bluuebunny/crossref_metadata_2025 huggingface dataset:
Yield Effects of Barley yellow dwarf virus in Soft Red Winter Wheat
Authors: Perry Keith L. , Kolb Frederic L. , Sammons Bernard , Lawson Clifford , Cisar Gordon , Ohm Herbert
Published: May 2007

Three cultivars of soft red winter wheat were evaluated to determine the relationship between the incidence and time of infection by Barley yellow dwarf virus (BYDV) and yield. Wheat was planted in 1995, 1996, and 1997 in a split-plot design with six replicates at sites in Indiana and Illinois. Yield plots were infested with different amounts of viruliferous aphids, and the incidence of BYDV in each plot was measured. In a 2-year study in Illinois with cv. Clark and the PAV-IL isolate of BYDV, yields were assessed following aphid infestation in fall, early spring, and late spring. Early spring infections resulted in larger yield reductions than late spring infections in both years and larger than fall infections in one year. Regression analyses to relate incidence of infection and yield with data from fall and early spring infections provided R2 values of 0.89 and 0.51 for the 1996 to 1997 and 1997 to 1998 seasons, respectively. An additional study at the same site in the 1996 to 1997 season compared the yield responses of cvs. Clark, Y88-3e, and PT8935b. Increases in the incidence of BYDV correlated with decreases in yield, with R2 values of 0.80, 0.78, and 0.90 for the three cultivars, respectively. Estimated yield losses in both studies and all cultivars ranged from 27 to 45 kg/ha or 0.34 to 0.55% for each percent increase in virus infection. In a third study over a 2-year period in Indiana with the same three wheat genot ypes and a second BYDV isolate (PAV-P), BYDV treatments resulted in significant reductions in yield, but yield loss and the incidence of BYDV were not linearly correlated. Given the differences in yield reductions caused by the two BYDV isolates, PAV-P may be an attenuated strain of BYDV and may cross-protect plants from naturally occurring strains of the virus.
A Field Method for Evaluating the Potential Durability of New Resistance Sources: Application to the Leptosphaeria maculans-Brassica napus Pathosystem
Authors: Brun H. , Levivier S. , Somda I. , Ruer D. , Renard M. , Chèvre A. M.
Published: May 2007

To increase the longevity of new resistance genes by avoiding a rapid change in pathogen populations, we established a new field method to determine, before the release of a resistant cultivar, whether and how rapidly the pathogen population is capable of responding to the selective pressure we impose. This method was applied to the Leptosphaeria maculans-Brassica napus pathosystem. The potential durability of two new major resistance genes introgressed into B. napus from the Brassica B genome was tested separately for each gene under field conditions for 4 years. Successive inoculations with residues of the resistant lines mixed with susceptible contaminated plant material recovered at harvest the previous year were performed in autumn. The Jlm1 resistance gene originating from B. juncea conferred complete resistance on the B. napus-B. juncea recombinant lines MX and MXS to inoculation of the cotyledons with a large diversity of L. maculans isolates. It also gave a high level of stem canker resistance in the field against natural populations of the pathogen. A similar level of resistance was obtained in the B. napus-B. nigra addition line LA4+, containing B. nigra chromosome 4 in a B. napus background. In the second year of the field experiment (i.e., the first in which residues from the resistant lines were included in the inoculation material), both MX and LA4+ maintained a high level of resistance. In the third and fourth years of the field experiment, the resistance of MX and MXS exposed to inoculum produced from their own residues broke down, but against fungal populations from susceptible B. napus or resistant B. nigra material remained effective. In contrast, LA4+ remained highly resistant to all sources of inoculum for the 4-year experiment.
Genetic Diversity of Panicum mosaic virus Satellite RNAs in St. Augustinegrass
Authors: Cabrera Over , Roossinck Marilyn J. , Scholthof Karen-Beth G.
Published: May 2007

St. Augustine decline is a viral disease caused by Panicum mosaic virus (PMV) alone or in combination with a satellite virus (SPMV) and/or satellite RNAs (satRNAs). A ribonuclease protection assay (RPA) was used to evaluate the genetic diversity of PMV satRNAs isolated from 100 naturally infected St. Augustinegrass plants (Stenotaphrum secundatum). Distinctive satRNA RPA profiles were observed for 40 of 52 samples from College Station (CS) and 37 of 48 samples from Corpus Christi (CC), Texas. A dendrogram constructed from the RPA data revealed that satRNAs were grouped in two distinct clusters based on their place of origin. From 100 samples, only 4 satRNAs from CS were placed in the CC group, and only 2 satRNAs from CC were placed in the CS group. The data show that there is genetic variability in PMV satRNAs in naturally occurring infections, and distinct geographically separate populations can be identified from CC and CS.
Environmental and Genetic Factors Influencing Self-Fertility in Phytophthora infestans
Authors: Smart C. D. , Mayton H. , Mizubuti E. S. G. , Willmann M. R. , Fry W. E.
Published: May 2007

