bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Host Species Specificity of Magnaporthe grisea Toward Foxtail Millet Using a Genetic Cross Between Isolates from Wheat and Foxtail Millet
Authors: Murakami J. , Tomita R. , Kataoka T. , Nakayashiki H. , Tosa Y. , Mayama S.
Published: May 2007

Host species specificity of Magnaporthe grisea toward foxtail millet was analyzed using F1 cultures derived from a cross between a Triticum isolate (pathogenic on wheat) and a Setaria isolate (pathogenic on foxtail millet). On foxtail millet cvs. Beni-awa and Oke-awa, avirulent and virulent cultures segregated in a 1:1 ratio, suggesting that a single locus is involved in the specificity. This locus was designated as Pfm1. On cv. Ki-awa, two loci were involved and one of them was Pfm1. The other locus was designated as Pfm2. Interestingly, Pfm1 was not involved in the pathogenic specificity on cv. Kariwano-zairai. These results suggest that there is no “master gene” that determines the pathogenic specificity on all foxtail millet cultivars and that the species specificity of M. grisea toward foxtail millet is governed by cultivar-dependent genetic mechanisms that are similar to gene-for-gene interactions controlling race-cultivar specificity.
Thrips Resistance in Pepper and Its Consequences for the Acquisition and Inoculation of Tomato spotted wilt virus by the Western Flower Thrips
Authors: Maris P. C. , Joosten N. N. , Peters D. , Goldbach R. W.
Published: May 2007

Different levels of thrips resistance were found in seven Capsicum accessions. Based on the level of feeding damage, host preference, and host suitability for reproduction, a thrips susceptible and a resistant accession were selected to study their performance as Tomato spotted wilt virus (TSWV) sources and targets during thrips-mediated virus transmission. Vector resistance did not affect the virus acquisition efficiency in a broad range of acquisition access periods. Inoculation efficiency was also not affected in short inoculation periods, but was significantly lower on plants of the thrips resistant accession during longer inoculation access periods. Under the experimental conditions used, the results obtained show that transmission of TSWV is little affected by vector resistance. However, due to a lower reproduction rate on resistant plants and a lower preference of thrips for these plants, beneficial effects of vector resistance might be expected under field conditions.
Identification, Phylogenetic Analysis, and Biological Characterization of Serratia marcescens Strains Causing Cucurbit Yellow Vine Disease
Authors: Rascoe J. , Berg M. , Melcher U. , Mitchell F. L. , Bruton B. D. , Pair S. D. , Fletcher J.
Published: May 2007

A serious vine decline of cucurbits known as cucurbit yellow vine disease (CYVD) is caused by rod-shaped bacteria that colonize the phloem elements. Sequence analysis of a CYVD-specific polymerase chain reaction (PCR)-amplified 16S rDNA product showed the microbe to be a γ-proteobacterium related to the genus Serratia. To identify and characterize the bacteria, one strain each from watermelon and zucchini and several noncucurbit-derived reference strains were subjected to sequence analysis and biological function assays. Taxonomic and phylogenetic placement was investigated by analysis of the groE and 16S rDNA regions, which were amplified by PCR and directly sequenced. For comparison, eight other bacterial strains identified by others as Serratia spp. also were sequenced. These sequences clearly identified the CYVD strains as Serratia marcescens. However, evaluation of metabolic and biochemical features revealed that cucurbit-derived strains of S. marcescens differ substantially from strains of the same species isolated from other environmental niches. Cucurbit strains formed a distinct cluster, separate from other strains, when their fatty acid methyl ester profiles were analyzed. In substrate utilization assays (BIOLOG, Vitek, and API 20E), the CYVD strains lacked a number of metabolic functions characteristic for S. marcescens, failing to catabolize 25 to 30 compounds that were utilized by S. marcescens reference strains. These biological differences may reflect gene loss or repression that occurred as the bacterium adapted to life as an intracellular parasite and plant pathogen.
Genetic Variability and Host Specialization in the Latin American Clade of Ceratocystis fimbriata
Authors: Baker Christine J. , Harrington Thomas C. , Krauss Ulrike , Alfenas Acelino C.
Published: May 2007

The Ceratocystis fimbriata complex includes many undescribed species that cause wilt and canker diseases of many economically important plants. Phylogenetic analyses of DNA sequences have delineated three geographic clades within Ceratocystis fimbriata. This study examined host specialization in the Latin American clade, in which a number of lineages were identified using sequences of the internal transcribed spacer (ITS) region of the rDNA. Three host-associated lineages were identified from cacao (Theobroma cacao), sweet potato (Ipomoea batatas), and sycamore (Platanus spp.), respectively. Isolates from these three lineages showed strong host specialization in reciprocal inoculation experiments on these three hosts. Six cacao isolates from Ecuador, Trinidad, and Columbia differed genetically from other cacao isolates and were not pathogenic to cacao in inoculation tests. Further evidence of host specialization within the Latin American clade of Ceratocystis fimbriata was demonstrated in inoculation experiments in growth chambers using sweet potato, sycamore, Colocasia esculenta, coffee (Coffea arabica), and mango (Mangifera indica) plants; inoculation experiments in Brazil using Brazilian isolates from cacao, Eucalyptus spp., mango, and Gmelina arborea; and inoculation experiments in Costa Rica using Costa Rican isolates from cacao, coffee, and Xantho-soma sp. Hosts native to the Americas appeared to be colonized by only select pathogen genotypes, whereas nonnative hosts were colonized by several genotypes. We hypothesize that local populations of Ceratocystis fimbriata have specialized to different hosts; some of these populations are nascent species, and some host-specialized genotypes have been moved to new areas by humans.
Isolation and Characterization of Rhizobacteria from Composts That Suppress the Severity of Bacterial Leaf Spot of Radish
Authors: Krause M. S. , De Ceuster T. J. J. , Tiquia S. M. , Michel F. C. , Madden L. V. , Hoitink H. A. J.
Published: May 2007

