Host specificity and genetic diversity of race 4 strains of Ralstonia solanacearum

Ralstonia solanacearum race 4 isolates were obtained from Zingiberaceae plants in India during bacterial wilt outbreaks. Polyphasic phenotypic and genotypic analysis revealed intraracial diversity and dominance of biovar 3 over biovar 4. Biovar 3 strains were isolated from very severely wilted Zingiberaceae plants in the field and found to be present across diverse geographical, host and seasonal boundaries. It was hypothesized that these isolates belong to a single, ‘fast wilting’, lineage. Using one ‘fast wilting’ isolate in controlled inoculations, rapid wilt was observed in ginger within 5–7 days. Wilting was also observed in several other closely and distantly related hosts such as turmeric (Curcuma longa), aromatic turmeric (Curcuma aromatica), black turmeric (Curcuma caesia), sand ginger (Kaempferia galanga), white turmeric (Curcuma zeodaria), awapuhi (Zingiber zerumbet), greater galangal (Alpinia galanga), globba (Globba sp.), small cardamom (Elettaria cardamomum) and large cardamom (Ammomum subulatum) of the Zingiberaceae family, and in tomato (Solanum lycopersicum). Molecular analysis, including multiplex PCR‐based phylotyping, sequence analysis of 16S rDNA, 16–23S intergenic spacer and the recN gene, and multilocus sequence typing, revealed minimal differences between fast wilting isolates, confirming that almost all belong to the same lineage. Biovar 4 was isolated from plants showing slow wilt progression and self‐limiting wilting in restricted geographical locations instead, and was identified to be genetically distinct from the fast wilting biovar 3 isolates. To the authors' knowledge, this is the first report of host range and genetic analysis of R. solanacearum race 4 in India.     

Bacterial wilt of sweet potato caused by <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> in Taiwan

During the last decade, a new bacterial disease has impaired the yield of vegetable sweet potato (30–80%) in Taiwan. Infected plants developed stunting, root and stem rot, vascular discoloration and wilting. Ten bacterial isolates that caused the same symptoms in sweet potatoes after inoculation were reisolated and classified as Ralstonia solanacearum phylotype I biovar 4 based on physical and molecular analyses. Moreover, these isolates also caused wilting in convolvulaceous, solanaceaous and cruciferous plants. This report is the first of bacterial wilt of sweet potato caused by R. solanacearum in Taiwan.     

Bacterial wilt of bellflower caused by Ralstonia solanacearum in Japan

A sudden wilt of bellflower (Campanula lactiflora) was observed in Japan in 1997. A bacterium that formed white fluidal and mucoid colonies resembling those of Ralstonia solanacearum was isolated from the infected plants. The bacterium was bacteriologically identified as biovar 3 of R. solanacearum. This is the first report of R. solanacearum affecting a plant species of the Campanulaceae family.     

Pathogenic and genetic variability of Ralstonia solanacearum strains from the Philippines

In the Philippines, bacterial wilt caused by Ralstonia solanacearum is one of the most important diseases affecting vegetables and banana. In this study, 89 strains of R. solanacearum isolated from various hosts were screened for their biovar, phylotype, pathogenicity, and genetic diversity. Foreign strains were included for comparison with these Philippine strains. Results of the biochemical and multiplex-PCR tests divided the Philippine strains into five biovars (1, 2, 3, 4, and N2) and three phylotypes (I, II, and IV). Three potato strains belonged to biovar N2/phylotype IV. Pathogenicity tests divided the strains into five pathogenicity types based on their virulence in tomato, potato, eggplant, sweet pepper, and tobacco. Strains classified as biovar N2 were weakly pathogenic to potato (pathogenicity type III) and almost all strains isolated from banana were not pathogenic to the test plants except potato (pathogenicity type V). The results of AFLP analysis divided the strains into four clusters. Cluster 1 was composed of strains isolated from solanaceous crops, ginger (Zingiber officinale), and Morus sp. from the Philippines and other Asian countries. Cluster 2 grouped the potato strains (biovar N2) from the Philippines and Japan and blood disease bacterium strains from Indonesia. Cluster 3 contained the local and foreign strains isolated from potato (biovar 2) and banana (biovar 1). Cluster 4 consisted only of the tomato strain from the USA.     

