bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular Mapping of Rsp1, Rsp2, and Rsp3 Genes Conferring Resistance to Septoria Speckled Leaf Blotch in Barley
Authors: Lee S. H. , Neate S. M.
Published: May 2007

Septoria speckled leaf blotch (SSLB) caused by Septoria passerinii is a common disease in barley. SSLB resistance genes Rsp1, Rsp2, and Rsp3 have previously been identified in the United States Department of Agriculture National Small Grains collection accessions CIho 14300, CIho 4780, and CIho 10644, respectively. Populations of 100 to 120 F2 individuals were evaluated for SSLB resistance in the greenhouse. Inheritance was evaluated in F2:3-derived families in the field. Partial molecular maps for three Rsp genes were constructed on F2 and F2:3 families derived from crosses between Robust and the resistant accessions CIho 14300, CIho 4780, and CIho 10644. The resistant locus Rsp1 was mapped to the short arm of chromosome 3H with two flanking diversity arrays technology (DArT) markers, bPb-6978 (8.9 cM) and bPb-9945 (16.3 cM), and two random amplified polymorphic DNA (RAPD) markers, OPC2441R (3.0 cM) and UBC285158R (4.3 cM). The genes Rsp2 and Rsp3 were positioned on the short arm of barley chromosome 1H with two restriction fragment length polymorphism (RFLP), six DArT, and three RAPD markers. An RFLP marker, MWG938, and an RAPD marker, OPAH5545C, were tightly associated with Rsp2 at a distance of 0 cM. Five DArT markers spanning the short arm of 1H surrounded Rsp3 at a distance of 2.3 and 5.8 cM, while two RAPD markers—OPBA12314C (2.4 cM) in coupling and OPB17451R (3.5 cM) in repulsion—flanked Rsp3. Molecular marker data associated with Rsp2 and Rsp3 indicated that the two genes are closely linked on chromosome 1HS. A total of 17 of 154 simple sequence repeats (SSRs) tested were associated with Rsp genes on chromosome 1H and 3H, and they were also integrated into genetic linkage maps of the three F2 Robust populations. Knowledge about the map position of Rsp genes on barley chromosomes will be useful for breeding for SSLB resistance in barley and eventual gene cloning.
Expression Analysis of Rice Defense-Related Genes in Turfgrass in Response to Magnaporthe grisea
Authors: Jo Young-Ki , Wang Guo-Liang , Boehm Michael J.
Published: May 2007

Magnaporthe grisea (anamorph = Pyricularia grisea) causes blast on rice (Oryza sativa) and gray leaf spot on turfgrass. Gray leaf spot is a serious disease on St. Augustinegrass (Stenotaphrum secundatum), perennial ryegrass (Lolium perenne), and tall fescue (Festuca arundinacea). Virulence assays performed in this study revealed that M. grisea collected from rice could also cause disease on St. Augustinegrass and tall fescue. One rice isolate, Che86061, caused similar disease reactions on susceptible cultivars of rice and St. Augustinegrass and an incompatible interaction on resistant cultivars of both species. To explore whether similar defense-related genes are expressed in rice and St. Augustinegrass, a rice cDNA library was screened using pooled cDNAs derived from M. grisea-infected St. Augustinegrass. Thirty rice EST (expressed sequence tag) clones showing differential expression in St. Augustinegrass following M. grisea inoculation were identified and classified into six putative functional groups. Northern blot analyses of seven EST clones that collectively represented each putative functional group confirmed that the expression of five out of seven EST clones was similar in both rice and St. Augustinegrass. This study represents one of the first attempts to use a broad-scale genomic approach and resources of a model monocot system to study defense gene expression in St. Augustinegrass following M. grisea infection.
The Genomic Sequence of Pseudomonas fluorescens Pf-5: Insights Into Biological Control
Authors: Loper Joyce E. , Kobayashi Donald Y. , Paulsen Ian T.
Published: May 2007

The complete sequence of the 7.07 Mb genome of the biological control agent Pseudomonas fluorescens Pf-5 is now available, providing a new opportunity to advance knowledge of biological control through genomics. P. fluorescens Pf-5 is a rhizosphere bacterium that suppresses seedling emergence diseases and produces a spectrum of antibiotics toxic to plant-pathogenic fungi and oomycetes. In addition to six known secondary metabolites produced by Pf-5, three novel secondary metabolite biosynthesis gene clusters identified in the genome could also contribute to biological control. The genomic sequence provides numerous clues as to mechanisms used by the bacterium to survive in the spermosphere and rhizosphere. These features include broad catabolic and transport capabilities for utilizing seed and root exudates, an expanded collection of efflux systems for defense against environmental stress and microbial competition, and the presence of 45 outer membrane receptors that should allow for the uptake of iron from a wide array of siderophores produced by soil microorganisms. As expected for a bacterium with a large genome that lives in a rapidly changing environment, Pf-5 has an extensive collection of regulatory genes, only some of which have been characterized for their roles in regulation of secondary metabolite production or biological control. Consistent with its commensal lifestyle, Pf-5 appears to lack a number of virulence and pathogenicity factors found in plant pathogens.
Pathogenicity of Pepper mild mottle virus Is Controlled by the RNA Silencing Suppression Activity of Its Replication Protein but Not the Viral Accumulation
Authors: Tsuda Shinya , Kubota Kenji , Kanda Ayami , Ohki Takehiro , Meshi Tetsuo
Published: May 2007

