Bacterial wilt resistance in tomato associated with tolerance of vascular tissues to Pseudomonas solanacearum

High populations of Pseudomonas solanacearum were detected in some, but not all stems of bacterial wilt resistant ('CRA 66', 'Hawaii 7996' and 'Caraibo') and susceptible ('Floradel') tomatoes. Latent infection, i.e. spread of P. solanacearum into xylem vessels, was confirmed in Caraibo, Hawaii 7996 and 'CRA 66' (the resistant parent of Caraibo). None of the plants within the resistant cultivars wilted and those cultivars were characterized by tolerance of the vascular tissues to high bacterial densities. In contrast, plants of cultivar Floradel showed consistent symptoms and wilted rapidly, with higher mean bacterial density than resistant cultivars. Bacterial wilt resistance was not associated with resistance to bacterial root invasion but with the capability of the plant to limit P. solanacearum colonization in the stem. The extent of bacterial colonization is proposed as a criterion to quantify tolerance, complementary to absence of external wilt symptoms used in breeding programmes for resistance.     

Resistance to bacterial wilt in tomato as discerned by spread of Pseudomonas (Burholderia) solanacearum in the stem tissues

Different criteria were compared for assessing bacterial wilt resistance in 13 tomato genotypes varying in disease susceptibility. Wilt severity and bacterial invasiveness at collar and midstem were compared in the field under cooler (March to May, 20–28°C) and warmer months (June to August, 23–29°C), which were unfavourable and favourable to wilt symptom expression, respectively. A model was proposed for determining resistance regardless of climatic conditions prevalent during field experimentation. This model was based on an estimate of bacterial invasiveness termed the colonization index. Using a qualitative imprint method we confirmed that the more resistant the genotype, the lower the bacterial colonization of the stem. The colonization index accounted both for wilted plants and for infected asymptomatic plants in which Pseudomonas solanacearum populations failed to produce wilt. The colonization index at midstem was the more useful indicator of resistance under favourable conditions. When environmental conditions were unfavourable to wilt, colonization index at collar level discerned resistant genotypes more clearly. The results formed the basis for a model for predicting the degree and stability of resistance in tomato.     

Assessment of resistance to bacterial wilt incited by Ralstonia solanacearum in tomato germplasm

Journal of Plant Diseases and Protection - Bacterial wilt incited by Ralstonia solanacearum has been found the most damaging and widespread diseases of tomato throughout the world and causes heavy...     

Effect of calcium nutrition on resistance of tomato against bacterial wilt induced by Ralstonia solanacearum

This study investigated the effect of calcium nutrition on tomato bacterial wilt caused by Ralstonia solanacearum and the regulation of resistance mechanisms. Plants cultured in nutrient solution with calcium concentrations of 0.5, 5.0, and 25.0 mM, were inoculated with R. solanacearum by the root dip method. Severity of disease development, Ca concentration in tomato root and shoot tissues, hydrogen peroxide (H₂O₂) concentration, peroxidase (POD, EC 1.11.1.7) and polyphenol oxidase (PPO, EC 1.10.3.2) in tomato leaves were analyzed. Disease severities of low, medium and high Ca treatments were 100 %, 77.1 % and 56.8 % respectively. Plant growth in high Ca treatment was significantly better than those in low Ca treatment in height, stem diameter and biomass. Tomato plants absorbed significantly more Ca in roots and shoots as the level of Ca in the nutrient solution increased. In addition, H₂O₂ level in high Ca treatment rose faster and reached a higher peak with 10.86 μM gFW^?1(31.32 % greater than medium Ca plants). The activities of POD and PPO also have a greater increase in high Ca treatment with 99.09 U gFW^?1 and 107.24 U gFW^?1 compared to 40.70 U gFW^?1 and 77.45 U gFW^?1 in low Ca treatment. A negative correlation was found between Ca concentration, level of H₂O₂, POD, PPO in tomato, and disease severity, indicating that they played an important role in resistance of tomato to this disease. These results suggested that Ca was involved in the regulation of H₂O₂ concentration, and activity of POD and PPO in tomato.     

Mechanisms of resistance to Ralstonia solanacearum in tomato rootstocks and integrated management of bacterial wilt using high grafting

Journal of General Plant Pathology -     

Development of resistance in tomato plants grafted onto Solanum torvum against bacterial wilt disease

Journal of Plant Diseases and Protection - The bacterial wilt disease caused by Ralstonia solanacearum is a major constraint in the cultivation of tomato in Saint Lucia. The limited options and...     