Phytophthora infestans is generally regarded as heterothallic-requiring physical proximity of two individuals of different mating type (A1 and A2) for oosporogenesis. Recent reports of limited selfing in young cultures of this oomycete stimulated us to investigate factors contributing to the phenomenon. The ability to produce oospores rapidly (within 2 weeks) in pure, single individual cultures (self-fertility) was tested in 116 individual isolates. The 116 isolates were from geographically diverse locations (16 countries) and were genetically diverse. Mating type and growth medium were the most prominent factors in determining if an isolate would be self-fertile. The majority of A2 isolates (45 of 47 tested) produced oospores when grown on a 50:50 mixture of V8 and rye B medium. In contrast, the majority of A1 isolates (65 of 69 tested) did not produce oospores on this medium. None of the 116 isolates produced oospores when grown on rye B medium (with no V8 juice). Further tests on representative A1 and A2 isolates revealed that oatmeal agar, tomato juice agar, and V8-juice agar all induced the A2 mating type isolate to produce oospores but did not induce the A1 mating type isolate to produce oospores. Calcium carbonate and pH did not alter the self-fertile oospore production in either A1 or A2 mating type isolates. For in vivo tests, the application of fungicide to potato or tomato leaf tissue either before or after inoculation did not stimulate any individual isolate (one A2 and one A1 isolate) to produce oospores in infected tissue. However, in all of the controls for all experiments (in vivo and in vitro), many oospores were produced rapidly if both strains grew in physical proximity.
Effects of Irrigation and Verticillium dahliae on Cauliflower Root and Shoot Growth Dynamics
Authors: Xiao C. L. , Subbarao K. V.
Published: May 2007

Cauliflower root and plant growth and Verticillium wilt development were evaluated under different moisture regimes in the presence or absence of V. dahliae. Treatments included two main plots (V. dahliae-infested and fumigated), two subplots (furrow and subsurface drip irrigation), and three sub-subplots (deficit, moderate, and excessive regimes) that were arranged in a split-split-plot design in the field. Soil cores with roots were periodically sampled at 5 and 25 cm distance from plants. Total roots in each soil core were extracted with a hydropneumatic root elutriator, and root length from each sample was determined with a digital image analysis system. Incidence and severity of Verticillium wilt, plant height, number of leaves, and dry weights of leaves and roots were determined on 10 plants sampled at 7- to 10-day intervals 1 month after cauliflower transplanting and continued until harvest. To evaluate the effects of Verticillium wilt-induced stress on cauliflower plants, stomatal resistance was measured in upper healthy and lower (or diseased) leaves. Root length density at 5 and 25 cm from plant was significantly (P < 0.05) higher in subsurface drip than in furrow irrigation. Root length density was significantly higher in excessive irrigation regime than in the other regimes. Concomitantly, there was higher wilt incidence and severity in excessive and moderate regimes than deficit regime regardless of the irrigation method. Plant height was affected by irrigation methods and deficit regime. Neither the method of irrigation nor the quantity of water affected the other variables. Stomatal resistance in lower diseased leaves was significantly higher in infested than in fumigated plots but it was not in the upper healthy leaves. In this study, cauliflower yield was not affected by V. dahliae and irrigation method, but the deficit irrigation regime resulted in reduced yield even though it suppressed wilt in cauliflower. Thus, higher moisture levels resulted in higher root length density in V. dahliae-infested plots that in turn lead to greater incidence of Verticillium wilt and severity. The pathogen also affected physiological processes such as hydraulic conductance of cauliflower leaves, but not shoot growth or yield under these experimental conditions.
The Epidemiology of Powdery Mildew on Concord Grapes
Authors: Gadoury David M. , Seem Robert C. , Ficke Andrea , Wilcox Wayne F.
Published: May 2007

Vitis labruscana ‘Concord’ is a grape cultivar widely grown in the United States for processing into juice and other grape products. Concord grapes are sporadically but sometimes severely damaged by the grape powdery mildew pathogen, Uncinula necator. Although the foliage is often reported to be moderately resistant to powdery mildew, severe fruit infection occurs in some years. We observed the seasonal development of powdery mildew on leaves, rachises, and berries of unsprayed Concord grapevines. Inoculations of flower and fruit clusters revealed a brief period of berry susceptibility and a protracted period of rachis susceptibility. The rachis remained highly susceptible to infection, and the severity of rachis infection increased throughout the growing season until the rachis formed a periderm shortly before harvest. In contrast, berries were nearly immune to infection within 2 weeks after fruit set. Rachis and berry infections were detected before the disease was observed on foliage, and the incidence of rachis and berry infection often exceeded disease incidence observed on foliage until after fruit acquired substantial ontogenic resistance. Excellent control of fruit infection, and adequate control of leaf infection, was achieved by two fungicide applications targeted at the peak period of fruit susceptibility. Although Concord is thought to be moderately resistant to powdery mildew, the rachis is highly susceptible, and may be the avenue by which prebloom infections make their way onto the developing fruit. Late-season infection of the rachis neither spread to the fruit, nor did it cause fruit to drop prematurely, and may be of little economic consequence on fruit destined for processing. Although fruit of V. vinifera cultivars have been reported to remain susceptible to infection until berry sugar levels reach 8 to 15%, Concord fruit become nearly immune to infection nearly 6 weeks before this stage of development. Because powdery mildew does not become conspicuous on foliage until late summer, it is generally regarded as a late-season problem on Concord grapes, and previous management programs have reflected this belief. However, the greatest contribution to control of fruit infection is due to fungicides applied during the peak period of fruit susceptibility, from bloom until shortly after fruit set, long before the disease is observed on foliage.
Investigating the Spatiotemporal Genetic Structure of Phytophthora capsici in Michigan
Authors: Lamour K. H. , Hausbeck M. K.
Published: May 2007