Composts can induce systemic resistance in plants to disease. Unfortunately, the degree of resistance induced seems highly variable and the basis for this effect is not understood. In this work, only 1 of 79 potting mixes prepared with different batches of mature, stabilized composts produced from several different types of solid wastes suppressed the severity of bacterial leaf spot of radish caused by Xanthomonas campestris pv. armoraciae compared with disease on plants produced in a nonamended sphagnum peat mix. An additional batch of compost-amended mix that had been inoculated with Trichoderma hamatum 382 (T382), which is known to induce systemic resistance in plants, also suppressed the disease. A total of 11 out of 538 rhizobacterial strains isolated from roots of radish seedlings grown in these two compostamended mixes that suppressed bacterial leaf spot were able to significantly suppress the severity of this disease when used as inoculum in the compost-amended mixes. The most effective strains were identified as Bacillus sp. based on partial sequencing of 16S rDNA. These strains were significantly less effective in reducing the severity of this disease than T382. A combined inoculum consisting of T382 and the most effective rhizobacterial Bacillus strain was less effective than T382 alone. A drench applied to the potting mix with the systemic acquired resistance-inducing chemical acibenzolar-S-methyl was significantly more effective than T382 in several, but not all tests. We conclude that systemic suppression of foliar diseases induced by compost amendments is a rare phenomenon. Furthermore, inoculation of compost-amended potting mixes with biocontrol agents such as T382 that induce systemic resistance in plants can significantly increase the frequency of systemic disease control obtained with natural compost amendments.
Joint Action of Disease Control Measures: A Case Study of Alternaria Leaf Blight of Carrot
Authors: Ben-Noon E. , Shtienberg D. , Shlevin E. , Dinoor A.
Published: May 2007

The efficacy of chemical (i.e., foliar fungicide sprays), genetic (i.e., moderately resistant cultivars), and cultural (i.e., drip-irrigation system) control measures was quantified individually and in combination in the management of Alternaria dauci, the causal agent of Alternaria leaf blight of carrot. Whereas host resistance and drip irrigation affected both the time of disease onset and the rate of disease progression, chemical control affected only the latter. In all cases, a single control measure did not provide an acceptable level of disease suppression. Control efficacy values (based on the relative area under the disease progress curve) for chemical, genetic, and cultural control were 58 ± 11, 39 ± 20, and 60 ± 22%, respectively (values are means ± standard error). By contrast, implementing two control measures concurrently always improved disease suppression significantly compared with the individual measures. Control efficacy values were 91 ± 8% for the integration of chemical and genetic measures and 82 ± 23% for the integration of chemical and cultural measures. Moreover, yields in plots protected by two control measures simultaneously were higher by 10.1 to 28.6 t/ha than those in the respective plots protected by single measures. The joint effect of chemical control and host resistance was additive, whereas that of chemical control and drip irrigation was synergistic in most cases. A literature review was performed to determine if these findings represent a general relationship between chemical and genetic, and chemical and cultural measures. Based on 19 reviewed cases, it was concluded that additive effects are the rule and synergistic or antagonistic effects are the exception. Synergistic effects of two control measures were observed when one control measure improved the efficacy of the other directly or when one control measure induced host resistance or predisposed the pathogen to increased susceptibility. These results may enable a more effective selection of candidate control measures for integration in the future.
Assessment of Dothistroma Needle Blight of Pinus radiata Using Airborne Hyperspectral Imagery
Authors: Coops N. , Stanford M. , Old K. , Dudzinski M. , Culvenor D. , Stone C.
Published: May 2007

Dothistroma needle blight is a serious foliar disease in Australian Pinus radiata plantations causing defoliation, decreased productivity and, in extreme cases, tree death. Conventional methods of monitoring forest health such as aerial survey and ground assessments are labor intensive, time consuming, and subjective. Remote sensing provides a synoptic view of the canopy and can indicate areas affected by damaging agents such as pests and pathogens. Hyperspectral airborne remote sensing imagery (CASI-2) was acquired over pine stands in southern New South Wales, Australia which had been ground assessed and ranked on an individual tree basis, according to the extent of Dothistroma needle blight. A series of spectral indices were tested using two different approaches for extracting crown-scale reflectance measurements and relating these to ground-based estimates of severity. Dothistroma needle blight is most severe in the lower crown and statistically significant relationships were found between crown reflectance values and ground estimates using a ‘halo’ approach (which ignored each tree crown's brightest central pixels). Independent accuracy assessment of the method indicated that the technique could successfully detect three levels of Dothistroma needle blight infection with an accuracy of over 70%.
Effect of Germination Initiation on Competitive Capacity of Trichoderma atroviride P1 Conidia
Authors: Hjeljord Linda Gordon , Tronsmo Arne
Published: May 2007

