Ralstonia Syzygii Research Articles

Characterization and Identification of Potential Lactic Acid Bacteria as Biological Control Agent against Ralstonia syzygii subsp. celebesensis of the Banana Blood Disease

Banana is one of the primary fruits cultivated in Malaysia and currently decimated by the emergence of a disease, known as banana blood disease (BBD) which caused by Ralstonia syzygii subsp. celebesensis (Rsc). The BBD has significantly affected the yield and profits of the worldwide banana industry. To date, various approaches including chemical and biological controls have been attempted to manage this disease but none of them succeed in controlling the disease. The uses of lactic acid bacteria (LAB) in managing plant diseases have been reported earlier but little information is available. Therefore, this project is designed to identify and investigate potential plant-associated LAB as biological control agent (BCA) against Rsc by using agar well diffusion method. The inhibition zones of each well were observed after 72h and the isolated LAB that showed inhibition zones were proceed for molecular characterization using PCR amplification followed by gel electrophoresis. The sequences were used for phylogenetic analysis. In addition, each of the potential LAB were used to identify their morphological characterizations and biochemical testing. Throughout the study, the highest inhibition zones of LAB from kimchi and fermented milk achieved a diameter of 21.30 mm and 28.70 mm, respectively. Kimchi isolates showed the highest similarity which is 97% as Lactiplantibacillus plantarum species. Among the fermented milk isolates, the highest similarity which is 98% identified as Lacticaseibacillus paracasei.

In-vitro endophytic bacteria activity against Ralstonia syzygii subsp. indonesiensis, the causal agent of bacterial wilt disease in tomato (Solanum lycopersicum) to achieve sustainable agriculture

The bacterial wilt disease transmitted through soil, identified as Ralstonia syzygii subsp. indonesiensis, is a substantial vascular disease and major constraint in various Solanaceae plants. The use endophytic bacteria-as Arthrobacter sp. and Bacillus thuringiensis-is one approach to control this pathogen. In vitro test using the disc paper method was conducted to evaluate the compatibility of Arthrobacter sp. and B. thuringiensis, along with their inhibitory capabilities through secondary metabolite production, in suppressing the growth of R. syzygii subsp. indonesiensis. The results showed incompatibility among isolates of endophytic bacterial, consequently, both endophytic bacteria were capable suppressing the growth of the R. syzygii subsp. indonesiensis significantly. Arthrobacter sp. formed a larger inhibition zone compared to B. thuringiensis, and both endophytic bacteria can be candidate biological control agents to reduce pesticide use to control R. syzygii subsp. indonesiensis, which therefore achieving more environmentally agriculture sustainability.

Consortium solid formula of bacillus spp. to control bacterial wilt on chili plants

Due to its high economic worth and ability to be consumed in both fresh and processed forms, chili (Capsicum annum L.) is a fruit that is frequently used as a vegetable. Pests and plant diseases, such as bacterial wilt caused by Ralstonia syzygii subsp. Indonesiensis, are the causes of reduced chili productivity. Plants will eventually perish as a result of symptoms like withering of the leaf tips that eventually spread to the undersides of the leaves. The lower vascular tissue and roots of the affected chili plants appear brown. Bacterial wilt is an important disease of chili caused by Ralstonia syzigii subsp. indonesiensis and can reduce yield by up to 90%. The research aimed to obtain the best solid formula consortium of Bacillus spp. for controlling R. syzygii subsp. indonesiensis on chili peppers. The study consists of 2 stages: 1). Production of a consortium combination of Bacillus spp. in solid waste 2). Test the ability of the consortium of Bacillus spp. in solid waste to control bacterial wilt in chili plants. The study used a completely randomized design that consisted of 19 treatments and four replications. Each treatment was introduced on chili seeds and seedlings, while the inoculation of R. syzygii subsp. indonesiensis was conducted on chili aged 35 days after planting. The results showed that the best solid formula consortium of the Bacillus spp. for controlling bacterial wilt (R. syzigii subsp. indonesiensis) on chili plants is the Bran + Bagasse formula for four weeks of storage.

