bluuebunny/crossref_metadata_2025 huggingface dataset:
Conidial Germination, Infection Structure Formation, and Early Colony Development of Powdery Mildew on Poinsettia
Authors: Celio G. J. , Hausbeck M. K.
Published: May 2007

Powdery mildew disease on poinsettias (Euphorbia pulcherrima) growing in commercial greenhouses was first observed in the United States in 1990 and has become an economically significant problem for poinsettia growers in the Midwest and northern United States since 1992. The temporal development of infection structures produced by conidial germ tubes of the pathogen (Oidium sp.) and the effect of high temperature on their development were investigated using poinsettia leaf disks placed in humidity chambers. Observations were made using light microscopy and scanning electron microscopy. At 20°C (85% relative humidity), conidia germinated and formed an appressorium within 6 h of inoculation. Germination over time followed a monomolecular curve (r2 = 0.77, P ≤ 0.0001). Within 24 h postinoculation, germinated conidia had formed secondary germ tubes and a haustorium. The percentage of germinated conidia with appressoria and one or more secondary germ tubes increased linearly with time (r2 = 0.92, P ≤ 0.0001), while the percentage of germinated conidia with appressoria and haustoria increased mono-molecularly (r2 = 0.93, P ≤ 0.0001). Conidia had an ornamented appearance, and all conidiophores had arced basal cells. When the incubation temperature was 30°C, conidial germination and development of secondary germ tubes and a haustorium were reduced.
Factors Affecting the Release of Ascospores of Anisogramma anomala
Authors: Pinkerton J. N. , Johnson K. B. , Stone J. K. , Ivors K. L.
Published: May 2007

Relationships between environmental factors and release of ascospores of Anisogramma anomala, the causal agent of eastern filbert blight, were examined in four European hazelnut (Corylus avellana) orchards during a 2-year period. In each orchard, Burkhard volumetric spore traps and automated weather-monitoring equipment were deployed for 12-week periods beginning at budbreak, when hazelnut becomes susceptible to infection. Ascospores of A. anomala were released when stromata on the surface of hazelnut branches were wet from rain but not from dew. Release of ascospores ceased after branch surfaces dried. The duration of free moisture on branch surfaces regulated the initiation and rate of ascospore release, but no significant effects of temperature, relative humidity, wind, or light on ascospore release were apparent. Most (>90%) ascospores were captured during precipitation events that exceeded 20 h in duration, which represented about 10% of the total precipitation events each season. Quantitative relationships between the hourly capture of A. anomala ascospores and hours since the beginning of a precipitation event were developed. With the onset of precipitation, the hourly rate of ascospore capture increased until the fifth hour of rain, remained relatively constant between the fifth and twelfth hours, and then declined gradually. During the 12-week spore-trapping periods, the likelihood and rates of ascospore release associated with precipitation were highest at budbreak and then declined through April and May until early June, when the reserve of ascospores in the perithecia was depleted. Large numbers of ascospores were captured in the volumetric spore traps, indicating that ascospores may be commonly dispersed long distances on air currents as well as locally by splash dispersal within the canopy, as reported previously. The results indicate that monitoring seasonal precipitation patterns may be useful for estimating the quantity and temporal distribution of airborne inoculum during the period that the host is susceptible to infection.
Ecological Studies of Transformed Trichoderma harzianum Strain 1295–22 in the Rhizosphere and on the Phylloplane of Creeping Bentgrass
Authors: Lo C.-T. , Nelson E. B. , Hayes C. K. , Harman G. E.
Published: May 2007

A β-glucuronidase (GUS) reporter gene and a hygromycin B (hygB) phosphotransferase gene were integrated separately into the Trichoderma harzianum strain 1295-22 genome, using biolistic transformation. The mycelial growth and biocontrol ability of the transformed strains did not differ from that of the original strain. The transformed Gus+-kanamycin-resistant (Gus+KanR) strains were used to monitor growth and interactions with Rhizoctonia solani on creeping bentgrass plants. The hygB-resistant (hygBR) strains were used to selectively recover strain 1295-22 from the rhizosphere soil and phylloplane of creeping bentgrass after spray applications. The population levels of two hygBR strains and the original strain were very similar for all treatments. All three strains persisted for the duration of the experiment (28 days) in both the rhizosphere soil and on leaves, although population levels declined somewhat over the course of the experiment in unautoclaved soils. In this study, the results demonstrated that hygBR strains remained dominant over time when assayed on Trichoderma-selective medium containing hygB. The hygBR strains were not displaced by strains that colonized untreated plants. Microscopic observation showed that the Gus+KanR strains colonized the rhizoplane, seed coat, and phylloplane of creeping bentgrass. These results supported our earlier observation that strain 1295-22 was rhizosphere and phyllo-plane competent. Interactions between T. harzianum and R. solani were readily observed in situ and changed over time. Two types of reactions were found in these experiments. In the first type, sections of hyphae of R. solani near the hyphae of T. harzianum appeared damaged, and the pathogen appeared necrotic when viewed with a microscope. The second type, observed less frequently than the first type, was typical of myco-parasitism. The findings in this study provide new insight into the interactions between R. solani and T. harzianum, providing a basis for future research.
A Small RNA Resembling the Beet Western Yellows Luteovirus ST9-Associated RNA Is a Component of the California Carrot Motley Dwarf Complex
Authors: Watson Michael T. , Tian Tongyan , Estabrook Elizabeth , Falk Bryce W.
Published: May 2007