Trichoderma biocontrol isolates are most effective as highly concentrated inocula. Their antagonism to other fungi may be a result of pregermination respiration. In a nutrient-rich medium, almost all Trichoderma atroviride P1 (P1) conidia initiated germination processes and increased respiration, even in dense suspensions. When 1 × 107 P1 conidia/ml were coinoculated with 1 × 105 Botrytis cinerea conidia/ml, dissolved oxygen fell to <1% within 2 h and the pathogen failed to germinate. More dilute P1 suspensions consumed oxygen slowly enough to allow coinoculated B. cinerea to germinate. On nutrient-poor media, fewer P1 conidia initiated germination. Oxygen consumption by the inoculum and inhibition of B. cinerea were enhanced when P1 conidia were nutrient activated before inoculation. Pregermination respiration also affected competitive capacity of the antagonist on solid substrates, where respiratory CO2 stimulated germination rate and initial colony growth. These parameters were directly correlated with inoculum concentration (R2 ≥ 0.97, P < 0.01). After initiating germination, Trichoderma conidia became more sensitive to desiccation and were killed by drying after only 2 h of incubation on a nutrient-rich substrate at 23°C. These results indicate that nutrient-induced changes preceding germination in Trichoderma conidia can either enhance or decrease their biological control potential, depending on environmental conditions in the microhabitat.
Microcycle Conidiation in Cercospora zeae-maydis
Authors: Lapaire Carrie L. , Dunkle Larry D.
Published: May 2007

Conidia of Cercospora zeae-maydis are the primary inoculum causing gray leaf spot of maize. On nutrient-deficient substrates, but not on water on the leaf surface, conidia germinate and develop secondary conidia on conidiophores produced from germ tubes or conidial cells. A population of conidia increases its numbers more than twofold by 2 days on the surface of a water droplet and by fourfold on trichomes. This microcycle conidiation is suppressed by hydrogen peroxide and ammonium compounds but not by nitrate compounds, amino acids, or simple sugars. Microcycle conidiation is sensitive to α-amanitin and cycloheximide, suggesting that new RNA and proteins must be synthesized. Upon transfer from a humid to a dry atmosphere, secondary conidia and conidiophores dehydrate and collapse. Mature, dehydrated, secondary conidia are liberated by wind speeds approximately one-third those required to liberate hydrated conidia. The dispersed secondary conidia can rehydrate and germinate normally. Because this microcycle conidiation occurs at the expense of endogenous reserves, the ability to produce secondary conidia is lost after four successive cycles without a period of growth on nutrient media. This alternative method of maintaining inoculum potential during periods of fluctuating relative humidity may have epidemiological consequences when primary conidia fail to infect.
Temporal Dynamics of Corn Flea Beetle Populations Infested withPantoea stewartii, Causal Agent of Stewart's Disease of Corn
Authors: Esker P. D. , Nutter F. W.
Published: May 2007

In order to better understand the epidemiology of the Stewart's disease of corn pathosystem, quantitative information concerning the temporal dynamics of the amount of pathogen inoculum present in the form of Pantoea stewartii-infested corn flea beetles (Chaetocnema pulicaria) is needed. Temporal changes in the proportion of P. stewartii-infested corn flea beetle populations were monitored by testing individual corn flea beetles for the presence of P. stewartii using a peroxidase-labeled, enzyme-linked immunosorbent assay. Approximately 90 corn flea beetles were collected each week from seven locations in Iowa from September 1998 through October 2000 using sweep nets. The proportion of P. stewartii-infested beetles at the end of the 1998 growing season ranged from 0.04 to 0.19. In spring 1999, the proportion of overwintering adult corn flea beetles infested with P. stewartii ranged from 0.10 to 0.11 and did not differ significantly from the previous fall based on χ2. During the 1999 corn-growing season, the proportion of infested corn flea beetles ranged from 0.04 to 0.86, with the highest proportions occurring in August. In fall 1999, the proportion of beetles infested with P. stewartii ranged from 0.20 to 0.77. In spring 2000, the proportion of overwintering adult corn flea beetles infested with P. stewartii ranged from 0.08 to 0.30; these proportions were significantly lower than the proportions observed in fall 1999 at Ames, Chariton, and Nashua. During the 2000 corn-growing season, the proportion of P. stewartii-infested corn flea beetles ranged from 0.08 to 0.53, and the highest observed proportions again occurred in August. Corn flea beetle populations sampled in late fall 2000 had proportions of infested beetles ranging from 0.08 to 0.20. This is the first study to quantify the temporal population dynamics of P. stewartii-infested C. pulicaria populations in hybrid corn and provides new quantitative information that should be useful in developing risk models to predict the seasonal and site-specific risks associated with Stewart's disease of corn.
An Oligonucleotide Array for the Identification and Differentiation of Bacteria Pathogenic on Potato
Authors: Fessehaie Anania , De Boer Solke H. , Lévesque C. André
Published: May 2007

