Showing papers in "Phytopathology in 1998"
TL;DR: There was a general trend across all experiments toward greater suppression and enhanced consistency against multiple cucumber pathogens using strain mixtures, and PGPR-mediated disease suppression was observed againstangular leaf spot in 1996 and against a mixed infection of angular leaf spot and anthracnose in 1997.
Abstract: Plant growth-promoting rhizobacteria (PGPR) strains INR7 (Bacillus pumilus), GB03 (Bacillus subtilis), and ME1 (Curtobacterium flaccumfaciens) were tested singly and in combinations for biological control against multiple cucumber pathogens. Investigations under greenhouse conditions were conducted with three cucumber pathogens-Colletotrichum orbiculare (causing anthracnose), Pseudomonas syringae pv. lachrymans (causing angular leaf spot), and Erwinia tracheiphila(causing cucurbit wilt disease)-inoculated singly and in all possible combinations. There was a general trend across all experiments toward greater suppression and enhanced consistency against multiple cucumber pathogens using strain mixtures. The same three PGPR strains were evaluated as seed treatments in two field trials over two seasons, and two strains, IN26 (Burkholderia gladioli) and INR7 also were tested as foliar sprays in one of the trials. In the field trials, the efficacy of induced systemic resistance activity was determined against introduced cucumber pathogens naturally spread within plots through placement of infected plants into the field to provide the pathogen inoculum. PGPR-mediated disease suppression was observed against angular leaf spot in 1996 and against a mixed infection of angular leaf spot and anthracnose in 1997. The three-way mixture of PGPR strains (INR7 plus ME1 plus GB03) as a seed treatment showed intensive plant growth promotion and disease reduction to a level statistically equivalent to the synthetic elicitor Actigard applied as a spray.
639 citations
TL;DR: Conclusive evidence is presented that silicon is involved in the increased resistance of cucumber to powdery mildew by enhancing the antifungal activity of infected leaves by activating low-molecular-weight metabolites.
Abstract: The controversial role of silicon in plant disease resistance, described mostly as a passive mechanical protection, has been addressed. Conclusive evidence is presented that silicon is involved in the increased resistance of cucumber to powdery mildew by enhancing the antifungal activity of infected leaves. This antifungal activity was attributable to the presence of low-molecular-weight metabolites. One of these metabolites, described here as a phytoalexin, was identified as a flavonol aglycone rhamnetin (3,5,3',4'-tetrahydroxy-7-O-methoxyflavone). This is the first report of a phytoalexin for this chemical group in the plant kingdom and of a flavonol phytoalexin in cucumber, a chemical defense long believed to be nonexistent in the family Cucurbitaceae. The antifungal activity of leaf extracts was better expressed after acid hydrolysis, extending to another plant species the concept that some phytoalexins are synthesized as glycosylated phytoalexins or their precursors.
398 citations
TL;DR: Introduction of pchBA into CHA0 increased the production of salicylic acid in vitro and in the rhizosphere of tobacco, but did not improve the ability ofCHA0 to induce systemic resistance in tobacco.
Abstract: Application of salicylic acid induces systemic acquired resistance in tobacco. pchA and pchB, which encode for the biosynthesis of salicylic acid in Pseudomonas aeruginosa, were cloned into two expression vectors, and these constructs were introduced into two root-colonizing strains of P. fluorescens. Introduction of pchBA into strain P3, which does not produce salicylic acid, rendered this strain capable of salicylic acid production in vitro and significantly improved its ability to induce systemic resistance in tobacco against tobacco necrosis virus. Strain CHA0 is a well-described biocontrol agent that naturally produces salicylic acid under conditions of iron limitation. Introduction of pchBA into CHA0 increased the production of salicylic acid in vitro and in the rhizosphere of tobacco, but did not improve the ability of CHA0 to induce systemic resistance in tobacco. In addition, these genes did not improve significantly the capacity of strains P3 and CHA0 to suppress black root rot of tobac...
345 citations
TL;DR: Results from a biocontrol agent-fortified compost mix induced systemic acquired resistance (SAR) in cucumber against anthracnose caused by Colletotrichum orbiculare and in Arabidopsis against bacterial speck caused by Pseudomonas syringae pv.
Abstract: A biocontrol agent-fortified compost mix, suppressive to several diseases caused by soilborne plant pathogens, induced systemic acquired resistance (SAR) in cucumber against anthracnose caused by Colletotrichum orbiculare and in Arabidopsis against bacterial speck caused by Pseudomonas syringae pv maculicola KD4326 A peat mix conducive to soilborne diseases did not induce SAR The population size of P syringae pv maculicola KD4326 was significantly lower in leaves of Arabidopsis plants grown in the compost mix compared to those grown in the peat mix Autoclaving destroyed the SAR-inducing effect of the compost mix, and inoculation of the autoclaved mix with nonautoclaved compost mix or Pantoea agglomerans 278A restored the effect, suggesting the SAR-inducing activity of the compost mix was biological in nature Topical sprays with water extract prepared from the compost mix reduced symptoms of bacterial speck and the population size of pathogenic KD4326 in Arabidopsis grown in the peat mix bu
289 citations
TL;DR: Findings clearly demonstrate that fungi are the dominant causal agents of apple replant disease in Washington state.