Virions were purified from Anthriscus cerefolium or Coriandrum sativum plants infected with the viruses that cause California carrot motley dwarf. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of virion preparations yielded a single prominent protein species of approximately 28,000 molecular weight; however, denaturing agarose gel electrophoresis showed that virions contained three prominent single-stranded RNAs of approximately 5.6, 4.2, and 2.8 kb. Northern hybridization analyses, using transcripts generated from cloned cDNAs that corresponded to each of the virion RNAs, showed that the 5.6- and 4.2-kb RNAs were the genomic RNAs of the carrot red leaf luteovirus (CRLV) and the carrot mottle umbravirus (CMoV), respectively. Virions also contained an approximately 1.3-kb RNA related to the CMoV genomic RNA. The 2.8-kb RNA did not hybridize with CRLV or CMoV cRNA probes. Analysis of naturally infected carrot (Daucus carota) plants showed that CRLV, CMoV, and the 2.8-kb RNA were always present in carrot motley dwarf—affected plants. Greenhouse aphid- and mechanical-transmission experiments showed that the 2.8-kb RNA was consistently present in plants also infected by both CRLV and CMoV, but never in plants infected by only CMoV. Near full-length cloned cDNAs corresponding to the 2.8-kb RNA were prepared, and the complete nucleotide sequence was determined to be 2,835 nucleotides. Two large open reading frames (ORFs), 1a and 1b, were present within the sequence and were separated by an amber (UAG) stop codon. A third ORF (ORF 2), capable of encoding a protein of 4,289 molecular weight, was located near the 3′ terminus. BLASTP results showed that the 2.8-kb RNA was most closely related to the beet western yellows luteovirus (BWYV) ST9-associated RNA. Based on its biological and molecular characteristics, we have named the 2.8-kb RNA the CRLV-associated RNA (CRLVaRNA).
Enhancement of Teliospore Germination in Wheat- and Wild Grass-Infecting Species of Tilletia on Activated Charcoal Medium
Authors: Boyd María L. , Carris Lori M.
Published: May 2007

The effect of activated charcoal as an amendment to water agar medium on teliospore germination was analyzed for two species of wheat-infecting bunts, Tilletia controversa and T. tritici, and two related wild-grass infecting species, T. bromi and T. fusca. Final percentages of teliospore germination, area under the germination progress curves (AUGPC), and a standardized AUGPC (SAUGPC) on carbon agar and water agar were compared among strains. Carbon agar (CA) significantly increased the final germination percentage of teliospores, AUGPC, and SAUGPC when compared with water agar (WA) for all taxa under study. Additionally, CA reduced significantly the incubation (i.e., lag) period when compared with WA for teliospores of T. bromi, T. controversa, and T. fusca. Bovine serum albumin and polyvinyl pyrrolidone were used as alternative chemical adsorbent amendments to WA to establish the role of activated charcoal in the medium. Only media amended with bovine serum albumin and activated charcoal improved the final germination percentage of all taxa. Polyvinyl pyrrolidone was not significantly better than water agar.
Protein(s) from the Gram-Positive Bacterium Clavibacter michiganensis subsp. michiganensis Induces a Hypersensitive Response in Plants
Authors: Alarcón C. , Castro J. , Muñoz F. , Arce-Johnson P. , Delgado J.
Published: May 2007

The gram-positive tomato pathogen Clavibacter michiganensis subsp. michiganensis induced a local necrotic response on four-o'clock (Mirabilis jalapa) and tobacco (Nicotiana tabacum) plants. This necrosis response was characteristic of the hypersensitive response (HR). The cell-free culture supernatant from strain CMM623 also induced a necrosis that was phenotypically similar to that induced by the bacteria. Inhibitors of plant metabolism suppressed the necrotic reaction of both M. jalapa and tobacco. The HR-inducing activity present in the supernatant was heat stable, sensitive to proteases, and had an apparent molecular mass in the range of 35 to 50 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The properties observed for the necrosis-inducing activity resembled harpin and PopA described from gram-negative phytopathogenic bacteria.
Resistance and Susceptibility of Arabidopsis thaliana to Bacterial Wilt Caused by Ralstonia solanacearum
Authors: Yang Chang-Hsien , Ho Gan-Der
Published: May 2007