Oligonucleotides, 16 to 24 bases long, were selected from the 3′ end of the 16S gene and the 16S–23S intergenic spacer regions of bacteria pathogenic on potato, including Clavibacter michiganensis subsp. sepedonicus, Ralstonia solanacearum, and the pectolytic erwinias, including Erwinia carotovora subsp. atroseptica and carotovora and E. chrysanthemi. Oligonucleotides were designed and formatted into an array by pin spotting on nylon membranes. Genomic DNA from bacterial cultures was amplified by polymerase chain reaction using conserved ribosomal primers and labeled simultaneously with digoxigenin-dUTP. Hybridization of amplicons to the array and subsequent serological detection of digoxigenin label revealed different hybridization patterns that were distinct for each species and subspecies tested. Hybridization of amplicons generally was restricted to appropriate homologous oligonucleotides and cross-hybridization with heterologous oligonucleotides was rare. Hybridization patterns were recorded as separate gray values for each hybridized spot and revealed a consistent pattern for multiple strains of each species or subspecies isolated from diverse geographical regions. In preliminary tests, bacteria could be correctly identified and detected by hybridizing to the array amplicons from mixed cultures and inoculated potato tissue.
A Temperature and Leaf Wetness Duration-Based Model for Prediction of Gray Leaf Spot of Perennial Ryegrass Turf
Authors: Uddin W. , Serlemitsos K. , Viji G.
Published: May 2007

Gray leaf spot is a serious disease of perennial ryegrass (Lolium perenne), causing severe epidemics in golf course fairways. The effects of temperature and leaf wetness duration on the development of gray leaf spot of perennial ryegrass turf were evaluated in controlled environment chambers. Six-week-old Legacy II ryegrass plants were inoculated with an aqueous conidial suspension of Pyricularia grisea (approximately 8 × 104 conidia per ml of water) and subjected to four different temperatures (20, 24, 28, and 32°C) and 12 leaf wetness durations (3 to 36 h at 3-h intervals). Three days after inoculation, gray leaf spot developed on plants at all temperatures and leaf wetness durations. Disease incidence (percent leaf blades symptomatic) and severity (index 0 to 10; 0 = leaf blades asymptomatic, 10 = >90% leaf area necrotic) were assessed 7 days after inoculation. There were significant effects ( α = 0.0001) of temperature and leaf wetness duration on disease incidence and severity, and there were significant interactions ( α = 0.0001) between them. Among the four temperatures tested, 28°C was most favorable to gray leaf spot development. Disease incidence and severity increased with increased leaf wetness duration at all temperatures. A shorter leaf wetness duration was required for disease development under warmer temperatures. Analysis of variance with orthogonal polynomial contrasts and regression analyses were used to determine the functional relationships among temperature and leaf wetness duration and gray leaf spot incidence and severity. Significant effects were included in a regression model that described the relationship. The polynomial model included linear, quadratic, and cubic terms for temperature and leaf wetness duration effects. The adjusted coefficients of determination for the fitted model for disease incidence and severity were 0.84 and 0.87, respectively. The predictive model may be used as part of an integrated gray leaf spot forecasting system for perennial ryegrass turf.
Distribution and Rate of Movement of the Curtovirus Beet mild curly top virus (Family Geminiviridae) in the Beet Leafhopper
Authors: Soto Maria J. , Gilbertson Robert L.
Published: May 2007

A polymerase chain reaction (PCR)-based method for the detection of the curtovirus Beet mild curly top virus (BMCTV, previously named the Worland strain of Beet curly top virus) was developed and used to investigate the BMCTV-beet leafhopper interaction. Using PCR and a BMCTV-specific primer pair, an ≈1.1-kb BMCTV DNA fragment was amplified from adult leafhoppers and from the organs involved in circulative transmission: the digestive tract, hemolymph, and salivary glands. The temporal distribution of BMCTV in the leafhopper was determined using insects given acquisition access periods (AAPs) ranging from 1 to 48 h on BMCTV-infected shepherd's purse plants. BMCTV was detected in the digestive tract after all AAPs, in the hemolymph after AAPs of 3 h or greater, and in the salivary glands after AAPs of 4 h or greater. The amount of virus detected in the hemolymph and salivary glands increased with AAP length. The virus persisted for up to 30 days in leafhoppers (given a 3-day AAP on BMCTV-infected plants) maintained on corn plants, a nonhost for BMCTV, but transovarial transmission was not detected. These results are consistent with a persistent but nonpropagative mode of circulative transmission.
Influence of Temperature and Wetness Duration on Conidia and Appressoria of Colletotrichum acutatum on Symptomless Strawberry Leaves
Authors: Leandro L. F. S. , Gleason M. L. , Nutter F. W. , Wegulo S. N. , Dixon P. M.
Published: May 2007

Strawberry leaves (cv. Tristar) inoculated with Colletotrichum acuta-tum conidia were incubated at 10, 15, 20, 25, 30, and 35°C under continuous wetness, and at 25°C under six intermittent wetness regimes. The number of conidia and appressoria was quantified on excised leaf disks. In order to assess pathogen survival, inoculated leaves were frozen and incubated to induce acervular development. Germination, secondary3 conidiation, and appressorial development were significantly (P ≤ 0.05) affected by temperature and wetness treatments. Under continuous wetness, the optimum temperature range for conidial germination was 23.0 to 27.7°C, whereas the optimum temperature for appressorial development ranged from 17.6 to 26.5°C. Secondary conidiation showed an optimum temperature range of 21.3 to 32.7°C and was most abundant between 12 and 36 h after inoculation. Conidial germination, appressorial production, and secondary conidiation were favored by increasing wetness duration and more than 4 h of wetness were required for secondary conidiation. In a greenhouse, C. acutatum survived up to 8 weeks on leaves. The number of acervuli formed on leaves after freezing and incubation was closely (r2 ≥ 0.95) related to appressorial populations prior to this treatment and was greatest following periods of continuous wetness. Production of secondary conidia and appressoria of C. acutatum on symptomless strawberry leaves under a range of environmental conditions suggests that these processes also occur under field conditions and contribute to inoculum availability during the growing season.
Ultrastructural and Cytochemical Aspects of Silicon-Mediated Rice Blast Resistance
Authors: Rodrigues Fabrício Á. , Benhamou Nicole , Datnoff Lawrence E. , Jones Jeffrey B. , Bélanger Richard R.
Published: May 2007