Pepper mild mottle virus (PMMoV) infects pepper plants, causing mosaic symptoms on the upper developing leaves. We investigated the relationship between a virus pathogenicity determinant domain and the appearance of mosaic symptoms. Genetically modified PMMoV mutants were constructed, which had a base substitution in the 130K replication protein gene causing an amino acid change or a truncation of the 3′ terminal pseudoknot structure. Only one substitution mutant (at amino acid residue 349) failed to cause symptoms, although its accumulation was relatively high. Conversely, the pseudoknot mutants showed the lower accumulation, but they still caused mosaic symptoms as severe as the wild-type virus. Therefore, the level of virus accumulation in a plant does not necessarily correlate with the development of mosaic symptoms. The activity to suppress posttranscriptional gene silencing (PTGS) was impaired in the asymptomatic mutant. Consequently, pathogenicity causing mosaic symptoms should be controlled by combat between host PTGS and its suppression by the 130K replication protein rather than virus accumulation.
Mechanism of Action and Efficacy of Seed Meal-Induced Pathogen Suppression Differ in a Brassicaceae Species and Time-Dependent Manner
Authors: Mazzola Mark , Brown Jack , Izzo Antonio D. , Cohen Michael F.
Published: May 2007

The effect of seed meals derived from Brassica juncea, B. napus, or Sinapis alba on suppression of soilborne pathogens inciting replant disease of apple was evaluated in greenhouse trials. Regardless of plant source, seed meal amendment significantly improved apple growth in all orchard soils; however, relative differences in pathogen suppression were observed. All seed meals suppressed root infection by native Rhizoctonia spp. and an introduced isolate of Rhizoctonia solani AG-5, though B. juncea seed meal often generated a lower level of disease control relative to other seed meal types. When introduction of the pathogen was delayed until 4 to 8 weeks post seed meal amendment, disease suppression was associated with proliferation of resident Streptomyces spp. and not qualitative or quantitative attributes of seed meal glucosinolate content. Using the same experimental system, when soils were pasteurized prior to pathogen infestation, control of R. solani was eliminated regardless of seed meal type. In the case of B. juncea seed meal amendment, the mechanism of R. solani suppression varied in a temporal manner, which initially was associated with the generation of allylisothiocyanate and was not affected by soil pasteurization. Among those tested, only B. juncea seed meal did not stimulate orchard soil populations of Pythium spp. and infection of apple roots by these oomycetes. Although application of B. napus seed meal alone consistently induced an increase in Pythium spp. populations, no significant increase in Pythium spp. populations was observed in response to a composite B. juncea and B. napus seed meal amendment. Suppression of soil populations and root infestation by Pratylenchus spp. was dependent upon seed meal type, with only B. juncea providing sustained nematode control. Collectively, these studies suggest that use of a composite B. juncea and B. napus seed meal mixture can provide superior control of the pathogen complex inciting apple replant disease relative to either seed meal used alone.
Genetic and Morphological Diversity of Temperate and Tropical Isolates of Phytophthora capsici
Authors: Bowers J. H. , Martin F. N. , Tooley P. W. , Luz E. D. M. N.
Published: May 2007

Phytophthora capsici is a diverse species causing disease on a broad range of both temperate and tropical plants. In this study, we used cultural characteristics, amplified fragment length polymorphism (AFLP), and DNA sequence analyses of the ribosomal internal transcribed spacer (ITS) region and mitochondrial cytochrome oxidase II (cox II) genes to characterize temperate and tropical isolates from a wide range of host species. All but one temperate isolate grew at 35°C, while all tropical isolates did not. All but two tropical isolates formed chlamydospores, while temperate isolates did not. There was strong bootstrap support for separation of temperate and tropical isolates using AFLP analysis; however, the temperate isolates appeared as a subgroup within the observed variation of the tropical isolates. The majority of temperate isolates clustered within a single clade with low variation regardless of host or geographical origin, while the tropical isolates were more variable and grouped into three distinct clades. Two clades of tropical isolates grouped together and were affiliated closely with the temperate isolates, while the third tropical clade was more distantly related. Phylogenetic analysis of the ITS regions resulted in similar groupings and variation within and between the temperate and tropical isolates as with the AFLP results. Sequence divergence among isolates and clades was low, with more variation within the tropical isolates than within the temperate isolates. Analysis of other species revealed shorter branch lengths separating temperate and tropical isolates than were observed in comparisons among other phylogenetically closely related species in the genus. Analysis of cox II sequence data was less clear. Although the temperate and tropical isolates grouped together apart from other species, there was no bootstrap support for separating these isolates. Restriction fragment length polymorphism (RFLP) analysis of the ITS regions separated the temperate and tropical isolates, as in the AFLP and ITS phylogenetic analyses. However, RFLP analysis of the cox I and II gene cluster did not distinguish between temperate and tropical isolates. The differences in grouping of isolates in these two RFLP studies should be helpful in identifying isolate subgroups. Our data do not fully clarify whether or not temperate and tropical isolates should be separated into different species. The available worldwide data are incomplete and the full range of variation in the species is not yet known. We suggest refraining from using the epithet P. tropicalis until more data are available.
Overexpression of a Gene Encoding a Catabolite Repression Element in Alternaria citri Causes Severe Symptoms of Black Rot in Citrus Fruit
Authors: Katoh H. , Ohtani K. , Yamamoto H. , Akimitsu K.
Published: May 2007