Nitric oxide-overproducing transformants of Pseudomonas fluorescens with enhanced biocontrol of tomato bacterial wilt

The relation between nitric oxide (NO) production and the protective ability of Pseudomonas fluorescens T5 against bacterial wilt disease in tomato was examined. The endogenous nitric oxide reductase gene of T5 was disrupted by homologous recombination using a suicide plasmid. Three disruptants were obtained, and all had higher levels of NO production. Infection with Ralstonia solanacearum was reduced in tomato plants treated with the NO-overproducing transformants compared with the wild type. These results suggest that the modification of pseudomonads to increase their level of NO production is a new approach to enhancing their biocontrol efficacy.     

Interaction between silicon amendment,bacterial wilt development and phenotype of Ralstonia solanacearum in tomato genotypes

Silicon amendment significantly reduced bacterial wilt incidence expressed as area under disease progress curve for tomato genotypes L390 (susceptible) by 26.8% and King Kong2 (moderately resistant) by 56.1% compared to non-treated plants grown in hydroponic culture. However, wilt incidence in silicon-treated plants of genotype L390 reached 100% at 13 days post-inoculation (dpi), while in genotype King Kong2, plant death was retarded by 6 days, with 20% reduction of final wilt incidence. Bacterial numbers were significantly lower in silicon-treated compared to non-treated plants in King Kong2 at 2 dpi in midstems and in all organs at 5 dpi, and in Hawaii 7998 (resistant) in all organs at 2 dpi. Differences between genotypes were obvious on midstem level (5 dpi), where bacterial populations were generally significantly lower compared to roots. Increased tolerance was observed in genotypes L390 and King Kong2 with silicon treatment.Silicon accumulated in roots and was low in stems and leaves. Inoculation with Ralstonia solanacearum did not significantly affect silicon uptake and distribution. Negative correlations between root silicon content and bacterial numbers of midstems in genotypes Hawaii 7998 and King Kong2 suggested an induced resistance. Indications for an influence of host genotype and silicon treatment on the phenotypic conversion of R. solanacearum strain To-udk2-sb from fluidal to non-fluidal colonies in planta were observed.This is the first report on the effect of silicon on a bacterial disease and in a silicon-non-accumulator plant.     

Effects of avirulent bacteriocin-producing strains of Pseudomonas solanacearum on the control of bacterial wilt of tobacco

Root systems of tobacco dipped in suspensions containing 2 × 10⁹ colony forming units (CFU)/ml of avirulent bacteriocin-producing strains (ABPS) of Pseudomonas solanacearum and assayed immediately after planting in steam-sterilized soil had 8 × 10⁶ CFU/root system of ABPS. The bacterial population declined to an average of 5·3 × 10⁵ CFU/root system after 30 days. Roots of seedlings dipped in bacterial suspensions of ABPS were more effectively protected against wilt caused by P. solanacearum than those dipped in suspensions of an avirulent nonbacteriocin-producing strain (ANBPS). Lipopolysaccharide (LPS) isolated from one ABPS (121) inhibited the attachment of bacteria on roots by 70% but had no effect on the reduction of wilt, whereas bacterial cells significantly reduced the disease severity as compared to LPS or water treatment. In steam-sterilized soil containing a 1:1 mixture (5 × 10⁵ CFU/g of oven-dried soil) of ABPS 121 or 237 and the virulent strain K-60, ABPS 121 reduced multiplication of the virulent strain in soil and in the rhizosphere of seedlings. When roots of seedlings were dipped in a suspension of 2 × 10⁹ CFU/ml of ABPS before planting, root colonization by the virulent strain added to steam-sterilized soil at 2 × 10⁶ CFU/g of oven-dried soil was significantly reduced. When roots were dipped in a suspension of ABPS and assayed 20 days after planting, 98% of the bacterial population was found in the original zone of inoculation and only 2% was detected in new growths of the root system. Plants which were grown in soil infested with ABPS 121 or K-60 had both strains present at variable populations along all sections of roots.     