Tomato bacterial wilt caused by Ralstonia solanacearum is a model system for studying plant-bacterial interactions, because it is genetically one of the best characterized plant diseases. We demonstrate here that four different strains of R. solanacearum, two from radishes (Rd4 and Rd15) and two from tomato (Ps21 and Ps95), can infect 27 different ecotypes of Arabidopsis thaliana, causing different responses. All ecotypes tested were highly susceptible to strain Rd15, which caused symptoms similar to those observed in tomato plants. For example, leaf drooping and discoloration developed just 3 days after inoculation, and plants completely wilted within 1 week. Strains Rd4 and Ps95 were less infectious than Rd15. With these two strains, a variety of disease responses were observed among different ecotypes at 2 weeks after inoculation; both susceptible and resistant ecotypes of A. thaliana were identified. Ps21 was the least infectious of the four strains and caused almost no symptoms in any of the ecotypes of Arabidopsis tested. Direct bacterial isolation and plant skeleton hybridization analysis from infected plants indicated that bacterial colonization was correlated with the severity of symptoms. Growth of bacteria was limited to the infection site in resistant plants, whereas the bacteria spread throughout susceptible plants by 1 week after inoculation.
Systemic Induction of Peroxidases, 1,3-β-Glucanases, Chitinases, and Resistance in Bean Plants by BinucleateRhizoctoniaSpecies
Authors: Xue L. , Charest P. M. , Jabaji-Hare S. H.
Published: May 2007

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-β-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-β-glucanases and the peroxidases were positively correlated with induced resistance.
Genetic Heterogeneity ofLeucostomaSpecies in Michigan Peach Orchards
Authors: Wang Dechun , Iezzoni Amy , Adams Gerard
Published: May 2007

The pathogens causing perennial canker of peach, Leucostoma spp., were characterized in Michigan orchards to identify which species and subgroups (cryptic species) were prevalent on a highly susceptible peach cultivar, Loring, and a less susceptible cultivar, Redhaven. Four hundred and three strains of Leucostoma were isolated from cankers in three southwest Michigan orchards where ‘Loring’ occurred adjacent to ‘Redhaven’ in side-by-side plots. Based on colony morphology and small nuclear rDNA size polymorphisms, three cryptic species were detected; 89% of the isolates were identified as L. persoonii phenetic group 1, 10% were L. cincta group 4, and 1% were L. persoonii group 2. Pathogen profiles differed significantly between cankers on small branches of ‘Loring’ and ‘Redhaven’, and between cankers on small branches and trunks. Of 1,232 random pairings among isolates of L. persoonii group 1, 95% were vegetatively incompatible. A minimum of 11 and a maximum of 17 maternal lines were identified based upon mitochondrial DNA restriction fragment length polymorphisms among 69 isolates of L. persoonii group 1 from one split-cultivar block of 72 trees. Vegetative compatibility loci were randomly associated with maternal lines. Evidence of clonality was absent in L. persoonii group 1, as no correlation occurred between these two sets of independent genetic markers and no genotype was over-represented. The number of cryptic species, the number of maternal lines, and the frequency of incompatibility within maternal lines indicate that considerable genetic variation exists within the Leucostoma populations in Michigan peach orchards, and that sexual recombination is common.
Compost and Compost Water Extract-Induced Systemic Acquired Resistance in Cucumber and Arabidopsis
Authors: Zhang W. , Han D. Y. , Dick W. A. , Davis K. R. , Hoitink H. A. J.
Published: May 2007

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 but not in the compost mix. The peat mix water extract applied as a spray did not control bacterial speck on plants grown in either mix. Topical sprays with salicylic acid (SA) reduced the severity of bacterial speck on plants in the peat mix but did not further reduce the severity of symptoms on plants in the compost mix. The activity of the compost water extract was heat-stable and passed through a 0.2-μm membrane filter. β-1,3-Glucanase activity was low in cucumber plants grown in either mix, but when infected with C. orbiculare, this activity was induced to significantly higher levels in plants grown in the compost mix than in plants grown in the peat mix. Similar results were obtained for β-D-glucuronidase (GUS) activity driven by a PR2 (β-1,3-glucanase) gene promoter in transgenic Arabidopsis plants grown in the compost or peat mix. GUS activity was induced with topical sprays of the compost water extract or SA in plants not inoculated with the pathogen, suggesting that compost-induced disease suppression more than likely involved the potentiation of resistance responses rather than their activation and that compost-induced SAR differed from SAR induced by pathogens, SA, or compost water extract.
Production of a Mating Inhibitor by Ustilago hordei
Authors: Sherwood John E. , Kosted Paula J. , Anderson Cynthia M. , Gerhardt Shirley A.
Published: May 2007