Abstract: Systematic studies were conducted to elucidate the role of different soil microbial groups in the development of apple replant disease. Populations of targeted microorganisms were reduced by the application of semiselective biocides and soil pasteurization. Bacteria were not implicated in the disease, because application of the antibiotic chloramphenicol reduced soil populations of bacteria but failed to improve growth of apple transplants, while enhanced growth was achieved at pasteurization temperatures that did not alter attributes of the bacterial community recovered from apple roots. Populations of Pratylenchus penetrans were below the damage threshold level in eight of nine orchards surveyed, and nematicide applications failed to enhance apple growth in four of five replant soils tested, indicating that plant parasitic nematodes have a minor role or no role in disease development. Application of the fungicide difenconazole or metalaxyl enhanced growth of apple in all five replant soils, as ...
270 citations
TL;DR: Ability to infect both potato and tomato apparently did not increase the fitness of this genotype relative to US-8, as predicted previously, and it generally was not possible to predict which genotypes would be present in a location from 1 year to the next.
Abstract: Dramatic changes occurred within populations of Phytophthora infestans in the United States and Canada from 1994 through 1996. Occurrence of the US-8 genotype, detected rarely during 1992 and 1993, increased rapidly and predominated in most regions during 1994 through 1996. US-7, which infected both potato and tomato and made up almost 50% of the sample during 1993, was detected only rarely among 330 isolates from the United States analyzed during 1994. It was not detected at all in more limited samples from 1996. Thus, ability to infect both potato and tomato apparently did not increase the fitness of this genotype relative to US-8, as predicted previously. US-1, the previously dominant genotype throughout the United States and Canada, made up 8% or less of the samples analyzed during 1994 through 1996. A few additional genotypes were detected, which could indicate the beginnings of sexual reproduction of P. infestans within the United States and Canada. However, clonal reproduction still predominated in all locations sampled; opportunities for sexual reproduction probably were limited, because the A1 and A2 mating types usually were separated geographically. The high sensitivity of the US-1 genotype to the fungicide metalaxyl also could have reduced opportunities for contact between the mating types in fields where this compound was applied. The previous correlation between metalaxyl sensitivity and genotype was confirmed and extended to a new genotype, US-17: all US-1 isolates tested were sensitive; all isolates of the US-7, US-8, and US-17 genotypes tested to date have been resistant. Isolates of P. capsici and P. erythroseptica, two other species often found on tomato and potato, could be easily distinguished from each other and from P. infestans using a simple allozyme assay for the enzyme glucose-6-phosphate isomerase. This technique could be useful for rapid identification of species, in addition to genotype of P. infestans. It generally was not possible to predict which genotypes would be present in a location from 1 year to the next. Long-distance movement of US-8 in seed tubers was documented, and this was probably the primary means for the rapid spread of this genotype from 1993 through 1996.
218 citations
TL;DR: Genomic diversity of phytoplasma groups appears to be correlated with their sharing common insect vectors, host plants, or both in nature.
Abstract: The recent development of molecular-based probes such as mono- and polyclonal antibodies, cloned phytoplasma DNA fragments, and phytoplasma-specific primers for polymerase chain reaction (PCR) has allowed for advances in detection and identification of uncultured phytoplasmas (formerly called mycoplasma-like organisms). Comprehensive phylogenetic studies based on analysis of 16S ribosomal RNA (rRNA) or both 16S rRNA and ribosomal protein gene operon sequences established the phylogenetic position of phytoplasmas as members of the class Mollicutes, and the revealed phylogenetic interrelationships among phytoplasmas formed a basis for their classification. Based on restriction fragment length polymorphism (RFLP) analysis of PCR-amplified 16S rRNA gene sequences, phytoplasmas are currently classified into 14 groups and 38 subgroups that are consistent with groups delineated based on phylogenetic analysis using parsimony of 16S rRNA gene sequences. In the past decades, numerous phyto-plasma strains a...
205 citations
TL;DR: Exoglc1 showed in vitro a stronger inhibitory effect on germ tube growth of B. cinerea than on conidia germination and caused morphological changes such as leakage of cytoplasm and cell swelling, strengthening the hypothesis that exo-beta-1,3-glucanase activity is one of the mechanisms of action involved in the suppression of B-cinerea.