Molecular identification of some African strains of Ralstonia solanacearum from eucalypt and potato

Ralstonia solanacearum is a known bacterial pathogen of eucalypt and potato plants in Africa. A survey was undertaken to detect this pathogen in eucalypt plantations in South Africa, the Democratic Republic of Congo, and Uganda. Numerous bacterial strains were isolated from trees with symptoms typical of bacterial wilt, but only seven were positively identified as R. solanacearum. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, based on the hrp (hypersensitive response and pathogenicity) gene region was used to determine and group the biovars of these R. solanacearum strains. The eucalypt isolates and one potato isolate formed a biovar 3 cluster, whereas the two other potato isolates formed a cluster that corresponded to biovar 2. Amplified fragment length polymorphism (AFLP) analysis confirmed these clusters. Therefore, PCR-RFLP can be used as a reliable diagnostic technique to enable researchers to rapidly identify the pathogen.     

Specific Detection of Biovars of Ralstonia solanacearum in Plant Tissues by Nested-PCR-RFLP

A sensitive and specific assay, based on a Nested-PCR-RFLP protocol, was developed for the detection of biovars of Ralstonia solanacearum, the causal agent of bacterial wilt. Oligonucleotide primer pairs were selected within the hrp gene region. Specific amplification of the hrp fragments was obtained for all R. solanacearum strains and also for two closely related species, Pseudomonas syzygii and the blood disease bacterium. No amplification was observed for a wide range of other bacterial species, including R. pickettii and Burkholderia cepacia. Digestion with HindII provided four distinct restriction profiles specific to biovars or groups of biovars of R. solanacearum: one for biovar 1 strains originating from the Southern part of Africa, one for American biovar 1 and biovars 2 and N2 strains, one for biovars 3 and 4 strains, and one for biovar 5 strains. When applied to either pure culture or infected plant tissues, Nested-PCR allowed detection as low as 10³cfu ml^–1, which corresponds to 1cfu per reaction. Amplification was partially or completely inhibited by compounds contained in plant extracts (potato plant and potato tuber, tomato, tobacco, eggplant, pepper and Pelargonium asperum). A combined PVPP/BSA treatment prior to amplification permitted reliable Nested-PCR detection of R. solanacearum strains in plant samples. Nested-PCR-RFLP, assessed with isolates from Reunion Island but also applicable to any R. solanacearum strain, provides a wide range of possible uses for identification, detection and epidemiological investigations.     

Behavior and mutation of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> in <Emphasis Type="Italic">Solanum toxicarium</Emphasis> grown in aseptic culture

To study the behavior and mutation of Ralstonia solanacearum in Solanum toxicarium, which is resistant to bacterial wilt, S. toxicarium was grown in aseptic culture and inoculated with R. solanacearum. Although 60%–80% of the inoculated plants were wilting after 2 to 3 days, most wilted plants had recovered by 20 days after inoculation. The pathogen was reisolated from over 98% of inoculated plant stems, but the percentage of recovery decreased the closer the isolation sites were toward the upper stem sections. Three colony types, characterized as fluidal white, nonfluidal red, and a mixture of fluidal white and nonfluidal red, were reisolated from the stems. Nonfluidal red colonies were less virulent on tomato plants than fluidal white colonies.     

