bluuebunny/crossref_metadata_2025 huggingface dataset:
Single Cystosorus Isolate Production and Restriction Fragment Length Polymorphism Characterization of the Obligate Biotroph Spongospora subterranea f. sp. subterranea
Authors: Qu Xinshun , Christ Barbara J.
Published: May 2007

Spongospora subterranea f. sp. subterranea causes powdery scab in potatoes and is distributed worldwide. Genetic studies of this pathogen have been hampered due, in part, to its obligate parasitism and the lack of molecular markers for this pathogen. In this investigation, a single cystosorus inoculation technique was developed to produce large amounts of S. subterranea f. sp. subterranea plasmodia or zoosporangia in eastern black nightshade (Solanum ptycanthum) roots from which DNA was extracted. Cryopreservation of zoosporangia was used for long-term storage of the isolates. S. subterranea f. sp. subterranea-specific restriction fragment length polymorphism (RFLP) markers were developed from randomly amplified polymorphic DNA (RAPD) fragments. Cystosori of S. subterranea f. sp. subterranea were used for RAPD assays and putative pathogen-specific RAPD fragments were cloned and sequenced. The fragments were screened for specificity by Southern hybridization and subsequent DNA sequence BLAST search. Four polymorphic S. subterranea f. sp. subterranea-specific probes containing repetitive elements, and one containing single copy DNA were identified. These RFLP probes were then used to analyze 24 single cystosorus isolates derived from eight geographic locations in the United States and Canada. Genetic variation was recorded among, but not within, geographic locations. Cluster analysis separated the isolates into two major groups: group I included isolates originating from western North America, with the exception of those from Colorado, and group II included isolates originating from eastern North America and from Colorado. The techniques developed in this study, i.e., production of single cystosorus isolates of S. subterranea f. sp. subterranea and development of RFLP markers for this pathogen, provide methods to further study the genetic structure of S. subterranea f. sp. subterranea.
Chitosan Oligomers and Copper Sulfate Induce Grapevine Defense Reactions and Resistance to Gray Mold and Downy Mildew
Authors: Aziz Aziz , Trotel-Aziz Patricia , Dhuicq Laurent , Jeandet Philippe , Couderchet Michel , Vernet Guy
Published: May 2007

Chitosan (CHN), a deacetylated derivative of chitin, was shown to be efficient in promoting plant defense reactions. CHN oligomers of different molecular weight (MW) and degree of acetylation (DA) triggered an accumulation of phytoalexins, trans- and cis-resveratrol and their derivatives ε-viniferin and piceid, in grapevine leaves. Highest phytoalexin production was achieved within 48 h of incubation with CHN at 200 μg/ml with an MW of 1,500 and a DA of 20% (CHN1.5/20), while oligomers with greater MW were less efficient, indicating that a specific MW threshold could be required for phytoalexin response. Treatment of grapevine leaves by highly active CHN1.5/20 also led to marked induction of chitinase and β-1,3-glucanase activities. CHN1.5/20 applied together with copper sulfate (CuSO4) strongly induced phytoalexin accumulation. CuSO4alone, especially at low concentrations also elicited a substantial production of phytoalexins in grapevine leaves. Evidence is also provided that CHN1.5/20 significantly reduced the infection of grapevine leaves by Botrytis cinerea and Plasmopara viticola, and in combination with CuSO4conferred protection against both pathogens.
Genetic Characterization and Diversity of Rathayibacter toxicus
Authors: Agarkova I. V. , Vidaver A. K. , Postnikova E. N. , Riley I. T. , Schaad N. W.
Published: May 2007

