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.     

Distribution of Pseudomonas solanacearum in the stem tissues of tomato plants with different levels of resistance to bacterial wilt

Ten tomato cultivars, from the USA, Taiwan and French Antilles, were compared for resistance to bacterial wilt as measured by disease and pathogen population invasiveness and density in the stem at the collar and midstem regions. On the basis of disease incidence, cultivars ranked from highly susceptible to totally resistant. By contrast, no significant difference was observed in bacterial population in wilting plants, regardless of the cultivar. All symptomless plants were latently infected at the collar level. Percentage of symptomless plants with bacteria at the midstem level was significantly correlated with the degree of resistance; the more resistant, the lower the stem colonization. Restriction of Pseudomonas solanacearum invasiveness in the vascular tissues of the stem is associated with resistance properties in tomato. This observation may be useful in developing improved criteria for selecting stable resistance to the disease.     

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.     

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.     

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.     

Enhanced control of bacterial wilt of tomato by DL-3-aminobutyric acid and the fluorescent Pseudomonas isolate CW2

Journal of Plant Diseases and Protection - Soil treatments with DL-3-aminobutyric acid (BABA) and the fluorescent Pseudomonas isolate CW2 significantly reduced bacterial wilt on tomato plants...     

Effects of para-aminobenzoic acid on bacterial speck symptom development and Pseudomonas syringae pv. tomato populations in tomato leaves

Para-aminobenzoic (PABA) is reported to induce resistance against a range of plant pathogens in different crops in a salicylic acid-dependent manner. However, factors affecting its efficacy are not well understood. Foliar PABA applications on tomato seedlings reduced lesion incidence caused by Pseudomonas syringae pv. tomato (Pst) in a dose-dependent manner in distal leaves up to 18 mM under controlled environment conditions, but only three out of six commercial processing tomato cultivars tested showed a response to PABA. Leaves in direct contact with 9 and 18 mM PABA of both PABA-responsive and PABA-nonresponsive cultivars showed phytotoxicity. In a PABA-responsive cultivar, one, two and three PABA applications were equally effective at reducing lesion incidence in distal leaves, but the duration of control only persisted for approximately 7 days. Although PABA application reduced lesion incidence in distal leaves, the Pst population in leaves was unaffected. Lesions on PABA-treated plants were larger than nontreated plants, and thus the proportion of leaf surface area with lesions was unaffected by PABA treatment. In in vitro assays, 18 and 72 mM PABA produced zones of inhibition against Pst 15 and 50% larger than the ethanol control, demonstrating direct antimicrobial effects of PABA. PABA application did not affect symptom development in a mixed infection of Pst or Xanthomonas spp. in one field experiment with a PABA-responsive cultivar. Further research is needed to understand why PABA was unsuccessful in the field before it is to be used as a practical disease management tool for foliar bacterial diseases of tomato.

     

温度和病原接种浓度对番茄青枯病菌侵染的影响

本试验以不同浓度的Pseudomovas solanacearum悬浮液接种番茄品种湘引79-1号(抗)和粤农2号(感),置于不同温度的光照培养箱内,观察发病情况.当菌液浓度为3×108CFU/mL、3×106CFU/mL在温度25℃以上时9天或更短时间就表现青枯病症状,发病的最低温度是22℃,但潜育期较长,需20天,温度30℃与25℃的病情指数没有显著差别,但30℃潜育期短,只需5天.25℃潜育期稍长,则要9天.病情的发展随着菌液浓度的增高而严重,3×108CFU/mL浓度的病情指数最高达90%,3×102CFU/mL浓度的病情指数最低为40%.     