Ustilago hordei, the cause of barley covered smut, was found to produce a factor that inhibited its own mating. The mating inhibition factor (MIF) specifically inhibited mating of U. hordei and other Ustilago spp., but not teliospore germination or sporidial growth. MIF did prevent teliospore germination of Tilletia caries and T. contraversa. MIF was found at low levels in culture supernatants of either mating type of U. hordei grown separately, but at higher levels when both mating types were grown together, in the supernatants of MAT-1 mating type cells transformed with the MAT-1 pheromone gene mfa1 and of MAT-2 cells transformed with either mfa1 or the MAT-1 pheromone receptor gene pra1. Diploid cells produced no detectable inhibitor, nor did MAT-1 cells with a disrupted mating type locus that deleted both mfa1 and pra1. MIF production was restored when mfa1, but not pra1, was added back to the MAT-1Δ cells. MIF activity was altered by protease treatment. Highly purified MIF from MAT-1 cells contained cysteine methyl ester, farnesyl cysteine, farnesyl cysteine methyl ester, and a dodecapeptide with a mass consistent with that of MAT-1 pheromone lacking the terminal cysteine. Since smut fungi must first mate to become pathogenic, mating inhibition has the potential to be an effective method of disease control for these pathogens.
Immunocytochemical Evidence that Secretion of Pectin Occurs During Gel (Gum) and Tylosis Formation in Trees
Authors: Rioux D. , Nicole M. , Simard M. , Ouellette G. B.
Published: May 2007

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.
Biological Control of Botrytis cinerea in Cyclamen with Ulocladium atrum and Gliocladium roseum Under Commercial Growing Conditions
Authors: Köhl J. , Gerlagh M. , De Haas B. H. , Krijger M. C.
Published: May 2007

The effect of treatments with conidial suspensions of Ulocladium atrum and Gliocladium roseum on leaf rot of cyclamen caused by Botrytis cinerea was investigated under commercial greenhouse conditions. Spraying U. atrum (1 × 106 conidia per ml) or G. roseum (2 × 106 conidia per ml and 1 × 107 conidia per ml) at intervals of 2 to 3 weeks during the production period and spraying U. atrum (1 × 106 conidia per ml) at intervals of 4 to 6 weeks resulted in a significant reduction of natural infections of petioles by B. cinerea. U. atrum or G. roseum (1 × 107conidia per ml) was as effective as the standard fungicide program. B. cinerea colonized senesced leaves within the plant canopy and infected adjacent petioles and leaves later. The antagonists colonized senesced leaves and reduced B. cinerea development on these leaves. Thus, the inoculum potential on petioles adjacent to necrotic leaf tissues was reduced. The fate of U. atrum conidia on surfaces of green cyclamen leaves during a 70-day period after application was studied. The number of conidia per square centimeter of leaf surface remained relatively constant during the entire experiment. Sixty percent of the conidia sampled during the experiments retained the ability to germinate. When green leaves were removed from the plants to induce senescence and subsequently were incubated in a moist chamber, U. atrum colonized the dead leaves. Senesced leaves also were colonized by other naturally occurring fungi including B. cinerea. On leaves treated with U. atrum from all sampling dates, sporulation of B. cinerea was significantly less as compared with the untreated control. Our results indicate that early applications of U. atrum before canopy closure may be sufficient to achieve commercially satisfactory control of Botrytis leaf rot in cyclamen.
Localization of Glucose Oxidase with Immunocytochemistry in the Biocontrol Fungus Talaromyces flavus
Authors: Stosz S. K. , Roy S. , Murphy C. , Wergin W. , Fravel D. R.
Published: May 2007

