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1.
Bacterial canker caused by Clavibacter michiganensis subsp. michiganensis is known to cause significant economic losses to tomato production worldwide. Biological control has been proposed as an alternative to current chemical containment methods, which are often inefficient and may leave adverse effects on the environment. However, only little headway has so far been made in developing biocontrol strategies against C. michiganensis subsp. michiganensis. To address this knowledge gap, we investigated the antagonistic capacity of PCA, produced by Pseudomonas sp. LBUM223, and DAPG and HCN, both produced by Pseudomonas sp. LBUM300, on C. michiganensis subsp. michiganensis under in vitro and in planta conditions. Nonsynthesizing isogenic mutants of the producer strains were also developed to further dissect the role of each individual metabolite on C. michiganensis subsp. michiganensis biological control. Novel specific quantitative polymerase chain reaction TaqMan assays allowed quantification of C. michiganensis subsp. michiganensis in tomato plants and rhizospheric soil. Pseudomonas spp. LBUM223 and LBUM300 significantly repressed C. michiganensis subsp. michiganensis growth in vitro, while their respective nonproducing mutants showed less or no significant antagonistic activity. In planta, only Pseudomonas sp. LBUM300 was capable of significantly reducing disease development and C. michiganensis subsp. michiganensis rhizospheric population, suggesting that the production of both DAPG and HCN was involved. In summary, simultaneous DAPG/HCN production by Pseudomonas sp. LBUM300 shows great potential for controlling bacterial canker of tomato. 相似文献
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3.
Clavibacter michiganensis subsp. michiganensis is an actinomycete, causing bacterial wilt and canker disease of tomato (Solanum lycopersicum). We used virus-induced gene silencing (VIGS) to identify genes playing a role in host basal defense response to C. michiganensis subsp. michiganensis infection using Nicotiana benthamiana as a model plant. A preliminary VIGS screen comprising 160 genes from tomato known to be involved in defense-related signaling identified a set of 14 genes whose suppression led to altered host-pathogen interactions. Expression of each of these genes and three additional targets was then suppressed in larger-scale VIGS experiments and the effect of silencing on development of wilt disease symptoms and bacterial growth during an N. benthamiana-C. michiganensis subsp. michiganensis compatible interaction was determined. Disease susceptibility and in planta bacterial population size were enhanced by silencing genes encoding N. benthamiana homologs of ubiquitin activating enzyme, snakin-2, extensin-like protein, divinyl ether synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase 2, and Pto-like kinase. The identification of genes having a role in the host basal defense-response to C. michiganensis subsp. michiganensis advances our understanding of the plant responses activated by C. michiganensis subsp. michiganensis and raises possibilities for devising novel and effective molecular strategies to control bacterial canker and wilt in tomato. 相似文献
4.
A new selective and highly sensitive medium was developed for isolation of Clavibacter michiganensis subsp. michiganensis (Cmm), the causal agent of bacterial canker of tomato, from seed and latently infected plants. The new medium (BCT) proved to be superior to all published semiselective media for Cmm and is denoted as selective medium because of (i) its mean plating efficiency, amounting to ≤89% within 7 days for all 30 Cmm strains from different sources tested; (ii) the high selectivity, because accompanying bacterial species occurring on tomato plants and seed or bacteria obtained from culture collections were inhibited to an extent of 98 to 100%; and (iii) the remarkable detection sensitivity. Thus, 8 CFU of Cmm in field plant homogenates containing 12,750 CFU of accompanying saprophytes were detected on BCT. Under these extreme conditions, all of the published semiselective media (D2, KBT, D2ANX, SCM, mSCM, CMM1, mCNS, and EPPO) gave false-negative results. Either some media were rather toxic and Cmm growth was also inhibited or the other, less toxic media allowed growth of high numbers of saprophytes, so that Cmm growth was suppressed. Exclusively, BCT also supported growth of the closely related C. michiganensis subsp. insidiosus, nebraskensis, and tessellarius. The new medium is recommended for Cmm detection in tomato seed, and in symptomless tomato plantlets, to improve disease control of bacterial canker of tomato. 相似文献
5.