Activation of tomato plant defence responses against bacterial wilt caused by Ralstonia solanacearum using DL-3-aminobutyric acid (BABA)

In this study, we investigated the ability of DL-3-aminobutyric acid (BABA) to protect tomato against bacterial wilt caused by Ralstonia solanacearum. This was combined with studies of accumulation of total phenolic compounds, free and total salicylic acid (SA), and activity of enzymes related to plant defence, i.e., polyphenol oxidase (PPO) and catalase (CAT). Under greenhouse conditions, tomato plants pre-treated by soil drenching with BABA profoundly reduced disease severity of bacterial wilt compared to plants receiving a soil drench with water. Thus, BABA reduced leaf wilting index by 75.3 % and vascular browning index by 69.9 %, without any in vitro inhibitory activity on the pathogen. BABA treatment significantly reduced the population of R. solanacearum in stems of tomato plants and additionally also significantly increased both fresh and dry weight of roots and shoots of tomato plants compared with the inoculated control. Application of BABA resulted in a high increase in PPO activity both in plants with and without inoculation. Compared to water-treated plants, treatment with BABA also induced a significant increase of total phenolic compounds as well as of free and total SA in leaves of both inoculated and non-inoculated tomato plants at all sampling times. CAT activity decreased in tomato plants treated with BABA in comparison with the water-treated control plants and the decrease in activity correlated with an increasing total SA accumulation. These findings suggest that BABA treatment resulted in induction of resistance to bacterial wilt in tomato.     

Molecular epidemiology of Ralstonia solanacearum strains from plants and environmental sources in Portugal

Several outbreaks of bacterial wilt disease caused by the quarantine bacterium Ralstonia solanacearum were identified in Portugal. Intensive surveys recognized the bacterium as endemic in the main irrigated agricultural ecosystems. Between 1999 and 2006 all isolates of R. solanacearum were characterized as belonging to biovar 2A. In 2007, biovar 1 strains were recorded in potato fields under a confined area. A panel of 101 Portuguese isolates obtained from biotic and environmental samples was studied. Following a polyphasic approach, these isolates were analysed by SDS-PAGE of the whole cell proteins, MSP-PCR (csM13), rep-PCR (BOXA1R and ERIC-2) and FAFLP (EcoRI?+?0/MseI?+?C). A 750?bp sequence of endoglucanase (egl) gene was studied for 17 representative isolates and 95 accessions retrieved from the GeneBank. Numerical analysis of protein profiles correlated quite well with biovar subphenotype, producing a unique megacluster (r?=?71.1%). MSP-PCR was more discriminative (r?=?62%). Rep-PCR approaches displayed higher polymorphism levels with ERIC 2 primer producing high diversity indexes (D and J′). FAFLP was the most reproducible method (95%) displaying 229 polymorphic characters and the highest evenness (J′). For all the methods small clusters disclosed a clonal origin for isolates with a common geographical origin/matrix. FAFLP identified an adaptative microevolution phenomenon for surface water strains. Polyphasic approach congruence highlighted the inability of individual methods to explain the whole diversity. Mr. Bayes egl-based phylogenetic tree allocated the 17 Portuguese isolates into the sub-clusters of narrow (nhr) and broad host range (bhr) of Phylotype II unveiling the epidemiological story of R. solanacearum in Portugal and identified different populations coexisting in the same habitats. This is the first report of the presence of R. solanacearum Phylotype II, bhr strains in Western Europe.     

