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1.
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.  相似文献   

2.
内生菌B47的定殖能力及其对番茄青枯病的防治作用   总被引：11，自引：4，他引：11  
从番茄茎分离的内生枯草芽孢杆菌菌株B47对番茄青枯病有较好的防治作用,利用该菌株的抗链霉素突变菌株,研究其在土壤和番茄植株根、茎中的定殖能力及其对番茄青枯病的防治作用。结果表明,枯草芽孢杆菌菌株B47可在土壤和番茄植株中定殖。B47施到土壤中后的15~45天,其数量逐步增加,45天后,其数量逐步下降。B47在土壤中的定殖能力随土壤的种类和土壤的处理情况而异。施入菜地土后的第45天,B47在非灭菌土中的数量是9.91×105cfu/g土壤干重,而在灭菌土中的数量是9.84×107cfu/g土壤干重。接种后,番茄植株根和茎中的B47数量,从苗期到结果期逐渐增加,但到了成熟期呈下降趋势。B47和番茄青枯病菌混合施入土壤后,随B47的数量增加番茄青枯病菌的数量显著降低。当番茄植株根和茎中B47的含量分别为1.17×104cfu/g鲜重和3.33×104cfu/g鲜重时,接种番茄青枯病菌后的第20天,对番茄青枯病的防治效果达79.79%。  相似文献   

3.
A method for detectingClavibacter michiganensis ssp.michiganensis in tomato seeds was evaluated. The method is based on rapid screening of tomato seed lots using indirect immunofluorescence staining (IF), followed by dilution plating of IF positive seed lots. Different polyclonal antisera, prepared againstC. michiganensis ssp.michiganensis were tested for their specificity using IF. All strains ofC. michiganensis ssp.michiganensis tested reacted with the polyclonal antisera. Two of nine saprophytic isolates from tomato seeds were positive with the antisera as well as with the control normal serum, but cells of these isolates were distinct in shape from cells ofC. michiganensis ssp.michiganensis.For extraction of the pathogen from the seed, seeds were either blended with a stomacher or soaked at 4–6 °C. The stomacher method yielded more fluorescent cells in IF than 24 h soaking of seed samples. However, soaking of seeds for 48 h generally yielded less saprophytes and overall higher numbers ofC. michiganensis ssp.michiganensis colonies in dilution plating when compared to blending by a stomacher. SCM medium was generally more selective than KBT and modified CNS medium. However, the efficacy of the medium was dependent on the seed lot and/or extraction method used. Confirmation of suspected colonies with YDC (yeast-dextrose-carbonate medium), IF and a pathogenicity test on tomato seedlings proved to be highly reliable (P>0.95). For routine testing of seed lots it is recommended to screen tomato seed lost after soaking seeds for 24 h at 4–6 °C with IF, followed by plating of IF-positive seed lots on modified CNS and SCM after soaking seeds for an additional 24 h.  相似文献   

4.
Corynebacterium flaccumfaciens, causing bacterial wilt in bean, is a quarantine organism for Europe. This paper compares two methods for detecting C. flaccumfaciens in seed lots on the market. Experimentally contaminated bean seeds were homogenized and a standard extraction and concentration procedure was used. The final pellets were used for immunofluorescence staining and indirect isolation (through bean seedlings). The immunofluorescence staining sensitivity threshold was 1.23 × 107, corresponding to 1.82 × 104 fluorescent cells per ml of final concentrate. The risk of false negative reactions to immunofluorescence staining caused by serologically unrelated C. flaccumfaciens strains was also examined. The indirect isolation sensitivity threshold was 1.23 × 106 producing 43% symptomatic plants and 53% positive reisolation. There was no serious interference between C. flaccumfaciens and Pseudomonas syringae pv. phaseolicola in mixed inoculations and both pathogens were successfully reisolated. The comparative results are discussed in relation to the applicability of the methods to commercial lots of bean seeds.  相似文献   