Effects of intercropping and soil amendment with urea and calcium oxide on the incidence of bacterial wilt of tomato and survival of soil-borne Pseudomonas solanacearum in Taiwan

Intercropping and soil amendment experiments were conducted to determine if they reduced populations of Pseudomonas solanacearum and bacterial wilt of tomato at the Asian Vegetable Research and Development Center (AVRDC) and at three other locations in Taiwan. At AVRDC, intercropping tomato with cowpea planted within the row significantly reduced bacterial wilt ( P  < 0.05) compared to when tomato was cropped alone. The P. solanacearum population in soil was not affected by intercropping with cowpea, soybean, or Welsh onion. At the same site, however, a preplanting soil amendment consisting of urea (200 kg ha⁻¹ N) and CaO (5000 kg ha⁻¹) significantly reduced the pathogen population and tomato bacterial wilt ( P  < 0.001). The effect of the soil amendment was not consistent when applied to soil from three other sites in Taiwan; in soil from two sites no reduction of the pathogen population occurred. At these sites, tomato bacterial wilt in the field was not reduced significantly after amending. In comparison with a non-amended control, the addition of only CaO reduced the P. solanacearum population in AVRDC soil significantly ( P  < 0.05), but the reduction was significantly greater when the complete soil amendment was added. In contrast, urea alone did not affect the survival of P. solanacearum in the soil. In a greenhouse experiment with AVRDC soil, P. solanacearum was undetectable 2 weeks after soil amendment, but in the same treatment tomato yield was significantly reduced by 48% ( P  < 0.05) compared with non-amended treatments. The suppressive effect of the soil amendment on the P. solanacearum population was probably due to the generation of one or several toxic substances during the transformation of urea in the presence of CaO.     

Evaluation of bacterial wilt resistance in tomato lines nearly isogenic for the Mi gene for resistance to root-knot

Resistance to bacterial wilt, caused by Ralstonia solanacearum , in tomato lines CRA 66 and Caraïbo is reported to be decreased by root-knot nematode galling and by introduction of the Mi gene for nematode resistance. The Mi gene is located on tomato chromosome 6, which also carries a major quantitative trait locus (QTL) for resistance to bacterial wilt. Bacterial wilt resistance was evaluated in F3-progenies derived from two crosses between near-isogenic lines, Caraïbo × Carmido and CRA 66 × Cranita, differing for small and large introgressions from Lycopersicon peruvianum that carry the Mi gene, respectively. These introgressed regions were mapped using RFLP markers. Plants homozygous Mi⁺/Mi⁺ (susceptible to the nematode) and homozygous Mi/Mi (resistant) for the Mi gene were selected in F2 and used to produce F3 progenies. Parents and F3-lines with Mi/Mi had resistance to bacterial wilt reduced by 30% in Caraïbo × Carmido and by 15% in CRA 66 × Cranita. Caraïbo and Carmido were demonstrated to be isolines and the small introgression from L. peruvianum resulted in loss of the QTL for bacterial wilt resistance, which is probably allelic or linked in repulsion to the Mi gene. In contrast, resistance to bacterial wilt segregated in the F3 lines from the cross CRA 66 × Cranita, giving families varying in resistance between the levels shown by the parents. Consequently, two hypotheses were considered: (i) after only four backcrosses, the parents were not isolines and the genes for resistance to bacterial wilt from CRA 66 were still segregating, and (ii) the parents were isolines and variation in resistance to bacterial wilt in F3 was due to recombination events among the large L. peruvianum introgressed chromosome region from Cranita.     

Magnesium oxide nanoparticles induce systemic resistance in tomato against bacterial wilt disease

Bacterial wilt is a serious problem affecting many important food crops. Recent studies have indicated that treatment with biotic or abiotic stress factors may increase the resistance of plants to bacterial infection. This study investigated the effects of magnesium oxide nanoparticles (MgO NP) on disease resistance in tomato plants against Ralstonia solanacearum, as well as its antibacterial activity. The roots of tomato seedlings were inoculated with R. solanacearum and then immediately treated with MgO NP; the treated plants showed very little inhibition of bacterial wilt. In contrast, when roots were drenched with a MgO NP suspension prior to inoculation with the pathogen, the incidence of disease was significantly reduced. Rapid generation of reactive oxygen species such as O₂^?˙ radicals was observed in tomato roots treated with MgO NP. Further O₂^?˙ was rapidly generated when tomato plant extracts or polyphenols were added to the MgO NP suspension, suggesting that the generation of O₂^?˙ in tomato roots might be due to a reaction between MgO NP and polyphenols present in the roots. Salicylic acid‐inducible PR1, jasmonic acid‐inducible LoxA, ethylene‐inducible Osm, and systemic resistance‐related GluA were up‐regulated in both the roots and hypocotyls of tomato plants after treatment of the plant roots with MgO NP. Histochemical analyses showed that β‐1,3‐glucanase and tyloses accumulated in the xylem and apoplast of pith tissues of the hypocotyls after MgO NP treatment. These results indicate that MgO NP induces systemic resistance in tomato plants against R. solanacearum.     