A gene (AcCreA) encoding a catabolite repression element (CreA) with (two zinc fingers of the Cys2His2 type was isolated from the postharvest fungal pathogen Alternaria citri. The AcCreA overexpression mutant AcOEC2 of A. citri showed normal growth on pectin medium and on segments of peel or the juice sac area from citrus fruit. Production of endopolygalacturonase, an essential virulence factor of this pathogen, was similar in AcOEC2 and the wild type in pectin-containing media. However, addition of glucose to the medium showed that carbon catabolite repression of endopolygalacturonase gene (Acpg1) expression, as well as endopolygalacturonase production, was lost in AcOEC2. The wild-type strain of A. citri causes rot mainly in the central axis of citrus fruit without development of rotting in the juice sac area; however, AcOEC2 caused severe black rot symptoms in both the central axis and juice sac areas. These results indicate that AcCreA-mediated catabolite repression controls the virulence or infection of this pathogen, and that the wild-type A. citri does not cause symptoms in the juice sac area due to carbon catabolite repression by sugars in the juice of the juice sac area.
Genetic and Physical Mapping of the Partial Resistance Gene, Pi34, to Blast in Rice
Authors: Zenbayashi-Sawata K. , Fukuoka S. , Katagiri S. , Fujisawa M. , Matsumoto T. , Ashizawa T. , Koizumi S.
Published: May 2007

Partial resistance to rice blast in the Oryza sativa japonica group cv. Chubu 32 is controlled by Pi34, a major quantitative trait locus (QTL) on chromosome 11, and several uncharacterized QTLs. The objectives of the study were (i) high-resolution genetic and physical mapping of Pi34 and (ii) identification of new QTL imparting resistance to rice blast. Chubu 32 was crossed with a susceptible chromosomal segment substitution line (CSSL) of cv. Koshihikari. From 4,012 of segregating individuals, 213 recombinants in the Pi34 region were screened by using polymerase chain reaction-based markers and tested resistance in the field and greenhouse. The Pi34 locus is located in the 54.1-kb region on the genomic sequence of cv. Nipponbare. We constructed a bacterial artificial chromosome (BAC) library of Chubu 32, selected the clone containing Pi34, and sequenced it. The Pi34 locus consequently was located on an interval of 65.3 kb containing 10 predicted open reading frames (ORFs). Two of these ORFs were predicted only in Chubu 32 and encoded transposable elements. The other eight ORFs were found in both Chubu 32 and Nipponbare and one of them, which encoded an unknown protein, showed significantly different amino acid sequences between two cultivars. The new QTL, Piq6(t), was detected on the short arm of chromosome 6 and the genetic distance of flanking markers was 16.9 centimorgans in Nipponbare. Pi34 and Piq6(t) acted additively on resistance to rice blast but the effect of Piq6(t) was relatively small compared with Pi34.
Defense Responses in Grapevine Leaves AgainstBotrytis cinereaInduced by Application of aPythium oligandrumStrain or Its Elicitin, Oligandrin, to Roots
Authors: Mohamed N. , Lherminier J. , Farmer M.-J. , Fromentin J. , Béno N. , Houot V. , Milat M.-L. , Blein J.-P.
Published: May 2007

Pythium oligandrum is known to display antagonistic activities against several species of pathogenic fungi. It also produces an elicitor of plant defense named oligandrin, which belongs to the elicitin family (10-kDa proteins synthesized by Phytophthora and Pythium species). Here, the potential of P. oligandrum or its purified elicitin to limit the progression of B. cinerea on grapevine leaf and the resulting plant-microorganism interactions are described. P. oligandrum or oligandrin were applied to roots, and changes in the ultrastructure and at the molecular level were examined. When B. cinerea was applied to leaves of pretreated plants, leaf invasion was limited and the protection level reached about 75%. On leaf tissues surrounding B. cinerea inoculation, modifications of cuticle thickness, accumulation of phenolic compounds, and cell wall apposition were observed, indicating that grapevine can be considered reactive to elicitins. No macroscopic hypersensitive reaction associated with the elicitation treatment was observed. At the molecular level, the expression of three defense-related genes (LTP-1, β-1,3-glucanase, and stilbene synthase) was studied. RNAs isolated from B. cinerea-infected leaves of grapevine challenged or not with P. oligandrum or oligandrin were analyzed by real-time reverse transcription-polymerase chain reaction. In grapevine leaves, LTP-1 gene expression was enhanced in response to oligandrin, and RNA transcript levels of β-1,3-glucanase and stilbene synthase increased in response to all treatments with different magnitude. Taken together, these results open new discussion on the concept of plant reactivity to elicitins, which has until now, been mainly based on plant hypersensitive responses.
Forecasting Sclerotinia Disease on Lettuce: A Predictive Model for Carpogenic Germination of Sclerotinia sclerotiorum Sclerotia
Authors: Clarkson John P. , Phelps Kath , Whipps John M. , Young Caroline S. , Smith Julie A. , Watling Martyn
Published: May 2007

A predictive model for production of apothecia by carpogenic germination of sclerotia is presented for Sclerotinia sclerotiorum. The model is based on the assumption that a conditioning phase must be completed before a subsequent germination phase can occur. Experiments involving transfer of sclerotia from one temperature regime to another allowed temperature-dependent rates to be derived for conditioning and germination for two S. sclerotiorum isolates. Although the response of each isolate to temperature was slightly different, sclerotia were fully conditioned after 2 to 6 days at 5°C in soil but took up to 80 days at 15°C. Subsequent germination took more than 200 days at 5°C and 33 to 52 days at 20°C. Upper temperature thresholds for conditioning and germination were 20 and 25°C, respectively. A predictive model for production of apothecia derived from these data was successful in simulating the germination of multiple burials of sclerotia in the field when a soil water potential threshold of between −4.0 and −12.25 kilopascals (kPa) was imposed. The use of a germination model as part of a disease forecasting system for Sclerotinia disease in lettuce is discussed.
Distribution of Phytophthora spp. in Field Soils Determined by Immunoassay
Authors: Miller S. A. , Madden L. V. , Schmitthenner A. F.
Published: May 2007