Production of the enzyme glucose oxidase has been shown to be involved in the biological control of Verticillium wilt by Talaromyces flavus. In this study, the location of glucose oxidase was determined in T. flavus by immunocytochemistry using glucose oxidase-specific polyclonal antibody. Immunostaining revealed that glucose oxidase was both intracellular and extracellular. Old, as well as young, hyphal cells contained glucose oxidase, but labeling of the cell wall-associated enzyme decreased as the cells aged. Exocytosis rather than cell lysis was the primary means of release of glucose oxidase from cells. Enzyme stability studies confirmed that the glucose oxidase of T. flavus is an extremely stable enzyme, retaining 13% of its original activity after 2 weeks at 25°C and retaining activity for several days at temperatures up to 50°C.
Hypovirus Transmission to Ascospore Progeny by Field-Released Transgenic Hypovirulent Strains of Cryphonectria parasitica
Authors: Anagnostakis Sandra L. , Chen Baoshan , Geletka Lynn M. , Nuss Donald L.
Published: May 2007

Strains of the chestnut blight fungus, Cryphonectria parasitica, have been genetically engineered to contain an integrated full-length cDNA copy of the prototypic virulence-attenuating hypovirus CHV1-EP713. Unlike natural hypovirulent C. parasitica strains, these transgenic hypovirulent strains are able to transmit virus to ascospore progeny under laboratory conditions. This ability provides the potential to circumvent barriers to cytoplasmic virus transmission imposed by the fungal vegetative incompatibility system. During July 1994, transgenic hypovirulent strains were introduced into a Connecticut forest site (Biotechnology Permit 94-010-01). Subsequent analysis of the release site confirmed hypovirus transmission from transgenic hypovirulent strains to ascospore progeny under field conditions. Additionally, it was possible to recover transgenic hypovirulent strains from the test site as long as 2 years after the limited, single-season release. Evidence also was obtained for cytoplasmic transmission of transgenic cDNA-derived hypovirus RNA, including transmission to mycelia of a virulent C. parasitica canker after treatment with conidia of a transgenic strain. Finally, a transgenic hypovirulent strain was recovered from a superficial canker formed on an untreated chestnut tree. Genetic characteristics of the recovered strain suggested that the canker was initiated by an ascospore progeny derived from a cross involving an input transgenic hypovirulent strain. The durability of a molecular marker for field-released cDNA-derived hypovirus RNA is discussed.
Secretion of a Fungal Extracellular Catalase by Claviceps purpurea During Infection of Rye: Putative Role in Pathogenicity and Suppression of Host Defense
Authors: Garre Victoriano , Tenberge Klaus B. , Eising Rainer
Published: May 2007

Hydrogen peroxide of the host origin accumulates in plant apoplasts in response to pathogen attack and probably functions directly in defense reactions or in signaling, according to a previous study. Since Claviceps purpurea produces compatible interactions with hundreds of host species, we hypothesized that the fungus might interfere with H2O2-mediated defense by means of secreted catalases. In axenic culture of C. purpurea, catalase activity accumulated in the medium and was inhibited by the catalase inhibitor aminotriazole. Polyacrylamide gel electrophoresis followed by diaminobenzidine (DAB)-mediated activity staining showed that one specific catalase found in culture filtrate was also present in rye ovaries infected with C. purpurea and in honeydew. This catalase form is probably induced during infection. In situ activity staining, using DAB-mediated enzyme-cytochemistry in electron microscopy, located catalase activity in hyphal walls during both axenic culture and infection of rye. Activity staining accumulated in periplasmic spaces and was especially strong at hyphal surfaces; control staining after aminotriazole inhibition was negative. Intracellular activity staining in organelles of the fungal secretory pathway substantiated that catalase was secreted by C. purpurea. With molecular cytology, anticatalase epitopes were localized with different heterologous catalase antibodies at sites corresponding to the activity staining pattern. In all infection phases, immunogold labeling indicated that the putative catalase was secreted via multivesicular bodies into the fungal wall and diffused into the host apoplast exclusively at the hostpathogen interface. The secretion of fungal catalase is a novel finding in phytopathology, and we discuss its role in the ubiquitous ergot disease.
Development of Potato Late Blight Epidemics: Disease Foci, Disease Gradients, and Infection Sources
Authors: Zwankhuizen Maarten J. , Govers Francine , Zadoks Jan C.
Published: May 2007

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 the field and regional levels, confirmed the role of the organic fields as mid-season infection sources. The mean slope of field gradients downwind of refuse piles (point sources) was significantly steeper (100-fold difference) than the mean slope of field gradients downwind of organic fields (area sources). The genotypic composition of the P. infestans populations along the gradient and of the source populations in the organic potato crops did not differ significantly. Analysis of the region gradient revealed genotype-specific disease gradients. Control measures are recommended.
Phylogenetic Relationships Within the Family Potyviridae: Wheat Streak Mosaic Virus and Brome Streak Mosaic Virus Are Not Members of the Genus Rymovirus
Authors: Stenger Drake C. , Hall Jeffrey S. , Choi Il-Ryong , French Roy
Published: May 2007

