Bacterial Leaf Blight Disease Of Rice Research Articles

Development of Spectral Features for Monitoring Rice Bacterial Leaf Blight Disease Using Broad-Band Remote Sensing Systems

Development of Spectral Features for Monitoring Rice Bacterial Leaf Blight Disease Using Broad-Band Remote Sensing Systems

Transcriptomic and proteomic profiling reveals toxicity and molecular action mechanisms of bioengineered chitosan‑iron nanocomposites against Xanthomonas oryzae pv. oryzae

Bacterial leaf blight (BLB) pathogen, Xanthomonas oryzae pv. oryzae (Xoo) is the most devastating bacterial pathogen, which jeopardizes the sustainable rice (Oryza sativa L.) production system. The use of antibiotics and conventional pesticides has become ineffective due to increased pathogen resistance and associated ecotoxicological concerns. Thus, the development of effective and sustainable antimicrobial agents for plant disease management is inevitable. Here, we investigated the toxicity and molecular action mechanisms of bioengineered chitosan‑iron nanocomposites (BNCs) against Xoo using transcriptomic and proteomic approaches. The transcriptomic and proteomics analyses revealed molecular antibacterial mechanisms of BNCs against Xoo. Transcriptomic data revealed that various processes related to cell membrane biosynthesis, antioxidant stress, DNA damage, flagellar biosynthesis and transcriptional regulator were impaired upon BNCs exposure, which clearly showing the interaction of BNCs to Xoo pathogen. Similarly, proteomic profiling showed that BNCs treatment significantly altered the levels of functional proteins involved in the integral component of the cell membrane, catalase activity, oxidation-reduction process and metabolic process in Xoo, which is consistent with the results of the transcriptomic analysis. Overall, this study suggested that BNCs has great potential to serve as an eco-friendly, sustainable, and non-toxic alternative to traditional agrichemicals to control the BLB disease in rice.

First report of rice bacterial leaf blight disease caused by Pantoea ananatis in the United States.

In August 2021, bacterial leaf blight-like symptoms were observed on 14 out of 570 rice genotypes (Oryza sativa) in research field plots of global rice germplasm grown in Arkansas (eXtra Figure S1. A & B). The disease was characterized by spreading lesions on leaves, panicle sterility and reduced yield in highly susceptible, mature rice germplasm. No spread of disease to nearby plants was observed. Isolations were performed at Colorado State University, where soakates from symptomatic leaves were spread onto nutrient agar. After 72 h at 28°C, uniform, distinct, yellow-colored bacterial colonies were observed. To screen for the presence of common rice bacterial pathogens, PCR amplification directly from colonies or from DNA isolated from symptomatic field-collected leaves was performed. Primers specific for Xanthomonas oryzae pvs. oryzae and oryzicola (Lang et al., 2010), Burkholderia glumae (Echeverri-Rico et al., 2021), and Pseudomonas fuscovaginae (Ash et al., 2014) did not amplify indicating these organisms were not present. Sequencing of 16S rRNA gene (Weisburg et al., 1991) amplicons suggested the bacteria belonged to the genera Pantoea and Sphingomonas (NCBI accession no. OP683332 and OP683333, respectively). Amplicons resulting from primers specific to the gyrB gene region of P. ananatis (Kini et al., 2021) were sequenced and the fragment was compared to the P. ananatis PA13 reference genome using a BLAST analysis. One candidate (AR358) showed 100% identity with the P. ananatis gyrB region. Primers specific for Sphingomonas sp. (Bangratz et al., 2020) confirmed the second candidate (AR359) as a Sphingomonas sp. The identity of P. ananatis was confirmed by the Plant Pathogen Confirmatory Diagnostics Laboratory (Beltsville, MD, USA). To determine pathogenicity, leaves from 7-day-old seedlings of rice (Oryza sativa) cultivar Kitaake were scissor-clip inoculated (Kauffman et al., 1973) with four different treatments and compared to control leaves inoculated with sterile water. Treatments for the experiment consisted of bacterial suspensions (108 CFU/ml) of the two candidate organisms, P. ananatis (strain AR358) or Sphingomonas sp. (strain AR359), individually or in a 1:1 ratio of P. ananatis:Sphingomonas sp., or soakate from infected field tissue. Lesions similar to those observed in the field were only detected on leaves inoculated with P. ananatis or infected field tissue soakate at 7-days post-inoculation (eXtra Figure S1. C). Bacteria were recovered from the leaves of the artificially inoculated seedlings from three treatments (P. ananatis, P. ananatis:Sphingomonas sp. and soakate from the infected field tissue) and were determined to be P. ananatis based on colony morphology, amplification of 16s rRNA, and gyrB sequence data. Our results confirm the pathogenicity of P. ananatis to rice and fulfill Koch's postulates. P. ananatis was also recovered from several similarly diseased rice breeding lines at the University of Arkansas System Division of Agriculture Rice Research and Extension Center. We conclude that P. ananatis is the causal pathogen for leaf blight-like symptoms observed in the global rice cultivars grown in Arkansas. P. ananatis was previously reported as a pathogen on rice in several rice growing regions, including China (Yu et al., 2021), India (Reshma et al., 2022), and Africa (Kini et al., 2017), however, this is the first report of P. ananatis as a pathogen of rice in the United States.