Phytophthora capsici isolates were recovered from pepper and cucurbit hosts at seven locations in Michigan from 1998 to 2000. Isolates were characterized for compatibility type (CT), mefenoxam sensitivity (MS), and amplified fragment length polymorphism (AFLP) marker profiles. In total, 94 AFLP bands were resolved. Individual populations were highly variable. Within populations, 39 to 49% of the AFLP bands were polymorphic and estimated heterozygosities ranged from 0.16 to 0.19. Of the 646 isolates fingerprinted, 70% (454) had unique AFLP profiles. No clones were recovered between years or locations. Pairwise F statistics (ΦST) between populations from different locations ranged from 0.18 to 0.40. A tree based on unweighted pair-group method with arithmetic average cluster analysis indicates discrete clusters based on location. Isolates from the same location showed no clustering based on the year of sampling. Analysis of molecular variance partitioned variability among (40%) and within populations (60%). The overall estimated ΦST was 0.34 (SD = 0.03). A1/A2 CT ratios were ≈1:1, and MS frequencies were similar between years for the two locations sampled over time. These data suggest that P. capsici persists in discrete outcrossing populations and that gene flow among locations in Michigan is infrequent.
Abundant Pathogenic Variation in the New Hybrid Rust Melampsora ×columbiana on Hybrid Poplar
Authors: Newcombe George , Stirling Brigid , Bradshaw H. D.
Published: May 2007

The recently described rust hybrid Melampsora ×columbiana was discovered as a result of its novel pathogenic variation on Populus trichocarpa × P. deltoides (T×D) hybrid poplar. To characterize this pathogenic variation, 10 commercial T×D clones, all F1 clones, were chosen as host differentials. Fourteen mononuredinial isolates of Pacific Northwestern field collections of M. ×columbiana, from 1996 to 1998 inclusive, were determined to be 13 distinct pathotypes. In contrast, four Southeastern isolates of M. medusae could not be distinguished on the same T×D host differentials, although they can be distinguished as pathotypes using P. deltoides differentials. The first three pathotypes of M. ×columbiana (Mxc1, Mxc2, and Mxc3) and a Mississippi isolate of M. medusae were inoculated onto a three-generation T×D pedigree, formerly used to characterize the Mmd1 gene for resistance to M. medusae. Resistance to the Mxc3 pathotype mapped to the same linkage group (group Q) as the Mmd1 gene. In contrast, linked genes for resistance to Mxc1 and to Mxc2 were located on linkage group O, and were unlike Mmd1 and Mxc3 in that they were inherited from P. deltoides. The latter two genes resembled Mmd1 and Mxc3 in that infection type was correlated with quantitative traits such as uredinial density and latent period. Pathogenic variation in M. ×columbiana matches resistance genes from both P. trichocarpa and P. deltoides and reveals the vulnerability to hybrid rust of commercial T×D hybrid poplar clones.
Isolation and Characterization of Derivative Lines of the Onion Yellows Phytoplasma that Do Not Cause Stunting or Phloem Hyperplasia
Authors: Oshima Kenro , Shiomi Toshiki , Kuboyama Tsutomu , Sawayanagi Toshimi , Nishigawa Hisashi , Kakizawa Shigeyuki , Miyata Shin-ichi , Ugaki Masashi , Namba Shigetou
Published: May 2007

Two lines of onion yellows phytoplasma producing milder symptoms were isolated from the original line (OY-W). One has an additional characteristic, non-insect-transmissibility (OY-NIM), compared with the other (OY-M). OY-M was established after maintaining OY-W for 11 years on a plant host (Chrysanthemum coronarium) with an insect vector (Macrosteles striifrons), and OY-NIM was isolated after subsequent maintenance of OY-M in plants by periodic grafting. Polymerase chain analysis suggested that OY-NIM cannot traverse the gut or survive in the hemolymph of the leafhopper. OY-W results in witches'-broom formation and stunted growth in the host plant. In contrast, OY-M and OY-NIM do not cause stunting in the host plant, although they result in witches'-broom. Histopathological analysis of these lines revealed that the hyperplastic phloem tissue and severe phloem necrosis seen in OY-W did not exist in OY-M and OY-NIM. This was attributed to a reduction in the population of phytoplasma in tissues in both OY-M- and OY-NIM-infected plants. The results suggest that the cause of stunting and phloem hyperplasia may be genetically different from the cause of witches'-broom. Pulsed field gel electrophoresis analysis showed that OY-M had a smaller genome size (≈870 kbp) than OY-W (≈1,000 kbp). Thus, some of the OY-W genes responsible for pathogenicity may not be present in OY-M.
Effects of Angular Leaf Spot and Rust on Yield Loss of Phaseolus vulgaris
Authors: de Jesus W. C. , do Vale F. X. R. , Coelho R. R. , Hau B. , Zambolim L. , Costa L. C. , Filho A. Bergamin
Published: May 2007