Abstract: The exo-beta-1,3-glucanase (EC 3.2.1.58) activity of Pichia anomala strain K, an antagonistic yeast of Botrytis cinerea on postharvest apples, was studied in a synthetic medium supplemented with laminarin, a cell wall preparation (CWP) of B. cinerea, or glucose. The highest enzyme activity was detected in culture media containing a CWP of B. cinerea as the sole carbon source, whereas the lowest activity was observed in culture media supplemented with glucose. Exoglc1, an exo-beta-1,3-glucanase, was purified to homogeneity from culture filtrates of strain K containing a CWP. The molecular mass of exoglc1 was estimated to be under 15 kDa. Optimum activity of exoglc1 was recorded at 50 degrees C and pH 5.5. The exoglc1 K(m) value was estimated at 22.4 mg/ml. Exoglc1 showed in vitro a stronger inhibitory effect on germ tube growth of B. cinerea than on conidia germination and caused morphological changes such as leakage of cytoplasm and cell swelling. Exo-beta-1,3-glucanase activity was detected on apples treated with strain K and was similar to exoglc1 on the basis of activity on native gel. Moreover, the addition of a CWP to a suspension of P. anomala stimulated both in situ exo-beta-1,3-glucanase activity and protective activity against the pathogen, strengthening the hypothesis that exo-beta-1,3-glucanase activity is one of the mechanisms of action involved in the suppression of B. cinerea by P. anomala strain K.
193 citations
TL;DR: Computer-assisted sequence analysis showed significant homology between ToCV clones that hybridize specifically with RNAs 1 and 2 and the lettuce infectious yellows virus methyltransferase of RNA 1 and the HSP70 heat shock protein homolog of RNA 2, respectively, indicating that ToCV is another member of the growing subgroup of bipartite closteroviruses transmitted by whiteflies.
Abstract: Tomato chlorosis virus (ToCV) is the second whitefly-transmitted, phloem-limited, bipartite closterovirus described infecting tomato. ToCV is distinct from tomato infectious chlorosis virus (TICV), based on lack of serological and nucleic acid cross-reactions and differences in vector specificity. TICV is transmitted only by the greenhouse whitefly (Trialeurodes vaporariorum), whereas ToCV is transmitted by the greenhouse whitefly, the banded-wing whitefly (T. abutilonea), and Bemisia tabaci biotypes A and B (B. argentifolii). Double-stranded (ds) RNA analyses of ToCV show two prominent dsRNAs of approximately 7,800 and 8,200 bp, with several small dsRNAs. Digoxigenin-11-UTP-labeled riboprobes derived from cDNA clones representing portions of RNAs 1 and 2 were used in Northern blot hybridizations to detect two large nonhomologous dsRNAs and a subset of smaller dsRNAs. These probes were used in dot blot hybridizations to detect ToCV in infected tomato. Inclusion bodies and cytoplasmic vesicles were consistently observed in phloem tissues of ToCV-infected Nicotiana clevelandii. Computer-assisted sequence analysis showed significant homology between ToCV clones that hybridize specifically with RNAs 1 and 2 and the lettuce infectious yellows virus methyltransferase of RNA 1 and the HSP70 heat shock protein homolog of RNA 2, respectively. Thus, ToCV is another member of the growing subgroup of bipartite closteroviruses transmitted by whiteflies.
192 citations
TL;DR: Compost made from organic household and garden waste was used to substitute part of the peat in potting mixtures used for growing woody ornamental nursery stock and suppressiveness to R. solani was associated with high population densities of cellulolytic and oligotrophic actinomycetes.
Abstract: Compost made from organic household and garden waste was used to substitute part of the peat in potting mixtures used for growing woody ornamental nursery stock The effects of amendment with compost on the colonization of potting mixture by Rhizoctonia solani (AG1) were studied in greenhouse experiments A bioassay was developed using cucumber as a sensitive herbaceous test plant as a substitute for woody ornamental cuttings Pathogen growth in the potting mixture was estimated by measuring the distance over which damping-off of seedlings occurred Compost from two commercial composting facilities suppressed growth of R solani in potting mixtures with 20% of the product when the compost was fresh (directly after delivery) or long matured (after 5 to 7 months of additional curing) In contrast, short-matured compost (1 month of additional curing) from the same batches stimulated pathogen growth In vitro mycelial growth of R solani on mixtures with mature compost was inhibited by microbial antagonism Compost-amended potting mixtures responded differentially to the addition of cellulose powder; the effect on suppressiveness depended on curing time and origin of the compost In long-matured compost, suppressiveness to R solani was associated with high population densities of cellulolytic and oligotrophic actinomycetes The ratio of the population density of actinomycetes to that of other bacteria was around 200-fold higher in mature suppressive compost than in conducive compost
188 citations
TL;DR: Analysis of segregation of susceptibility, tolerance, and resistance during the BC1F1 toBC1F4 crosses indicated that tolerance is controlled by a dominant major gene and resistance by two to three additive recessive genes.
Abstract: Vidavsky, F., and Czosnek, H. 1998. Tomato breeding lines resistant and tolerant to tomato yellow leaf curl virus issued from Lycopersicon hirsutum. Phytopathology 88:910-914. Two tomato yellow leaf curl virus (TYLCV)-resistant plants from accessions LA1777 and LA386 of the wild tomato species Lycopersicon hirsutum have been crossed. The resulting resistant F1 plants were crossed with the domesticated tomato L. esculentum, and a series of selfing was performed. At each generation, individuals were selected for resistance (no symptoms and undetectable viral DNA) and tolerance (no symptoms but with detectable viral DNA) following controlled massive and repeated inoculations with viruliferous whiteflies. A stable BC1F4 line (denominated 902) that does not segregate for resistance was obtained. This line does not support virus accumulation, even upon extensive whitefly-mediated inoculation of young seedlings, and does not need protection with nets or insecticides. Another stable BC1F4 line (denominated 908) was tolerant to the virus. Both lines have good horticultural characteristics and bear 80- to 120-g red fruits. Analysis of segregation of susceptib ility, tolerance, and resistance during the BC1F1 to BC1F4 crosses indicated that tolerance is controlled by a dominant major gene and resistance by two to three additive recessive genes. The resistant and tolerant lines do not need to be protected by insecticides or nets.