Phenotypic characterization of Pseudomonas fluorescens PfG32R and its spontaneous gacS mutants and biocontrol activity against bacterial wilt disease of tomato

Pseudomonas fluorescens strain PfG32R actively suppresses the occurrence of bacterial wilt disease of tomato caused by Ralstonia solanacearum. To determine the characteristics affected by spontaneous mutations in the gacS gene, three spontaneous mutants NR1, NR9, and ASW6 were tested for production of several enzymes, antimicrobial activity, and biocontrol activity against bacterial wilt disease of tomato in a greenhouse. Production of biosurfactant and of enzymes such as Tween-80 lipase, gelatin protease, and lecithinase; swarming activity; and antifungal activities against Fusarium oxysporum f. sp. radicis-lycopersici and F. graminearum were inactivated by mutations in the gacS gene, indicating that they are under the regulation of the GacS/GacA two-component regulatory system. Levan production, swimming, antifungal activity against Magnaporthe grisea, and antibacterial activity against R. solanacearum and Erwinia carotovora ssp. carotovora were not controlled by the GacS/GacA system. Biocontrol activities of the three spontaneous mutants varied from strain to strain, indicating that the biocontrol activities of PfG32R are influenced not only by the GacS/GacA system but also by other complex factors, which may not be regulated by the system.     

Induction of transient ethylene and reduction in severity of tomato bacterial wilt by Pythium oligandrum

Pythium oligandrum (PO) is a mycoparasite on a wide range of fungi and suppresses diseases caused by fungal pathogens when colonizing the rhizosphere. PO and its cell wall proteins (CWPs) have elicitor activity that induces defence responses in plants. The potential of a mycelial homogenate of PO to suppress bacterial diseases was studied in roots of tomato ( Lycopersicon esculentum cv. Micro-Tom) inoculated with Ralstonia solanacearum . PO-treated plants showed enhanced resistance to R. solanacearum and reduction in severity of wilt symptoms. As ethylene often acts as one of the signal molecules for induced resistance, its production following treatment of tomato roots with the mycelial homogenate or CWP of PO was measured. The level of ethylene in PO- and CWP-treated plants was transiently elevated six- to 11-fold at 4–8 h after treatment, followed by high expression of three basic ethylene-inducible defence-related genes ( PR-2b , PR-3b and PR-5b ). Analysis of PR-5b gene expression in the leaves of PO- and CWP-treated plants suggested that PR gene expression was induced systemically. The expression of LeERF2 and LeETR4 , which confer an ethylene-dependent signalling pathway, was also significantly accelerated by such treatments. These results indicate that PO has the potential to control bacterial wilt disease and that CWP may play an important role in the induction of resistance to R. solanacearum accompanying the activation of the ethylene-dependent signalling pathway.     

Occurrence of bacterial wilt of peanut (Arachis hypogaea) caused by Pseudomonas solanacearum and opportunistic infection of aibika (Abelmoschus manihot) in Papua New Guinea

Pseudomonas solanacearum , the cause of bacterial wilt was isolated from stems, roots and developing cotyledons of peanut and the stem of a single aibika plant growing in a subsistence garden in Port Moresby. The garden had been cleared and planted to peanut after a fallow of at least 2 years. The bacterium could not be isolated from roots or rhizosphere soil of locally growing grasses and weeds, nor could seed transmission be demonstrated in peanut.     

Molecular characterization of Ralstonia solanacearum strains from Ethiopia and tracing potential source of bacterial wilt disease outbreak in seed potatoes

Bacterial wilt, caused by Ralstonia solanacearum, is emerging as a major threat to potato production in Ethiopia, reaching epidemic proportions in the Chencha district recently, with a prevalence of 97% of potato fields in 2015. The recent disease outbreak in the district coincided with a significant introduction of seed potatoes. This research was therefore initiated to genetically characterize the pathogen so as to trace its source, identify its relationship with outbreaks in the rest of the country, and make intervention recommendations. Ralstonia solanacearum isolates were sampled both from seed and ware potato fields in Chencha and from seed potato fields in production regions suspected of being potential sources of the pathogen. Multiplex PCR and phylogenetic analysis of partial endoglucanase gene sequences identified all of the isolates as phylotype IIB sequevar 1. VNTR sequence analysis distinguished 11 different haplotypes, nine of which were unique to the Chencha district. However, one of the haplotypes was common to all seed potato producer regions of Ethiopia except for the Shashemene area. The unique and diverse VNTR haplotypes of the pathogen in Chencha indicates that it is well established in the district. When a geographical map of the VNTR haplotypes was superimposed with the main cross‐regional seed potato distribution pattern of the country, it became evident that the pathogen was being disseminated via latently infected seed from the Holeta‐Jeldu area in the Central Highlands of Ethiopia. Identification of largely uninfected highland districts and multiplication of high‐grade seed potato exclusively in those districts should be given priority.     

