong to Bacillus sp. Additionally, the antagonists KY16 and KY17 were identified as Bacillus siamensis based on 16S rRNA, gyrA, and rpoB gene sequences. Antimicrobial biosynthesis genes of the antagonistic bacteria were also examined. The results revealed that iturin A and surfactin biosynthesis genes were detectable. From this study, the selected antagonistic bacteria can be used to control the bacterial leaf blight disease of purple rice in rice paddy fields.

Phosphorylation of AGO1a by MAP kinases is required for miRNA mediated resistance against Xanthomonas oryzae pv. oryzae infection in rice

Bacterial leaf blight is a devastating disease caused by Xanthomonas oryzae pv. oryzae (Xoo) which causes severe crop loss in rice. The molecular mechanism that initiates defense against such pathogens remains unexplored. Reports have suggested crucial role of several miRNAs in regulating immune responses in plants. Argonaute (AGO) proteins have been implicated in imparting immunity against pathogens by using small RNAs as guide molecules. Here, we show that phosphorylation of rice AGO1a by MAP kinases is required for miRNA expression regulation during Xoo infection. AGO1a is induced in response to pathogen infection and is under the control of SA signaling pathway. The pathogen responsive MAP kinases MPK3, MPK4 and MPK6, interact with AGO1a in planta and can phosphorylate the protein in vitro. Overexpression of AGO1a extends disease resistance against Xoo in rice and leads to a higher accumulation of miRNAs. Conversely, overexpression of a non phosphorylatable mutant protein aggravates disease susceptibility and remarkably suppresses the miRNA expression levels. At a molecular level, phosphorylation of AGO1a by MAP kinase is required for increased accumulation of miRNAs during pathogen challenge. Taken together, the data suggests that OsAGO1a is a direct phosphorylation target of MAP kinases and this phosphorylation is crucial for its role in imparting disease resistance.

Action mechanism of the potential biocontrol agent Brevibacillus laterosporus SN19-1 against Xanthomonas oryzae pv. oryzae causing rice bacterial leaf blight.

The causal agent of rice bacterial leaf blight (BLB) is Xanthomonas oryzae pv. oryzae (Xoo), which causes serious damage to rice, leading to yield reduction or even crop failure. Brevibacillus laterosporus SN19-1 is a biocontrol strain obtained by long-term screening in our laboratory, which has a good antagonistic effect on a variety of plant pathogenic bacteria. In this study, we investigated the efficacy and bacterial inhibition of B. laterosporus SN19-1 against BLB to lay the theoretical foundation and research technology for the development of SN19-1 as a biopesticide of BLB. It was found that SN19-1 has the ability to fix nitrogen, detoxify organic phosphorus, and produce cellulase, protease, and siderophores, as well as IAA. In a greenhouse pot experiment, the control efficiency of SN19-1 against BLB was as high as 90.92%. Further investigation of the inhibitory mechanism of SN19-1 on Xoo found that the biofilm formation ability of Xoo was inhibited and the pathogenicity was weakened after the action of SN19-1 sterile supernatant on Xoo. The activities of enzymes related to respiration and the energy metabolism of Xoo were significantly inhibited, while the level of intracellular reactive oxygen species was greatly increased. Scanning electron microscopy observations showed folds on the surface of Xoo. A significant increase in cell membrane permeability and outer membrane permeability and a decrease in cell membrane fluidity resulted in the extravasation of intracellular substances and cell death. The results of this study highlight the role of B. laterosporus SN19-1 against the pathogen of BLB and help elucidate the underlying molecular mechanisms.

Evaluation of Equisetin as an Anti-Microbial and Herbicidal Agent from Endophytic Fungus Fusarium sp. JDJR1

Equisetin was isolated from endophytic fungus Fusarium sp. JDJR1 via bioassay-guided isolation, and it was evaluated as an anti-microbial and herbicidal agent. Bioactivity assessments revealed that equisetin exhibited a broad spectrum of fungicidal and anti-bacterial activity against various plant pathogens. The EC50 values against Botrytis cinerea, Fusarium graminearum, Sclerotinia sclerotiorum, and Rhizoctonia solani ranged from 10.7 to 21.0 μg/mL. Notably, equisetin demonstrated activity against Xanthomonas oryzae pv. oryzicola, Xanthomonas oryzae pv. oryzae and Pseudomonas solanacearum with an MIC range of 4–16 μg/mL, surpassing the efficacy of the positive control: streptomycin sulfate. Equisetin, at a concentration of 100 μg/mL, could completely inhibit the spore germination of F. graminearum. In in vivo protection experiments, the protective efficacy of equisetin against B. cinerea on tomato fruits at 200 μg/mL reached 72.9%. Furthermore, in herbicidal activity tests that use the Petri dish bioassay method, equisetin had a good inhibitory effect on the root growth of weeds. At a concentration of 100 μg/mL, the inhibition rates for Echinochloa crusgalli and Eclipta prostrata root were 98.8 and 94.4%, respectively.