The effect of pH on the survival of Clavibacter michiganensis ssp. michiganensis and its transmission via roots of tomato in hydroponic culture was studied in laboratory and greenhouse experiments. In a laboratory experiment, C . m. ssp. michiganensis could not survive for 24 h in nutrient solutions with a pH of 4·0 or 4·5, while 1, 14, 51 and 62% of inoculum survived at pH 5·0, 5·5, 6·0 and 6·5, respectively. When tomato plants were inoculated with C . m. ssp. michiganensis through wounds on the stems, the bacteria moved downward from the inoculation site to the roots and infectious bacteria were released from the roots into the nutrient solution. Of two pH regimes tested in greenhouse nutrient-film technique (NFT) culture, the C . m. ssp. michiganensis population was significantly lower in pH 5·0 than in pH 6·5 in most sampling data. In treatments in which C . m. ssp. michiganensis was introduced by transplanting two root-inoculated plants, significantly more plants developed canker at pH 6·5 (34 out of 48 plants) than at pH 5·0 (11 out of 48 plants). When the bacterium was introduced by transplanting two stem-inoculated plants at pH 6·5, seven out of 24 plants developed canker. The potential of pH manipulation in controlling tomato bacterial canker in hydroponic culture is discussed. 相似文献
6.
Adaptation of soil solarization to the integrated management of soilborne pests of tomato under humid conditions 总被引：1，自引：0，他引：1
ABSTRACT Soil solarization was shown to be cost effective, compatible with other pest management tactics, readily integrated into standard production systems, and a valid alternative to preplant fumigation with methyl bromide under the tested conditions. Solarization using clear, photoselective, or gas-impermeable plastic was evaluated in combination with metham sodium, 1,3-dichloropropene + chloropicrin, methyl bromide + chloropicrin, pebulate, or cabbage residue. Strip solarization, applied to 20-cm-high, 0.9-m-wide beds, was conducted to achieve compatibility with standard production practices and resulted in soil temperatures 2 to 4 degrees C above those temperatures resulting when using conventional flatbed solarization. Soil temperatures were 1 to 2 degrees C higher at the edges of the raised beds, eliminating any border effects associated with solarization. Following a 40- to 55-day solarization period, the plastic was painted white and used as a production mulch for a subsequent tomato crop. The incidence of Southern blight and the density of Paratrichodorus minor and Criconemella spp. were lower (P < 0.05) in solarized plots. No differences (P < 0.05) in the incidence of Fusarium wilt and the density of nutsedge and Helicotylenchus spp. were observed between plots receiving solarization and plots fumigated with a mixture of methyl bromide + chloropicrin. The severity of root galling was lower (P < 0.05) when soil solarization was combined with 1,3-dichloropropene + chloropicrin (16.2 + 3.4 g/m(2)) and a gas-impermeable film. The incidence of bacterial wilt was not affected by soil treatments. Marketable yields in plots using various combinations of soil solarization and other tactics were similar (P < 0.05) to yields obtained in plots fumigated with methyl bromide + chloropicrin. The results were validated in several large scale field experiments conducted by commercial growers. 相似文献
7.
rep-PCR-Mediated Genomic Fingerprinting: A Rapid and Effective Method to Identify Clavibacter michiganensis 总被引：1，自引：0，他引：1
Louws FJ Bell J Medina-Mora CM Smart CD Opgenorth D Ishimaru CA Hausbeck MK de Bruijn FJ Fulbright DW 《Phytopathology》1998,88(8):862-868
ABSTRACT The genomic DNA fingerprinting technique known as repetitive-sequence-based polymerase chain reaction (rep-PCR) was evaluated as a tool to differentiate subspecies of Clavibacter michiganensis, with special emphasis on C. michiganensis subsp. michiganensis, the pathogen responsible for bacterial canker of tomato. DNA primers (REP, ERIC, and BOX), corresponding to conserved repetitive element motifs in the genomes of diverse bacterial species, were used to generate genomic fingerprints of C. michiganensis subsp. michiganensis, C. michiganensis subsp. sepedonicus, C. michiganensis subsp. nebraskensis, C. michiganensis subsp. tessellarius, and C. michiganensis subsp. insidiosum. The rep-PCR-generated patterns of DNA fragments observed after agarose gel electrophoresis support the current division of C. michiganensis into five subspecies. In addition, the rep-PCR fingerprints identified at least four types (A, B, C, and D) within C. michiganensis subsp. michiganensis based on limited DNA polymorphisms; the ability to differentiate individual strains may be of potential use in studies on the epidemiology and host-pathogen interactions of this organism. In addition, we have recovered from diseased tomato plants a relatively large number of naturally occurring avirulent C. michiganensis subsp. michiganensis strains with rep-PCR fingerprints identical to those of virulent C. michiganensis subsp. michiganensis strains. 相似文献
8.