Foliar Application or Seed Priming of Cholic Acid-Glycine Conjugates can Mitigate/Prevent the Rice Bacterial Leaf Blight Disease via Activating Plant Defense Genes.

Xanthomonas Oryzae pv. oryzae (Xoo) causes bacterial blight and Rhizoctonia solani (R. solani) causes sheath blight in rice accounting for >75% of crop losses. Therefore, there is an urgent need to develop strategies for the mitigation of these pathogen infections. In this study, we report the antimicrobial efficacy of Cholic Acid-Glycine Conjugates (CAGCs) against Xoo and R. solani. We show that CAGC C6 is a broad-spectrum antimicrobial and is also able to degrade biofilms. The application of C6 did not hamper plant growth and showed minimal effect on the plant cell membranes. Exogenous application of C6 on pre-infection or post-infection of Xoo on rice susceptible genotype Taichung native (TN1) can mitigate the bacterial load and improve resistance through upregulation of plant defense genes. We further demonstrate that C6 can induce plant defense responses when seeds were primed with C6 CAGC. Therefore, this study demonstrates the potential of CAGCs as effective antimicrobials for crop protection that can be further explored for field applications.

Isolation and Characterization of Novel Phages Targeting Xanthomonas oryzae: Culprit of Bacterial Leaf Blight Disease in Rice.

Background: Bacterial leaf blight (BLB) disease caused 80% of disease incidence in paddy in Kedah and Selangor states of Malaysia. The pathogenic bacterium, Xanthomonas oryzae pv. oryzae (Xoo), is one of the destructive pathogens infecting lowland irrigated and rainfed paddy in Asia's tropical and temperate environments. Bacteriophages (or phages) have been proposed to control the pathogen due to their efficacy and safety aspects. Material and Methods: In this study, a total of 70 Xoo-phages were isolated from termite which living in rice-growing area. Results: 2 lytic phages NΦ-1 and NΦ-3 were selected due to the high titer of the virus. Electron microscopic analysis showed that those phages belonged to the family Podoviridae, order Caudovirales with short noncontracted tails. Moreover, these phages have a narrow host range specifically target Xoo with a higher burst size. Whole-genome sequencing showed that the Xoo-phage NΦ-1 and NΦ-3 consists of a linear double-stranded DNA molecule of length 41,151 and 38,454 bp, respectively. Conclusion: This study successfully characterized two novel Xanthomonas phages and their potential as antimicrobial agents against BLB disease in rice.

Novel Ferulic Amide Ac6c Derivatives: Design, Synthesis, and Their Antipest Activity.