Although exogenous application of silicon (Si) confers efficient control of rice blast, the probable hypothesis underlying this phenomenon has been confined to that of a mechanical barrier resulting from Si polymerization in planta. However, in this study, we provide the first cytological evidence that Si-mediated resistance to Magnaporthe grisea in rice correlates with specific leaf cell reaction that interfered with the development of the fungus. Accumulation of an amorphous material that stained densely with toluidine blue and reacted positively to osmium tetroxide was a typical feature of cell reaction to infection by M. grisea in samples from Si+ plants. As a result, the extent of fungal colonization was markedly reduced in samples from Si+ plants. In samples from Si- plants, M. grisea grew actively and colonized all leaf tissues. Cytochemi-cal labeling of chitin revealed no difference in the pattern of chitin localization over fungal cell walls of either Si+ or Si- plants at 96 h after inoculation, indicating limited production of chitinases by the rice plant as a mechanism of defense response. On the other hand, the occurrence of empty fungal hyphae, surrounded or trapped in amorphous material, in samples from Si+ plants suggests that phenolic-like compounds or phytoalexins played a primary role in rice defense response against infection by M. grisea. This finding brings new insights into the complex role played by Si in the nature of rice blast resistance.
Resistance of Postharvest Biocontrol Yeasts to Oxidative Stress: A Possible New Mechanism of Action
Authors: Castoria Raffaello , Caputo Leonardo , De Curtis Filippo , De Cicco Vincenzo
Published: May 2007

We detected the generation of the reactive oxygen species (ROS) superoxide anion ( O·- 2) and hydrogen peroxide (H2O2) in apple wounds 2 immediately after wounding, and assessed the relationships between (i) timely colonization of apple wounds by biocontrol yeasts, (ii) resistance of these microorganisms to oxidative stress caused by ROS, and (iii) their antagonism against postharvest wound pathogens. We analyzed a model system consisting of two yeasts with higher (Cryptococcus laurentii LS-28) or lower (Rhodotorula glutinis LS-11) antagonistic activity against the postharvest pathogens Botrytis cinerea and Penicillium expansum. LS-28 exhibited faster and greater colonization of wounds than LS-11. In contrast to LS-28, the number of LS-11 cells dropped 1 and 2 h after application, and then increased only later. In vitro, LS-28 was more resistant to ROS-generated oxidative stress. The combined application of biocontrol yeasts and ROS-deactivating enzymes in apple wounds prevented the decrease in number of LS-11 cells mentioned above, and enhanced colonization and antagonistic activity of both biocontrol yeasts against B. cinerea and P. expansum. Polar lipids of LS-11 contained the more unsaturated and oxidizable α-linolenic acid, which was absent in LS-28. Resistance to oxidative stress could be a key mechanism of biocontrol yeasts antagonism against postharvest wound pathogens.
Characterization of Colletotrichum Isolates from Tamarillo, Passiflora, and Mango in Colombia and Identification of a Unique Species from the Genus
Authors: Afanador-Kafuri Lucia , Minz Dror , Maymon Marcel , Freeman Stanley
Published: May 2007

This study was conducted to identify the species of Colletotrichum infecting tamarillo, mango, and passiflora in Colombia and to assess whether cross-infection between host species is occurring. Isolates of Colletotrichum spp. from tamarillo (n = 54), passiflora (n = 26), and mango (n = 15) were characterized by various molecular methods and by morphological criteria. Morphological characterization grouped the tamarillo isolates as C. acutatum and the passiflora and mango isolates as C. gloeosporioides. Species-specific primer analysis was reliable and confirmed grouping of the tamarillo isolates (besides Tom-6) as C. acutatum and the mango isolates (besides Man-76) as C. gloeosporioides. However, DNA of the passiflora isolates was not amplified by either C. acutatum- or C. gloeosporioides-specific primers, but reacted with a new primer, Col1, designed according to the internal transcribed spacer (ITS) 1 region of these isolates. Isolates Tom-6 and Man-76 also reacted positively with the Col1 primer. All the isolates reacting with the C. acutatum- and C. gloeosporioides-specific primers failed to react with primer Col1. Isolate Pass-35 from passiflora did not react with any of the taxon-specific primers. Arbitrarily primed polymerase chain reaction (ap-PCR), random amplified polymerase DNA (RAPD)-PCR, and A+T-rich DNA analyses delineated representative isolates into subgroups within the designated species. Molecular analyses indicated that the C. acutatum tamarillo isolates were uniform or clonal, whereas the C. gloeosporioides mango isolates and Colletotrichum passiflora isolates were heterogeneous. Likewise, sequence analysis of the complete ITS (ITS1-5.8S-ITS2) region identified certain isolates to their respective species: tamarillo isolates as C. acutatum; mango isolates as C. gloeosporioides; passiflora, Tom-6, and Man-76 isolates as a Colletotrichum sp. as yet undefined; and the Pass-35 isolate as an additional undefined Colletot-richum sp. Molecular analyses of the population of Colletotrichum isolates from passiflora, Tom-6 from tamarillo, and Man-76 from mango indicate that this population may not be host specific.
Molecular Mapping of the Leaf Rust Resistance Gene Rph6 in Barley and Its Linkage Relationships with Rph5 and Rph7
Authors: Zhong Shaobin , Effertz Roger J. , Jin Yue , Franckowiak Jerome D. , Steffenson Brian J.
Published: May 2007