Primer Design of Volatile Synthesis Coding Genes in Ralstonia syzygii subsp. celebesensis

Microbes produce various types of volatile organic compounds (VOCs) through metabolism, which can be used for diagnostic purposes. Microbes' types and classes of VOCs are very wide, including fatty acid derivatives (hydrocarbons, alcohols, and ketones), aromatic compounds, nitrogen-containing compounds, and volatile sulfur compounds. Microbial volatile organic compounds (VOCs) can also be divided into several chemical classes: alkenes, alcohols, ketones, benzos, pyrazines, sulfides, acids, esters, and terpenes. This study aimed to design primers for genes encoding volatile synthesis in Ralstonia syzygii subsp. celebesensis, which causes blood disease in the banana plant. Some of the genes involved are adc (acetone synthesis), adhP (ethanol synthesis), ilvA, nirBD (ammonia synthesis), mdcA (propionic acid synthesis), cysI (hydrogen sulfide synthesis), and speBC (putrescine synthesis). Primers were designed and examined for specificity in silico using Primer3Plus, Geneious Prime, and BLAST programs. The numbers of nine pairs designed primers were successfully amplifying the related nine VOC genes of R. syzygii subsp. celebesensis for qPCR.

A Study on the Distribution Pattern of Banana Blood Disease (BBD) and Fusarium Wilt Using Multispectral Aerial Photos and a Handheld Spectrometer in Subang, Indonesia

Knowledge of the health of banana trees is critical for farmers in order to profit from banana cultivation. Fusarium wilt and banana blood disease (BBD), two significant diseases that infect banana trees, are caused by Fusarium oxysporum and Ralstonia syzygii, respectively. They have caused a decline in crop yield, as they destroy trees, starting sequentially from the pseudostem to the fruit. The entire distribution of BBD and fusarium on a plantation can be understood using advanced geospatial information obtained from multispectral aerial photographs taken using unmanned aerial vehicles (UAVs) and a reliable data field for infected trees. Vegetation and soil indices derived from multispectral aerial photographs, such as the normalized difference vegetation index, the modified chlorophyll absorption ratio index, the normalized difference water index (NDWI), and soil pH, may have to be relied upon to explain the precise location of these two diseases. This study used a random forest algorithm to handle a large dataset consisting of multispectral and spectral models. The results show that the soil indices, soil pH, and NDWI are the most important variables for predicting the spatial distribution of these two diseases. Simultaneously, the plantation area affected by BBD is more extensive than that affected by fusarium if variations in planted banana cultivars are not considered.

Evaluation of banana cultivars and the pathogenesis-related class 3 and 10 proteins in defense against Ralstonia syzygii subsp. celebesensis, the causal agent of banana blood disease

Evaluation of banana cultivars and the pathogenesis-related class 3 and 10 proteins in defense against Ralstonia syzygii subsp. celebesensis, the causal agent of banana blood disease

Pathogenesis study of blood disease bacterium in local bananas of Malaysia

Banana blood disease (BBD), a banana wilt disease caused by blood disease bacterium (BDB), Ralstonia syzygii subsp. celebesensis (RSC), was first reported in Malaysia in 2007. BDB results in wilting appearance and internal bleeding-like symptoms, which eventually leads to entire crop loss including ready-harvested fruit bunch. Understanding the pathogenesis of BDB in local common banana cultivars provides good foundation to tailor disease management plan to mitigate BBD. To address this need, our study was performed to characterize BDB isolated from locally grown banana plants and to compare the pathogenesis of the bacterium in four most common local banana cultivars. In this study, BDB isolated from Malaysia's plantation was identified and characterized via the molecular and phenotypic method. The pathogenicity of characterized virulent BDB isolate was determined in four local banana cultivars via disease rating that was newly developed specifically for BDB-infected bananas. The identity of the isolated strains was first confirmed as Blood Disease Bacterium using Sanger sequencing analysis of 16s rRNA gene. Standard biochemical tests and growth curve study showed that the BDB isolates were Gram-negative, non-motile rod-shaped, catalase-positive, oxidase-positive bacterium, and had an optimum growth rate at 35 °C. The pathogenicity of a selected virulent BDB isolate was then determined via Koch's Postulates in four local banana cultivars. Recognizing the lack of a specific disease rating scale to quantify the pathogenesis of BDB in bananas, we developed a disease rating scale specifically to BDB-infected bananas in this study. Based on the pathogenicity test, local banana cultivars (cv.) in Malaysia included cv. Berangan, cv. Nipah, cv. Awak and cv. Nangka demonstrated disease severity index (DSI) higher than 95%, indicating their susceptibility towards to BDB. In conclusion, our results showed that the four most common banana cultivars in Malaysia were highly susceptible to infection by virulent BDB.