C. J. LOPEZ-HERRERA R. M. PEREZ-JIMENEZ M. J. BASALLOTE-UREBA T. ZEA BONILLA & J. M. MELERO-VARA 《Plant pathology》1997,46(3):329-340
Phytophthora root rot is of paramount importance in avocado orchards of southern Spain. Soil solarization has been demonstrated to control the pathogen in infested areas from which infected trees had been removed. We aimed to determine whether soil solarization in established avocado orchards controls the disease. Soil solarization increased average maximum hourly soil temperatures by 6.5–6.9°C in unshaded areas of avocado orchards in coastal areas of southern Spain, depending on depth and year. The corresponding temperatures in shaded areas were c. 2–3°C lower. P. cinnamomi in soil, on infected avocado rootlets, and in a nutrient substrate buried at 30–60 cm depth was reduced to negligible amounts after 6–8 weeks of solarization in both unshaded and shaded locations of avocado orchards. P. cinnamomi could not be detected in avocado rootlets up to 14 months later, suggesting a long-term effect. Soil solarization did not affect growth of the trees, and fruit yields were increased as compared with control plots. Following soil solarization for 3 weeks from mid-July 1994, when maximum hourly temperatures reached 33–36°C, P. cinnamomi could not be recovered from a depth of up to 45 cm in unshaded areas or from a depth of up to 30 cm in shaded areas after the initial 10-day period. The viability of inoculum of the pathogen buried at depths between 15 and 60 cm in bare soil was determined by sequential sampling in two solarization experiments starting 12 June and 4 July 1995, respectively. In the first experiment, P. cinnamomi could not be detected at any depth after 4–8 weeks of solarization in unshaded areas but could be recovered at all depths except 15 cm in shaded areas. In the second experiment, where temperatures were higher and the soil surface not shaded, P. cinnamomi could not be recovered after 2 weeks at 15 and 30 cm. 相似文献
9.
Journal of Plant Diseases and Protection - Clavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial canker in tomatoes representing a significant threat to worldwide tomato production.... 相似文献
10.
Tomato bacterial canker, caused by Clavibacter michiganensis subsp. michiganensis, is transmitted by infected or infested seed and mechanically from plant to plant. Wounds occurring during seedling production and crop maintenance facilitate the dissemination of the pathogen. However, the effects of environmental factors on C. michiganensis subsp. michiganensis translocation and growth as an endophyte have not been fully elucidated. A virulent, stable, constitutively bioluminescent C. michiganensis subsp. michiganensis strain BL-Cmm 17 coupled with an in vivo imaging system allowed visualization of the C. michiganensis subsp. michiganensis colonization process in tomato seedlings in real time. The dynamics of bacterial infection in seedlings through wounds were compared under low (45%) and high (83%) relative humidity. Bacteria multiplied rapidly in cotyledon petioles remaining after clip inoculation and moved in the stem toward both root and shoot. Luminescent signals were also observed in tomato seedling roots over time, and root development was reduced in inoculated plants maintained under both humidity regimes. Wilting was more severe in seedlings under high-humidity regimes. A strong positive correlation between light intensity and bacterial population in planta suggests that bioluminescent C. michiganensis subsp. michiganensis strains will be useful in evaluating the efficacy of bactericides and host resistance. 相似文献
11.
ABSTRACT Hydathodes of tomato leaves served as extremely efficient infection courts for the bacterial canker pathogen, Clavibacter michiganensis subsp. michiganensis. Chlorotic lesions developed at the tips of leaflet lobes about 2 weeks after inoculation of guttation droplets. Lesions expanded along the leaflet margins and became necrotic. Movement of C. michiganensis subsp. michiganensis from the inoculated leaflet into the rachis was slow and erratic. Histological observations revealed that pathogen populations first developed within large intercellular spaces lying beneath the stomata, which serve as water pores in tomato hydathodes. Bacteria were first observed within vessels of the large marginal fimbriate veins 7 days after inoculation. By 14 days after inoculation, large populations could be seen within the vessels; and by 21 days after inoculation, tissue collapse was widespread and masses of bacteria could be seen in the intercellular spaces and within necrotic cells. 相似文献
12.
Clavibacter michiganensis subsp. michiganensis (CMM) strains collected between 2005-2008 from greenhouses in different locations in Okayama Prefecture, Japan, were fingerprinted by repetitive sequence-based polymerase chain reaction (rep-PCR) with ERIC and BOX primers. One hundred and eighty strains from eight different locations in Okayama were differentiated into four haplotypes (A to D) based on rep-PCR. Regardless of the year of isolation, location or cultivar of tomato, the strains in each greenhouse and location belonged to the same haplotype, suggesting the strains originated from the previous greenhouse population. Based on Morisita's index of dispersion ( I δ ), the distribution of diseased plants in the greenhouses, where disbudding and defoliation using either scissors or by hand were carried out in the same direction to promote the spread of CMM, occurred in an aggregated distribution in a quadrant along a row of plants, but the distribution of diseased plants indicated a random distribution in a quadrant along a furrow of plants (two adjoining rows of plants). These results showed that disbudding and defoliation contribute highly to the secondary spread of bacterial canker in commercial greenhouses. 相似文献
13.