The barley cv. Bolivia carries two leaf rust (Puccinia hordei) resistance genes, Rph2 and Rph6, and is the only known source of the latter gene. A resistant line (Bolivia-Rph6) carrying Rph6 only was obtained in the F4 generation of a cross between cv. Bolivia and the susceptible cv. Bowman via progeny testing with differential isolates of the leaf rust pathogen. Genetic analyses and bulk segregant analysis using amplified fragment length polymorphism (AFLP) and restriction fragment length polymorphism (RFLP) markers localized Rph6 on the short arm of barley chromosome 3H at a distance of 4.4 centimorgans (cM) distal from RFLP marker MWG2021 and 1.2 cM proximal from RFLP marker BCD907. The allelic relationship of Rph6 to other leaf rust resistance genes mapping to this region of chromosome 3H (namely Rph5 and Rph7) were tested using crosses among cvs. Magnif 102 (carrying Rph5), Bolivia-Rph6 (Rph6), and Cebada Capa (Rph7). Segregation analyses indicated that Rph6 is allelic to Rph5 and closely linked to Rph7. The data generated from this study will facilitate breeding for leaf rust resistance via marker-assisted selection and provide a starting point for positional gene cloning.
Green Fluorescent Detection of Fungal Colonization and Endopolygalacturonase Gene Expression in the Interaction of Alternaria citri with Citrus
Authors: Isshiki A. , Ohtani K. , Kyo M. , Yamamoto H. , Akimitsu K.
Published: May 2007

Alternaria citri, a postharvest pathogen, produces endopolygalacturonase (endoPG) and causes black rot on citrus fruit. We previously described that an endoPG-disrupted mutant of Alternaria citri was significantly reduced in its ability to macerate plant tissue and cause black rot symptoms on citrus. In order to investigate colonization of citrus fruit tissues by Alternaria citri, pTEFEGFP carrying a green fluorescent protein (GFP) gene was introduced into wild-type Alternaria citri and its endoPG-disrupted mutant (M60). Green fluorescence was observed in spores, germ tubes, appressoria, and infection hyphae of transformants G1 (derived from wild type) and GM4 (derived from M60). Hyphae of G1 but not GM4 vertically penetrated the peel, but the hyphae of both G1 and GM4 spread equally in the juice sac area of citrus fruit. Green fluorescence of Alternaria citri transformant EPG7 carrying a GFP gene under control of the endoPG gene promoter of Alternaria citri was induced by pectin in the peel during the infection stage, but repressed completely in the juice sac area, likely by carbon catabolite repression by sugars in the juice.
Melon Resistance toCucurbit yellow stunting disorder virusIs Characterized by Reduced Virus Accumulation
Authors: Marco Cristina F. , Aguilar Juan M. , Abad Jesús , Gómez-Guillamón María Luisa , Aranda Miguel A.
Published: May 2007

The pattern of accumulation of Cucurbit yellow stunting disorder virus (CYSDV; genus Crinivirus, family Closteroviridae) RNA has been analyzed in several cucurbit accessions. In susceptible accessions of melon (Cucumis melo), cucumber (Cucumis sativus), marrow (Cucurbita maxima), and squash (Cucurbita pepo), CYSDV RNA accumulation peaked during the first to second week postinoculation in the first to third leaf above the inoculated one; younger leaves showed very low or undetectable levels of CYSDV. Three melon accessions previously shown to remain asymptomatic after CYSDV inoculation under natural conditions were also assayed for their susceptibility to CYSDV. Hybridization and reverse transcription-polymerase chain reaction (RT-PCR) analysis of noninoculated leaves showed that only one of these, C-105, remained virus-free for up to 6 weeks after whitefly inoculation. In this accession, very low CYSDV levels were detected by RT-PCR in whitefly-inoculated leaves, and therefore, multiplication or spread of CYSDV in C-105 plants appeared to remain restricted to the inoculated leaves. When C-105 plants were graft inoculated, CYSDV RNA could be detected in phloem tissues, but the systemic colonization of C-105 by CYSDV upon graft inoculation seemed to be seriously impeded. Additionally, in situ hybridization experiments showed that, after C-105 graft inoculation, only a portion of the vascular bundles in petioles and stems were colonized by CYSDV and virus could not be found in leaf veins. RT-PCR experiments using primers to specifically detect negative-sense CYSDV RNA were carried out and showed that CYSDV replication took place in graft-inoculated C-105 scions. Therefore, the resistance mechanism may involve a restriction of the virus movement in the vascular system of the plants and/or prevention of high levels of virus accumulation.
Amplified Fragment Length Polymorphism Diversity in Cephalosporium maydis from Egypt
Authors: Saleh Amgad A. , Zeller Kurt A. , Ismael Abou-Serie M. , Fahmy Zeinab M. , El-Assiuty Elhamy M. , Leslie John F.
Published: May 2007