Valorization of Chlorella thermophila biomass cultivated in dairy wastewater for biopesticide production against bacterial rice blight: a circular biorefinery approach

Biopesticides offer a sustainable and efficient alternative to synthetic pesticides, providing a safer and more eco-friendly solution to pest management. The present work proposes an innovative approach that integrates crop protection and wastewater treatment using thermophilic microalgal strain Chlorella thermophila (CT) cultivated in nutrient-rich dairy wastewater as a growth medium. The microalgae was cultivated mixotrophically and was able to reduce both organic carbon as well as nutrient load of the dairy wastewater efficiently. The integrated circular biorefinery approach combines biomass cultivation, extraction of biopesticide compounds, and conversion to biocrude. The antimicrobial activity of the biopesticidal extracts against Xanthomonas oryzae and Pantoea agglomerans, the causative agent of bacterial rice blight, is assessed through in vitro studies. The biomass extract obtained is able to inhibit the growth of both the above-mentioned plant pathogens successfully. Mass spectroscopy analysis indicates the presence of Neophytadiene that has previously been reported for the inhibition of several pathogenic bacteria and fungi. Several other value-added products such as linoleic acid and nervonic acids were also been detected in the microalgal biomass which have extremely high nutraceutical and medicinal values. Furthermore, the study investigates the potential for co-production of biocrude from the biorefinery process via hydrothermal liquefaction. Overall, the findings of this present work represent an innovative and sustainable approach that combines wastewater treatment and crop protection using microalgal biomass.

Mutational analysis of predicted active site residues of an exoglucanase secreted by Xanthomonas oryzae pv. oryzae to determine their role in catalysis and in virulence on rice

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight disease in rice. As a part of its virulence repertoire, Xoo secretes a cell wall degrading enzyme Cellobiosidase (CbsA), which is a critical virulence factor and also a determinant of tissue specificity. CbsA protein is made up of an N-terminal catalytic domain and a C-terminal fibronectin type III domain. According to the CAZy classification, the catalytic domain of CbsA protein belongs to the glycosyl hydrolase-6 (GH6) family that performs acid–base catalysis. However, the identity of the catalytic acid and the catalytic base of CbsA is not known. Based on the available structural and biochemical data, we identified putative catalytic residues and probed them by site-directed mutagenesis. Intriguingly, the biochemical analysis showed that none of the mutations abolishes the catalytic activity of CbsA, an observation that is contrary to other GH6 family members. All the mutants exhibited altered enzymatic activity and caused significant virulence deficiency in Xoo emphasising the requirement of specific exoglucanase activity of wild-type CbsA for virulence on rice. Our study highlights the need for further studies and the detailed characterisation of bacterial exoglucanases.

Seed Soaking Using Methanol Kalanchoe pinnata Leaf Extracts Induces Rice Resistance against Bacterial Leaf Blight

Bacterial leaf blight (Xanthomonas oryzae pv. oryzae) is a significant rice disease. Aqueous crude extracts of Kalanchoe pinnata were shown to induce rice resistance against the disease. This study aims at testing the disease-reducing effects of K. pinnata leaf extracts using the liquid–liquid extraction method with three different solvents (dichloromethane, methanol, and water). This serves as a basis to select appropriate extracts for effective disease control. Three concentrations (1, 1.5, and 2%) of each extract were tested using seed soaking. The extracts did not show adverse effects on seed germination and seedling growth. Methanol extracts showed significantly different effects compared to those of the untreated control. The involvement of induced resistance in the disease reduction was shown through activities of the four defense-related and antioxidant enzymes, i.e., peroxidase (POX), catalase (CAT), polyphenol oxidase (PPO), and phenylalanine ammonia lyase (PAL). Using 1% methanol extract, activities of POX and CAT involved in hydrogen peroxide production in rice tissues increased 1–4 days after pathogen inoculation (DAI) and remained at high levels until 6 DAI. Activities of PPO and PAL involved in resistance signaling pathways significantly increased after pathogen inoculation. Activities of the four enzymes generally increased after pathogen inoculation and reached higher levels with extract applications.