Prevalence and Incidence of Bacterial Wilt Disease (Ralstonia syzygii subsp. indonesiensis) on Tomato in Simpang Empat District Karo

The major disease known as bacterial wilt, which affects tomato plants in Indonesia, is brought on by Ralstonia syzygii subsp. indonesiensis. In Simpang Empat District, Karo Regency, Indonesia, R. syzygii subsp. indonesiensis causes bacterial withery in tomato plants. The purpose of this study is to map its spread. This study is an experimental one that used the survey method to determine the prevalence of disease, the incidence of disease, and the pathogenic isolation of tomato plants at nine different locations throughout Simpang Empat District. The pathogen was isolated from the sample and identified in the lab as R. syzygii subsp. indonesiensis, according to the results. The outcome of the presence of bacterial wilt brought on by R. syzygii subsp. indonesiensis bacteria with various illness occurrences. At a proportion of disease incidence of 57.07%, Ndokum Siroga Location 1 in the community experienced the most disease.

Field and Laboratory Detection of Clove Sumatra Disease Caused by Ralstonia syzygii subsp. syzygii in Java, Indonesia

The significant decrease in clove production in Indonesia is mainly due to the Sumatra disease caused by bacterial Ralstonia syzygii subsp. syzygii. It is necessary to manage the disease broadly based on disease detection in the field and the laboratory. This study aims to determine the technique for detecting the distribution pattern of Sumatra disease using geographic information systems and validate the presence of R. syzygii subsp. syzygii in clove plant tissues by molecular analyses based on the endoglucanase gene. This research was conducted by acquiring aerial photos using uncrewed aerial vehicles processed using photogrammetric techniques to produce geographic information system outputs as a thematic map of the clove Sumatra disease distribution pattern. The plant samples were collected for molecular analysis of the pathogens causing clove Sumatra disease in the laboratory. DNA extraction was performed and amplified by polymerase chain reaction (PCR) using UGMRss-specific primers followed by Sanger sequencing. The aerial photo images showed that the distribution of clove Sumatra disease has a random pattern, which tends to spread between healthy and diseased plants. Furthermore, the PCR and phylogenetic analyses revealed that all symptomatic clove plant samples (leaves, twigs, and roots) were positively infected by R. syzygii subsp. syzygii.

Copper and iron metal resistant rhizospheric bacteria boost the plant growth and bacoside A content in Bacopa monnieri under stress conditions.

Bacteria that enhance plant growth and development and are found in the vicinity of roots are referred to as plant growth-promoting rhizobacteria. Some beneficial bacteria help plant tolerance to many hazardous chemical elements. In this context, Cupriavidus basilensis, Novosphingobium humi, Bacillus zanthoxyli, Bacillus sp., Paenibacillus alvei, Ancylobacter aquaticus and Ralstonia syzygii metal-tolerant rhizospheric bacteria were isolated from rhizospheric soil associated with Bacopa monnieri. The beneficial effects of rhizospheric bacteria on B. monnieri plant physiology and biochemical responses were investigated under pot conditions at two levels (100μM and 500μM) of CuSO4 or FeCl3. N. humi, A. aquaticus and R. syzygii bacterial strains were associated with significantly increased height and biomass under normal and stress conditions. An assay for indole acetic acid in isolated rhizospheric bacteria found differential secretion except Bacillus zanthoxyli. Bacoside A is a major phytocompound in B. monnieri with medicinal value; maximum induction was observed in the R. syzygii treatment. High concentration of copper and iron salts negatively influenced height, biomass and photosynthetic pigments; however N. humi, A. aquaticus, Bacilllus sp. and R. syzygii beneficial bacterial helped plants under stress conditions. Moreover, a significant enhancement in chlorophyll a and b was noticed in C. basilensis, B. zanthoxyli, Bacilllus sp., P. alvei and R. syzygii treatments, without much influence on carotenoid levels. Therefore, the present study emphasises the importance of isolating plant growth-promoting rhizobacteria for use in bacopa plants exposed to metals such as copper and iron in soil.

Identification and characterization of resistance quantitative trait loci against bacterial wilt caused by the Ralstonia solanacearum species complex in potato.

The online version contains supplementary material available at 10.1007/s11032-022-01321-9.

Transmission of Blood Disease in Banana.