Three field experiments were conducted in 1997, 1998, and 1999 to investigate the effects of angular leaf spot and rust, separately or combined, on host growth and yield of individual bean plants (Phaseolus vulgaris). In each experiment, three treatments were established by inoculating cv. Carioca with Phaeoisariopsis griseola, Uromyces appendiculatus, or with both pathogens. An additional control treatment was not inoculated, but was sprayed with a fungicide. In the 1997 and 1999 experiments, angular leaf spot reached higher disease levels than rust, whereas in 1998, rust was more severe than angular leaf spot. Host growth, expressed as healthy leaf area duration (HAD), and yield were the highest in 1997 and lowest in 1998. In each experiment, the treatments did not differ significantly to the area under leaf area progress curve, HAD, and healthy leaf area absorption (HAA). All inoculated treatments had significantly more severe disease and less yield than the control treatment. Based on the analysis of 60 plants in each experiment, yield was not related to the areas under disease progress curve for either or both diseases. In 1997 and 1999, yield was related to HAD (R2 = 0.57 and 0.43) and HAA(R2 = 0.60 and 0.55). Based on the combined analysis of all 36 plots, angular leaf spot reduced the leaf area because of defoliation, whereas rust did not affect the leaf area. Rust reduced yield more than four times that of angular leaf spot, although the decrease in photosynthesis to angular leaf spot was twice that of rust.
Fine Mapping and Identification of Nucleotide Binding Site/Leucine-Rich Repeat Sequences at theMERLocus inPopulus deltoides‘S9-2’
Authors: Zhang J. , Steenackers M. , Storme V. , Neyrinck S. , Van Montagu M. , Gerats T. , Boerjan W.
Published: May 2007

Melampsora larici-populina is the most damaging leaf pathogen for poplar in Europe. Previous genetic analyses have revealed both qualitative and quantitative resistance to this fungus. As a starting point for positional cloning of the gene or genes conferring qualitative resistance to M. larici-populina races E1, E2, and E3, a local genetic map of the Melampsora resistance (MER) locus was constructed based on amplified fragment length polymorphism (AFLP) markers. Eleven AFLP markers were identified by bulked segregant analysis. These markers were used to identify 17 recombinants at the MER locus, from a total of 512 progenies derived from three interspecific crosses involving the same resistant female parent, Populus deltoides ‘S9-2’. The local genetic map covered a 3.4-centimorgan interval encompassing the target locus. Sequence analysis of these AFLP markers revealed similarities to the nucleotide binding site/leucine-rich repeat class of disease resistance genes.
Seasonal Availability of Inoculum for Constriction Canker of Peach in New Jersey
Authors: Lalancette N. , Robison D. M.
Published: May 2007

The seasonal sporulation ability of Phomopsis amygdali was evaluated over a 2-year period by sampling twig cankers from commercial ‘Encore’ and ‘Jerseyglo’ peach orchards. Canker size, pycnidia per canker, percent pycnidia forming cirri, and spore production were evaluated once each month from January 1997 through December 1998. Average canker size and number of pycnidia per canker, which followed a sinusoidal pattern of change, were at lowest values in spring and at maximum values in fall. In 1997, the ability of pycnidia to sporulate (produce cirri) increased significantly from a minimum during winter to a maximum during summer. In contrast, pycnidia sporulation in 1998 increased slowly throughout the year with a maximum in the fall. Inoculum potential, measured as number of spores per canker, peaked between June and August and was lowest in winter. Although pycnidia numbers in 1997 were about 50% of that observed in 1998, approximately twice as many conidia per canker were produced in 1997; consequently, the number of conidia produced per canker was equivalent in both years. These results indicated that P. amygdali acts as an r strategist by maintaining abundant pycnidia and sporulation capability throughout much of the year. Therefore, infection is most likely not limited by the availability of inoculum.
Occurrence and Molecular Characterization of Strobilurin Resistance in Cucumber Powdery Mildew and Downy Mildew
Authors: Ishii H. , Fraaije B. A. , Sugiyama T. , Noguchi K. , Nishimura K. , Takeda T. , Amano T. , Hollomon D. W.
Published: May 2007

Between 1998 and 1999, control failure of powdery mildew (Podosphaera fusca) and downy mildew (Pseudoperonospora cubensis) by the strobilurin fungicides azoxystrobin and kresoxim-methyl was observed in cucumber-growing areas of Japan. Results from inoculation tests carried out on intact cucumber plants and leaf disks clearly showed the distribution of pathogen isolates highly resistant to azoxystrobin and kresoximmethyl. Fragments of the fungicide-targeted mitochondrial cytochrome b gene were polymerase chain reaction amplified from total pathogen DNA and their sequences analyzed to elucidate the molecular mechanism of resistance. A single point mutation (GGT to GCT) in the cytochrome b gene, resulting in substitution of glycine by alanine at position 143, was found in resistant isolates of downy mildew. This substitution in cytochrome b seemed to result in high resistance to strobilurins in this pathogen. The same mutation was found in some but not all resistant isolates of powdery mildew. This study suggests that a mutation at position 143 in the target-encoding gene, resulting in an amino acid substitution, was probably a major cause of the rapid development of high strobilurin resistance in these two pathogens.
Quantifying the Rate of Release and Escape of Phytophthora infestans Sporangia from a Potato Canopy
Authors: Aylor Donald E. , Fry William E. , Mayton Hilary , Andrade-Piedra Jorge L.
Published: May 2007