5.
A rapid glasshouse‐based bioassay method to screen large numbers of cotton plants for responses to Fusarium oxysporum f. sp. vasinfectum (Fov) was developed. Different Fov inoculum concentrations and methods of inoculation were assessed using resistant and susceptible cotton cultivars. Cotton seeds were planted directly into Fov‐inoculated soil. Studies of seed germination, seedling establishment, seedling mortality and fusarium wilt symptoms (i.e. stunting, foliar symptoms and vascular browning) were performed to optimize the bioassay parameters. Growing seedlings in Fov‐inoculated soils at 5 × 104 or 1 × 105 CFU g?1 soil, in individual seedling tubes with 12 h at 28–30°C and 12 h at 15–18°C, gave consistent results when assessing Fov disease responses 6 weeks after inoculation. When fusarium wilt resistance ranks (FWRRs) and vascular browning index (VBI) means of 18 Australian and other cotton cultivars from the Fov glasshouse bioassay were compared against their fusarium field performance ranks (F‐ranks), assessed on adult plants for cotton cultivar release, Pearson’s correlation was highly significant for both comparisons. The level of congruence between field and glasshouse data indicated that this protocol should be an effective tool for large‐scale screening for Fov‐resistance responses in diverse germplasm and breeding populations and for advancing genetic research to develop molecular markers for Fov resistance in cotton.  相似文献   

6.
Ralstonia solanacearum causes a lethal bacterial wilt disease in many plants by colonizing the vascular tissues of the hosts. Upon inoculation of tomato seedlings through either leaf or root, the wilting symptoms occur first at the apical region and then proceed downward along the shoot. The systemic order of the disease initiation and progression in the host, independent of the site of pathogen inoculation, is yet to be investigated. To understand the disease progression more clearly, we have carried out a systematic study of the pathogen localization by GUS staining of inoculated tomato seedlings, at 24-hour intervals from 0 days post-inoculation (dpi) to 5 dpi. In both inoculation methods, pathogen colonization was observed at 1 dpi at the apical meristem as well as the cotyledon leaves, where the disease initiates. As the disease progressed, colonization by the pathogen towards the lower region of the shoot was observed. Disease consistency and pathogenicity magnitude were observed to be higher using the leaf inoculation method than the root inoculation method. Several R. solanacearum transposon-induced mutants that were reduced in virulence by root inoculation but virulent by leaf inoculation were obtained. Using GUS staining, it was observed that these mutants were unable to localize in the shoot region when inoculated in the root. Our study indicates that the apical meristem and the cotyledon leaves are the first regions to be colonized in inoculated tomato seedlings, which might explain the disease initiation from this region.  相似文献   

7.
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.  相似文献   

8.
Clavibacter michiganensis subsp. michiganensis (Cmm), the causal agent of bacterial canker and wilt, causes severe economic losses in tomato net‐houses and greenhouses worldwide. In this study, seedlings which were transplanted and inoculated monthly over 2 years wilted and died earlier in the spring (21–24°C) and autumn (18–23°C) than in the winter (15–18°C) and summer (28–31°C): T50 (the time taken for 50% of the plants to wilt or die) was 2 and 3–4 months after inoculation, respectively. A highly significant correlation was found between the average temperatures during the first month after inoculation and T50; the shortest T50 mortality (70 days) was observed for an average temperature of 26°C. Expression of virulence genes (pat‐1, celA, chpC and ppaA) by Cmm was higher in plants inoculated in the spring than in those inoculated in the summer. In another set of experiments, seedlings were inoculated and maintained in controlled‐environment growth chambers for 2 weeks. Subsequently, they were transplanted and maintained in commercial‐type greenhouses for 4–5 months. The temperatures prevailing in the first 48 h after inoculation were found to affect Cmm population size and virulence gene expression and to have season‐long effects on bacterial canker development.  相似文献   

9.
An organic hydroponic system that we developed has potential to control root diseases including bacterial wilt of hydroponically grown tomato. In inoculation tests with Ralstonia solanacearum during tomato plant cultivation in conventional inorganic hydroponics and in our organic system, many of the tomato seedlings in the conventional system wilted and died, but none of the seedlings in the organic hydroponics wilted or developed any symptoms, suggesting that the organic system can suppress this bacterial wilt disease. Interestingly, a rhizosphere biofilm, formed only on roots in the organic hydroponic system, may be responsible for the suppression of the bacterial wilt.  相似文献   

10.
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.  相似文献   

11.