TL;DR: This study confirms that C. polonica, an associate of the aggressive bark beetle Ips typographus, is pathogenic to Norway spruce, and shows that some nonaggressive bark beetles may vector phytopathogenic fungi.
Abstract: The pathogenicity of two isolates of each of four bark beetle-associated blue-stain fungi was evaluated after mass inoculation of about 40-year-old Norway spruce trees (Picea abies) Trees
TL;DR: An assay was developed that can identify unknown isolates of Pythium or Phytophthora species in a single hybridization based on arrays of species-specific amplified fragments or oligonucleotides derived from the internal transcribed spacer region, which are blotted as dots on a nylon membrane.
Abstract: Levesque, C. A., Harlton, C. E., and de Cock, A. W. A. M. 1998. Identification of some oomycetes by reverse dot blot hybridization. Phytopathology 88:213-222. An assay was developed that can identify unknown isolates of Pythium or Phytophthora species in a single hybridization. This reverse dot blot system is based on arrays of species-specific amplified fragments or oligonucleotides derived from the internal transcribed sp acer (ITS) region, which are blotted as dots on a nylon membrane. By using total DNA from a sample as the template, universal primers, and digoxigenindUTP, the ITS was amplified and labeled simultaneously by the polymerase chain reaction (PCR). A small aliquot of the resultant labeled and amplified product was used as a probe for hybridization to a dot blot membrane that contained the immobilized species-specific oli gonucleotides or amplified PCR fragments. The reverse dot blot system based on arrays of oligonucleotides showed far fewer cro ss-hybridizations than one based on entire amplified ITS I fragments. Unknown species can be identified simply by visualizing the positive hybridization reaction between the DNA labeled directly from the sample and the immobilized specific oligonucleotide. Currently, the assay can be used to identify Pythium aphanidermatum, P. ultimum, P. acanthicum, and Phytophthora cinnamomi. An oligonucleotide that was originally designed to identify Phytophthora hybridized to 10 of the 14 Phytophthora species tested. Another oligonucleotide designed to identify oomycetes hybridized to the 68 species tested, which represented two of the four orders of this phylum. Additional keywords: chemiluminescence, detection, diagnosis, NIH IMAGE.
TL;DR: Bootstrap analysis and determination of confidence intervals showed these geographic groupings to be extremely robust and classified into two major groups according to the type of cultivar or system of cultivation from which they originated.
Abstract: Gonzalez, M., Rodriguez, R., Zavala, M. E., Jacobo, J. L., Hernandez, F., Acosta, J., Martinez, O., and Simpson, J. 1998. Characterization of Mexican isolates of Colletotrichum lindemuthianum by using differential cultivars and molecular markers. Phytopathology 88:292-299. Differential cultivars and molecular markers were used to analyze 59 isolates of the bean anthracnose pathogen, Colletotrichum lindemuthianum, from different regions of Mexico. Ten distinct races were determined, three of which had not been reported previously in Mexico. Isolates were found to infect only a narrow range of the differential cultivars used and were restricted to cultivars of Middle American origin. A comparison of random amplified polymorphic DNA and amplified fragment length polymorphism (AFLP) analyses was carried out on a subset of the fungal isolates. Determination of genetic distances based on AFLP data and production of a dendrogram demonstrated two levels of association: i) isolates classified into two major groups according to the type of cultivar or system of cultivation from which they originated, and ii) isolates could be classified into smaller subgroups generally associated with the geographic location from which they were obtained. Bootstrap analysis and determination of confidence intervals showed these geographic groupings to be extremely robust.
TL;DR: The ability of C. saitoana to prevent the necrotrophic growth of the pathogen and stimulate structural defense responses may be the basis of its biocontrol activity.
Abstract: Biocontrol activity of Candida saitoana and its interaction with Botrytis cinerea in apple wounds were investigated. When cultured together, yeast attached to Botrytis sp. hyphal walls. In wounded apple tissue, C. saitoana restricted the proliferation of B. cinerea, multiplied, and suppressed disease caused by either B. cinerea or Penicillium expansum. In inoculated apple tissue without the yeast, fungal colonization caused an extensive degradation of host walls and altered cellulose labeling patterns. Hyphae in close proximity to the antagonistic yeast exhibited severe cytological injury, such as cell wall swelling and protoplasm degeneration. Colonization of the wound site by C. saitoana did not cause degradation of host cell walls. Host cell walls in close contact with C. saitoana cells and B. cinerea hyphae were well preserved and displayed an intense and regular cellulose labeling pattern. In addition to restricting fungal colonization, C. saitoana induced the formation of structural defense responses in apple tissue. The ability of C. saitoana to prevent the necrotrophic growth of the pathogen and stimulate structural defense responses may be the basis of its biocontrol activity.