Rathayibacter toxicus is a nematode-vectored gram-positive bacterium responsible for a gumming disease of grasses and production of a highly potent animal and human toxin that is often fatal to livestock and has a history of occurring in unexpected circumstances. DNA of 22 strains of R. toxicus from Australia were characterized using amplified fragment length polymorphism (AFLP) and pulsed-field gel electrophoresis (PFGE). AFLP analysis grouped the 22 strains into three genetic clusters that correspond to their geographic origin. The mean similarity between the three clusters was 85 to 86%. PFGE analysis generated three different banding patterns that enabled typing the strains into three genotypic groups corresponding to the same AFLP clusters. The similarity coefficient was 63 to 81% for XbaI and 79 to 84% for SpeI. AFLP and PFGE analyses exhibited an analogous level of discriminatory power and produced congruent results. PFGE analysis indicated that the R. toxicus genome was represented by a single linear chromosome, estimated to be 2.214 to 2.301 Mb. No plasmids were detected.
Analyses of Lettuce Drop Incidence and Population Structure ofSclerotinia sclerotiorumandS. minor
Authors: Wu B. M. , Subbarao K. V.
Published: May 2007

To understand the geographical distribution of lettuce drop incidence and the structure of Sclerotinia minor and S. sclerotiorum populations, commercial lettuce fields were surveyed in the Salinas, San Joaquin, and Santa Maria Valleys in California. Lettuce drop incidence, pathogen species, and mycelial compatibility groups (MCGs) were determined and analyzed using geostatistic and geographical information system tools. Lettuce drop incidence was lowest in the San Joaquin Valley, and not significantly different between the other two valleys. Semivariogram analysis revealed that lettuce drop incidence was not spatially correlated between different fields in the Salinas Valley, suggesting negligible field-to-field spread or influence of inoculum in one field on other fields. Lettuce drop incidence was significantly lower in fields with a surface drip system than in fields with furrow or sprinkler irrigation systems, suggesting that the surface drip system can be a potential management measure for reducing lettuce drop. In the San Joaquin Valley, S. sclerotiorum was the prevalent species, causing drop in 63.5% of the fields, whereas S. minor also was identified in 25.4% of the fields. In contrast, in the Salinas Valley, S. minor was the dominant species (76.1%) whereas S sclerotiorum only observed in only 13.6% fields, in which only a few plants were infected by S. sclerotiorum. In the Santa Maria Valley, both species frequently were identified, with S. minor being slightly more common. Although many MCGs were identified in S. minor, most of them consisted of only one or two isolates. In all, ≈91.4% of the isolates belonged to four MCGs. Among them, MCG-1 was the most prevalent group in all three valleys, accounting for 49.8% of total isolates. It was distributed all over the surveyed areas, whereas other MCGs were distributed more or less locally. Populations of S. sclerotiorum exhibited greater diversity, with 89 isolates collected from the Salinas and San Joaquin Valleys belonging to 37 different MCGs. Among them, the most recurrent MCG-A contained 16 isolates, and 30 MCGs contained only 1 isolate each. Many MCGs occurred within only one or a part of the two valleys. Potential reasons for this abundant diversity are discussed.
Distribution and Pathogenic Characterization of Pyrenophora tritici-repentis and Stagonospora nodorum in Ohio
Authors: Engle Jessica S. , Madden Laurence V. , Lipps Patrick E.
Published: May 2007

To determine the distribution of Stagonospora nodorum and Pyrenophora tritici-repentis on wheat in Ohio, flag leaves with lesions were collected from wheat-producing counties in 2002 and 2003. Counties were arbitrarily grouped into seven regions. Log-linear analysis of pathogen presence within regions indicated that the presence of S. nodorum was independent of the presence of P. tritici-repentis. A logistic analysis revealed that the occurrence of both pathogens varied by region in one or both years. The aggressiveness of S. nodorum isolates was determined by inoculating two susceptible genotypes with a subsample of isolates from each region from both years. S. nodorum isolates obtained from northeast Ohio, with fewer wheat fields, were less aggressive than those from other regions. Isolates obtained from west-central Ohio, surrounded by regions with high wheat production annually, were significantly more aggressive than those obtained in the remaining five regions. Isolates from the five other regions did not differ significantly (P > 0.05) in aggressiveness. Races 1 and 2, and a few race 3 isolates, of P. tritici-repentis were detected in Ohio. The distribution of P. tritici-repentis races 1 and 2 was not associated with any region, although the prevalence of race 1 was three times greater than race 2. The rarer race 3 was associated with three dispersed regions. Results indicate that S. nodorum was the major wheat leaf-blotching pathogen. There were no positive or negative associations of S. nodorum and P. tritici-repentis or individual races of P. tritici-repentis in any of the tested regions, which indicates that neither pathogen can be used to predict the presence of the other. The isolated northeastern corner of Ohio appeared to contain isolates of S. nodorum with unique characteristics and potentially only one race of P. tritici-repentis, indicating that this area may be genetically isolated from the remaining tested areas of the state.
Development of a Scale for Evaluation of Tomato yellow leaf curl virus Resistance Level in Tomato Plants
Authors: Lapidot M. , Ben-Joseph R. , Cohen L. , Machbash Z. , Levy D.
Published: May 2007