Synthesis, X-ray Crystal Structure, and Antimicrobial Studies of New Quinazolin-4(3H)-one Derivatives Containing the 1,2,4-Triazolo[3,4-b][1,3,4]thiadiazole Moiety and 4-Piperidinyl Linker.

A total of 35 new quinazolinone derivatives bearing the 1,2,4-triazolo[3,4-b][1,3,4]thiadiazole scaffold and the 4-piperidinyl linker were designed, prepared, and assessed for their antibacterial and antifungal activities. Among these derivatives, the chemical structure of compound F5 was clearly verified via single-crystal X-ray diffraction analysis. The experimental results revealed that some of the compounds displayed good even excellent inhibitory effects toward the tested phytopathogenic bacteria. For instance, compound F33 was capable of strongly inhibiting Xanthomonas oryzae pv. oryzae (Xoo) in vitro with an EC50 (half-maximal effective concentration) value of 4.1 μg/mL, about 16-fold more effective than the commercialized bactericide bismerthiazol. Significantly, this compound also effectively suppressed the proliferation of Xoo in the potted rice plants, showing a good in vivo protection efficacy of 47.6% at 200 μg/mL. Subsequently, the antibacterial mechanisms of compound F33 were explored by means of different biophysical and biochemical methods. Last, some of the compounds were found to possess relatively good antifungal activities in vitro, like compound F19 against Phytophthora nicotianae (with an inhibition rate of 67.2% at 50 μg/mL). In a word, the current experimental results imply that the 4-piperidinyl-bridged quinazolinone-1,2,4-triazolo[3,4-b][1,3,4]thiadiazole derivatives possess potential as lead compounds for developing more efficient anti-Xoo bactericides.

Bioinspired total synthesis and biological activity of Pegaharine A.

Phytopathogens cause various diseases by parasitizing crops, reducing crop yield and resulting in substantial economic losses in agricultural production. A novel type isolated from the perennial herbaceous Peganum harmala L. seeds, β-carboline alkaloids pegaharine A (PA), has become a hot topic in developing plant-originated green pesticides owing to their significant physiological activities. A scalable bioinspired total synthesis of PA is accomplished in the present work. The systematical biological assay study showed that PA exhibited moderate inhibitory activity against nine tested plant pathogenic fungi and showed significant inhibitory activity in vitro against the three tested plant pathogenic bacteria. Most noteworthy is the inhibitory rates of PA on Xanthomonas oryzae pv. oryzae (Xoo), X. oryzae pv. oryzicola (Xoc) and X. axonopodis pv. citri (Xac) of 93.6%, 92.1% and 86.1%, respectively, which are better than the control drug, bismerthiazol (63.4%, 61.2% and 53.7% at 100 μg mL-1 concentration). Furthermore, the EC50 value of PA against Xoo, Xoc and Xac was 52.2, 60.0 and 65.1 μg mL-1 , respectively, superior to 72.9, 64.2 and 70.1 μg mL-1 of the control drug. Moreover, the anti-Xoo mechanistic studies revealed that PA exerted its antibacterial effects by increasing the permeability of the bacterial membrane, reducing the extracellular polysaccharide content and inducing morphological changes in bacterial cells. A novel β-carboline alkaloid, PA, was prepared by biomimetic total synthesis. Its significant antibacterial activity was closely related to the permeation of bacterial cell membranes, which was confirmed by anti-Xoo mechanistic studies. More importantly, the structure could be regarded as a model for developing novel bactericides. © 2023 Society of Chemical Industry.

Elevating plant immunity by translational regulation of a rice WRKY transcription factor.

Plants have intricate mechanisms that tailor their defence responses to pathogens. WRKY transcription factors play a pivotal role in plant immunity by regulating various defence signalling pathways. Many WRKY genes are transcriptionally activated upon pathogen attack, but how their functions are regulated after transcription remains elusive. Here, we show that OsWRKY7 functions as a crucial positive regulator of rice basal immunity against Xanthomonas oryzae pv. oryzae (Xoo). The activity of OsWRKY7 was regulated at both translational and post-translational levels. Two translational products of OsWRKY7 were generated by alternative initiation. The full-length OsWRKY7 protein is normally degraded by the ubiquitin-proteasome system but was accumulated following elicitor or pathogen treatment, whereas the alternate product initiated from the downstream in-frame start codon was stable. Both the full and alternate OsWRKY7 proteins have transcriptional activities in yeast and rice cells, and overexpression of each form enhanced resistance to Xoo infection. Furthermore, disruption of the main AUG inrice increased the endogenous translation of the alternate stabilized form of OsWRKY7 and enhanced bacterial blight resistance. This study provides insights into the coordination of alternative translation and protein stability in the regulation of plant growth and basal defence mediated by the OsWRKY7 transcription factor, and also suggests a promising strategy to breed disease-resistant rice by translation initiation control.