Thirty-eight novel ferulic amide 1-aminocyclohexane carboxylic acid (Ac6c) derivatives D1-D19 and E1-E19 were designed and synthesized, and their antibacterial, antifungal, and insecticidal activities were tested. Most of the synthesized compounds displayed excellent activity againstXanthomonas oryzae pv. oryzae (Xoo), with EC50 values ranging from 11.6 to 83.1 μg/mL better than that of commercial bismerthiazol (BMT, EC50 = 84.3 μg/mL), as well as much better performance compared to that of thiediazole copper (TDC, EC50 = 137.8 μg/mL). D6 (EC50 = 17.3 μg/mL), D19 (EC50 = 29.4 μg/mL), E3 (EC50 = 29.7 μg/mL), E9 (EC50 = 27.0 μg/mL), E10 (EC50 = 18.6 μg/mL), and E18 (EC50 = 20.8 μg/mL) showed much higher activity on Xanthomonas oryzae pv. oryzicola compared with BMT (EC50 = 80.1 μg/mL) and TDC (EC50 = 124.7 μg/mL). In relation to controlling the fungus, Rhizoctonia solani, E1, E10, and E13 had much lower EC50 values of 0.005, 0.140, and 0.159 μg/mL compared to hymexazol at 74.8 μg/mL. Further in vivo experiments demonstrated that E6 and E12 controlled rice bacterial leaf blight disease better than BMT and TDC did. Scanning electron microscopy (SEM) studies revealed that E12 induced the Xoo cell membrane collapse. Moreover, D13 (73.7%), E5 (80.6%), and E10 (73.4%) also showed moderate activity against Plutella xylostella. These results indicated that the synthesized ferulic amide Ac6c derivatives showed promise as candidates for treating crop diseases.

English

Bacterial leaf blight (BLB) is a disease caused by Xanthomonas oryzae pv. oryzae (Xoo) of rice in rice-producing countries including Indonesia and attack rice in all stages of growth. In the advanced, crop production will be decreased by up to 50–70%. Recently, the practical efforts to overcome the problem by using resistant varieties, antibiotics, and sanitation; however, the ability of the pathogen to forms the new virulent pathotypes is noteworthy. Alternatively, the pathogen could be environmental-friendly controlled by utilizing bacteriophages as biological control agents because of their specific characteristics to their bacterial hosts. This research aimed to obtain information about the characteristic of the first isolated bacteriophages from Indonesia. The result showed that two bacteriophages had been isolated from soil in Arjasa Jember and soil in Gadingan Situbondo, namely phage XooX1IDN and phage XooX2IDN, respectively. The two phages were inactivated at 80ºC and stable at pH within the range of 6 to 8. The phage XooX1IDN has a genome size of approximately 39 kb, while phage XooX2IDN had a genome size 38 kb. Morphologically, both phages possessed the family of Myoviridae. Phage biocontrol in vitro assay showed that both phages significantly reduced the growth of BLB pathogen, indicating that both phages potentially, as biological control agents for BLB disease in rice. © 2021 Friends Science Publishers

Fluorescence-based immunoassay for the detection of Xanthomonas oryzae pv. oryzae in rice leaf

The identification of rice bacterial leaf blight disease requires a simple, rapid, highly sensitive, and quantitative approach that can be applied as an early detection monitoring tool in rice health. This paper highlights the development of a turn-off fluorescence-based immunoassay for the early detection of Xanthomonas oryzae pv. oryzae (Xoo), a gram-negative bacterium that causes rice bacterial leaf blight disease. Antibodies against Xoo bacterial cells were produced as specific bio-recognition molecules and the conjugation of these antibodies with graphene quantum dots and gold nanoparticles was performed and characterized, respectively. The combination of both these bio-probes as a fluorescent donor and metal quencher led to changes in the fluorescence signal. The immunoreaction between AntiXoo-GQDs, Xoo cells, and AntiXoo-AuNPs in the immuno-aggregation complex led to the energy transfer in the turn-off fluorescence-based quenching system. The change in fluorescence intensity was proportional to the logarithm of Xoo cells in the range of 100–105 CFU mL−1. The limit of detection was achieved at 22 CFU mL−1 and the specificity test against other plant disease pathogens showed high specificity towards Xoo. The detection of Xoo in real plant samples was also performed in this study and demonstrated satisfactory results.