番茄青枯病病原菌拮抗菌株的筛选及其田间防控作用研究

从番茄青枯病发病严重田块的健康植株根际土壤中分离筛选得到2株高效拮抗菌株,命名为W12和W118,经16SrDNA基因鉴定均属芽胞杆菌属;用PCR扩增的方法扩增脂肽类抗生素合成基因,结果表明W12和W118含有合成bacillomycin、iturin和fengycin三种抗生素的基因;将2株拮抗菌用于田间试验,结果表明混合菌株防控效果最好,3次灌菌后防控效果达到62.3%,单独施用菌株W118较单独施用W12防控效果好,3次灌菌后防控效果达到56.7%。     

番茄青枯病生物防治研究进展

番茄青枯病是番茄生产上最严重的病害之一,化学防治效果不佳,且对环境不友好。本文概述了国内外应用无致病力青枯菌菌株、拮抗细菌和生物技术等方法对番茄青枯病进行生物防治的研究进展,并对生物防治存在的相关问题及应用前景进行了讨论。     

青枯雷尔氏菌无致病力突变菌株的构建及其防效评价模型分析

 利用青枯雷尔氏菌(Ralstonia solanacearum)无致病力菌株防治番茄青枯病具有很好的应用潜力。作者通过分离筛选自然弱毒株、⁶⁰Co辐射诱变和EZ-Tn5插入诱变,分别获得3、12和40株青枯雷尔氏菌无致病力突变菌株。经盆栽番茄苗致病性检测,15 d后均未发病,证实均为无致病力青枯雷尔氏菌。进一步对番茄青枯病的防治试验表明,从番茄青枯病发病田块分离的无致病力突变菌株FJAT1458的防治效果最好,防效达100%。该菌株能定殖番茄植株根系土壤、根部和茎部,定殖数量均表现为“先增后减”的趋势,并且接种浓度越大、苗龄越小,定殖数量越大。从构建的防效模型可以看出,不同接种浓度条件下,植株发病率随时间变化符合的回归方程不同,相关系数R值也不同,接种浓度越大,R值越小。本研究获得的青枯雷尔氏菌无致病力突变菌株FJAT1458对番茄青枯病具有很好的防病效果。     

林木青枯病研究进展

青枯病是由青枯劳尔氏菌引起的一种严重的植物土传病害,我国南方多种树种被其侵染发病。本文介绍了我国林木青枯病的发生情况,概括了青枯病检测与防治方面的研究进展,并对其中的一些问题进行了探讨。     

Compost enhances plant resistance against the bacterial wilt pathogen Ralstonia solanacearum via up-regulation of ascorbate-glutathione redox cycle

The interactions between the pathogen Ralstonia solanacearum and potato Solanum tuberosum plants were studied to investigate the reactive oxygen species metabolic system and ascorbate (ASC)-glutathione (GSH) redox cycle in response to compost application. Single potato eyepieces were germinated and grown in pots containing sandy soil with or without compost at a rate of 7.5 g kg^?1 soil. Non-compost- and compost-treated plants (CTP) were inoculated with R. solanacearum 25 days after planting and then analyzed after 10 days, unless otherwise stated. The present results revealed that pathogen infection caused a remarkable decrease in plant growth related parameters and productivity and an increase in disease incidence. However, under these conditions compost had substantially improved plant growth and decreased disease incidence and bacterial population. R. solanacearum resulted in significant enhancement in the activities of NADPH oxidase, lipoxygenase, the production rate of superoxide and hydroxyl radicals, levels of hydrogen peroxide, membrane lipid peroxidation, and protein oxidation indicating the induction of oxidative stress in potato roots. However, the pathogen-mediated enhancement in indices of oxidative stress was considerably decreased by compost application, which enhanced the activities of ascorbate peroxidase (APX, EC 1.11.1.11), monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), dehydroascorbate reductase (DHAR, EC 1.8.5.1) and glutathione reductase (GR, EC 1.6.4.2) in infected potato plants, implying a better ROS-scavenging activity. Data also indicated that there were general increases in ASC and GSH content in infected compost treated plants, but non-compost treated ones significantly had lower levels of such redox metabolites. In addition, significantly higher ratios of ASC/DHA (dehydroascorbate) and GSH/GSSG (glutathione disulphide) were generally found in CTP than in non-compost treated-ones. The obtained results suggest that compost provides effective protection against the Ralstonia bacterial pathogen via up-regulation of the capacity of the ASC-GSH cycle and modulation of the cellular redox status, thereby eliminating ROS damage and sustaining membrane stability.