Banana Blood disease is a bacterial wilt caused by Ralstonia syzygii subsp. celebesensis and is an economically important disease in Indonesia and Malaysia. Transmission of this pathogen is hypothesized to occur through insects mechanically transferring bacteria from diseased to healthy banana inflorescences and other pathways involving pruning tools, water movement, and root-to-root contact. This study demonstrates that the ooze from the infected male bell and the sap from various symptomatic plant parts are infective, and the cut surfaces of a bunch peduncle, petiole, corm, pseudostem, and the rachis act as infection courts for R. syzygii subsp. celebesensis. In addition, evidence is provided that R.syzygii subsp. celebesensis is highly tool transmissible, that the bacterium can be transferred from the roots of a diseased plant to the roots of a healthy plant and transferred from the mother plant to the sucker. We provide evidence that local dispersal of Blood disease occurs predominantly through mechanical transmission by insects, birds, bats, or human activities from diseased to healthy banana plants and that long-distance dispersal occurs through the movement of contaminated planting material. Disease management strategies to prevent crop losses associated with this emerging disease are discussed based on our findings.

Xylem Limited Bacteria (XLB) Clove Disease Control Technology Through Utilization of Endophytic Bacteria

Xylem Limited Bacterium (XLB) disease (Ralstonia syzygii) is the most seriuos disease on clove plant. Biological control using Endophytic Bacteria (Bacillus sp. Bc25 and Fluorescentpseudomonad Pf22) is expected to control clove XLB disease.The research aim to get Bacillus sp. Bc25 and Fluorescent pseudomonad Pf22product are effective to control XLB disease on clove plant.The study of Bacillus sp. Bc25 and Fluorescent pseudomonad Pf22 was done in watering on clove plant field were infected by XLB disease in Solok West Sumatera on January to December 2017.The study used a randomized block design (RBD) with four replications. The parameters observed were the disease attack level and plant growth.The study results showed thatBacillus sp.Bc25 andFluorescentpseudomonadPf 22 can control XLB disease and increase clove growth.Bacillus sp. Bc25 and Fluorescentpseudomonad Pf22 combinationdemonstrate XLB disease control capabilities and promote the best clove plants growthby intensity suppressing of XLB disease from 86.80% to 20.59% and increasing the clove plants growth from the stem circle 20.00 cm to 46.00 cm.Combination product of biological agents Bacillus sp. Bc25 and FluorescentpseudomonadPf22 can be developed to control XLB diseases and increase clove plant growth.Keywords : Clove, Xylem Limited Bacteria Disease (Ralstonia syzygii), Endophytic Bacteria (Bacillus sp. Bc25 and Fluorescent pseudomonad Pf22.

Evaluation of the Effect of PGPR Strains on Tomato Growth and Suppression of Ralstonia Wilt Disease

The tomato (Lycopersicum esculentum Mill.) has substantial scope for development due to its high economic value and large export potential. Ralstonia syzygii subsp. Indonesiensis (RSI) is the cause of bacterial wilt disease which attacks the vascular system in Solanaceae. It can cause large losses in yield and has caused global concern because of its widespread distribution and attack on many important crops. The aim of this research was to identifiy and characterize the ability of indigenous rhizobacterial isolates to control RSI and promote tomato growth. The PGPR traits studied were production of hydrogen cyanide, siderophores, biosurfactant, and ammonia, and protease activity. Bacterial identification was performed using 16S rRNA. Our findings revealed that the strains identified shared some similarities with Bacillus thuringiensis strain ATCC 10792 (IR.2.3.5), B. mycoides strain ATCC 6462 (IR.1.3.4), Bacillus thuringiensis strain IAM 12077 (IR.2.2. 1), Serratia ficaria strain DSM 4569 (IR.3.1.4), Enterobacter oryzendophyticus strain REICA_082 (IR.2.2.7), Cronobacter dublinensis subsp. lausannensis strain E515 (IR.2.2.5) and S. rubidaea strain DSM 4480 (IR.2.2.6). All of the isolates were tested for a variety of abilities related to growth promotion and biocontrol.
 Keywords: Ralstonia wilt; 16S rRNA identification; PGPR traits

Draft Genome Sequence of Ralstonia syzygii subsp. celebesensis from Indonesia, the Causal Agent of Blood Disease of Banana.

Draft Genome Sequence of Ralstonia syzygii subsp. celebesensis from Indonesia, the Causal Agent of Blood Disease of Banana.