Populations of Phytophthora spp. were determined by enzyme-linked immunosorbent assay (ELISA) in field soils used for pepper and soybean production in Ohio. Soybean fields were sampled extensively (64 fields, n = 6 samples per field over 2 years) and intensively (4 fields, n = 64 samples per field in 1 year) to assess heterogeneity of P. sojae populations. Four pepper fields (n = 64), three of which had a history of Phytophthora blight (caused by P. capsici), also were sampled intensively during a 6-month period. Mean (m), variance (v), and measures of aggregation (e.g., variance-to-mean ratio [v/m]) of immunoassay values, translated to Phytophthora antigen units (PAU), were related to the disease history in each of the pepper and soybean fields. Mean PAU values for fields in which Phytophthora root rot (soybean) or blight (pepper) had been moderate to severe were higher than in fields in which disease incidence had been low or not observed. A detection threshold value of 11.3 PAU was calculated with values for 64 samples from one pepper field, all of which tested negative for Phytophthora by bioassay and ELISA. Seven of the eight intensively sampled fields contained at least some detectable Phytophthora propagules, with the percentage of positive samples ranging from 1.6 to 73.4. Mean PAU values ranged from 1 to 84 (extensive soybean field sampling), 6 to 24 (intensive soybean field sampling), and 4 to 30 (intensive pepper field sampling); however, variances ranged from 0 to 7,774 (extensive sampling), 30 to 848 (intensive soybean field sampling), and 5 to 2,401 (intensive pepper field sampling). Heterogeneity of PAU was high in most individual soybean and pepper fields, with values of v/m greater than 1, and log(v) increasing with log(m), with a slope of about 2.0. Spatial autocorrelation coefficients were not significant, indicating there was no relationship of PAU values in neighboring sampling units (i.e., field locations) of the intensively sampled fields. Combined results for autocorrelations and v/m values indicate that Phytophthora was highly aggregated in these fields but that the scale of the aggregation (e.g., average focus size) was less than the size of the sampling units. Because of the observed variability, we calculated that sample sizes of 20 or more would be needed to estimate precisely the mean density of Phytophthora in most cases.
Influence of Crop Management on Take-All Development and Disease Cycles on Winter Wheat
Authors: Colbach Nathalie , Lucas Philippe , Meynard Jean-Marc
Published: May 2007

Wheat was assessed at four crop growth stages for take-all (Gaeumannomyces graminis var. tritici) in a series of field trials that studied the effects of five wheat management practices: sowing date, plant density, nitrogen fertilizer dose and form, and removal/burial of cereal straw. An equation expressing disease level as a function of degree days was fitted to the observed disease levels. This equation was based on take-all epidemiology and depended on two parameters reflecting the importance of the primary and secondary infection cycles, respectively. Early sowing always increased disease frequency via primary infection cycle; its influence on the secondary cycle was variable. Primary infection and earliness of disease onset were increased by high density; however, at mid-season take-all was positively correlated to the root number per plant, which was itself negatively correlated to plant density. At late stages of development, neither plant density nor root number per plant had any influence on disease. A high nitrogen dose increased both take-all on seminal roots and severity of primary infection cycle but decreased take-all on nodal roots and secondary infection cycle. Ammonium (versus ammonium nitrate) fertilizer always decreased disease levels and infection cycles, whereas straw treatment (burial versus removal of straw from the previous cereal crop) had no influence.
Toxicity of Glucosinolate Degradation Products from Brassica napus Seed Meal Toward Aphanomyces euteiches f. sp. pisi
Authors: Smolinska U. , Morra M. J. , Knudsen G. R. , Brown P. D.
Published: May 2007

Brassica tissues are potentially useful in the control of Aphanomyces root rot of peas (Pisum sativum), but identity of the responsible compounds and specific impacts of those compounds on the pathogen's infection potential remain uncertain. Brassica napus seed meals and water extracts from these meals were used to determine the effect of glucosinolate hydrolysis products on Aphanomyces euteiches f. sp. pisi. B. napus meal (‘Dwarf Essex’) containing glucosinolates and intact myrosinase, the enzyme responsible for glucosinolate hydrolysis, completely inhibited infection by A. euteiches f. sp. pisi oospores. Water extracts from this meal, likewise, severely inhibited infection by oospores, as well as mycelial growth. Extracts from autoclaved ‘Dwarf Essex’ meal, in which myrosinase was denatured, and a low glucosinolate B. napus variety (‘Stonewall’) produced little disease reduction and had less impact on mycelial growth. Gas chromatographic analysis of Brassica tissues and water extracts confirmed that glucosinolates remained in autoclaved ‘Dwarf Essex’ meal and that ‘Stonewall’ meal contained low glucosinolate concentrations. 5-Vinyloxazolidine-2-thione was identified by mass spectrometry as a dominant glucosinolate hydrolysis product in aqueous extracts of the inhibitory meal. Bioassays conducted with aqueous solutions of this compound reduced mycelial growth, but not to the extent of those from intact ‘Dwarf Essex’ meal. Water-soluble compounds produced from the hydrolysis of glucosinolates in B. napus tissues reduced A. euteiches oospore infection and inhibited mycelial growth, thus, demonstrating potential utility of Brassica species in the control of A. euteiches.
Clonality in Sclerotinia sclerotiorum on Infected Cabbage in Eastern North Carolina
Authors: Cubeta M. A. , Cody B. R. , Kohli Y. , Kohn L. M.
Published: May 2007