Effects of seed soaking and foliar spraying of Kalanchoe pinnata aqueous leaf extracts against rice bacterial leaf blight

This study aims at evaluating the disease-reducing effects against rice bacterial leaf blight (Xanthomonas oryzae pv. oryzae). Under greenhouse conditions, the activities of the four enzymes [peroxidase (POX), catalase (CAT), polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL)] after application of Kalanchoe pinnata aqueous leaf extracts using the combination of seed soaking and foliar spraying were studied. Overall, two extract concentrations [1 and 2% (w/v)] applied as seed soaking combined with the five extract concentrations [1, 2, 3, 4 and 5% (w/v)] applied as foliar spraying were tested. Three application methods were furthermore used for foliar spraying (7 days before pathogen inoculation (DBI), 14 DBI and their combination). Results showed the effects increased with the increase of extract concentrations and durations from application time points prior to pathogen inoculation. The combination of foliar spraying at 7 and 14 DBI provided stronger protection compared to single sprays. The effects involved induced resistance. Indeed, the activities of POX and CAT increased until 4 days after inoculation (DAI) and remained until 7 DAI, while those of PPO and PAL increased similarly then decreased until 7 DAI. Activities of these enzymes increased after pathogen inoculation and reached higher levels with extract applications.

Discovery of tryptanthrin analogues bearing F and piperazine moieties as novel phytopathogenic antibacterial and antiviral agents.

Plant bacterial infections and plant viruses seriously affect the yield and quality of crops. Based on the various activities of tryptanthrin, a series of tryptanthrin analogues bearing F and piperazine moieties were designed, synthesized, and evaluated for their biological activities against three plant bacteria and tobacco mosaic virus (TMV). Bioassay results indicated that compounds 6a-6l displayed excellent antibacterial activities in vitro and 6a-6c and 6g exhibited better antiviral activities against TMV than commercial ribavirin. In particular, 6b showed the most effect on Xanthomonas oryzae pv. oryzae (Xoo) with a half-maximal effective concentration (EC50 ) of 1.26 μg mL-1 , compared with the commercial pesticide bismerthiazol (BT; EC50 = 34.3 μg mL-1 ) and thiodiazole copper (TC; EC50 = 73.3 μg mL-1 ). Meanwhile, 6a also had the best antiviral activity at 500 μg mL-1 for curative, protection, and inactivation purposes, compared with ribavirin in vivo. Compound 6b could cause changes in bacterial morphology, induce the accumulation of reactive oxygen species, promote apoptosis of bacterial cells, inhibit the formation of biofilm, and block the growth of Xoo cells. Proteomic analysis revealed major differences in the bacterial secretory system pathways T2SS and T6SS, which inhibited membrane transport. Molecular docking revealed that 6a and 6g could interact with TMV coat protein preventing virus assembly. These results suggest that tryptanthrin analogues bearing F and piperazine moieties could be promising candidate agents for antibacterial and antiviral use in agricultural production. © 2023 Society of Chemical Industry.

MiR2118 Negatively Regulates Bacterial Blight Resistance through Targeting Several Disease Resistance Genes in Rice.

Plant miRNAs are a class of noncoding RNA with a length of 21-24 nt that play an important role in plant responses to biotic and abiotic stresses. Bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most serious bacterial diseases in rice. Our previous work showed that osa-miR2118b/n was induced by Xoo infection. However, the biological function of miR2118 has not yet been characterized in experiments. Herein, we constructed MIR2118b OE, as well as single and double mutants of MIR2118b/n using CRISPR/Cas9. Further results showed that osa-MIR2118b OE plants exhibited longer lesion lengths than the wild type after Xoo inoculation, while MIR2118 CRISPR plants exhibited shorter lesion lengths than the wild type after Xoo inoculation. Co-transformation experiments in rice protoplasts indicated that osa-miR2118 negatively regulated the transcripts of three nucleotide-binding sites and leucine-rich repeat (NLR) genes (LOC_Os08g42700.1, LOC_Os01g05600.1, and LOC_Os12g37290.1) which are predicted target genes of miR2118, but not the mutated NLR genes with a 3 bp insertion at the center of the binding sites. The transcriptional level of the three NLR genes was reversed relative to osa-miR2118 in the MIR2118b OE and MIR2118b CRISPR plants. The above results demonstrate that osa-miR2118b/n negatively regulates the resistance to bacterial blight through negatively regulating several NLR genes.