Effect of cold-water irrigation on bacterial wilt pathogen of tomato

Bacterial wilt caused by Ralstonia solanacearum is one of the most devastating bacterial diseases of plants worldwide. Management of bacterial wilt in tomato and other crops has been difficult, and so novel but easily implemented control methods are being sought. To evaluate the effect of cold-water irrigation on bacterial wilt of tomato, four treatments were used in which CF (chemically fertilized) soil and CF + FYM (chemical fertilizer + farmyard manure [FYM]) soil were inoculated with a bacterial suspension (R. solanacearum strain YU1Rif43) at 10⁶ colony forming units (CFU) g^?1 soil. Tomato seedlings were grown in Agri-pots in a plant growth chamber. The soil was irrigated with water that was kept at the same temperature in each treatment: 4, 10, 20, or 30°C. Incidence and severity of wilt, counting of the colonies of the culturable population of pathogen, and dry-mass and height of the plants were examined. After 45 days and in both kinds of soil, most of the plants had wilted in soil irrigated at 30°C. Wilt incidence was substantially reduced when transplanted seedlings were irrigated at lower temperatures (4–20°C). Survival of R. solanacearum was also reduced after being irrigated with water at lower temperatures, indicating that the reduced incidence of wilt was linked to reduced survival of the pathogen. Dry-mass and plant height were slightly higher under control conditions than in soils irrigated at lower temperatures. This study suggests that cold-water irrigation could significantly reduce bacterial wilt of tomato and have an adverse effect on survival of the wilt pathogen.     

Diversity of Brazilian biovar 2 strains of Ralstonia solanacearum

Ralstonia solanacearum is responsible for bacterial wilt disease. Specific and accurate identification of this pathogen is essential for protection of susceptible crops as well as breeding resistant varieties. Historically, R. solanacearum has been classified into biovars based on the use of sugar and alcohol as carbon sources, into races based on its ability to infect different hosts, more recently into phylotypes based on the intergenic transcribed sequence of the ribosomal RNA genes 16S and 23S and into sequevars based on the endoglucanase gene (egl) sequence. Race 3 biovar 2 (R3Bv2) is widespread in South and Central America, and in Brazil it is present in all potato-producing regions as the most prevalent strain. In this study, we classified 53 Brazilian R. solanacearum biovar 2 (Bv2) strains by traditional and molecular methods. PCR with specific primers confirmed all 53 bacterial strains as belonging to the R. solanacearum species complex, and all were classified as biovar 2A or 2T based on acidification of sugars and alcohols. Multiplex phylotype PCR assigned all strains to phylotype II. Phylogenetic analysis of egl sequences showed that most Bv2 strains from Brazil analyzed in this study did not cluster with known sequevars and are less clonal than the R3Bv2 strains reported for other countries. This is the first study to address the diversity of a collection of Brazilian R. solanacearum strains using the phylotype and sequevar classification scheme.     

Bacterial wilt-resistant tomato rootstock suppresses migration of <Emphasis Type="Italic">ralstonia solanacearum</Emphasis> into soil

Ralstonia solanacearum, the causal agent of bacterial wilt of tomato, grows in infected plants and migrates from the roots into the soil. We investigated the effectiveness of bacterial wilt-resistant tomato rootstock in reducing the migration of R. solanacearum from susceptible scions into the soil. Rootstock stems were either 3–5 cm tall (low-grafted, LG) or ≥?10 cm tall (high-grafted, HG). After inoculation of scions of the susceptible cultivar (SC) with R. solanacearum below the first flower, there was no difference in disease progression among LG, HG, and ungrafted SC plants, and plants had wilted by 2 weeks. However, the rate of detection of R. solanacearum in the soil of wilted plants was reduced by grafting. The size of the R. solanacearum population in the soil of fully wilted plants increased in the order of HG?<?LG?<?SC. These results show that grafting onto resistant rootstock strongly suppressed the migration of R. solanacearum into the soil by the time of full wilting, and the effect was stronger with a longer rootstock. Migration of R. solanacearum into soil increased with increasing disease severity in SC, LG and HG. These facts suggest that early uprooting of slightly infected plants could control the spread of the bacteria into the soil.     