We have developed a scale of differential hosts that enables the determination and comparison of level of resistance to Tomato yellow leaf curl virus (TYLCV) expressed by resistant tomato lines or by individual plants in a segregating population. The scale is composed of seven different homozygous tomato genotypes that exhibit different levels of TYLCV resistance, ranging from fully susceptible to highly resistant. The differential hosts composing the scale were inoculated with TYLCV under greenhouse conditions. Four weeks after inoculation the plants were evaluated for disease symptom severity, and virus DNA titer was determined. The different genotypes were arranged in the scale according to symptom severity score. The different genotypes were then tested under different environmental conditions, inoculated at different ages, and tested in a field experiment assaying TYLCV-induced yield reduction. While the symptom severity score of each individual resistant genotype changed under different environmental conditions, the relative position on the scale did not alter, except for one genotype. Thus, to evaluate disease resistance of a given tomato genotype, the genotype in question should be inoculated alongside the differential hosts composing the scale, and within 4 weeks one can determine the relative level of resistance of the tested genotype.
Interactive Effects of Two Soilborne Pathogens, Phytophthora capsici and Verticillium dahliae, on Chile Pepper
Authors: Sanogo S.
Published: May 2007

Phytophthora capsici and Verticillium dahliae are two mycelial microorganisms associated with wilt symptoms on chile pepper (Capsicum annuum). Both pathogens occur in the same field and can infect a single plant. This study examined the nature of the co-occurrence of P. capsici and V. dahliae. Chile pepper plants were inoculated with each pathogen separately or with both pathogens concomitantly or sequentially. In concomitant inoculations, plants were inoculated with a mixture of zoospores of P. capsici and conidia of V. dahliae. In sequential inoculations, plants were inoculated with zoospores of P. capsici 4 days prior to inoculation with conidia of V. dahliae, or plants were inoculated with conidia of V. dahliae 4 days prior to inoculation with zoospores of P. capsici. Stem necrosis and leaf wilting were visible 3 to 4 days earlier in plants inoculated with both P. capsici and V. dahliae than in plants inoculated with P. capsici alone. Stem necrosis and generalized plant wilting were observed in plants inoculated with P. capsici alone, and stem necrosis, generalized plant wilting, and vascular discoloration were observed in plants inoculated with both P. capsici and V. dahliae by 21 days after inoculation. These symptoms were not observed in control plants or plants inoculated with V. dahliae alone. The frequency of recovery of V. dahliae from stems was ≈85 to 140% higher across inoculum levels when plants were inoculated with both P. capsici and V. dahliae than when plants were inoculated by V. dahliae alone. Similarly, the frequency of recovery of V. dahliae from roots was ≈13 to 40% higher across inoculum levels when plants were inoculated with both P. capsici and V. dahliae than when plants were inoculated by V. dahliae alone. There was no apparent antagonism between the two pathogens when they were paired on growth media. In general, when P. capsici and V. dahliae were paired on growth media, mycelial growth of each pathogen grown alone was not significantly different from mycelial growth when the pathogens were paired. Results suggest that wilt development is hastened by the presence of both P. capsici and V. dahliae in the same plants. The presence of P. capsici and V. dahliae in the same inoculum court enhanced infection and colonization of chile pepper by V. dahliae.
Symptom Remission and Specific Resistance of Pepper Plants After Infection by Pepper golden mosaic virus
Authors: Carrillo-Tripp Jimena , Lozoya-Gloria Edmundo , Rivera-Bustamante Rafael F.
Published: May 2007