Cephalosporium maydis, the causal agent of late wilt of maize, was first described in Egypt in the 1960s, where it can cause yield losses of up to 40% in susceptible plantings. We characterized 866 isolates of C. maydis collected from 14 governates in Egypt, 7 in the Nile River Delta and 7 in southern (Middle and Upper) Egypt, with amplified fragment length polymorphism (AFLP) markers. The four AFLP primer-pair combinations generated 68 bands, 25 of which were polymorphic, resulting in 52 clonal haplotypes that clustered the 866 isolates into four phylogenetic lineages. Three lineages were found in both the Nile River Delta and southern Egypt. Lineage IV, the most diverse group (20 haplotypes), was recovered only from governates in the Nile River Delta. In some locations, one lineage dominated (up to 98% of the isolates recovered) and, from some fields, only a single haplotype was recovered. Under field conditions in Egypt, there is no evidence that C. maydis reproduces sexually. The nonuniform geographic distribution of the pathogen lineages within the country could be due to differences in climate or in the farming system, because host material differs in susceptibility and C. maydis lineages differ in pathogenicity.
Characterization and Distribution of Two Races of Phialophora gregata in the North-Central United States
Authors: Harrington T. C. , Steimel J. , Workneh F. , Yang X. B.
Published: May 2007

Genetic variation and variation in aggressiveness in Phialophora gregata f. sp. sojae, the cause of brown stem rot of soybean, was characterized in a sample of 209 isolates from the north-central region. The isolates were collected from soybean plants without regard to symptoms from randomly selected soybean fields. Seven genotypes (A1, A2, A4, A5, A6, M1, and M2) were distinguished based on DNA fingerprinting with microsatellite probes (CAT)5 and (CAC)5, with only minor genetic variation within the A or M genotypes. Only the A1, A2, and M1 genotypes were represented by more than one isolate. The A genotypes dominated in the eastern Iowa, Illinois, and Ohio samples, whereas the M genotypes were dominant in samples from western Iowa, Minnesota, and Missouri. In growth chamber experiments, isolates segregated into two pathogenicity groups based on their aggressiveness toward soybean cvs. Kenwood and BSR101, which are relatively susceptible and resistant, respectively, to brown stem rot. In both root dip inoculation and inoculation by injecting spores into the stem near the ground line (stab inoculations), isolates of the A genotypes caused greater foliar symptoms and more vascular discoloration than isolates of the M genotypes on both cultivars of soybean. All isolates caused foliar symptoms in both cultivars and in three additional cultivars of soybean with resistance to brown stem rot. Greater differences between the A and M genotypes were seen in foliar symptoms than in the linear extent of xylem discoloration, and greater differences were seen in Kenwood than in BSR101. Inoculation of these genotypes into five cultivars of soybean with different resistance genes to brown stem rot showed a genotype × cultivar interaction. A similar distinction was found in an earlier study of the adzuki bean pathogen, P. gregata f. sp. adzukicola, and consistent with the nomenclature of that pathogen, the soybean pathogens are named the aggressive race (race A) and the mild race (race M) of P. gregata f. sp. sojae.
Genetic and Molecular Analyses in Crosses of Race 2 and Race 7 of Albugo candida
Authors: Adhikari Tika B. , Liu Jean Q. , Mathur Snehlata , Wu Chunren X. , Rimmer S. Roger
Published: May 2007

The inheritance of avirulence and polymorphic molecular markers in Albugo candida, the cause of white rust of crucifers, was studied in crosses of race 2 (Ac2), using isolates MiAc2-B1 or MiAc2-B5 (metalaxyl-insensitive and virulent to Brassica juncea cv. Burgonde) with race 7 (Ac7), using isolate MsAc7-A1 (metalaxyl-sensitive and virulent to B. rapa cv. Torch). Hybrids were obtained via co-inoculation onto a common susceptible host. Putative F1 progeny were selfed to produce F2 progeny. The parents and F1 progeny were examined for virulence on the differential cultivars B. juncea cv. Burgonde and B. rapa cv. Torch. Segregation of avirulence or virulence of F2 populations was analyzed on cv. Torch. Putative F1 hybrids were confirmed by random amplified polymorphic DNA markers specific for each parent. Avirulence or virulence of F 2 progeny to B. rapa cv. Torch suggested 3:1 in each of three populations, supporting the hypothesis of a single dominant avirulence gene. Amplified fragment length polymorphism markers also segregated in regular Mendelian fashion among F2 progeny derived from two F1 hybrids (Cr2-5 and Cr2-7) of Cross-2. This first putative avirulence gene in A. candida was designated AvrAc1. These results suggest that a single dominant gene controls avirulence in race Ac2 to B. rapa cv. Torch and provides further evidence for the gene-for-gene relationship in the Albugo-Brassica pathosystem.
Comparison of Spray and Point Inoculation To Assess Resistance to Fusarium Head Blight in a Multienvironment Wheat Trial
Authors: Miedaner T. , Moldovan M. , Ittu M.
Published: May 2007