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.
Suppression of Dollar Spot by Hypovirulent Isolates of Sclerotinia homoeocarpa
Authors: Zhou Ting , Boland Greg J.
Published: May 2007

Selected hypovirulent isolates of Sclerotinia homoeocarpa were evaluated for efficacy in suppressing dollar spot of turfgrass under growth room and field conditions. Under growth room conditions, hypovirulent isolates Sh12B, Sh09B, or Sh08D of S. homoeocarpa caused 3.4 to 30.4% diseased turf in comparison to virulent isolates Sh48B and Sh14D, which caused 80.2 to 90.2% disease. In treatments that received both virulent and hypovirulent isolates, only hypovirulent isolate Sh12B significantly reduced disease as compared with the control with virulent isolates alone. In a field experiment in 1993 on swards of creeping bentgrass artificially inoculated with a virulent isolate of the pathogen, all treatments containing hypovirulent isolate Sh12B applied as a mycelial suspension, granular mix, or alginate pellets developed significantly less disease (6.3 to 20.8% diseased turf) compared with their respective formulation controls (23.8 to 31.2%). Suppression of dollar spot by treatment with mycelial suspensions of isolate Sh12B was evident up to 45 days postinoculation, and disease suppression was still significant 1 year after application when compared with the water control. Applications of hypovirulent isolate Sh09B did not reduce dollar spot in any treatments. Significant suppression of dollar spot by isolate Sh12B was also observed in the experiment conducted in 1994. In addition, suppression of dollar spot by hypovirulent isolate Sh12B was evaluated on swards with naturally occurring inoculum during 1994. Treatments with a mycelial suspension and alginate pellets of hypovirulent isolate Sh12B significantly reduced dollar spot compared with their respective formulation controls. With few exceptions, there was no statistical difference between treatments with hypovirulent isolate Sh12B and the fungicide chlorothalonil (Daconil 2787). Multiple applications of the hypovirulent isolate did not result in greater suppression of dollar spot as compared with a single application. The results indicate that hypovirulence has potential as an effective strategy for the management of dollar spot.
Asymmetrical Distribution of Barley Yellow Dwarf Virus PAV Variants Between Host Plant Species
Authors: Mastari Jamila , Lapierre Hervé , Dessens Johannes T.
Published: May 2007

A large epidemiological study of the genetic variation of barley yellow dwarf virus (BYDV) serotype PAV involving different host plant species was conducted. French BYDV PAV isolates were collected from barley and ryegrass, and their capsid protein gene sequences characterized using restriction fragment length polymorphism, single-strand conformation polymorphism, and sequence analyses. The data show that BYDV PAV isolates from five different continents are separated into two distinct groups named cpA and cpB, which are distributed irrespective of geographical location. Amino acid identity of the capsid proteins ranged from 93 to 99.5% in group cpA and from 95 to 99.5% in group cpB, while this value was only from 82 to 88% between the groups. Moreover, isolates from each group were found preferentially (up to 98%) in one of the two plant species examined. These results show that host plant species play a role in isolate selection and maintenance and that they contribute to the genetic diversity of BYDV PAV.
Description of the Bacterium Causing Blight of Leek as Pseudomonas syringae pv. porri (pv. nov.)
Authors: Samson R. , Shafik H. , Benjama A. , Gardan L.
Published: May 2007

Forty bacterial strains isolated from leek blight (Allium porrum) in France and other countries were studied by conventional biochemical methods, serological reactions, numerical taxonomy, DNA-DNA hybridization, and ice nucleation activity, as well as by pathogenicity on leek and other host plants. They were compared with reference strains of Pseudomonas, mainly pathotype strains of P. syringae pathovars and strains of P. syringae pv. syringae isolated from various host plants including onions. Leek strains sorted with P. syringae species (sensu lato) by LOPAT tests (production of levan-sucrase, oxidase, pectinase, arginine dihydrolase, and hypersensitive reaction on tobacco). Leek strains were pathogenic to leek and produced symptoms identical to those observed in the field. They were the only strains in our study that could cause blight of leek. Thus, our results justify the creation of a new pathovar. Leek strains constituted a highly homogeneous DNA group and a discrete phenon by numerical taxonomy, and they belonged to O-serogroup POR. The name of P. syringae pv. porri is proposed for the bacterium causing leek blight. Criteria for routine identification are presented and taxonomic status is discussed.
Genetic Change Within Populations of Phytophthora infestans in the United States and Canada During 1994 to 1996: Role of Migration and Recombination
Authors: Goodwin Stephen B. , Smart Christine D. , Sandrock Robert W. , Deahl Kenneth L. , Punja Zamir K. , Fry William E.
Published: May 2007