A means for determining the rate of release, Q (spores per square meter per second), of spores from a source of inoculum is paramount for quantifying their further dispersal and the potential spread of disease. Values of Q were obtained for Phytophthora infestans sporangia released from an area source of diseased plants in a potato canopy by comparing the concentrations of airborne sporangia measured at several heights above the source, with the concentrations predicted by a Lagrangian Stochastic simulation model. An independent estimate of Q was obtained by quantifying the number of sporangia per unit area of source at the beginning of each sampling day by harvesting diseased plant tissue and enumerating sporangia from these samples. This standing spore crop was the potential number of sporangia released per area of source during the day. The standing spore crop was apportioned into time segments corresponding to sporangia concentration measurement periods using the time trace of sporangia sampled above the source by a Burkard continuous suction spore sampler. This apportionment of the standing spore crop yielded potential release rates that were compared with modeled release rates. The two independent estimates of Q were highly correlated (P = 0.003), indicating that the model has utility for predicting release rates for P. infestans sporangia and the spread of disease between fields.
Cocultures of Peronospora tabacina and Nicotiana Species to Study Host-Pathogen Interactions
Authors: Heist E. P. , Nesmith W. C. , Schardl C. L.
Published: May 2007

Long-term cocultures of the tobacco blue mold pathogen, Peronospora tabacina, with Nicotiana tabacum and N. repanda callus were derived from infected host plant tissue. In this apparently contaminant-free system, sporulation occurred under similar conditions as in intact plants. Sporangia were collected from cocultures and used to complete Koch's postulates. The cocultures were grown under two light regimes. One consisted of 23 h of light followed by 1 h of darkness and the second comprised total darkness. Sporulation occurred frequently in the 23 h light-grown cocultures but resulted in production of abnormal sporangiophores and sporangia. Production of normal sporangiophores and sporangia was achieved by transferring light-grown cocultures to overnight darkness and resulted in necrosis of the callus. Cocultures of Peronospora tabacina with either host species, grown in total darkness, frequently sporulated with minimal necrosis over the course of 1 year. Thus, cocultures should prove useful as a source of Peronospora tabacina over extended periods of time at low risk of pathogen release, for studying the physiology of Peronospora tabacina- Nicotiana interactions, maintaining Peronospora tabacina lines for genetic studies, and providing a reliable source of axenic inoculum for research.
Host Specialization in the Charcoal Rot Fungus, Macrophomina phaseolina
Authors: Su G. , Suh S.-O. , Schneider R. W. , Russin J. S.
Published: May 2007

To investigate host specialization in Macrophomina phaseolina, the fungus was isolated from soybean, corn, sorghum, and cotton root tissue and soil from fields cropped continuously to these species for 15 years in St. Joseph, LA. Chlorate phenotype of each isolate was determined after growing on a minimal medium containing 120 mM potassium chlorate. Consistent differences in chlorate sensitivity were detected among isolates from different hosts and from soil versus root. To further explore genetic differentiation among fungal isolates from each host, these isolates were examined by restriction fragment length polymorphism and random amplified polymorphic DNA (RAPD) analysis. No variations were observed among isolates in restriction patterns of DNA fragments amplified by polymerase chain reaction covering the internal transcribed spacer region, 5.8S rRNA and part of 25S rRNA, suggesting that M. phaseolina constitutes a single species. Ten random primers were used to amplify the total DNA of 45 isolates, and banding patterns resulting from RAPD analysis were compared with the neighbor-joining method. Isolates from a given host were genetically similar to each other but distinctly different from those from other hosts. Chlorate-sensitive isolates were distinct from chlorate-resistant isolates within a given host. In greenhouse tests, soybean, sorghum, corn, and cotton were grown separately in soil infested with individual isolates of M. phaseolina that were chosen based on their host of origin and chlorate phenotype. Root colonization and plant weight were measured after harvesting. More colonization of corn roots occurred when corn was grown in soil containing corn isolates compared with isolates from other hosts. However, there was no host specialization in isolates from soybean, sorghum, or cotton. More root colonization in soybean occurred with chlorate-sensitive than with chlorate-resistant isolates.
A Compromised Mlo Pathway Affects the Response of Barley to the Necrotrophic Fungus Bipolaris sorokiniana (Teleomorph: Cochliobolus sativus) and Its Toxins
Authors: Kumar Jagdish , Hückelhoven Ralph , Beckhove Ulrich , Nagarajan Subrahmaniam , Kogel Karl-Heinz
Published: May 2007

In search of new durable disease resistance traits in barley to control leaf spot blotch disease caused by the necrotrophic fungus Bipolaris sorokiniana (teleomorph: Cochliobolus sativus), we developed macroscopic and microscopic scales to judge spot blotch disease development on barley. Infection of barley was associated with cell wall penetration and accumulation of hydrogen peroxide. The latter appeared to take place in cell wall swellings under fungal penetration attempts as well as during cell death provoked by the necrotrophic pathogen. Additionally, we tested the influence of a compromised Mlo pathway that confers broad resistance against powdery mildew fungus (Blumeria graminis f. sp. hordei). Powdery mildew-resistant genotypes with mutations at the Mlo locus (mlo genotypes) showed a higher sensitivity to infiltration of toxic culture filtrate of Bipolaris sorokiniana as compared with wild-type barley. Mutants defective in Ror, a gene required for mlo-specified powdery mildew resistance, were also more sensitive to Bipolaris sorokiniana toxins than wild-type barley but showed less symptoms than mlo5 parents. Fungal culture filtrates induced an H2O2 burst in all mutants, whereas wild-type (Mlo) barley was less sensitive. The results support the hypothesis that the barley Mlo gene product functions as a suppresser of cell death. Therefore, a compromised Mlo pathway is effective for control of biotrophic powdery mildew fungus but not for necrotrophic Bipolaris sorokiniana. We discuss the problem of finding resistance traits that are effective against both biotrophic and necrotrophic pathogens with emphasis on the role of the anti-oxidative system of plant cells.
Latency- and Defense-Related Ultrastructural Characteristics of Apple Fruit Tissues Infected with Botryosphaeria dothidea
Authors: Kim Ki Woo , Park Eun Woo , Kim Young Ho , Ahn Kyung-Ku , Kim Pan Gi , Kim Kyung Soo
Published: May 2007