TL;DR: Both serological comparisons and sequence determination of the S RNA demonstrate that this virus represents a new and distinct species, belonging to a separate serogroup, and for which the name iris yellow spot virus (IYSV) is proposed.
Abstract: A new tospovirus was identified in iris cultivations in the Netherlands. Both serological comparisons and sequence determination of the S RNA demonstrate that this virus represents a new and distinct species, belonging to a separate serogroup, and for which the name iris yellow spot virus (IYSV) is proposed. The disease symptoms on iris are characterized by yellow spots on the leaves. Its experimental host range is very narrow and, in addition to iris, only includes Nicotiana benthamiana and Datura stramonium. The nucleoprotein of IYSV shows only 30 to 44% sequence identity with those of other tospoviruses identified so far; the highest homology being found with the tospovirus species of serogroup IV.
TL;DR: There was a strong correlation between tolerance toalpha-tomatine, the ability to degrade this compound, and pathogenicity on tomato, and these breakdown products were inhibitory to some of the saprophytes and nonpathogens of tomato, suggesting that tomato pathogens may have multiple tolerance mechanisms to alpha-Tomatine.
Abstract: alpha-Tomatine, synthesized by Lycopersicon and some Solanum species, is toxic to a broad range of fungi, presumably because it binds to 3beta-hydroxy sterols in fungal membranes. Several fungal pathogens of tomato have previously been shown to be tolerant of this glycoalkaloid and to possess enzymes thought to be involved in its detoxification. In the current study, 23 fungal strains were examined for their ability to degrade alpha-tomatine and for their sensitivity to this compound and two breakdown products, beta(2)-tomatine and tomatidine. Both saprophytes and all five non-pathogens of tomato tested were sensitive, while all but two tomato pathogens (Stemphylium solani and Verticillium dahliae) were tolerant of alpha-to-matine (50% effective dose > 300 muM). Except for an isolate of Botrytis cinerea isolated from grape, no degradation products were detected when saprophytes and nonpathogens were grown in the presence of alpha-tomatine. All tomato pathogens except Phytophthora infestans and Pythium aphani-dermatum degraded alpha-tomatine. There was a strong correlation between tolerance to alpha-tomatine, the ability to degrade this compound, and pathogenicity on tomato. However, while beta(2)-tomatine and tomatidine were less toxic to most tomato pathogens, these breakdown products were inhibitory to some of the saprophytes and nonpathogens of tomato, suggesting that tomato pathogens may have multiple tolerance mechanisms to alpha-tomatine.
TL;DR: The genetic isolation and limited geographic distribution of four of the lineages of F. oxysporum f.
Abstract: Genetic variation within a worldwide collection of 208 isolates of Fu-sarium oxysporum f. sp. cubense, representing physiological races 1, 2, 3, and 4 and the 20 reported vegetative compatibility groups (VCGs), was analyzed using modified DNA amplification fingerprinting. Also characterized were 133 isolates that did not belong to any of the reported VCGs of F. oxysporum f. sp. cubense including race 3 isolates from a Heliconia species and isolates from a symptomatic wild banana species growing in the jungle in peninsular Malaysia. The DNA fingerprint patterns were generally VCG specific, irrespective of geographic or host origin. A total of 33 different genotypes were identified within F. oxysporum f. sp. cu-bense; 19 genotypes were distinguished among the isolates that belonged to the 20 reported VCGs, and 14 new genotypes were identified among the isolates that did not belong to any of the existing VCGs. DNA fingerprinting analysis also allowed differentiation of nine clonal lineages within F. oxysporum f. sp. cubense. Five of these lineages each contained numerous closely related VCGs and genotypes, and the remaining four lineages each contained a single genotype. The genetic diversity and geographic distribution of several of these lineages of F. oxysporum f. sp. cubense suggests that they have coevolved with edible bananas and their wild diploid progenitors in Asia. DNA fingerprinting analysis of isolates from the wild pathosystem provides further evidence for the coevolution hypothesis. The genetic isolation and limited geographic distribution of four of the lineages of F. oxysporum f. sp. cubense suggests that the pathogen has also arisen independently, both within and outside of the center of origin of the host.
TL;DR: Natural potato late blight epidemics were studied to assess the relative impact of various inoculum sources of Phytophthora infestans in Southern Flevoland (the Netherlands) from 1994 through 1996 and confirmed the role of the organic fields as mid-season infection sources.
Abstract: Natural potato late blight epidemics were studied to assess the relative impact of various inoculum sources of Phytophthora infestans in Southern Flevoland (the Netherlands) from 1994 through 1996. Disease surveys were combined with characterization of isolates for mating type and DNA fingerprint pattern using probe RG57. Seventy-four percent of the commercial potato fields with early foci were clearly associated with nearby infested refuse piles. Characterization of isolates from refuse piles and fields confirmed the association. Infected seed tubers, volunteer plants, and infested allotment gardens appeared to be of minor importance for late blight development in potato fields. Several foci in refuse piles, potato fields, and allotment gardens contained more than one genotype. Due to favorable weather in August 1994, infested organic potato fields became major inoculum sources, resulting in the spread of P. infestans to adjacent conventional potato fields. Analyses of disease gradients, both at...