Genome Sequence Resource for Xanthomonas oryzae pv. oryzae (Causal Agent of Bacterial Leaf Blight of Rice) Isolated from Pakistan

Bacterial leaf blight, one of the most destructive diseases of rice, is prevalent in the major rice-growing regions of the world. Bacterial leaf blight is caused by Xanthomonas oryzae pv. oryzae ( Xoo) and greatly affects the quantity and quality of the rice crop. We sequenced the whole genome of Xoo isolated from a Basmati rice-growing region of Pakistan using long-read Oxford Nanopore sequencing technology. About 637 MB of data was retrieved, corresponding to 133× genome sequencing coverage. The final assembly yields a circular chromosome of 4,930,682 bp. The complete genome of the Xoo isolate from Pakistan will enhance sequence resources for global comparison of Xoo diversity across the region. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

A unique role of the pyrimidine de novo synthesis enzyme ODCase in Lysobacter enzymogenes

Bacterial species of the genus Lysobacter are environmentally ubiquitous with strong antifungal biocontrol potential. Heat-stable antifungal factor (HSAF) secreted by the biocontrol bacterium Lysobacter enzymogenes OH11 has broad-spectrum and highly efficient antifungal activity. Studying the biosynthetic regulations of HSAF would lay an important foundation for strain engineering toward improved HSAF production. In this work, we demonstrate that Le0752, an orotidine-5´-phosphate decarboxylase enzyme (ODCase) catalyzing a pivotal step of the UMP de novo biosynthesis pathway, is vital for HSAF-mediated antimicrobial activities and growth of L. enzymogenes OH11, but not for twitching motility. This gene regulates the production of HSAF by affecting the expression of lafB, a key gene in the HSAF biosynthesis operon, through the transcription factor Clp. Interestingly, bioinformatics analysis revealed that Le0752 belongs to the Group III ODCases, whereas its homologs in the closely related genera Xanthomonas and Stenotrophomonas belong to Group I, which contains most ODCases from Gram-positive bacteria, Gram-negative bacteria and cyanobacteria. Moreover, the Group I ODCase PXO_3614 from the Xanthomonas oryzae pv. oryzae PXO99A strain complemented the Le0752 mutant in regulating HSAF-mediated antagonistic activity. Together, these results highlight the important requirement of de novo pyrimidine biosynthetic enzymes for antibiotic HSAF production in L. enzymogenes, which lays an important foundation for improving HSAF production via metabolic flow design and for dissecting the regulatory functions of bacterial ODCases.

IN VITRO STUDY OF PHYLLOSPHERE BACTERIA AS PROMISING BIOCONTROL AGENTS AGAINST BACTERIAL LEAF BLIGHT DISEASE (Xanthomonas oryzae pv. oryzae) IN RICE PLANTS

This study aimed to explore the potential of phyllosphere bacterial isolates as biological control agents against Xanthomonas oryzae pv. oryzae (Xoo). We isolated a strain exhibiting typical morphological and biochemical characteristics of Xoo, such as a yellowish, spherical colony, Gram-negative, and starch hydrolyzing ability. This isolate was shown to cause necrotic lesions on rice leaves, corroborating its pathogenic nature. Subsequently, we isolated ten diverse phyllosphere bacteria from rice plants. These isolates were characterized based on various morphological and biochemical attributes including colony shape, margin appearance, color, growth time, and the ability to fluoresce under UV light when cultivated on King's B medium. Crucially, none of these isolates induced a hypersensitive response in tobacco leaves, an initial indicator of their potential as safe biological control agents. In an antagonistic assay against Xoo, all the phyllosphere isolates demonstrated varying levels of inhibition, suggesting their potential role in biological control. Two isolates, PGM1 and PGM4, exhibited the highest antagonistic effects against Xoo. These findings provide preliminary evidence for the potential use of these phyllosphere isolates in managing bacterial leaf blight in rice, warranting further research to confirm their efficacy and safety under field conditions.

Repurposing Salicylamides to Combat Phytopathogenic Bacteria and Induce Plant Defense Responses.