Genetic diversity of Ralstonia solanacearum species complex strains obtained from Guangxi,China and their pathogenicity on plants in the Cucurbitaceae family and other botanical families

Bacterial wilt, caused by the Ralstonia solanacearum species complex (RSSC), is a destructive plant disease in Guangxi, China. However, the diversity of RSSC populations in the area is unknown. To this end, we performed an extensive bacterial wilt survey from 2015 to 2018. Using phylotype-specific multiplex PCR (Pmx-PCR) and an egl-based tree, 189 strains collected from 20 plant species were identified as R. pseudosolanacearum phylotype I, which included 14 sequevars (12, 13, 14, 15, 16, 17, 18, 30, 34, 44, 48, 54, 70, and 71); two strains isolated from potato plants belonged to R. solanacearum phylotype II, sequevar 1. Sequevars 13, 17, and 44 were prevalent in Guangxi, and sequevar 13 dominated the RSSC sequevars of four Cucurbitaceae plants. The susceptibility of different Cucurbitaceae species to bacterial wilt and the host range of 16 representative strains were further tested. Members of the Cucurbita, Momordica, and Luffa genera were susceptible to bacterial wilt, with wilt incidence ranging from 73% to 100%. Most strains were pathogenic to solanaceous plants, mulberry, and ginger plants but not to melon crops; however, the strains from kidney bean, pepper, and Cucurbitaceae plants were highly virulent to melon crops. This is the first comprehensive report on the genetic and host range diversity of the RSSC in Guangxi and the susceptibility of different Cucurbitaceae species to bacterial wilt, which can provide valuable information for the development of bacterial wilt control strategies.     

花生青枯病防治的研究

花生青枯病(Pseudomonas solanacearum E.F.Smith)是六十年代以来,在山东省逐渐蔓延起来的主要病害,面积已达20万亩以上。一般发病率5～10%,重者达50%以上。该病每年6月初始见,6月下旬至7月中旬为发病盛期,累计病株数符合y=de~(-(b/x))曲线。本病的发生与土壤含砂率有密切关系。含砂量越高,发病越重(r=0.8956~*)。在相同土壤中,土壤田间持水量与发病呈负相关(r=-0.905~*)。试验示范证实,采用以抗病良种为主的综合防病措施,是防治本病经济有效的方法,推广面积在3万亩以上。氯化苦处理土壤也有良好的效果。     

番茄枯萎病菌和青枯病菌拮抗细菌的评价

为筛选出对番茄枯萎病和青枯病有较好防效的生防菌,采用平板对峙法,以番茄枯萎病菌Fusarium oxysporum和番茄青枯病菌Ralstonia solanacearum为靶标菌,从江苏沭阳、宿迁、溧水及内蒙古海拉尔分离到的2 062株细菌菌株中筛选拮抗菌株,并采用平板对峙法、拮抗菌液灌根法、分子生物学方法进行拮抗物质检测、盆栽试验及种属鉴定。结果表明:从2 062株细菌中共筛选到21株对番茄枯萎病和青枯病具有很强拮抗作用的菌株,均能分泌蛋白酶,具有解磷作用;不能分泌几丁质酶和纤维素酶,仅4株细菌能分泌嗜铁素。拮抗细菌SY290对番茄枯萎病和番茄青枯病防效最高,分别达到74.2%和75.0%,SQ728和LS536次之,但防效均大于60%。结合各菌株形态特征、16S r DNA与gyr-B序列分析结果,菌株SY177、SY290和SQ728鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens,菌株LS536为枯草芽胞杆菌B.subtilis。     

Immunofluorescence Colony-staining (IFC) for Detection and Quantification of Ralstonia (Pseudomonas) solanacearum Biovar 2 (Race 3) in Soil and Verification of Positive Results by PCR and Dilution Plating