TL;DR: Inoculation of bean hypocotyls with a nonpathogenic binucleate Rhizoctonia (BNR) species induced systemic resistance and protection of the roots and cotyledons to later challenge with the root rot pathogen Rhiz octonia solani or the anthracnose pathogen Colletotrichum lindemuthianum.
Abstract: Inoculation of bean hypocotyls with a nonpathogenic binucleate Rhizoctonia (BNR) species induced systemic resistance and protection of the roots and cotyledons to later challenge with the root rot pathogen Rhizoctonia solani or the anthracnose pathogen Colletotrichum lindemuthianum. Bean seedlings that were treated with BNR 48 h prior to their challenge with R. solani or C. lindemuthianum had few necrotic lesions and reduced disease severity as compared with seedlings not treated with BNR. Treatment with BNR 48 h prior to their challenge also elicited a significant and systemic increase in all cellular fractions of peroxidases, 1,3-beta-glucanases, and chitinases compared with the diseased and control plants. Compared with control plants, total peroxidases and glucanases increased twofold and eightfold, respectively, in all protected bean tissues. BNR 232-CG could not be recovered from the challenged hypocotyls or cotyledons, indicating that there was no contact between the inducer and the pathogen. Both the 1,3-beta-glucanases and the peroxidases were positively correlated with induced resistance.
TL;DR: This is the first demonstration of antifungal activity of a corn 14-kDa trypsin inhibitor protein, and the expression of this protein among tested genotypes may be related to their difference in resistance to A. flavus.
Abstract: Corn genotypes resistant or susceptible to Aspergillus flavus were extracted for protein analysis using a pH 2.8 buffer. The profile of protein extracts revealed that a 14-kDa protein is present in relatively high concentration in kernels of seven resistant corn genotypes, but is absent or present only in low concentration in kernels of six susceptible ones. The N-terminal sequence of this 14-kDa protein showed 100% homology to a corn trypsin inhibitor. The 14-kDa protein purified from resistant varieties also demonstrated in vitro inhibition of both trypsin activity and the growth of A. flavus. This is the first demonstration of antifungal activity of a corn 14-kDa trypsin inhibitor protein. The expression of this protein among tested genotypes may be related to their difference in resistance to A. flavus infection and subsequent aflatoxin contamination.
TL;DR: Current observations support the view that partial to complete embolism, which almost always accompanies these factors, might be the main cause triggering the formation of vessel occlusions.
Abstract: During gel (gum) formation in angiosperm trees, fibrillar material accumulated in protective layers of xylem parenchyma cells before being secreted across half-bordered pit membranes into vessel elements. Immunogold labeling demonstrated that this fibrillar material was mainly composed of partially esterified pectic polysaccharides. The primary wall of expanding tyloses, an extension of the parenchyma protective layer, secreted similar pectic substances to completely block vessel elements. In most studies, these occluding structures were reported to be formed in response to causative factors such as aging processes, injuries, or infections. Current observations support the view that partial to complete embolism, which almost always accompanies these factors, might be the main cause triggering the formation of vessel occlusions. Whereas pectin seems to be the basic component of gels (gums) and of the external layer of tyloses, other substances, such as phenols, were also detected either as a part of these plugs or as accumulations beside them in vessels. Finally, it is proposed that the term 'gel' instead of 'gum' be used in future studies to describe the occluding material secreted by ray and paratracheal parenchyma cells.
TL;DR: The degree of gametic disequilibrium was higher in the mid-season than in the late-season population, and it is estimated that 66% of the isolates in the inoculated plots were asexual progeny of the 10 inoculated isolates, and 24% were sexual recombinants.
Abstract: Zhan, J., Mundt, C. C., and McDonald, B. A. 1998. Measuring immigration and sexual reproduction in field populations of Mycosphaerella graminicola. Phytopathology 88:1330-1337. A field experiment was conducted to determine the relative contributions of immigration and sexual reproduction to the genetic structure of Mycosphaerella graminicola populations during the course of an epidemic. The genetic structure of M. graminicola populations sampled from wheat plots inoculated artificially with 10 isolates was compared with control plots infected naturally by airborne ascospores. Restriction fragment length polymorphisms (RFLPs) were used to test the randomness of associations among loci, and DNA fingerprints were used to identify clones. All isolates in the control plots had unique genotypes and RFLP loci were at gametic equilibrium, findings consistent with ra ndom mating. The proportion of isolates in the inoculated plots with DNA fingerprints that differed from the 10 inoculated isolates increased from 3% in the early to 39 and 34% in the mid- and late season, respectively. The degree of gametic disequilibrium was higher in the mid-season than in the late-season population. By the end of the growing season, we estimate that 66% of the isolates in the inoculated plots were asexual progeny of the 10 inoculated isolates, 10% were immigrants, and 24% were sexual recombinants. The proportion of infections caused by ascospores increased over the growing season.