Apple fruit tissues infected with Botryosphaeria dothidea were examined by transmission electron microscopy using susceptible cv. Fuji and resistant cv. Jonathan. Immature (green) and mature (red) fruits of cv. Fuji with restricted or expanding lesions were also examined to reveal subcellular characteristics related with latent and restricted disease development. In infected susceptible mature fruits, cytoplasmic degeneration and organelle disruption commonly occurred, accompanying cell wall dissolution around invading hyphae. Cell wall dissolution around invading hyphae in subepidermis was rare in immature, red halo-symptomed cv. Fuji and resistant cv. Jonathan fruits. In infected immature fruits of cv. Fuji, presumably at the latent state of disease development, cellular degeneration was less severe, and invading hyphae contained prominent microbody-lipid globule complexes or the deposition of thin electron-dense outer layer around cell wall of intercellular hyphae. Both mature fruits with red halos and resistant apple fruits formed cell wall protuberances at the outside of cell walls. In addition, electron-dense extramural layers were formed in the resistant apple fruits. Aberrant hyphal structures such as intrahyphal hyphae were found only in resistant fruit tissues, indicating the physiologically altered fungal growth. These ultrastructural changes of host tissues and fungal hyphae may reflect the pathogenesis of apple white rot under varying conditions of apple fruits.
Rate of Tomato yellow leaf curl virus Translocation in the Circulative Transmission Pathway of its Vector, the Whitefly Bemisia tabaci
Authors: Ghanim Murad , Morin Shai , Czosnek Henryk
Published: May 2007

Whiteflies (Bemisia tabaci, biotype B) were able to transmit Tomato yellow leaf curl virus (TYLCV) 8 h after they were caged with infected tomato plants. The spread of TYLCV during this latent period was followed in organs thought to be involved in the translocation of the virus in B. tabaci. After increasing acquisition access periods (AAPs) on infected tomato plants, the stylets, the head, the midgut, a hemolymph sample, and the salivary glands dissected from individual insects were subjected to polymerase chain reaction (PCR) without any treatment; the presence of TYLCV was assessed with virus-specific primers. TYLCV DNA was first detected in the head of B. tabaci after a 10-min AAP. The virus was present in the midgut after 40 min and was first detected in the hemolymph after 90 min. TYLCV was found in the salivary glands 5.5 h after it was first detected in the hemolymph. Subjecting the insect organs to immunocapture-PCR showed that the virus capsid protein was in the insect organs at the same time as the virus genome, suggesting that at least some TYLCV translocates as virions. Although females are more efficient as vectors than males, TYLCV was detected in the salivary glands of males and of females after approximately the same AAP.
Potential for Integrated Control of Sclerotinia sclerotiorum in Glasshouse Lettuce Using Coniothyrium minitans and Reduced Fungicide Application
Authors: Budge Simon P. , Whipps John M.
Published: May 2007

All pesticides used in United Kingdom glasshouse lettuce production (six fungicides, four insecticides, and one herbicide) were evaluated for their effects on Coniothyrium minitans mycelial growth and spore germination in vitro agar plate tests. Only the fungicides had a significant effect with all three strains of C. minitans tested, being highly sensitive to iprodione (50% effective concentration [EC50] 7 to 18 μg a.i. ml-1), moderately sensitive to thiram (EC50 52 to 106 μg a.i. ml-1), but less sensitive to the remaining fungicides (EC50 over 200 μg a.i. ml-1). Subsequently, all pesticides were assessed for their effect on the ability of C. minitans applied as a solid substrate inoculum to infect sclerotia of Sclerotinia sclerotiorum in soil tray tests. Despite weekly applications of pesticides at twice their recommended concentrations, C. minitans survived in the soil and infected sclerotia equally in all pesticide-treated and untreated control soil trays. This demonstrated the importance of assessing pesticide compatibility in environmentally relevant tests. Based on these results, solid substrate inoculum of a standard and an iprodione-tolerant strain of C. minitans were applied individually to S. sclerotiorum-infested soil in a glasshouse before planting lettuce crops. The effect of a single spray application of iprodione on disease control in the C. minitans treatments was assessed. Disease caused by S. sclerotiorum was significantly reduced by C. minitans and was enhanced by a single application of iprodione, regardless of whether the biocontrol agent was iprodione-tolerant. In a second experiment, disease control achieved by a combination of C. minitans and a single application of iprodione was shown to be equivalent to that of prophylactic sprays with iprodione every 2 weeks. The fungicide did not affect the ability of C. minitans to spread into plots where only the fungicide was applied and to infect sclerotia. These results indicate that integrated control of S. sclerotiorum with soil applications of C. minitans and reduced foliar iprodione applications was feasible, did not require a fungicide tolerant isolate, and that suppression of Sclerotinia disease by C. minitans under existing chemical control regimes has credence.
Phenotype Instability in Botrytis cinerea in the Absence of Benzimidazole and Dicarboximide Fungicides
Authors: Yourman L. F. , Jeffers S. N. , Dean R. A.
Published: May 2007