Eighty-four isolates of Sclerotinia sclerotiorum from four cabbage production fields in North Carolina and 16 isolates from an experimental cabbage field plot in Louisiana were DNA-fingerprinted and tested for mycelial compatibility. In a comparison with 594 unique DNA fingerprints of S. sclerotiorum from Canadian canola, no fingerprints were shared among Canadian, North Carolina, and Louisiana populations. DNA fingerprints from the North Carolina sample were distinctive from those of the Canadian and Louisiana samples, with significantly more hybridizing fragments in the 7.7- to 18-kilobase range. Forty-one mycelial compatibility groups (MCGs) and 50 unique DNA fingerprints were identified from the North Carolina sample. Three MCGs and three fingerprints were identified from the Louisiana sample. From the North Carolina sample, 32 MCGs were each associated with a unique fingerprint; of these, there were 11 clones (i.e., cases in which two or more isolates belonged to the same MCG and shared the same DNA fingerprint). Six clones sampled from two or more fields represented approximately 29% of the total sample (24 of 84 isolates), with six clones recovered from fields 75 km apart. There were 10 cases in which one MCG was associated with more than one DNA fingerprint and two cases in which one DNA fingerprint was associated with more than one MCG. The small sample from Louisiana was strictly clonal. The North Carolina sample had a clonal component, but deviated from one-to-one association of MCG with DNA fingerprint to an extent consistent with more recombination or transposition than the other two populations sampled.
Reduced Fusarium Ear Rot and Symptomless Infection in Kernels of Maize Genetically Engineered for European Corn Borer Resistance
Authors: Munkvold G. P. , Hellmich R. L. , Showers W. B.
Published: May 2007

Field experiments were conducted in 1994, 1995, and 1996 to evaluate the incidence and severity of Fusarium ear rot and the incidence of symp-tomless Fusarium infection in kernels of maize hybrids genetically engineered with Bacillus thuringiensis genes encoding for the δ-endotoxin CryIA(b). Treatments included manual infestation with European corn borer (ECB) larvae and insecticide applications to limit ECB activity to specific maize growth stages or mimic standard ECB control practices. Fusarium symptoms and infection were affected by the specific cryIA(b) transformation used in each hybrid that determines tissue-specific expression of CryIA(b). In hybrids expressing CryIA(b) in kernels, incidence and severity of Fusarium ear rot and incidence of symptomless kernel infection were reduced compared with near-isogenic hybrids lacking cryIA(b) genes. In plants that were manually infested with ECB, ear rot incidence was reduced by 87, 58, and 68%; severity was reduced by 96, 54, and 64%; and incidence of kernel infection by Fusarium species was reduced by 17, 38, and 38% in 1994, 1995, and 1996, respectively. Results were similar in treatments that were not manually infested, but differences between transgenic and nontransgenic hybrids were smaller. Most kernel infection was due to F. moniliforme, F. proliferatum, and F. subglutinans (section Liseola) collectively, and it was within this group that transgenic hybrids exhibited reduced infection. Expression of CryIA(b) in plant tissues other than kernels did not consistently affect Fusarium symptoms or infection. Disease incidence was positively correlated with ECB damage to kernels. Insecticide applications also reduced Fusarium symptoms and infection when applied to nontransgenic plants.
Crab Apple Blossoms as a Model for Research on Biological Control of Fire Blight
Authors: Pusey P. L.
Published: May 2007

Nonseasonal availability of pomaceous flowers could improve laboratory detection and prefield testing of biocontrol agents for fire blight of pear and apple. Crab apple was selected as a model because of its high flower productivity on 1-year-old wood, high susceptibility to fire blight, and availability from nurseries. Cultivars Manchurian and Snowdrift were manipulated to bloom once by transferring dormant nursery trees from a cold room to a greenhouse and a second time by defoliating trees and applying 1% cytokinin and 0.1% gibberellins to the buds with a brush. Different sets of trees were induced at different times to bloom, so that flowers were produced 12 months in the year. When known bacterial antagonists (Erwinia herbicola strain C9-1 and Pseudomonas fluorescens strain A506) were applied alone or in combination to the stigmas of detached crab apple blossoms prior to inoculation with the pathogen (E. amylovora strain Ea153), population interactions over time were comparable to those reported in previous studies involving pear or apple. In a subsequent series of experiments, the relative effects of 12 bacterial strains on stigmatic populations of strain Ea153 were similar for detached blossoms of crab apple in the laboratory, blossoms of intact crab apple trees in the greenhouse, and blossoms of pear and apple in the field. Additionally, when stigmas of detached crab apple blossoms were inoculated with antagonists (strains C9-1 and A506) and the pathogen, and later subjected to a 24-h wetting period, bacterial populations in the flower hypanthium increased and disease was suppressed. These studies indicate that crab apple blossoms can serve as a suitable model for year-round evaluation and study of biocontrol agents for fire blight.
Xanthomonas translucens from Small Grains: Diversity and Phytopathological Relevance
Authors: Bragard C. , Singer E. , Alizadeh A. , Vauterin L. , Maraite H. , Swings J.
Published: May 2007