Fusarium head blight (FHB, scab), caused by Fusarium graminearum or F. culmorum, results in yield and quality reductions and accumulation of mycotoxins. Two inoculation methods are commonly used. Spraying a spore suspension on the head (spray inoculation) will detect resistance to initial infection (type I) and to disease spread within the spike (type II). Injecting a spore suspension into individual florets (point inoculation) will detect type II resistance only. To analyze the association of spray and point inoculation, 20 elite winter wheat cultivars from Romania, Germany, and Switzerland were inoculated in factorial field experiments in seven environments (location × year combinations) in Germany and Romania. Response to FHB was assessed by the percentage of visually infected spikelets and head weight relative to the noninoculated control. Point and spray inoculations resulted in a mean disease severity varying from 52 to 63%. Significant (P = 0.01) genotypic variation was found within and across the environments. Genotype-environment interaction was important also. Estimates of entry-mean heritability were higher for spray than for point inoculation as assessed by percent infected spikelets (0.81 versus 0.77) and relative head weight (0.77 versus 0.52). Significant (P = 0.01) interaction was found between inoculation methods. Consequently, coefficients of phenotypic correlation between both methods were low to medium for percent infected spikelets (0.40, P > 0.1) and relative head weight (0.52, P = 0.05). We conclude that the application of both inoculation methods should provide additional information for selection and scientific studies. Spray inoculation, however, is less laborious for large-scale routine screening of breeding materials.
Induced Resistance as a Mechanism of Biological Control by Lysobacter enzymogenes Strain C3
Authors: Kilic-Ekici Ozlem , Yuen Gary Y.
Published: May 2007

Induced resistance was found to be a mechanism for biological control of leaf spot, caused by Bipolaris sorokiniana, in tall fescue (Festuca arundinacea) using the bacterium Lysobacter enzymogenes strain C3. Resistance elicited by C3 suppressed germination of B. sorokiniana conidia on the phylloplane in addition to reducing the severity of leaf spot. The pathogen-inhibitory effect could be separated from antibiosis by using heat-inactivated cells of C3 that retained no antifungal activity. Application of live or heat-killed cells to tall fescue leaves resulted only in localized resistance confined to the treated leaf, whereas treatment of roots resulted in systemic resistance expressed in the foliage. The effects of foliar and root applications of C3 were long lasting, as evidenced by suppression of conidial germination and leaf spot development even when pathogen inoculation was delayed 15 days after bacterial treatment. When C3 population levels and germination of pathogen conidia was examined on leaf segments, germination percentage was reduced on all segments from C3-treated leaves compared with segments from non-treated leaves, but no dose-response relationship typical of antagonism was found. Induced resistance by C3 was not host or pathogen specific; foliar application of heat-killed C3 cells controlled B. sorokiniana on wheat and also was effective in reducing the severity of brown patch, caused by Rhizoctonia solani, on tall fescue. Treatments of tall fescue foliage or roots with C3 resulted in significantly elevated peroxidase activity compared with the control.
Suppression of Rhizoctonia solani on Impatiens by Enhanced Microbial Activity in Composted Swine Waste-Amended Potting Mixes
Authors: Diab H. G. , Hu S. , Benson D. M.
Published: May 2007

Peat moss-based potting mix was amended with either of two composted swine wastes, CSW1 and CSW2, at rates from 4 to 20% (vol/vol) to evaluate suppression of pre-emergence damping-off of impatiens (Impatiens balsamina) caused by Rhizoctonia solani (anastomosis group-4). A cucumber bioassay was used prior to each impatiens experiment to monitor maturity of compost as the compost aged in a curing pile by evaluating disease suppression toward both Pythium ultimum and R. solani. At 16, 24, 32, and 37 weeks after composting, plug trays filled with compost-amended potting mix were seeded with impatiens and infested with R. solani to determine suppression of damping-off. Pre-emergence damping-off was lower for impatiens grown in potting mix amended with 20% CSW1 than in CSW2-amended and nonamended mixes. To identify relationships between disease suppression and microbial parameters, samples of mixes were collected to determine microbial activity, biomass carbon and nitrogen, functional diversity, and population density. Higher rates of microbial activity were observed with increasing rates of CSW1 amendment than with CSW2 amendments. Microbial biomass carbon and nitrogen also were higher in CSW1-amended mixes than in CSW2-amended potting mixes 1 day prior to seeding and 5 weeks after seeding. Principal component analysis of Biolog-GN2 profiles showed different functional diversities between CSW1- and CSW2-amended mixes. Furthermore, mixes amended with CSW1 had higher colony forming units of fungi, endospore-forming bacteria, and oligotrophic bacteria. Our results suggest that enhanced microbial activity, functional and population diversity of stable compost-amended mix were associated with suppressiveness to Rhizoctonia damping-off in impatiens.
Characterization of the Exoglucanase-Encoding Gene PaEXG2 and Study of Its Role in the Biocontrol Activity of Pichia anomala Strain K
Authors: Grevesse Cathy , Lepoivre Philippe , Jijakli Mohamed Haïssam
Published: May 2007

The PaEXG2 gene, encoding an exo-β-1,3-glucanase, was isolated from the biocontrol agent Pichia anomala strain K. PaEXG2 has the capacity for coding an acidic protein of 427 amino acids with a predicted molecular weight of 45.7 kDa, a calculated pI of 4.7, and one potential N-glycosylation site. PaEXG2 was disrupted by the insertion of the URA3 marker gene, encoding orotidine monophosphate decarboxylase in strain KU1, a uracil auxotroph derived from strain K. Strain KU1 showed inferior biocontrol activity and colonization of wounds on apples, compared to the prototrophic strain. Antagonism and colonization were recovered after the restoration of prototrophy by transformation with the URA3 gene. Integrative transformation was shown to be mostly ectopic in strain K descendants (only 4% of integration by homologous recombination). PaEXG2 disruption abolished all detectable extracellular exo-β-1,3-glucanase activity in vitro and in situ but did not affect biocontrol of Botrytis cinerea on wounded apples.