TL;DR: Results support the putative role of the TSWV GPs as viral attachment proteins and identified potential cellular receptor(s) in thrips.
Abstract: Interactions between viral and cellular membrane fusion proteins mediate virus penetration of cells for many arthropod-borne viruses. Electron microscope observations and circumstantial evidence indicate insect acquisition of tomato spotted wilt virus (TSWV) (genus Tospovirus, family Bunyaviridae) is receptor mediated, and TSWV membrane glycoproteins (GP1 and GP2) serve as virus attachment proteins. The tospoviruses are plant-infecting members of the family Bunyaviridae and are transmitted by several thrips species, including Frankliniella occidentalis. Gel overlay assays and immunolabeling were used to investigate the putative role of TSWV GPs as viral attachment proteins and deter mine whether a corresponding cellular receptor may be present in F. occidentalis. A single band in the 50-kDa region was detected with murine monoclonal antibodies (MAbs) to the TSWV-GPs when isolated TSWV or TSWV-GPs were used to overlay separated thrips proteins. This band was not detected when blots were probed with antibody to the non-structural protein encoded by the small RNA of TSWV or the TSWV nucleocapsid protein, nor were proteins from nonvector insects labeled. Anti-idiotype antibodies prepared to murine MAbs against GP1 or GP2 specifically labeled a single band at 50 kDa in Western blots and the plasmalemma of larval thrips midguts. These results support the putative role of the TSWV GPs as viral attachment proteins and identified potential cellular receptor(s) in thrips.
TL;DR: DNA fingerprinting by Pot2 rep-PCR provides an efficient means to monitor the population dynamics of the blast pathogen and is now feasible to conduct large-scale population studies to understand the impact of host genotypes on pathogen evolution.
Abstract: DNA samples from Magnaporthe grisea isolates were fingerprinted by using repetitive element-based polymerase chain reaction (rep-PCR) with two outwardly directed primer sequences from Pot2, an element found in approximately 100 copies in the fungal genome. Variable length fragments, defining the sequences lying between these elements, were generated, and fingerprint patterns specific for individual strains were established. "Long PCR" conditions, including higher pH (9.2) and increased extension time (10 min) were used to amplify DNA fragments ranging from 400 bp to longer than 23 kb. Polymorphisms specific to M. grisea strains were generated, allowing inference of their genetic relationships. Segregation analysis was used to confirm single-locus inheritance for the fragments amplified by rep-PCR. Cluster analysis revealed robust groupings that corresponded to previously determined MGR586 restriction fragment length polymorphism lineages of the rice-infecting strains of the pathogen. We have also demonstrated the utility of rep-PCR to differentiate isolates that infect rice from those that infect nonrice hosts. DNA fingerprinting by Pot2 rep-PCR provides an efficient means to monitor the population dynamics of the blast pathogen. Because of the method's low cost and ease in application, it is now feasible to conduct large-scale population studies to understand the impact of host genotypes on pathogen evolution.
TL;DR: It is demonstrated that maize residue can act as a long-term source of inoculum for infection of maize plants by these three Fusarium species because of significant interactions among strain, depth, residue size, and time.
Abstract: Cotten, T. K., and Munkvold, G. P. 1998. Survival of Fusarium moniliforme, F. proliferatum, and F. subglutinans in maize stalk residue. Phytopathology 88:550-555. The roles of residue size and burial depth were assessed in the survival of Fusarium moniliforme, F. proliferatum, and F. subglutinans in maize stalk residue. Stalk pieces (small or large sizes) were soaked in a spore suspension of F. moniliforme, F. proliferatum, or F. subglutinans and placed in a field on the soil surface or buried at 15- or 30-cm depths. Residue pieces were recovered periodically, cultured on a selective medium, and microscopically examined for the presence of the inoculated Fusarium species. After 630 days, the inoculated Fusarium species were recovered from 0 to 50% of the inoculated stalk pieces in a l ong-term, continuous maize field, from 0 to 28% of the inoculated stalk pi eces placed in a maize/soybean/oat rotation field, and from 0 to 25% of the noninoculated stalk pieces at both locations. Residue size and residue depth had significant effects on survival, but there were significant interactions among strain, depth, residue size, and time. Up to 343 days after placement in the field, survival of the three Fusarium species was not consistently different between buried residues and surface residues, but after 630 days, survival was greater from surf ace residues. Overall, fungus survival decreased more slowly in the surface residues than in the buried residues. Linear coefficients of determination ranged from 0.35 to 0.82 for the surface residues and from 0.81 to 0.98 for the buried residues. Decline in survival over time followed a more linear pattern in buried residues than in surface residues. Vegetative compatibility tests confirmed that F. moniliforme, F. proliferatum, and F. subglutinans strains can survive at least 630 days in surf ace or buried maize residue. These results demonstrate that maize residue can act as a long-term source of inoculum for infection of maize plants by these three Fusarium species.
TL;DR: Results indicate that wheat streak mosaic virus and BrSMV should be classified within a new genus of the family Potyviridae and should not be considered species of the genus Rymovirus.