Sixty-eight presumptive Xanthomonas translucens strains isolated from 15 small grains or grass species were studied by pathogenicity tests on barley, bread wheat, oat, and bromegrass species, and also by AFLP, analysis of fatty acid methyl esters (FAME), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of protein extracts. The X. translucens strains were divided into three pathogenicity types based on differences observed on barley and bread wheat. Two unspeciated strains producing atypical symptoms formed a fourth pathogenicity type. Pathogenicity on oat and bromegrass species varied within these types. Clusterings observed by AFLP analysis and, to a lesser extent, by FAME analysis were consistent with these pathogenicity groupings. The current results, as well as those of previous restriction fragment length polymorphism analyses of the same strains, support the recent reclassification of X. translucens pv. translucens and X. translucens pv. hordei as true synonyms. X. translucens pv. cerealis, X. translucens pv. translucens, and X. translucens pv. undulosa cluster in different groups by AFLP and FAME analyses. Even though distinction by simple biochemical tests is not clear-cut, the data indicate that the pathovars cerealis, translucens, and undulosa correspond to true biological entities.
Seedborne Fungal Contamination: Consequences in Space-Grown Wheat
Authors: Bishop Deborah L. , Levine Howard G. , Kropp Bradley R. , Anderson Anne J.
Published: May 2007

Plants grown in microgravity are subject to many environmental stresses that may promote microbial growth and result in disease symptoms. Wheat (cv. Super Dwarf) recovered from an 8-day mission aboard a NASA (National Aeronautics and Space Administration) space shuttle showed disease symptoms, including girdling of leaf sheaths and chlorosis and necrosis of leaf and root tissues. A Neotyphodium species was isolated from the seed and leaf sheaths of symptomatic wheat used in the spaceflight mission. Certain isozymes of a peroxidase unique to extracts from the microgravity-grown plants were observed in extracts from earth-grown Neotyphodium-infected plants but were not present in noninfected wheat. The endophytic fungus was eliminated from the wheat seed by prolonged heat treatment at 50°C followed by washes with water at 50°C. Plants from wheat seed infected with the Neotyphodium endophyte were symptomless when grown under greenhouse conditions, whereas symptoms appeared after only 4 days of growth in closed containers. Disease spread from an infected plant to noninfected plants in closed containers. Dispersion via spores was found on asymptomatic plants at distances of 7 to 18 cm from infected plants. The size and shape of the conidia, mycelia, and phialide-bearing structures and the ability to grow rapidly on carbohydrates, especially xylose, resembled the characteristics of N. chilense, which is pathogenic on orchard grass, Dactylis glomerata. The Neotyphodium wheat isolate caused disease symptoms on other cereals (wheat cv. Malcolm, orchard grass, barley, and maize) grown in closed containers.
Isolation and Characterization of an Endopolygalacturonase from Cochliobolus sativus and a Cytological Study of Fungal Penetration of Barley
Authors: Clay Ronald P. , Bergmann Carl W. , Fuller Melvin S.
Published: May 2007

Endopolygalacturonase (EPG) of Cochliobolus sativus was produced in shake culture and purified by high-performance liquid chromatography. The enzyme had a molecular mass of 34,000 Da, an isoelectric point in the range of 9.0 to 9.5, exhibited endo activity, was nongly-cosylated, and was inhibited by polygalacturonase-inhibiting proteins from bean, pear, and tomato. The amino terminus contained a 14 amino acid region homologous to a region at the N terminus of an EPG of C. carbonum. C. sativus EPG-specific monoclonal antibodies (MAbs) were generated. Western blot analysis confirmed the specificity of the antibodies for the EPG and detected the enzyme in an extract from Hordeum vulgare (cv. Golden Promise) leaf segments infected with C. sativus. Using conventional immunogold and enzyme-gold cytochemical methods, homogalacturonan, esterified pectin, and cellulose were localized in healthy and infected barley leaf epidermis at the electron microscope level. Additionally, the leaf cell wall polysaccharides recognized by purified C. sativus EPG were localized at the electron microscope level, using the purified enzyme as a primary cytochemical reagent, followed by a gold-labeled MAb specific for the enzyme. Loss of polygalacturonic acid in the vicinity of the invading pathogen was visualized cytochemically at the electron microscope level. These observations suggest the involvement of EPG during host penetration by the fungus.
Virulence and Molecular Diversity in Colletotrichum lindemuthianum from South, Central, and North America
Authors: Balardin R. S. , Jarosz A. M. , Kelly J. D.
Published: May 2007

Isolates of Colletotrichum lindemuthianum (138 total) from Argentina, Brazil, the Dominican Republic, Honduras, Mexico, and the United States were characterized into 41 races based on virulence to 12 differential cultivars of Phaseolus vulgaris. These 41 races were categorized into two groups: those found over a wide geographic area and those restricted to a single country. Races 7, 65, and 73 were widespread. Race 73 was the most common (28%). Race 7 was found once in Argentina and Mexico but at a higher frequency in the United States. Race 65 was found repeatedly in Brazil and the United States. Although 39% of the races were detected repeatedly and three races were widespread, no race was isolated from both P. vulgaris gene pools. Phenetic analyses showed no obvious patterns correlated with virulence clusters. No geographic pattern was evident. Molecular polymorphism generated by random amplified polymorphic DNA confirmed the extensive variability in virulence of C. lindemuthianum. Virulence phenotypes were grouped into 15 clusters. The two largest clusters contained isolates from all the geographic regions sampled. Molecular polymorphism was observed among isolates from races 65 and 73 within and among countries, except among Bra-zilian isolates of race 65. The genetic diversity of C. lindemuthianum was greatest in Mexico and Honduras. Our data suggest that C. lindemuthianum may not be highly structured to specific Phaseolus gene pools.
Variation at the b Mating Type Locus of Ustilago maydis
Authors: Zambino Paul , Groth James V. , Lukens Lewis , Garton James R. , May Georgiana
Published: May 2007