Pepper golden mosaic virus (PepGMV) is an important begomovirus infecting solanaceous crops in Mexico and Central America. Under controlled conditions for growth and inoculation with a low-pressure biolistic device, PepGMV-infected pepper plants consistently showed symptom remission or host recovery 12 to 15 days postinoculation (dpi). Inoculated plants initially developed the characteristic PepGMV symptoms; however, newer leaves presented a significant decrease or disappearance of symptoms. Younger asymptomatic, recovered leaves accumulated lower quantities of viral DNA and transcripts than the ones found in the symptomatic tissue. Nonetheless, viral DNA did not disappear during the evaluation period (up to 35 dpi), suggesting that a population of viral molecules escape from plant defensive mechanisms to maintain a subliminal, symptomless infection. Recovery was correlated with a specific resistance to PepGMV but not to Pepper huasteco yellow vein virus, a different gemi-nivirus commonly found in mixed infections with PepGMV. Virus-related small interfering RNAs were detected in practically all tissues (from symptomatic to recovered leaves) but it was not possible to establish a correlation between concentration and symptom severity. The participation of a posttranscriptional gene silencing mechanism in the recovery process and specific resistance is discussed.
Effects of Phenylpropanoid and Energetic Metabolism Inhibition on Faba Bean Resistance Mechanisms to Rust
Authors: Del Mar Rojas-Molina María , Rubiales Diego , Prats Elena , Sillero Josefina Carmen
Published: May 2007

Effects on penetration and hypersensitive resistance of the cinnamyl acid dehydrogenase (CAD) suicide inhibitor ([(2-hydroxyphenyl) amino] sulphinyl) acetic acid, 1.1 dimethyl ester, which suppresses phenylpro-panoid biosynthesis, and of D-mannose, which sequesters phosphate and reduces energy available in host cells, were studied in faba bean (Vicia faba) genotypes with differing resistance mechanisms to faba bean rust (Uromyces viciae-fabae). Inhibition of CAD reduced penetration resistance in lines 2N-34, 2N-52, V-1271, and V-1272, revealing an important role for phenylpropanoid biosynthesis in the resistance of these lines. Inhibition of CAD also inhibited hypersensitive cell death in these lines. D-mannose had little or no effect on resistance. By contrast, CAD inhibition did not affect penetration resistance of line BPL-261, which has a high degree of penetration resistance not associated with hypersensitive cell death. In BPL-261, D-mannose inhibited penetration resistance. The parallelism between the faba bean genotype responses to rust observed here and the response of barley genotypes with differing resistance mechanisms to powdery mildew after similar inhibitor treatments is analyzed and discussed.
Effect of Seed Quality and Combination Fungicide-Trichoderma spp. Seed Treatments on Pre- and Postemergence Damping-Off in Cotton
Authors: Howell Charles R.
Published: May 2007

Good quality seeds of cotton cultivars often escaped pre-emergence damping-off incited by Pythium spp. and Rhizopus oryzae, and they were resistant to postemergence damping-off incited by Rhizoctonia solani. Poor quality seeds, however, were highly susceptible to both phases of seedling disease and required seed treatment in order to survive. Pre-emergence damping-off incited by Pythium spp. and Rhizopus oryzae could be controlled by seed treatment with biocontrol preparations of a number of Trichoderma spp., but these treatments were much less effective in controlling postemergence disease incited by Rhizoctonia solani. Postemergence seedling disease can be controlled by fungicides, but they were much less effective in controlling the pre-emergence phase of the disease. Combination seed treatments of poor quality cotton seeds with fungicides and Trichoderma spp. preparations, followed by planting in pathogen-infested soil, indicated that this technique will control both phases of seedling disease. Seed treatment with either the fungicides or the biocontrol agents alone did not achieve this goal. The optimum combination treatment for disease control was that of chloroneb plus Trichoderma spp., followed by chloroneb plus metalaxyl (Deltacoat AD) plus T. virens strain G-6.
Novel Naturally Occurring Bean pod mottle virus Reassortants with Mixed Heterologous RNA1 Genomes
Authors: Gu Hongcang , Zhang Chunquan , Ghabrial Said A.
Published: May 2007