Stability of phenotypes of isolates of Botrytis cinerea that were sensitive or resistant to benzimidazole and dicarboximide fungicides was examined in the absence of fungicides in laboratory and growth room experiments. Twelve greenhouse isolates of B. cinerea were subcultured on potato dextrose agar (PDA) for 20 generations and on geranium seedlings for 15 generations. Three isolates of each of the following four phenotypes were used: sensitive to the fungicides thiophanate-methy1 (a benzimidazole) and vinclozolin (a dicarboximide) (STSV), resistant to both fungicides (RTRV), resistant to thiophanate-methy1 and sensitive to vinclozolin (RTSV), and sensitive to thiophanate-methy1 and resistant to vinclozolin (STRV). In three trials on PDA, 36 populations were subcultured; 8 populations changed phenotypes by the end of 20 generations, as determined by conidium germination on fungicide-amended medium. Five of the eight initially were STRV; the resulting phenotypes were STSV, RTSV, and RTRV. Populations from eight other isolates exhibited temporary changes in phenotype during intermediate generations on PDA but reverted to initial phenotypes by the twentieth generation; five of these populations changed to phenotype RTRV. In two geranium seedling trials, each of the 12 greenhouse isolates was inoculated onto a set of three seedlings for each generation, and diseased tissue that developed was used to initiate the next generation. Therefore, a total of 72 populations of B. cinerea were subcultured in the two trials; 5 of these populations changed phenotype at the end of 15 generations. Three of the five initially were STRV; these changed to phenotypes STSV or RTRV. In each of the two trials on geranium seedlings, a population subcultured from one STSV isolate changed phenotype one to phenotype RTRV and one to phenotype RTSV. In all trials, no population resistant to thiophanate-methy1 changed to a thiophanate-methy1-sensitive phenotype, and no population changed to phenotype STRV. Random amplified polymorphic DNA (RAPD) fingerprints were generated with the 12 initial isolates and 49 isolates subcultured on PDA or geranium seedlings. Cluster analyses of RAPD markers showed that subcultured isolates exhibiting the same phenotype clustered together and that subcultured isolates derived from a common greenhouse isolate but with different phenotypes were in different clusters. Some populations that did not change phenotype exhibited considerable differences in RAPD marker patterns. The results of this study indicate that, in the absence of fungicides, sensitive populations of B. cinerea can develop resistance to thiophanate-methy1 and vinclozolin, and this resistance can be maintained in populations through multiple generations. Populations resistant only to vinclozolin (STRV) exhibited a high frequency of phenotype change, and populations resistant to both fungicides (RTRV) were stable.
Detection of Rice Panicle Blast with Multispectral Radiometer and the Potential of Using Airborne Multispectral Scanners
Authors: Kobayashi T. , Kanda E. , Kitada K. , Ishiguro K. , Torigoe Y.
Published: May 2007

Rice reflectance was measured to determine the spectral regions most sensitive to panicle blast infection. Reflectance increased in the 430- to 530-, 580- to 680-, and 1,480- to 2,000-nm regions at the dough stage both in the laboratory and the field as the percentage of diseased spikelets increased. The wavebands of the greatest sensitivity were in the visible region, located near 485 and 675 nm. After the yellow-ripe growth stage, near-infrared rather than visible reflectance responded to panicle blast infections. Ratios of rice reflectance were evaluated as indicators of panicle blast. R470/R570 (reflectance at 470 nm divided by reflectance at 570 nm), R520/R675, and R570/R675 decreased significantly as the incidence of panicle blast increased at the dough stage. At the yellow-ripe stage, R550/R970 and R725/R900 were used to estimate panicle blast severity as measured in terms of the percentage of diseased spikelets. According to the simulation that uses ground-based sensor data, airborne multispectral scanners may be effective in detecting the occurrence of panicle blast using a band combination of 530- to 570- and 650- to 700-nm regions at the dough stage.
Completion of the Genome Sequence of Watermelon silver mottle virus and Utilization of Degenerate Primers for Detecting Tospoviruses in Five Serogroups
Authors: Chu Fang-Hua , Chao Chia-Hung , Chung Min-Hsun , Chen Ching-Chung , Yeh Shyi-Dong
Published: May 2007