Molecular docking and molecular dynamics simulation approach to screen natural compounds for inhibition of Xanthomonas oryzae pv. Oryzae by targeting peptide deformylase

Xanthomonas oryzae pv. Oryzae (Xoo) causes bacterial leaf blight (BLB) of rice which results in a huge loss in production. Many chemicals are used to control BLB disease. However, these chemicals are toxic to the environments, animals and human beings. Thus, there is a demand to discover potential and safe natural pesticides to manage BLB disease successfully. Therefore, we screened a library of phytochemicals of different plants having antibacterial activity by targeting Peptide Deformylase (PDF) of Xoo using in silico techniques. A library of 318 phytochemicals was prepared and subjected to rigid and flexible molecular docking against PDF followed by molecular dynamics simulation and free energy analysis of protein-ligand complexes. The results of virtual screening showed that 14 compounds from different plants have good binding energy as compare to reference molecule (3 R)-2,3-dihydro[1,3] thiazolo [3,2 a]benzimidazol-3-ol) (-7.7 kcal mol−1). Out of 14 hit compounds, eight compounds that were selected based on binding energy were analyzed by Molecular dynamic (MD) simulation. Analysis of MD simulation revealed that eight compounds namely; Bisdemethoxycurcumin, Rosmarinic acid, Piperanine, Dihydropiperlonguminine, Piperdardine, Dihydrocurcumin and Lonhumosides B achieved good stability during the 80 ns MD simulation at 300 K in term of the RMSD. Further, we calculated RMSF, RG, SASA, and interaction energy after 40 ns due to showing the stability of complexes. From our results, we conclude that these natural compounds could inhibit Xoo by targeting PDF receptor and can be used as potential bactericidal candidates against BLB disease of rice against Xoo and other bacteria. Communicated by Ramaswamy H. Sarma

Interaction of Antagonistic Bacteria that Effective to Control Rice Bacterial Leaf Blight Disease with Agricultural Chemicals and Bio-products

Interaction of Antagonistic Bacteria that Effective to Control Rice Bacterial Leaf Blight Disease with Agricultural Chemicals and Bio-products

Bacterial leaf blight resistance in rice: a review of conventional breeding to molecular approach.

Breeding for disease resistant varieties remains very effective and economical in controlling the bacterial leaf blight (BLB) of rice. Breeders have played a major role in developing resistant rice varieties against the BLB infection which has been adjudged to be a major disease causing significant yield reduction in rice. It would be difficult to select rice crops with multiple genes of resistance using the conventional approach alone. This is due to masking effect of genes including epistasis. In addition, conventional breeding takes a lot of time before a gene of interest can be introgressed. Linkage drag is also a major challenge in conventional approach. Molecular breeding involving markers has facilitated the characterization and introgression of BLB disease resistance genes. Biotechnology has brought another innovation in form of genetic engineering (transgenesis) of rice. Although, molecular breeding cannot be taken as a substitute for conventional breeding, molecular approach for combating BLB disease in rice is worthwhile given the demand for increased production of rice in a fast growing population of our society. This present article highlights the recent progress from conventional to molecular approach in breeding for BLB disease resistant rice varieties.

Controlling and Defence‐related Mechanisms of Bacillus Strains Against Bacterial Leaf Blight of Rice

AbstractBacillus strains are broadly studied for their beneficial role in plant growth and biological control of plant disease and pest; however, little is known about their underlying mechanisms. In this study, we assessed the controlling and defence‐related mechanisms of three Bacillus strains including rice seed‐associated strain B. subtilis A15, rhizobacterial strains B. amyloliquefaciens D29 and B. methylotrophicus H8, all of which are against bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae. Results indicated that all three strains showed strong biofilm formation ability. The culture filtrates of each strain significantly suppressed the growth and biofilm formation of X. oryzae, while changes in bacterial cell morphology such as cell swell and severe cell wall alterations were observed through the transmission electron microscopy images. PCR analysis revealed that all three strains harbour the antimicrobial‐associated genes that are responsible for biosynthesis of bacillomycin, fengycin, iturin and surfactin. Subsequent real‐time qPCR analysis revealed the upregulated expression of fenD and srfAA genes in D29 and H8, and fenD and ituC genes in A15 during their in vitro interaction with X. oryzae. It suggests that the antibacterial mechanisms of the three strains may be at least partially associated with their ability to secrete corresponding lipopeptides. Interestingly, the applications of the three strains in greenhouse conditions were found to be effective in controlling the BLB disease, which was achieved through the activation of inducing systemic resistance resulted from the enhanced activities of defence‐related enzymes. This is the first report of demonstration of the mode of antibacterial effect of Bacillus strains against X. oryzae. Overall, data from the current study provide valuable information for biological control of BLB disease in rice.