Abstract: Stenger, D. C., Hall, J. S., Choi, I.-R., and French, R. 1998. Phylogenetic relationships within the family Potyviridae: Wheat streak mosaic virus and brome streak mosaic virus are not members of the genus Rymovirus. Phytopathology 88:782-787. The complete nucleotide sequence of wheat streak mosaic virus (WSMV) has been determined based on complementary DNA clones derived from the 9,384-nucleotide (nt) RNA of the virus. The genome of WSMV has a 130-nt 5′ leader and 149-nt 3′-untranslated region and is polyadenylated at the 3′ end. WSMV RNA encodes a single polyprotein of 3,035 amino acid residues and has a deduced genome organization typical for a member of the family Potyviridae (5′-P1/HC-Pro/P3/6K1/CI/6K2/VPg-NIa/NIb/CP-3′). Because WSMV shares with ryegrass mosaic virus (RGMV) the biological property of transmission by eriophyid mites, WSMV has been assigned to the genus Rymovirus, of which RGMV is the type species. Phylogenetic analyses were conducted with complete polyprotein or NIb protein sequences of 11 members of the family Potyviridae, including viruses of monocots or dicots and viruses transmitted by aphids, whiteflies, and mites. WSMV and the monocot-infecting, mite-transmitted brome streak mosaic virus (BrSMV) are sister taxa and share a most recent common ancestor with the whitefly-transmitted sweet potato mild mottle virus, the type species of the proposed genus “Ipomovirus.” In contrast, RGMV shares a most recent common ancestor with aphid-transmitted species of the genus Potyvirus. These results indicate that WSMV and BrSMV should be classified within a new genus of the family Potyviridae and should not be considered species of the genus Rymovirus.
TL;DR: In vitro assays reliably distinguished DON- and NIV-producing types of F. culmorum; however, these assays could not predict production of DON by these isolates in the field, and Toxin production seemed to be a common feature in F. Culmorum.
Abstract: A susceptible synthetic winter rye population was inoculated with 42 isolates of Fusarium culmorum, originating from nine European countries and Australia, at two field locations in Germany. Significant (P = 0.01) genetic variation in aggressiveness of isolates of F. culmorum was observed across both field locations. Field samples were used to determine deoxynivalenol (DON), nivalenol (NIV), and ergosterol (ERG) contents. The 42 isolates also were incubated on rye grain in vitro, and DON and NIV contents were analyzed. Thirty-four isolates produced DON, and seven isolates produced NIV at both field locations and in vitro. Mean DON contents ranged from 0.5 to 64.6 mg/kg in grain from field trials and from 0.3 to 376.3 mg/kg in grain incubated in vitro; mean NIV contents ranged from 17.6 to 30.4 mg/kg in grain from field trials and from 0.8 to 381.0 mg/kg in grain incubated in vitro. No correlation was found between the DON content of field-grown grain and grain incubated in vitro. NIV-producing is...
TL;DR: The rep-PCR-generated patterns of DNA fragments observed after agarose gel electrophoresis support the current division of C. michiganensis into five subspecies 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.
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 polymorphism...
TL;DR: A large-scale comparison of two typing approaches for the analysis of Plum pox potyvirus isolates shows an overall excellent correlation between the results obtained in indirect double-antibody sandwich enzyme-linked immunosorbent assay and those derived from either specific PCR assays or restriction fragment length polymorphism analysis of PCR fragments.
Abstract: Candresse, T., Cambra, M., Dallot, S., Lanneau, M., Asensio, M., Gorris, M. T., Revers, F., Macquaire, G., Olmos, A., Boscia, D., Quiot, J. B., and Dunez, J. 1998. Comparison of monoclonal antibodies and polymerase chain reaction assays for the typing of isolates belonging to the D and M serotypes of plum pox potyvirus. Phytopathology 88:198-204. Plum pox potyvirus (PPV) isolates may be divided into four groups separated by serological, molecular, and epidemiological differences. Monoclonal antibodies specific for the two major groups of isolates, represented by the D and M serotypes of the virus, have been obtained. Polymerase chain reaction (PCR)-based assays allowing the direct detection and differentiation of PPV isolates have also been developed. We now report on a large-scale comparison of these two typing approaches. The results obtained show an overall excellent correlation between the results obtained in indirect double-antibody sandwich enzyme-linked immunosorbent assay using PPV-D‐ and PPV-M‐specific monoclonal antibodies and those derived from either specific PCR assays or restriction fragment length polymorphism analysis of PCR fragments. Without exception, all isolates reacting positively with the PPV-M‐specific m onoclonal antibody were found to belong to the M serotype using the PCRbased assays, while 51 out of 53 isolates recognized by the D-specific monoclonal antibodies belonged to the D serotype according to the PCR typing results. However, failure to react with a specific m onoclonal antibody did not prove as effective a predictor of the serotype of the isolate analyzed. In a few cases, the results obtained with the various techniques diverged, indicating low level variability of the epitopes rec ognized by the serotype-specific monoclonal antibodies. Isolates belonging to the two minor groups of PPV (El Amar and Cherry) also gave divergent results, indicating that the current typing assays are not suited for the analysis of such isolates.