Clavibacter michiganensis subsp. michiganensis is a very important pathogen that causes bacterial wilt of tomato (BWT). Biological control of plant diseases is a critical tool for protecting the environment from chemical pollution. Twenty-five isolates of the genus Trichoderma were obtained from a healthy tomato root. Of the 25 isolates, KABOFT4 showed highly antagonistic activity that controlled the growth of C. michiganensis subsp. michiganensis (Cmm7) under in vitro conditions. The 5.8S ribosomal RNA gene and internal transcribed spacer identified the isolate as Trichoderma harzianum KABOFT4. The effect of this isolate as a soil drench and/or foliar application on bacterial wilt under greenhouse conditions was studied. The germination percentage of tomato seed treated with KABOFT4 increased by 36.7% compared to infected seed treated with only the pathogen Cmm7. Under greenhouse conditions, tomato seedlings treated with KABOFT4 as a soil drench, foliar and soil treatment, and foliar treatment had a 61.3, 26.7, and 40% reduced disease severity relative to the infected control, respectively. All treatments had a positive effect on tomato plants that presented as greater vegetative growth and accumulation of dry matter. The best fresh and dry weight was recorded when plants were treated with KABOFT4 as a soil and foliar application. Tomato plants treated with KABOFT4 also had increased total phenol and flavonoid contents in inoculated and non-inoculated plants compared to untreated plants. Under greenhouse conditions, T. harzianum strains can be used as an environmentally friendly way to manage the most economically important tomato disease. The results showed that a native endophytic strain of T. harzianum was a potent biocontrol agent against C. michiganensis subsp. michiganensis. Application of this strain to tomatoes in the greenhouse resulted in a decrease in disease severity and an increase in crop biomass.

  相似文献   

12.
Ascochyta blight of chickpea (Cicer arietinum), caused by the fungus Didymella rabiei, has the potential to cause 100% crop loss in severe epiphytotics. Management of this disease often involves reducing sources of inoculum. The influence of sowing depth, host resistance, seed infection level and soil temperature on disease transmission was investigated in a series of glasshouse and growth room trials using seed artificially inoculated with D. rabiei. A positive correlation (R2=0.9992) was observed between rate of seed infection and the incidence of disease on seedlings. Disease transmission to seedlings was not significantly influenced by sowing depth (1, 3 and 6 cm) in separate trials on two cultivars. Susceptibility of the host showed no obvious influence on the frequency of disease transmission in two trials conducted using four cultivars ranging from highly susceptible to moderately susceptible/moderately resistant. Trials conducted in controlled conditions showed that there was no obvious relationship between soil temperature (5, 9, 14 and 19 °C) and the incidence of disease on seedlings.  相似文献   

13.
Routine laboratory testing of 710 bean seed lots from various origins forPseudomonas syringae pv. phaseolicola (Psp) with immunofluorescence microscopy (IF) showed that 27.5% of the seed lots (five subsamples of 1000 seeds tested per sample) contained two or more IF-positive cells in a total of 500 microscope fields (magnification 500×). Simultaneously performed dilution-platings of IF-positive subsamples on King's medium B confirmed presence of Psp for one-third of these IF-positive seed lots. The grey area of disagreement between both laboratory tests was studied by comparison of test data and by field trials.The number of IF-positive cells per subsample was positively correlated with isolation and identification of Psp (R=0.85). The detection level of IF was ca. 102 Psp cells per ml of undiluted subsample extract. The detection level of Psp by isolation on King's medium B was variable, being inversely related with the saprophyte to Psp ratio. The high sensitivity of IF was in part due to high percentages of dormant or dead IF-positive cells in the sample extract. Field trials over two years with 10 000 seeds per seed lot, showed disease incidence for 9 of the 22 seed lots. Of ten IF-positive lots with five positive subsamples per sample, nine were positive in the field test plot (the negative lot gave primary infection spots of Psp when used for commercial growing). By isolation, seven of these ten IF-positive lots were positive. Of the five IF-positive lots with two or less positive subsamples, isolation and field trial were both negative. Based on data on seed transmission from literature, field incidence was unlikely for these five samples in a 10 000 seeds field trial. All seven IF-negative lots were negative in the field trial. The value of IF and isolation for indexing bean seed lots for Psp is discussed.This study was carried out at the Centre for Plant Breeding and Reproduction Research (CPRO-DLO), Binnenhaven 1, P.O. Box 16, 6700 AA Wageningen the Netherlands, to which address correspondence should be addressed.  相似文献   