Population level diversity at the Ustilago maydis b mating type locus was determined in samples from four Minnesota locations using a combination of plate mating techniques and a polymerase chain reaction (PCR)-based assay. The PCR method allows rapid identification of b types from samples of natural populations and utilizes the hypervariable regions of the b locus that determine mating type specificity. Results demonstrated high levels of b diversity within populations, with one population yielding 17 of the total 18 b types found in the study. Pairwise GST values were in the range of 0.02 to 0.05, and common b mating types were found across broad geographic distances. These data demonstrated that very low levels of differentiation among U. maydis populations occur with respect to b locus variation. Consistent with frequency-dependent selection models, b types were represented at approximately equal frequencies within the entire Minnesota population. However, neutral evolutionary models for patterns of geographic distribution and variation at b cannot be entirely excluded. The importance to agricultural practices of understanding population genetic processes is discussed.
Acquisition of Resistance to Sterol Demethylation Inhibitors by Populations of Venturia inaequalis
Authors: Kunz Stefan , Deising Holger , Mendgen Kurt
Published: May 2007

Acquisition of resistance to sterol demethylation inhibitors (DMIs) by populations of Venturia inaequalis was investigated using a microscopical method developed by C. Siebels and K. Mendgen. Microscopical analysis of conidiophore formation enabled the earlier detection of resistance and a clearer distinction between DMI-resistant and DMI-sensitive populations than other in vivo methods commonly used to analyze inhibitory effects of fungicides. In addition, because observations were made on the level of individuals, quantitative measures of the composition of conidial populations were obtained. The development of DMI sensitivity was followed over a period of 3 years in control apple orchards that had never been treated with fungicides and in orchards with DMI history. The 50% effective dose values determined by microscopical evaluation of conidio-phore development for untreated populations revealed the baseline sensitivities of 0.3, 0.96, 0.09, 1.22, and 1.92 mg/liter for flusilazole, fenarimol, difenoconazole, tebuconazole, and pyrifenox, respectively. As compared with the baseline sensitivity, all populations with DMI history showed significant resistance to flusilazole. A strong nonlinear correlation (R = 0.96) was found between the resistance factors and the sum of all DMI treatments of the 3 years before taking the sample. According to this correlation, resistance can be expected in all apple orchards of the fruit-growing area along Lake Constance, Germany, in which more than two DMI treatments per season have been applied. Due to cross-resistance, the recently introduced DMI fungicides difenoconazole, tebuconazole, and pyrifenox did not allow the control of V. inaequalis populations resistant to flusilazole.
Structural and Physical Properties of a Necrosis-Inducing Toxin from Pyrenophora tritici-repentis
Authors: Zhang Hui-Fen , Francl Leonard J. , Jordahl James G. , Meinhardt Steven W.
Published: May 2007

Cultivar-specific toxic metabolites of Pyrenophora tritici-repentis are involved in the appearance of necrotic and chlorotic foliar lesions characteristic of tan spot. A P. tritici-repentis necrosis-inducing toxin, Ptr necrosis toxin, was purified from isolate 86-124, sequenced by gas-phase amino acid microsequencing, and characterized by circular dichroism (CD) spectroscopy and isoelectric focusing. The purified protein had a similar amino acid composition and molecular weight as previously reported. Analysis of the CD spectrum from 178 to 250 nm indicated a protein consisting of 13% α-helix, 36% antiparallel β-sheet, 25% turns, and 25% other structures. The Ptr necrosis toxin from isolate 86-124 has an isoelectric point near pH 10. Using overlapping proteolytic fragments obtained from the toxin, a sequence of 101 continuous amino acids was obtained, but the amino terminus was blocked and 9 to 16 amino acids could not be sequenced. Secondary structure prediction based on the amino acid sequence indicated a β-sheet protein with little α-helix, which is in agreement with the structure determined by CD spectroscopy. Sequence analysis indicated the presence of a possible membrane adhesion site and several possible phosphorylation sites that may be involved in phytotoxicity.
Inducers of Aflatoxin Biosynthesis from Colonized Maize Kernels Are Generated by an Amylase Activity from Aspergillus flavus
Authors: Woloshuk C. P. , Cavaletto J. R. , Cleveland T. E.
Published: May 2007

Aflatoxin biosynthesis was induced by compounds in filtrates (EF) obtained from cultures consisting of ground maize kernels colonized by Aspergillus flavus. The inducing activity increased to a maximum at 4 days of incubation and then decreased. Amylase activity was detected in the EF, suggesting that the inducers are products of starch degradation (glucose, maltose, and maltotriose). Analysis of the enzyme by isoelectric focusing electrophoresis indicated a single α-amylase with a pI of 4.3. No maltase or amyloglucosidase was detected in the EF. High-pressure liquid chromatography analysis of the EF indicated the presence of glucose, maltose, and maltotriose in near-equal molar concentrations (about 15 mM). With a β-glucuronidase (GUS) reporter assay consisting of A. flavus transformed with an aflatoxin gene promoter-GUS reporter gene fusion to monitor induction of aflatoxin biosynthesis, the minimum concentration of glucose, maltose, or maltotriose that induced measurable GUS activity was determined to be 1 mM. These results support the hypothesis that the best inducers of aflatoxin biosynthesis are carbon sources readily metabolized via glycolysis. They also suggest that α-amylase produced by A. flavus has a role in the induction of aflatoxin biosynthesis in infected maize kernels.
Adaptation of Soil Solarization to the Integrated Management of Soilborne Pests of Tomato Under Humid Conditions
Authors: Chellemi D. O. , Olson S. M. , Mitchell D. J. , Secker I. , McSorley R.
Published: May 2007