Development of a Highly Sensitive Loop-Mediated Isothermal Amplification Incorporated with Flocculation of Carbon Particles for Rapid On-Site Diagnosis of Blood Disease Bacterium Banana

Bananas are one of the most crucial fruit crops worldwide and significantly contribute to food security in developing countries. However, blood disease of bananas caused by Ralstonia syzygii subspecies celebensensis has become a threat to banana production. Rapid and accurate diagnosis of BDB for on-site detection is pivotal at an early stage for an effective disease control strategy. This study developed LAMP with specific primers targeting BDB, followed by a flocculation assay for visualising positive amplification in the LAMP assay. The assay was sensitive to picogram amounts of gDNA (0.5 pg). LAMP assay on BDB gDNA showed flocculation, but negative results on Fusarium oxysporus cubense and Ralstonia solanacearum confirming the specificity of the assays. Field testing conducted at MARDI headquarters and Taman Pertanian Universiti discovered that the LAMP-flocculation assays were successful in detecting BDB on symptomatic samples as well as on samples from a healthy plot with no symptom observed at the sampling stage, revealing that this assay can detect BDB at an early infection stage. The validation results showed that the LAMP-flocculation assay was comparable with the PCR technique. This newly developed technique is highly specific and sensitive for the early detection of BDB for the adoption of precautionary control measures.

Susceptibility of the Banana Inflorescence to Blood Disease.

The bacterium Ralstonia syzygii subsp. celebesensis causes Blood disease of banana, a vascular wilt of economic significance in Indonesia and Malaysia. Blood disease has expanded its geographic range in the last 20 years and is an emerging threat to Southeast Asian banana production. Many aspects of the disease cycle and biology are not well understood, including the ability of different parts of the female and male inflorescence of banana to act as infection courts. This study confirms that the banana varieties of Cavendish, and Kepok 'Kuning' are susceptible to Blood disease and that an inoculum concentration of 102 CFU/ml of R. syzygii subsp. celebesensis is adequate to initiate disease after pseudostem inoculation. Data show that infection occurs through both the male and female parts of a banana inflorescence and the rachis when snapped to remove the male bell. The infection courts are the female flowers, the male bell bract scar, the male bell flower cushion, the snapped rachis, and deflowered fingers. The location of these infection courts concurs with the dye studies demonstrating that dye externally applied to these plants parts enters the plant vascular system. Thus, the hypothesis is supported that infection of R. syzygii subsp. celebesensis occurs through open xylem vessels of the male and female parts of the banana inflorescence.

Ralstonia syzygii subsp. syzygii (Sumatra disease bacterium)

This datasheet on Ralstonia syzygii subsp. syzygii covers Identity, Overview, Distribution, Dispersal, Hosts/Species Affected, Diagnosis, Biology & Ecology, Impacts, Prevention/Control, Further Information.

Ralstonia syzygii subsp. celebesensis (banana blood disease)

Ralstonia syzygii subsp. celebesensis (banana blood disease)

Role of Trehalose Synthesis in Ralstonia syzygii subsp. indonesiensis PW1001 in Inducing Hypersensitive Response on Eggplant (Solanum melongena cv. Senryo-nigou).

Ralstonia syzygii subsp. indonesiensis (Rsi, former name: Ralstonia solanacearum phylotype IV) PW1001, a causal agent of potato wilt disease, induces hypersensitive response (HR) on its non-host eggplant (Solanum melongena cv. Senryo-nigou). The disaccharide trehalose is involved in abiotic and biotic stress tolerance in many organisms. We found that trehalose is required for eliciting HR on eggplant by plant pathogen Rsi PW1001. In R. solanacearum, it is known that the OtsA/ OtsB pathway is the dominant trehalose synthesis pathway, and otsA and otsB encode trehalose-6-phosphate (T6P) synthase and T6P phosphatase, respectively. We generated otsA and otsB mutant strains and found that these mutant strains reduced the bacterial trehalose concentration and HR induction on eggplant leaves compared to wild-type. Trehalose functions intracellularly in Rsi PW1001 because addition of exogenous trehalose did not affect the HR level and ion leakage. Requirement of trehalose in HR induction is not common in R. solanacearum species complex because mutation of otsA in Ralstonia pseudosolanacearum (former name: Ralstonia solanacearum phylotype I) RS1002 did not affect HR on the leaves of its non-host tobacco and wild eggplant Solanum torvum. Further, we also found that each otsA and otsB mutant had reduced ability to grow in a medium containing NaCl and sucrose, indicating that trehalose also has an important role in osmotic stress tolerance.