A procedure was developed for specific and sensitive quantitative detection of Ralstonia (Pseudomonas) solanacearum biovar 2 (race 3) in soil. It is based on immunofluorescence colony-staining (IFC) followed by confirmation of the identity of fluorescent colonies by PCR-amplification or dilution plating on a semi-selective medium, SMSA. Addition of sucrose and the antibiotics cycloheximide and crystal violet to the non-selective trypticase soy broth agar resulted in increased colony size and staining intensity of R. solanacearum in IFC. Verification of IFC-results by picking cells from IFC-positive colonies followed by dilution plating of the suspended cells on SMSA was highly efficient. The success rate was 92% and 96% with spiked and naturally contaminated soils respectively. Several other bacterial species which cross-reacted with polyclonal antibodies in IFC also grew on SMSA and were difficult to distinguish from R. solanacearum, thereby necessitating confirmation of the results. Rapid verification of IFC-positive results directly by PCR-amplification with primers D2/B specific to division 2 of R. solanacearum had a success rate of 86% and 96% with spiked and naturally contaminated soil samples, respectively. Primers D2/B reacted with all R. solanacearum division 2 strains, and strains of R. syzygii and the banana blood disease bacterium, but not with saprophytic bacteria cross-reacting in IFC with R. solanacearum antibodies. In comparative tests, IFC was able to detect consistently ca. 100 cfu g^–1 of soil, a detection level similar to that found with direct plating on SMSA, but less laboriously, whereas detection level with a bioassay on tomato plants was only 10⁴–10⁵ cfu g^–1 of soil.     

Validation and use of a qPCR protocol to quantify the spread of Ralstonia solanacearum in susceptible and resistant eucalypt plants

Bacterial wilt caused by Ralstonia solanacearum is a serious disease of eucalypt in humid and high temperature areas worldwide. Spreading of the bacterium in the field or to other nurseries occurs mainly by symptomless infected plant material. The use of pathogen-free propagating material as well as planting of resistant genotypes are currently the only strategies used for disease control. Therefore, a reliable and sensitive method for detection of low titres of R. solanacearum in infected plant tissue is essential for the success of management programmes. In this work, we adapted an efficient intercalating dye-based real-time PCR protocol to detect the bacterium in symptomless eucalypt plants as well as to investigate its movement in eucalypt clones CLR172 and CLR371, which exhibit resistant and susceptible phenotypes, respectively. We found that the bacterium translocates acropetally and basipetally in inoculated but symptomless cuttings of the resistant clone, as in cuttings of the susceptible clone displaying symptoms. Nevertheless, a smaller concentration of bacterial DNA was detected in tissues of the resistant clone. Mature biofilms occluding the xylem vessels were present in the susceptible clone whereas only single cells or small aggregates were observed in the resistant clone. This work contributes to improve our knowledge of the colonization process of R. solanacearum in eucalypt clones with different levels of susceptibility and to understand how the defence mechanisms against bacterial wilt in Eucalyptus work. Our findings could aid in the selection of the most resistant eucalypt clones to be used in wilt disease management programmes.     

烟草青枯病菌与其拮抗菌解淀粉芽胞杆菌的代谢表型差异分析

为探究解淀粉芽胞杆菌X60作为烟草青枯病生防菌剂的潜力,采用Biolog代谢表型技术比较了2种细菌的不同代谢表型。结果表明,烟草青枯病菌和解淀粉芽胞杆菌分别能代谢19%、41%的碳源,43%、77%的氮源,95%、86%的磷源以及100%、69%的硫源,分别有94、91种生物合成途径,49、95种渗透压表型以及19、94种pH代谢表型;解淀粉芽胞杆菌比烟草青枯病菌代谢显著的碳源有L-果胶糖、D-甘露糖等34种,氮源有腺苷、胞苷等29种;烟草青枯病菌比解淀粉芽胞杆菌代谢显著的碳源有D-糖二酸、半乳糖醇等9种,氮源有缩二脲、葡萄糖苷酸等11种;解淀粉芽胞杆菌的渗透压和pH环境适应力比烟草青枯病菌强;解淀粉芽胞杆菌具有脱羧酶和脱胺酶的活性。研究表明,2种细菌的代谢表型间存在较大差异,解淀粉芽胞杆菌的碳源、氮源、渗透压及pH代谢表型较烟草青枯病菌的丰富,烟草青枯病菌的磷源、硫源和生物合成途径代谢表型较解淀粉芽胞杆菌的丰富。     