The comovirus Bean pod mottle virus (BPMV) is widespread in the soybean-growing regions in the United States. It has a bipartite genome consisting of RNA1 and RNA2, which are encapsidated separately. We previously have reported the occurrence in nature of two distinct subgroups of BPMV strains (subgroups I and II), as well as reassortants between the two subgroups. Here, we report the isolation and molecular characterization of naturally occurring partial diploid reassortant strains, which are diploid for RNA1 and haploid for RNA2. Whereas the RNA1s of the partial diploids are derived from two distinct strain subgroups (I and II), the RNA2 is derived from either subgroup I or II. The partial diploid strains induced strikingly severe symptoms on soybean, which could be explained based on the presence of two distinct RNA1s in the same plant. This conclusion was supported by the finding that pseudo-recombinants constructed with two diverse RNA1s induced very severe symptoms on soybean that mimicked those produced by the naturally occurring partial diploids. No enhancement of symptom severity was observed with pseudorecombinants constructed with closely related RNA1s. Likewise, no enhancement of symptom severity was noted with pseudo-recombinants that are diploid for RNA2 and haploid for RNA1. The potential role of genetic reassortment in the epidemiology and pathogenesis of BPMV is discussed.
Phylogeny of Fusarium oxysporum f. sp. lactucae Inferred from Mitochondrial Small Subunit, Elongation Factor 1-α, and Nuclear Ribosomal Intergenic Spacer Sequence Data
Authors: Mbofung Gladys Y. , Hong Soon Gyu , Pryor Barry M.
Published: May 2007

Fusarium oxysporum f. sp. lactucae, causal agent of Fusarium wilt of lettuce, is a serious pathogen recently reported in Arizona. Sequence analysis of the mitochondrial small subunit (mtSSU), translation elongation factor 1-α (EF-1α) gene, and the nuclear ribosomal DNA intergenic spacer (IGS) region was conducted to resolve relationships among f. sp. lactucae isolates, F. oxysporum isolates from other hosts, and local non-pathogenic isolates. Analysis of mtSSU sequences provided limited phylogenetic resolution and did not differentiate the lactucae isolates from 13 other F. oxysporum isolates. Analysis of EF-1α sequences resulted in moderate resolution, grouping seven formae speciales with the lactucae isolates. Analysis of the IGS region revealed numerous sequence polymorphisms among F. oxysporum formae speciales consisting of insertions, deletions, and single nucleotide transitions and substitutions. Repeat sequence analysis revealed several duplicated subrepeat units that were distributed across much of the region. Based on analysis of the IGS sequence data, lactucae race 1 isolates resolved as a monophyletic group with three other formae speciales of F. oxysporum. In all analyses, lactucae race 2 isolates composed a separate lineage that was phylo-genetically distinct and distantly related to the lactucae race 1 isolates.
Risk Factors for the Phytophthora-Induced Decline of Alder in Northeastern France
Authors: Thoirain B. , Husson C. , Marçais B.
Published: May 2007

A lethal disease of common alder caused by Phytophthora alni, a new hybrid pathogen, has been spreading in Europe since the early 1990s. In 2004, we conducted an epidemiological survey in northeastern France to determine disease frequency and to investigate the impact of environmental factors on disease prevalence. Seventy-eight plots in the Rhin-Meuse basin were investigated. The survey was structured to enable critical examination of the possible impact of nitrogen pollution of the river water on disease prevalence. P. alni-induced alder decline was common throughout northeastern France. Altogether, disease was found in 80% of the plots containing alder, with 16% of all the alders affected. Striking differences existed between watercourse types. Lower proportions of diseased alders were found in watercourse types with rapid water flow, such as mountain streams of the Vosges and piedmont or watercourses on steep calcareous slopes, than in the slow watercourses of the low-lying valleys of the calcareous plateaus and of the clayey plains. Disease prevalence was not related to the total oxidized nitrogen concentration of the water. However, prevalence increased with the mean summer temperature of the river water and where clayey soils were found in the river banks. The results of this work can be used for the assessment of P. alni-induced alder decline risks in affected European countries and in areas where the disease could be introduced.
Genetic Analysis of Soybean Plant Introductions with Resistance toPhytophthora sojae
Authors: Gordon S. G. , Berry S. A. , St. Martin S. K. , Dorrance A. E.
Published: May 2007