The nucleotide sequence of the L RNA of Watermelon silver mottle virus (WSMoV) was determined. Combined with the previous work on M and S RNAs, the whole genomic sequence of this member of the genus Tospovirus was completed. The L RNA is 8,917 nucleotides in length, with one large open reading frame encoding a translation product of 2,878 amino acids (331.8 kDa) on the viral complementary strand. The L protein shares amino acid identities of only 44.3 and 46.5% with Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus, respectively; but an amino acid identity of 91.3% with Peanut bud necrosis virus. Among the sequenced tospoviruses, L protein was the most conserved gene product, whereas the nonstructural S protein was generally the most variable. Comparison of the deduced L protein of WSMoV with those of other members of the family Bunyaviridae revealed that its amino acid sequence includes the reported conserved motifs of RNA-dependent RNA polymerases. To develop a method for detecting tospo-viruses by reverse transcription-polymerase chain reaction (RT-PCR), two pairs of degenerate primers were designed from conserved regions of the L genes and used to amplify the corresponding regions of the L genes from total RNAs extracted from plant tissues infected with five serologically distinct tospoviruses. The DNA fragments obtained were identified as those of tospoviruses by restriction enzyme digestion and DNA sequencing. For field samples, watermelon and wax gourd infected with WSMoV, and lisianthus infected with TSWV were also successfully detected by these two pairs of degenerate primers, with a sensitivity similar to N-gene-specific primers. The results indicated that the RT-PCR with the degenerate primers is a fast and reliable method for detecting tospoviruses in different serogroups.
Genetic Variability of Canadian Populations of the Sapstain Fungus Ophiostoma piceae
Authors: Gagné Pierre , Yang Dian-Qing , Hamelin Richard C. , Bernier Louis
Published: May 2007

Genetic diversity was studied in seven Canadian populations of Ophiostoma piceae, the most prevalent sapstain fungus in Canadian softwoods. A total of 239 single-spore isolates were recovered following a systematic survey of sapstain fungi in logs and lumber at seven selected sawmills in six Canadian provinces (British Columbia, Alberta, Saskatchewan, Ontario, Québec, and New Brunswick). Sampling was carried out on five commercially important softwood species: balsam fir (Abies balsamea), white spruce (Picea glauca), black spruce (Picea mariana), jack pine (Pinus banksiana), and lodgepole pine (Pinus con-torta var. latifolia). The A and B mating types occurred at equal frequency (MAT A/ MAT B = 1.00:1.13) over all populations. Pseudo-allelic frequencies were estimated at each of 24 putative genetic loci by scoring for presence or absence of random amplified polymorphic DNA fragments generated by five primers. A total of 237 haplotypes were found among the 239 isolates, revealing a high level of genotypic diversity among isolates. Total gene diversity (HT = 0.414) was mostly attributable to diversity within populations (HS = 0.369). Thus, only 11.2% of the total variability was attributable to frequency differences among populations. An analysis of molecular variance revealed that most genetic variability occurred within subpopulations within mills (84.3%; P < 0.001), whereas low but statistically significant levels of genetic differentiation were also observed among subpopulations within populations (5.4%; P < 0.001) and among populations (10.3%; P < 0.001). Estimates of Nei' genetic distances were not correlated with geographic distances among sampling locations (r = -0.092; P = 0.310), although principal component analysis indicated that subpopulations located east of Saskatchewan were grouped on the same side of the second principal component axis. Overall, results suggest moderate genetic differentiation of O. piceae in Canada, which is consistent with the observation that sexual reproduction is frequently observed in this fungus.
Genetic Diversity of Japanese Strains of Ralstonia solanacearum
Authors: Horita Mitsuo , Tsuchiya Kenicki
Published: May 2007

The genetic diversity of 74 Japanese strains of Ralstonia solanacearum was assessed by pathogenicity tests and the repetitive sequencebased polymerase chain reaction (rep-PCR) fingerprint method. Based on their genomic fingerprints, biovar N2 strains were divided into two distinct groups, one consisting of potato isolates belonging to race 3, and the other consisting of tomato, eggplant, pepper, and tobacco isolates belonging to race 1. Biovar 3 strains had low average similarity and were divided into five groups that differed in original host or pathogenicity. Biovar 4 strains consisted of only one group at the 80% similarity level. Comparative analysis of the rep-PCR fingerprints of 78 strains, including six biovars from Japan and various countries, revealed two main clusters. Cluster 1 comprised all biovar 3, 4, and 5 strains, biovar 1 strains from Reunion, and some biovar N2 strains from Japan. Cluster 2 included most of the biovar 1, 2, and N2 strains. The fingerprints showed low average similarity with biovar N2 strains from Japan and Brazil.
Endophytic Colonization of Plants by the Biocontrol Agent Rhizobium etli G12 in Relation to Meloidogyne incognita Infection
Authors: Hallmann J. , Quadt-Hallmann A. , Miller W. G. , Sikora R. A. , Lindow S. E.
Published: May 2007

The external and internal colonization of potato and Arabidopsis roots by the biocontrol strain Rhizobium etli G12 containing a plasmidborne trp promoter green fluorescent protein transcriptional fusion, pGT-trp, was studied in the presence and absence of the root-knot nematode Meloidogyne incognita. Plant colonization behavior and biocontrol potential of the marked strain G12(pGT-trp) was not altered compared with the parental strain. Plasmid pGT-trp was stable for more than 80 generations without selection and conferred sufficient fluorescence to detect single bacterial cells in planta. Although bacteria were found over the entire rhizoplane, they preferentially colonized root tips, the emerging lateral roots, and galled tissue caused by Meloidogyne infestation. Internal colonization of potato roots was mainly observed in epidermal cells, especially root hairs. G12(pGT-trp) colonization was also observed in inner Arabidopsis root tissues in areas of vascularization. In the presence of M. incognita, G12(pGT-trp) colonized the interior of nematode galls in high numbers. In some cases, bacterial colonization even extended from the galled tissue into adjacent root tissue. The internally colonized sites in roots were often discontinuous. Fluorescence microscopy of gfp-tagged rhizobacteria was a sensitive and a rapid technique to study external and internal colonization of plant roots by bacteria interacting with nematodes.