Effect of variety choice, resistant rootstocks and chitin soil amendments on soil-borne diseases in soil-based, protected tomato production systems

Soil-borne diseases are the most significant crop protection problem in soil-based, low-input and especially organic glasshouse production systems in Europe. While chemical soil disinfestation has been the control method of choice in conventional farming systems, soil steaming has been the main strategy for the control of soil-borne diseases in organic production. Both methods are extremely expensive and have been increasingly restricted for environmental reasons by governments, and integrated and organic farming standard-setting bodies. The use of disease-tolerant varieties, grafting onto resistant rootstocks and chitin soil amendments were evaluated as potential replacements for soil steaming in organic and other low-input tomato production systems. When only Pyrenochaeta lycopersici and/or Meloidogyne spp. were present in soil, grafting and/or chitin soil amendment were found to be as effective in reducing root disease and/or increasing yield as soil steaming, but the efficacy of both treatments was reduced when Verticillum albo-atrum was also present in soil. No additive effects of combining grafting and chitin soil amendments could be detected. A more widespread use of grafting and/or chitin soil amendments may therefore allow significant reductions in the use of steam and chemical soil disinfestation in glasshouse crops. It will also allow integrated and organic farming standard-setting bodies to impose further restrictions on the use of soil disinfestation treatments.     

Effects of soil inoculants on the growth,transpiration and wilt disease of chickpea

Journal of Plant Diseases and Protection - Glasshouse experiments were conducted twice to assess the effects of soil inoculants (Rhizobium sp., Glomus fasciculatum and Trichoderma harzianum) and...     

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.     

Diversity of Ralstonia solanacearum causing potato bacterial wilt in Iran and the first record of phylotype II/biovar 2T strains outside South America

The diversity of 40 strains of Ralstonia solanacearum causing bacterial wilt of potato in the major potato-growing areas of Iran was assessed. Based on rep-PCR genomic fingerprinting, strains fell into two distinct groups. The first group contained 37 of the 40 strains and the second consisted of three strains from a narrow tropical region in Iran. The three strains from the narrow tropical region were found to be phenotypically and genotypically most similar to R. solanacearum biovar 2T strains, whereas all other strains were phenotypically and genotypically identified as being R. solanacearum biovar 2/race 3. Phylogenetic analysis of endoglucanase gene sequence information of two of the strains from the tropical region revealed that they belonged to phylotype II of the R. solanacearum species complex and had 100% sequence similarity to a biovar 2T strain from potato in Peru. This is the first report of the presence of R. solanacearum phylotype II/biovar 2T in Iran and the first report of the existence of this group of R. solanacearum outside South America.     

Influence of application rate and manure amendment on cnloridazon dissipation in the soil

Plots of a sugar beet field in Belgium were treated pre-emergence with 1.3, 1.95 or 2.6 kg a.i. ha^-1 chloridazon. Some of these plots had been amended 1 month before sowing with 50 1 ha^-1 cow manure. Soil samples were taken al regular intervals and analysed by gas-liquid chromatography and gas-liquid chromatography combined with mass spectrometry. During the first month following chloridazon application, soil dissipation followed apparent first-order kinetics with soil half-lives being independent of the dose, and were 37 days in the non-manured plots and 96 days in the manured plots. After the first month, rates of chloridazon soil dissipation increased, giving the same residual chloridazon soil concentration of c. 0.25 mg a.i, kg^-1 in all plots 1.8 months after application. Residual levels remained at this concentration up to the third month after application, and then disappeared, leaving no delectable soil residues at harvest. Chloridazon soil dissipation thus occurred according to unusual kinetics.