Plant growth-promoting rhizobacteria mediate induced systemic resistance in rice against bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae

Seven Bacillus plant growth-promoting rhizobacteria spp. were evaluated for growth promotion and induced systemic resistance in rice against Xanthomonas oryzae pv. oryzae (Xoo). The identities of colonies of X. oryzae pv. oryzae grown on mXOS and PSA medium were confirmed by PCR employing specific primers TXTF and TXT4R. Among the seven strains tested as fresh suspensions, talc and sodium alginate formulations under laboratory and green house conditions, maximum germination of 86% was recorded after seed treatments with fresh suspension of Bacillus subtilis GBO3 followed by 85% germination treated with Bacillus pumilus SE34 in comparison to only 71% germination in the untreated controls. Similarly, the maximum vigor index of 1374 was obtained by seed treatment with fresh suspensions of B. subtilis strain GBO3 followed by treatments with strain SE34 with vigor index of 1323 in contrast to an index of only 834 observed in untreated controls. Among the treatments, seed treatments with fresh suspension of seven strains resulted in better germination and vigor assessments than talc based or sodium alginate formulations. Seed treatments with fresh suspension of strain SE34 gave 71% protection, followed by B. subtilis GBO3 and B. pumilus T4 with 58% and 52% protection, respectively, compared to the untreated controls. Seed treatments with talc based formulation of SE34 gave 66% protection, while GBO3 and T4 resulted in 52% and 50% protection, respectively, with similar formulation. Seed treatment with talc and sodium alginate formulations of strain SE34 gave 58% protection followed by GBO3 with 40% protection. Seed treatment with fresh suspensions of strains SE34 and GBO3 followed by challenge inoculations with Xoo increased accumulation of phenylalanine ammonia lyase, peroxidase and polyphenol oxidase compared to untreated control seedlings. Thus, the results of the present study suggest that the PGPR strains used as fresh suspensions and powdered formulations may have commercial potential in plant growth promotion and in management of rice bacterial leaf blight disease.

벼 생육시기별 흰잎마름병 발병이 쌀 수량감소 및 미질에 미치는 영향

2002년부터 2005년까지 4년간 벼 흰잎마름병 초발병 조사결과 2002년보다 초발병일이 20<TEX>$\sim$</TEX>30일 이상 빨라지고 있음을 확인하였다. 벼 흰잎마름병 발병시기 및 병반면적율에 따라 수량이 감소하였는데 병반면적률이 10% 이하에서는 피해가 적었으나, 병반적율이 증가함에 따라서 수량감소가 점점 많아졌고, 25% 이상일 때부터 수량감소 및 현미천립중 등에서 현저히 낮아지고 병반면적율이 50% 이상이고 유수형성기 발병시 수량 29%, 현미천립중 2.3g, 현미완전립률 18%까지 감소되었다. 밥맛의 기준이 되는 식미치인 점도에서도 병반면적율이 증가함에 따라 낮은 식미치를 보였다. We investigated the first symptom emerging date of rice bacterial leaf blight disease during four years from 2002 to 2005. The disease occurrence date was earlier 20<TEX>$\sim$</TEX>30 days in 2005 than that of 2002. The damage in different rice growth stage by the bacterial disease on rice yield and grain quality in southern part of Korea was examined. The disease decreased rice yield following by increased infection rates. Slight loss in rice yield and brown head rice rate were observed at below 10% level of infected leaf area, while over 25% infected leaf area caused the significant decrease in rice yield and brown head rice rate. More than 50% of the infected leaf area rate caused 29% yield reduction in case of infection at panicle formation stage and 18% of brown head rice rate. Cooked rice quality was also affected in the diseased rice by damage in palatability score and viscosity.