Chalupowicz L Zellermann EM Fluegel M Dror O Eichenlaub R Gartemann KH Savidor A Sessa G Iraki N Barash I Manulis-Sasson S 《Phytopathology》2012,102(1):23-31
The vascular pathogen Clavibacter michiganensis subsp. michiganensis is responsible for bacterial wilt and canker of tomato. Pathogenicity of this bacterium is dependent on plasmid-borne virulence factors and serine proteases located on the chromosomal chp/tomA pathogenicity island (PAI). In this study, colonization patterns and movement of C. michiganensis subsp. michiganensis during tomato infection was examined using a green fluorescent protein (GFP)-labeled strain. A plasmid expressing GFP in C. michiganensis subsp. michiganensis was constructed and found to be stable in planta for at least 1 month. Confocal laser-scanning microscopy (CLSM) of inoculated stems showed that the pathogen extensively colonizes the lumen of xylem vessels and preferentially attaches to spiral secondary wall thickening of the protoxylem. Acropetal movement of the wild-type strain C. michiganensis subsp. michiganensis NCPPB382 (Cmm382) in tomato resulted in an extensive systemic colonization of the whole plant reaching the apical region after 15 days, whereas Cmm100 (lacking the plasmids pCM1 and pCM2) or Cmm27 (lacking the chp/tomA PAI) remained confined to the area surrounding of the inoculation site. Cmm382 formed biofilm-like structures composed of large bacterial aggregates on the interior of xylem walls as observed by CLSM and scanning electron microscopy. These findings suggest that virulence factors located on the chp/tomA PAI or the plasmids are required for effective movement of the pathogen in tomato and for the formation of cellular aggregates. 相似文献
14.
Using tomato seedlings, the plant defence activator acibenzolar-S-methyl (benzo-[1,2,3]-thiadiazole-7-carbothioic acid-S-methyl ester, ASM; Bion 50 WG) was assayed for its ability to induce resistance against Clavibacter michiganensis ssp. michiganensis , the causal agent of bacterial canker of tomato. In ASM-pretreated plants, reduction in disease severity (up to 76·3%) was correlated with lower bacterial growth (up to 68·2% lower) during the time course of infection. To understand the possible mechanism of action of ASM, alterations in the activities of peroxidase (POX) and chitinase were assessed as markers of resistance. The enhanced resistance of ASM-treated plants was associated with significant increases in the activities of POX and chitinase 相似文献
15.
Viable cells of Clavibacter michiganensis subsp. michiganensis (CMM), the causal agent of bacterial canker of tomato, were discriminated from the dead cells by quantitative real-time polymerase chain reaction (PCR), after the bacterial solution was treated with the DNA binding dye ethidium monoazide (EMA). The primers and TaqMan probe, based on the 16S-23S rDNA spacer sequences, were highly specific for CMM at the subspecies level. The detection limit of the direct real-time PCR was 103 colony forming units per mL (cfu mL−1 ) in samples and with an apparent sensitivity of 2 cfu of target cells in PCR reaction solution. Application of this method allows for selective quantification of viable cells of CMM and facilitates monitoring the pathogen in tomato seeds. 相似文献
16.
The survival and half-life of Clavibacter michiganensis ssp. michiganensis ( C. michiganensis ), the causal agent of bacterial canker of tomato, were determined in infected plant debris under natural field conditions in California, Ohio and Morocco using a semiselective agar medium. The organism survived significantly longer in tomato stems left on the soil surface than in stems buried in the soil at all locations studied. The pathogen was recovered in high amounts from tomato stems left on the soil surface for 314 days in Ohio and California, USA, and for 194 and 132 days in Melk Zhar and Aït Melloul, Morocco, respectively; it was recovered from stems buried in the soil for up to 314 days in Ohio, up to 240 days in California, and up to 60 days in Aït Melloul and Melk Zhar. The half-life of the pathogen in stems left on the soil surface ranged from 23·2 to 24·8 days in the USA, and from 7·8 to 12·3 days in Morocco, whereas the half-life in buried stems ranged from 14·0 to 16·7 days in the USA and from 3·7 to 9·5 days in Morocco. Based on the half-life data, the predicted survival times of C. michiganensis in stems on the soil surface in Ohio, California, Melk Zhar and Aït Melloul would be up to 822, 770, 424 and 261 days, respectively, while the predicted survival times in stems buried in the soil would be 541, 497, 305 and 128 days, respectively. These results show that the survival and half-life of C. michiganensis in plant debris are relatively long and are influenced by both tissue exposure and geographic location. 相似文献
17.