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.
Enzymatic Activity of the Mycelium Compared with Oospore Development During Infection of Pea Roots byAphanomyces euteiches
Authors: Kjøller Rasmus , Rosendahl Søren
Published: May 2007

To describe the disease cycle of the root pathogen Aphanomyces euteiches, enzymatic activity in the mycelium was compared with the development of oospores in pea roots. Plants were inoculated with two zoospore concentrations to achieve different disease levels. Hyphae were stained for fungal alkaline phosphatase activity in the roots. Additionally, enzyme activity was measured after electrophoresis of an A. euteiches-specific glucose-6-phosphate isozyme. Development of oospores in the roots was measured after staining the oospores with trypan blue. In plants inoculated with the higher zoospore concentration, the enzymatic activity of the pathogen mycelium peaked 10 to 14 days after inoculation, when oospore formation was initiated. Oospore formation was associated with a gradual increase in disease symptoms. At the last harvest, plants inoculated with the higher zoospore concentration had died. In these plants, oospores were found in 90% of the root length, while the enzymatic activity of the mycelium was low. This suggests that the pathogen mycelium is only active on living plants and does not grow saprophytically on dead plant material.
Primary Disease Gradients of Bacterial Blight of Rice
Authors: Mundt Christopher C. , Ahmed Hafiz U. , Finckh Maria R. , Nieva Lorna P. , Alfonso Rizal F.
Published: May 2007

The dispersal potential of Xanthomonas oryzae pv. oryzae, causal agent of bacterial blight of rice (Oryzae sativa), was investigated through measurement of primary disease gradients. Plants within individual hills of rice were inoculated, and the number of new lesions in the primary generation of dispersal from these inoculated sources was counted. Two dispersal models that can describe the number of infections at the source (the Kiyosawa and Shiyomi and the modified Gregory) were fit to the lesion counts. Estimated gradient slopes were similar in the 2 years of the study for both gradient models. However, the Kiyosawa and Shiyomi model gave a better fit in both years, as indicated by higher coefficients of determination and significances of slopes and by a more random pattern of residuals. Primary disease gradients were very steep, with half-distances (distances over which lesion numbers are calculated to decrease by half) estimated from the Kiyosawa and Shiyomi model of 0.091 and 0.081 m in 1994 and 1995, respectively. Splash dispersal of X. oryzae pv. oryzae is the most likely explanation for both the steep slopes measured and the superior fit of the Kiyosawa and Shiyomi model over the modified Gregory model.
A Major Gene for Resistance to White Pine Blister Rust in Western White Pine from the Western Cascade Range
Authors: Kinloch Bohun B. , Sniezko Richard A. , Barnes Gerald D. , Greathouse Tom E.
Published: May 2007

A dominant gene for resistance to white pine blister was indicated by Mendelian segregation in full-sib families of western white pine parent trees selected for phenotypic resistance in six heavily infected stands in the Western Cascades of Oregon and Washington. Seedlings were artificially inoculated three times between 1959 and 1964 and observed for development of stem infection. Segregation at this locus (Cr2) occurred in only two of the six parent populations sampled: one a natural stand, Champion Mine (CM), and the other a plantation of unknown seed origin. At CM, reduced penetrance of this gene was expressed by altered Mendelian ratios (mostly less-than-expected resistant phenotypes) in families of specific combinations of certain parents, indicating the presence of modifier genes with effects that ranged from mild to almost complete suppression of Cr2. Between 1968 and 1994, an apparent shift in virulence at CM caused all of the resistant selections to become infected and die. Recent inoculations of many of the same or related families from these parents, made from grafted ramets in a seed orchard, showed that Cr2 conditions a classical hypersensitive reaction (HR) in needle tissues, the primary infection courts. In the latter tests, seedlings were challenged with wild-type and four other sources of inoculum at and near CM that were also suspected of having wider virulence than wild type. No seedlings segregating for HR that were inoculated with wild type subsequently developed stem symptoms, but the other inocula induced both susceptible and HR needle spots on Cr2- genotypes, and many of these seedlings did develop stem infections. This implied that spore genotypes with specific virulence to Cr2 are carried in these inocula.
Amy1, the α-Amylase Gene ofAspergillus flavus: Involvement in Aflatoxin Biosynthesis in Maize Kernels
Authors: Fakhoury A. M. , Woloshuk C. P.
Published: May 2007