14.
芝麻枯萎病病原菌致病力室内鉴定方法   总被引：3，自引：0，他引：3  
 芝麻枯萎病(Sesame Fusarium wilt, SFW)是由尖孢镰刀菌芝麻专化型(Fusarium oxysporum Schl. f. sp. sesami (Zap.), FOS)引起的一种土传真菌病害, 是世界芝麻生产上的主要病害之一。为测定FOS致病力, 本文选用郑芝98N09等4个芝麻品种, 在苗期对15个FOS菌株的致病力强弱进行了室内鉴定和评价。结果表明, 采用1×106个/mL分生孢子悬浮液与无菌蛭石和无菌土壤按V1∶V2∶V6比例混合接菌(即最终接菌浓度为1.4×105孢子/g土壤), 在接菌后第7 d幼苗开始出现枯萎病症状, 调查菌株致病性的最佳时间为接菌后第25 d~28 d;在供试15个FOS菌株中, 对4个品种均表现为强致病力的菌株有8个(DI>50), 均表现为弱致病力的菌株有5个(DI<20);不同芝麻品种对不同菌株的抗性有一定差异。该方法可应用于芝麻枯萎病病原菌致病性测定和芝麻种质抗枯萎病特性评价, 并为后续的机理研究提供了技术支持。  相似文献   

15.
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.  相似文献   

16.
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.  相似文献   

17.
Verticillium dahliae Klebahn is the causal agent of tomato wilt disease. Isolates of V. dahliae can be classified based on pathogenicity to tomato, but the pathotypes are indistinguishable in morphology. We designed PCR primers for specific detection of isolates pathogenic to tomato (tomato pathotype) from the sequences of a pathotype-specific gene, vdt1. With the primer pair Tg5/Tc3, a PCR product (approximately 3.2 kb) specific to tomato pathotype was amplified from the genomic DNA of isolates. Using the primer pair, a tomato pathotype isolate was specifically detected from hypocotyls of inoculated tomato and eggplant. On the other hand, no amplification was observed from non-tomato pathotype isolates of V. dahliae, some other wilt pathogens of tomato and a healthy host plant. Therefore, the primer pair can be useful for pathotype-specific detection of V. dahliae as well as for diagnosis of wilt disease of tomato plant. Received 7 September 2001/ Accepted in revised form 3 December 2001  相似文献   

18.
Bacterial wilt is a serious disease of Iucerne. Although there are several ways to transmit the pathogen, seed transmission is practically the only way to introduce the bacteria to previously free areas. In this work, naturally infected seed material and commercial seed lots have been screened by a combination of immunofluorescence (IF) staining and dilution plating. Isolates were confirmed by biochemical characterization and finally by artificial infection of lucerne seedlings. For seed lots free from the pathogen, a diagnosis can mostly be achieved within 2 days. The culture step is necessary only if IF staining is not unequivocally negative, and the full procedure has to be run only for a small portion of the samples. The proposed seed assay provides a reliable alternative to the visual inspection of lucerne fields to fulfil the phytosanitary requirements for Clavibacter michiganensis ssp. insidiosus.  相似文献   

19.
A method for detection and quantitative estimation of tomato seedborne pathogenic bacteria has been developed. It enables detection in a 7 g tomato seed sample of as few as ten colony-forming units per gram tomato seeds of the following seedborne pathogens of tomato:Pseudomonas syringae pv. tomato,Pseudomonas corrugata, Xanthomonas campestris pv.vesicatoria, andClavibacter michiganense subsp.michiganense. With representative seed samples, the method employs dry grinding, weighing, bacterial extraction and quantitative calculation on selective or semi-selective medium. The efficiency of this method was tested by diluting pathogen-free seed lots with naturally or artificially infested tomato seeds. This procedure enables one to determine the minimal threshold of pathogen which can be detected by this method on media, in comparison with the percentage of diseased seedlings developed from the same seed lots in the growth chamber or in the greenhouse.  相似文献   

20.
Fifty-nine Ralstonia solanacearum isolates from diverse crops and regions were collected and characterized to determine the distribution and diversity of this soilborne pathogen in Guatemala. Three distinct types were present: a phylotype I, sequevar 14 strain, probably originating from Asia, infecting tomatoes and aubergines at moderate elevations; a phylotype II, sequevar 6 strain of American origin causing Moko disease in lowland banana plantations; and a phylotype II, sequevar 1 (race 3 biovar 2) strain causing brown rot on potatoes, Southern wilt of Pelargonium spp. and bacterial wilt of greenhouse tomatoes at high elevations. These data on strain diversity will inform effective regional efforts to breed for wilt resistance. A sensitive enrichment method did not detect the pathogen in fruits from naturally infected commercial tomato plants in Guatemalan fields and greenhouses, although it was detected in 6% of fruits from a wilt-resistant hybrid. Low numbers of R. solanacearum cells were also infrequently detected in fruits from plants artificially inoculated in the growth chamber with either race 3 biovar 2 or a phylotype II tomato strain.  相似文献   

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