Soil solarization was shown to be cost effective, compatible with other pest management tactics, readily integrated into standard production systems, and a valid alternative to preplant fumigation with methyl bromide under the tested conditions. Solarization using clear, photoselective, or gas-impermeable plastic was evaluated in combination with metham sodium, 1,3-dichloropropene + chloropicrin, methyl bromide + chloropicrin, pebulate, or cabbage residue. Strip solarization, applied to 20-cm-high, 0.9-m-wide beds, was conducted to achieve compatibility with standard production practices and resulted in soil temperatures 2 to 4°C above those temperatures resulting when using conventional flatbed solarization. Soil temperatures were 1 to 2°C higher at the edges of the raised beds, eliminating any border effects associated with solarization. Following a 40- to 55-day solarization period, the plastic was painted white and used as a production mulch for a subsequent tomato crop. The incidence of Southern blight and the density of Paratrichodorus minor and Criconemella spp. were lower (P < 0.05) in solarized plots. No differences (P < 0.05) in the incidence of Fusarium wilt and the density of nutsedge and Helicotylenchus spp. were observed between plots receiving solarization and plots fumigated with a mixture of methyl bromide + chloropicrin. The severity of root galling was lower (P < 0.05) when soil solarization was combined with 1,3-dichloropropene + chloropicrin (16.2 + 3.4 g/m2) and a gas-impermeable film. The incidence of bacterial wilt was not affected by soil treatments. Marketable yields in plots using various combinations of soil solarization and other tactics were similar (P < 0.05) to yields obtained in plots fumigated with methyl bromide + chloropicrin. The results were validated in several large scale field experiments conducted by commercial growers.
Ultrastructure, Morphology, and Sporogenesis of Pasteuria penetrans
Authors: Chen Z. X. , Dickson D. W. , Freitas L. G. , Preston J. F.
Published: May 2007

Pasteuria penetrans is a bacterial parasite of root-knot nematodes that shows great potential as a biocontrol agent. Scanning and transmission electron microscopy were used to study the ultrastructure, morphology, and sporogenesis of four isolates of P. penetrans. The effects of different Meloidogyne spp. and tobacco cultivars on sporangium size and morphology of endospores attached to the cuticle of second-stage juveniles (J2) of root-knot nematodes also were investigated. The P. penetrans isolates and their origins were P-20 from M. arenaria race 1 in Levy County, FL; P-100 from Meloidogyne sp. in Pasco County, FL; B-4 from Pratylenchus scribneri in Seminole County, FL; and P-120 from Meloidogyne spp. in Alachua County, FL. Sporangia of the four isolates were identical morphologically and similar in their dimensions, ranging from 2.39 to 3.42 μm in diameter and from 1.38 to 2.38 μm in height. Different Meloidogyne spp. and tobacco cultivars had no effect on sporangium size. Endospores attached to J2 were visualized in three forms: endospores retaining the sporangium wall, endospores covered with a thin exosporium, and endospores without covering. Sporogenesis of P. penetrans was similar to that of other gram-positive bacteria and generally matched the seven-stage scheme reported for Bacillus thuringiensis.
Influence of CaCl2onPenicillium digitatum, Grapefruit Peel Tissue, and Biocontrol Activity ofPichia guilliermondii
Authors: Droby S. , Wisniewski M. E. , Cohen L. , Weiss B. , Touitou D. , Eilam Y. , Chalutz E.
Published: May 2007

Interactions between CaCl2, grapefruit peel tissue, Penicillium digitatum, and the yeast antagonist Pichia guilliermondii strain US-7 were investigated. Application of 68 or 136 mM CaCl2to grapefruit surface wounds reduced the incidence of green mold caused by Penicillium digitatum by 43 to 52%. In laboratory tests, a cell suspension (107cells/ml) of Pichia guilliermondii containing either 68 or 136 mM CaCl2reduced the incidence of green mold from 27 to 3%. In large scale tests, dip application of 136 mM CaCl2with US-7 (107cells/ml) significantly decreased the number of wounds infected by Penicillium digitatum. CaCl2, with or without yeast cells, stimulated ethylene production in grapefruit tissue. Increasing concentrations of CaCl2resulted in decreased spore germination and germ tube elongation of Penicillium digitatum. Pectinolytic activity of crude enzyme preparations of Penicillium digitatum was also inhibited by the presence of increasing concentrations of CaCl2. US-7 exhibited a strong ability to maintain cytosolic Ca2+homeostasis at levels that did not exceed 1.4 μM when exposed to 150 mM CaCl2. On the other hand, strain 114 of Debaryomyces hansenii, which failed to give any protection against infection by Penicillium digitatum, showed reduced capacity to maintain Ca2+homeostasis. The effect of calcium in reducing infection of grapefruit wounds by Penicillium digitatum could be due to direct effects on host tissue (making cell walls more resistant to enzymatic degradation) or the pathogen (interfering with spore germination, growth, and inhibition of fungal pectinolytic enzymes). Alternatively, the ability of US-7 to maintain calcium homeostasis may allow it to grow or assist in its competitive ability in a microenvironment that, because of high levels of calcium ions, is inhibitory to growth of the green mold pathogen.