Based on the research strategy of "drug repurposing", a series of derivatives and marketed drugs that containing salicylic acid skeleton were tested for their antibacterial activities against phytopathogens. Salicylic acid can not only regulate some important growth metabolism of plants, but also induce plant disease resistance. The bioassay results showed that the salicylamides exhibited excellent antibacterial activity. Especially, oxyclozanide showed the best antibacterial effect against Xanthomonas oryzae, Xanthomonas axonopodis pv. citri and Pectobacterium atroseptica with MICs of 0.78, 3.12 and 12.5 μg.mL-1, respectively. In vivo experiments with rice bacterial leaf blight had further demonstrated that oxyclozanide exhibited stronger antibacterial activity than the commercial bactericide, thiodiazole copper. Oxyclozanide could induce plant defense responses through the determination of salicylic acid content and the activities of defense-related enzymes including CAT, POD, and SOD in rice. The preliminarily antibacterial mechanism study indicated that oxyclozanide exhibited the antibacterial activity by disrupting cell integrity and reducing bacterial pathogenicity. Additionally, oxyclozanide could induce plant defense responses through the determination of salicylic acid content.

Design, synthesis and antibacterial activity of coumarin-3-carboxylic acid derivatives containing acylhydrazone moiety

Plant diseases caused by bacteria lead to enormous yield and economic losses in agricultural production. To develop innovative bactericides for controlling plant disease, a series of coumarin-3-carboxylic acid derivatives containing acylhydrazone thioether/sulfoxide were designed and synthesized by utilizing the principles of bioisosterism and active splicing. The bactericidal activities of these derivatives were subsequently evaluated. The result indicated that E2 exhibited the highest antibacterial activities in vitro against Xanthomonas oryzae pv. oryzae, Ralstonia solanacearum, and Acidovorax citrulli, with EC50 values of 2.97, 1.17, and 1.23 µg/mL, respectively. Moreover, in vivo experiments showed that E2 exhibited robust protective properties against Ralstonia solanacearum, and Acidovorax citrulli, with efficacies of 72.52 % and 63.90 %, respectively, at a concentration of 100 µg/mL, which was comparable to the positive control kasugamycin at the same concentration. Scanning electron microscopy analysis revealed that E2 induced changes in the cellular morphology of Acidovorax citrulli, such as shrinkage and collapse. Subsequent investigation revealed that E2 had the capacity to compromise the structural stability of the bacterial membrane of Ralstonia solanacearum. This study represents the first report on the antibacterial activities of this series of coumarin-3-carboxylic acid derivatives containing the acylhydrazone thioether/single-sulfoxide moiety, and the results suggest that coumarin-3-carboxylic acid derivatives containing the acylhydrazone single-sulfoxide may hold potential as effective antibacterial agents. And coumarin-3-carboxylic acid derivatives exhibited promise as plant bacterial agent to prevent rice bacterial leaf blight, tomato bacterial wilt, and melon bacterial fruit blotch.

Resistance of Six Lines from Farmers against Bacterial Leaf Blight (Xanthomonas oryzae pv. oryzae)

Penyakit hawar daun bakteri (HDB) yang disebabkan oleh Xanthomonas oryzae pv. oryzae merupakan salah satu penyakit yang merugikan bagi produksi padi Indonesia. Salah satu upaya untuk mengendalikan penyakit tersebut ialah menggunakan varietas tahan HDB. Penelitian ini bertujuan untuk mengevaluasi perkembangan penyakit HDB pada enam galur hasil rakitan dari petani (TCIPB202101, TCIPB202102, TCIPB202103, TCIPB202104, TCIPB202105, dan TCIPB202106) dan dua varietas pembanding, yaitu Ciherang dan Inpari 30. Penelitian dilakukan di rumah kaca Cikabayan Dramaga dengan inokulasi buatan. Pengamatan meliputi masa inkubasi, panjang lesio dan intensitas penyakit pada 7 dan 14 hari setelah inokulasi (HSI). Hasil penelitian menunjukkan semua galur yang diuji merupakan galur yang tahan terhadap penyakit HDB dan memiliki ketahanan yang lebih baik dibandingkan dengan varietas pembandingnya. Galur TCIPB202105 dan TCIPB202106 memiliki respons ketahanan terbaik dengan rata-rata intensitas penyakit dibawah 1%, sedangkan TCIPB202101 dan TCIPB202102 memiliki intensitas penyakitnya yang paling tinggi namun masih dalam kategori agak tahan.