Aspergillus flavus is the causal agent of an ear and kernel rot in maize. In this study, we characterized an α-amylase-deficient mutant and assessed its ability to infect and produce aflatoxin in wounded maize kernels. The α-amylase gene Amy1 was isolated from A. flavus, and its DNA sequence was determined to be nearly identical to Amy3 of A. oryzae. When Amy1 was disrupted in an aflatoxigenic strain of A. flavus, the mutant failed to produce extracellular α-amylase and grew 45% the rate of the wild-type strain on starch medium. The mutant produced aflatoxin in medium containing glucose but not in a medium containing starch. The α-amylase-deficient mutant produced aflatoxin in maize kernels with wounded embryos and occasionally produced aflatoxin only in embryos of kernels with wounded endosperm. The mutant strain failed to produce aflatoxin when inoculated onto degermed kernels. In contrast, the wild-type strain produced aflatoxin in both the endosperm and embryo. These results suggest that α-amylase facilitates aflatoxin production and growth of A. flavus from a wound in the endosperm to the embryo. A 14-kDa trypsin inhibitor associated with resistance to A. flavus and aflatoxin in maize also inhibited the α-amylase from A. flavus, indicating that it is a bifunctional inhibitor. The inhibitor may have a role in resistance, limiting the growth of the fungus in the endosperm tissue by inhibiting the degradation of starch.
Genetic Structure of Cronartium ribicola Populations in Eastern Canada
Authors: Et-touil K. , Bernier L. , Beaulieu J. , Bérubé J. A. , Hopkin A. , Hamelin R. C.
Published: May 2007

The genetic structure of populations of Cronartium ribicola was studied by sampling nine populations from five provinces in eastern Canada and generating DNA profiles using nine random amplified polymorphic DNA markers. Most of the total gene diversity (Ht = 0.386) was present within populations (Hw = 0.370), resulting in a low level of genetic differentiation among populations in northeastern North America (Fst = 0.062). A hierarchical analysis of genetic structure using an analysis of molecular variance (AMOVA) revealed no statistically significant genetic differentiation among provinces or among regions. Yet, genetic differentiation among populations within regions or provinces was small (AMOVA φst = 0.078) but statistically significant (P < 0.001) and was several orders of magnitude larger than differentiation among provinces. This is consistent with a scenario of subpopulations within a metapopulation, in which random drift following migration and new colonization are major evolutionary forces. A phenetic analysis using genetic distances revealed no apparent correlation between genetic distance and the province of origin of the populations. The hypothesis of isolation-by-distance in the eastern populations of C. ribicola was rejected by computing Mantel correlation coefficients between genetic and geographic distance matrices (P > 0.05). These results show that eastern Canadian provinces are part of the same white pine blister rust epidemiological unit. Nursery distribution systems are controlled provincially, with virtually no seedling movement among provinces; therefore, infected nursery material may not play an important role in the dissemination of this disease. Long-distance spore dispersal across provincial boundaries appears to be an epidemiologically important factor for this pathogen.
Transformation of Soil Microbial Community Structure and Rhizoctonia-Suppressive Potential in Response to Apple Roots
Authors: Mazzola Mark
Published: May 2007

Changes in the composition of soil microbial communities and relative disease-suppressive ability of resident microflora in response to apple cultivation were assessed in orchard soils from a site possessing trees established for 1 to 5 years. The fungal community from roots of apple seedlings grown in noncultivated orchard soil was dominated by isolates from genera commonly considered saprophytic. Plant-pathogenic fungi in the genera Phytophthora, Pythium, and Rhizoctonia constituted an increasing proportion of the fungal community isolated from seedling roots with increasing orchard block age. Bacillus megaterium and Burkholderia cepacia dominated the bacterial communities recovered from noncultivated soil and the rhizosphere of apple seedlings grown in orchard soil, respectively. Populations of the two bacteria in their respective habitats declined dramatically with increasing orchard block age. Lesion nematode populations did not differ among soil and root samples from orchard blocks of different ages. Similar changes in microbial communities were observed in response to planting noncultivated orchard soil to five successive cycles of ‘Gala’ apple seedlings. Pasteurization of soil had no effect on apple growth in noncultivated soil but significantly enhanced apple growth in third-year orchard block soil. Seedlings grown in pasteurized soil from the third-year orchard block were equal in size to those grown in noncultivated soil, demonstrating that suppression of plant growth resulted from changes in the composition of the soil microbial community. Rhizoctonia solani anastomosis group 5 (AG 5) had no effect on growth of apple trees in noncultivated soil but significantly reduced the growth of apple trees in soil from third-year orchard soil. Changes in the ability of the resident soil microflora to suppress R. solani AG 5 were associated with reductions in the relative populations of Burkholderia cepacia and Pseudomonas putida in the rhizosphere of apple.