N. Ioannou 《Phytoparasitica》2000,28(3):248-256
Preplant soil fumigation with methyl bromide (MB) is presently standard practice in greenhouse tomato production. Since this
compound is scheduled to be phased out by 2005, the possibility of using solarization as an alternative soil disinfestation
method was examined in four greenhouse tomato trials. Solarization was applied for 8 weeks in July-August, using transparent
polyethylene sheets for soil mulching, and compared with MB fumigation applied in September, before planting, at 80 g/m2. Solarization raised the maximum soil temperature by 9°C and reduced the population density ofFusarium spp. in soil by 91–98%. Similar reductions of soil inoculum (95–99%) were obtained with MB fumigation. Both methods provided
effective control of Fusarium wilt, Verticillium wilt and corky root rot on tomato plants. MB fumigation was in addition highly
effective against root-knot nematodes, whereas nematode control with solarization did not exceed 50%. Both treatments resulted
in similar fruit yield increases, ranging within 90–140% compared with plants grown in untreated soil. During the second cropping
season following soil treatment, solarization exhibited two times higher residual effectiveness against vascular wilt diseases
compared with MB fumigation. The latter treatment, however, was superior to solarization in its residual effectiveness against
root-knot nematodes and to a lesser extent against corky root rot. Fruit yields from solarized and MB-fumigated soil during
the second cropping season were higher than those obtained from untreated soil by approximately 35% and 60%, respectively.
In Cyprus, solarization appears to be an effective alternative to MB fumigation in greenhouse tomato production, especially
if integrated with other approaches enabling more effective nematode control. 相似文献
18.
B. E. ABU-IRMAILEH 《Weed Research》1991,31(3):125-133
Black (BPE) and clear polyethylene mulches (CPE), 0.08 and 0.06 mm thick, respectively, were compared for their effectiveness for soil solarization over three seasons during 1986–1989 in weedy fields. Solarization for 6 weeks reduced weed growth and enhanced crop yields. However, further mulching with BPE after solarization with either BPE or CPE gave the best results. Not all weeds were sensitive to solarization. Some weed species were completely controlled; others were reduced to varying degrees; yet other weed species seemed to be enhanced by solarization. Solarization without further mulching was no better than farmer-practice in reducing weed growth or in increasing crop yield. Weeds required further removal after the middle of the growing season. Any soil disturbance after solarization reduced the weed control effect of solarization. Crops grew best in plots after solarization with BPE if they were planted in the same mulch after it was perforated. 相似文献
19.
Solarization trials were carried out over 3 years and in two countries to control crown gall disease on fruit trees and eliminate Agrobacterium . In 1992, agrobacteria in naturally infested soils of two Italian nurseries were monitored before and after solarization. Agrobacteria populations decreased by 99% and 92% after the treatment; however, crown gall incidence did not decrease. In 1993 and 1994 solarization was tested in Oregon in fields artificially infested with two marked strains of A. tumefaciens . In sandy loam soil, the target bacteria were eliminated in 4 weeks, while in silty clay soil the populations were markedly reduced after 2 months of treatment. Crown gall incidence on cherry rootstocks transplanted to the field at the end of 1993 was 3.7% in the sandy loam soil control plots, while no tumours were observed on plants from solarized plots. The use of solarization in combination with reduced doses of metham-sodium was also evaluated. 相似文献
20.
Verticillium dahliae, the causal agent of Verticillium wilt of tomato, causes serious damage to crops grown in unheated greenhouses. To control this disease, growers are obliged to employ strong soil disinfestants. The possibility of controllingV. dahliae by using soil solarization during the months of June — August was examined. The soil was covered with transparent polyethylene sheets for 10 weeks. The pathogen could not be isolated from the solarized soil, whereas the inoculum level in the nonsolarized soil remained high (1379–1806 propagules/g soil). The yield from the solarized soil was increased by 112.4% in comparison with the control, and no infected plants were observed. The percentage of infected roots was very low (0.3–0.4%) in relation to the nonsolarized soil (66.7–67.1%). From these results it was concluded that solarization can effectively control Verticillium wilt of greenhouse-grown tomato under the summer conditions in Crete. 相似文献