Potato bacterial wilt in India caused by strains of phylotype I, II and IV of Ralstonia solanacearum

Bacterial wilt or brown rot is one of the most devastating diseases of potato caused by a bacterium Ralstonia solanacearum (Smith 1986) Yabuuchi et al. (Microbiol Immunol 39:897–904 1995). Traditionally, R. solanacearum is classified into five races (r) on the basis of differences in host range and six biovars (bvs) on the basis of biochemical properties. Recently using molecular methods, R.?solanacearum has been classified into phylotypes based on the intergenic transcribed sequence of the ribosomal RNA genes 16S and 23S and into sequevars based on the endoglucanase gene (egl) sequence. In the present study, 75 bacterial strains, isolated from wilt infected potatoes from various potato growing regions of India, were classified by traditional and molecular methods. The identity of all the strains was confirmed as R. solanacearum as expected single 280-bp fragment resulted in all the strains following PCR amplification using R. solanacearum specific universal primer pair 759/760. Biovar (bv) analysis, based on utilization of disaccharide sugars and hexose alcohols, categorised the 75 strains into bv2 (78.7 %), 2 T (5.3 %), 3 (5.3 %) and 4 (10.7 %). The phylotype specific multiplex PCR assigned 78.7 % strains to phylotype II, 16.0 % to phylotype I and 5.3 % to phylotype IV. Phylogenetic analysis of egl gene sequences clustered all fifty nine phylotype II (bv2) strains with reference strain IPO1609 (IIB-1), all four phylotype IV (bv2T) strains with reference strain MAFF301558 (IV-8), three phylotype I (bv3) strains with reference strain MAFF211479 (I-30) and all eight phylotype I (bv4) and one phylotype I (bv3) strain with reference strain CIP365 (I-45). The study concluded that the Indian potato strains of R. solanacearum belong to three out of four phylotypes namely: the Asian phylotype I, the American phylotype II, and the Indonesian phylotype IV. This is the first study to address the diversity of R. solanacearum from potato in India using phylotype and sequevar scheme. We also report here for the first time the occurrence of phylotype IV sequevar 8 (bv2T) strain of R. solanacearum causing potato bacterial wilt in mid hills of Meghalaya in India.     

Involvement of jasmonic acid signalling in bacterial wilt disease resistance induced by biocontrol agent Pythium oligandrum in tomato

When the biocontrol agent Pythium oligandrum (PO) colonizes the rhizosphere, it suppresses bacterial wilt disease in tomato (Solanum lycopersicum cv. Micro‐Tom) caused by Ralstonia solanacearum, and a homogenate of its mycelia exhibits elicitor activity, inducing an ethylene (ET)‐dependent defence response in Micro‐Tom. Since salicylic acid (SA) and jasmonic acid (JA) play an important role in plant defence responses to pathogens, the involvement of SA‐ and JA‐dependent signal transduction pathways in resistance to R. solanacearum was investigated in tomato roots treated with a mycelial homogenate of PO. Bacterial wilt disease was also suppressed in tomato cv. Moneymaker treated with the PO homogenate. However, the SA‐inducible PR‐1(P6) gene was not up‐regulated in either Micro‐Tom or Moneymaker. SA did not accumulate in homogenate‐treated roots in comparison with distilled water‐treated controls, even 24 h after inoculation. Induced resistance against R. solanacearum was not compromised in SA‐non‐accumulating NahG transgenic plants treated with the PO homogenate. On the other hand, the expression of the JA‐responsive gene for the basic PR‐6 protein was induced in both tomato cultivars treated with the PO homogenate. Furthermore, quantitative disease assays showed that the induced resistance against R. solanacearum was compromized in PO homogenate‐treated jai1‐1 mutant plants defective in JA signalling. These results indicated that the JA‐dependent signalling pathway is required for PO‐induced resistance against R. solanacearum in tomato.