Phytophthora sojae, which causes Phytophthora root and stem rot of soybean, is a serious disease worldwide and is managed primarily by deploying cultivars with resistance. Thirty-two soybean plant introductions (PIs), all but three of which were from South Korea, were proposed as new sources of single-gene resistance to P. sojae. The objective of this study was to characterize the inheritance of resistance to P. sojae in these PIs. Twenty-two soybean populations from crosses of these PIs and the susceptible cv. Williams were inoculated with P. sojae OH17 (vir 1b, 1d, 2, 3a, 3b, 3c, 4, 5, 6, 7), and OH25 (vir 1a, 1b, 1c, 1k, 7). These isolates were selected because they are virulent on soybeans with all known Rps genes and many Rps gene combinations. Thirteen of the twenty-two populations had consistent segregation responses following inoculations between the two generations. In two PIs, resistance was conferred by two genes to OH17 and three genes to OH25. Resistance to both isolates was conferred by a single gene in PI 398440 although the individual families were not resistant to the same isolates. The data suggest that six of the populations have three-Rps gene combinations as previously proposed, while another four may have either a novel Rps gene or a four-Rps gene combination. Based on this phenotypic analysis, novel and uncharacterized Rps genes may be present in this material. More importantly, these PIs may serve as sources of novel Rps genes that can be used to more effectively manage Phytophthora root and stem rot.
Molecular Typing and Presence of Genetic Markers Among Strains of Banana Finger-Tip Rot Pathogen, Burkholderia cenocepacia, in Taiwan
Authors: Lee Yung-An , Chan Chih-Wen
Published: May 2007

Burkholderia cenocepacia (genomovar III of B. cepacia complex), the causal agent of banana finger-tip rot, is a common plant-associated bacterium but also an important opportunistic pathogen of humans. To better understand the nature of B. cenocepacia from banana, the genetic variation among B. cenocepacia isolates from various banana-growing regions in southern Taiwan was examined. Forty-four serial isolates recovered from diseased banana stigmata from three banana-growing regions during the periods ranging from 2002 to 2004 were investigated. All B. cenocepacia isolates picked from quinate-yeast extract tetracycline-polymyxin semiselective medium could cause onion maceration and were polymerase chain reaction (PCR) positive for bcscV, which is a type III secretion gene present in all members of the B. cepacia complex except B. cepacia (formerly genomovar I). Genetic diversity was assessed using recA PCR restriction fragment length polymorphism, recA nucleotide sequence analysis, and pulsed-field gel electrophoresis assays. The assays revealed the genetic variability among the isolates and also allowed us to trace the relationship among isolates. The isolates all were assigned to genomovar III and consisted of two groups, A and B, which corresponded to recA lineage IIIA and IIIB. The group B strains were separated into B1 and B2 subgroups and the B1 strains were further divided into distinct lineages. The B1 strains were the most frequently detected and occurred in all regions tested. There was no significant difference between strains from each subgroup in the virulence on banana fingers of cv. Cavendish. PCR assays were further used to determine whether B. cenocepacia from banana contained the cable pilus subunit gene (cblA), IS1356, and B. cepacia epidemic strain marker (BCESM), which are DNA markers associated with epidemic B. cepacia clinic strains. The results indicated that cblA and IS1356 were absent but the BCESM was found in all isolates. The present study revealed that banana is a natural reservoir of genetically diversified B. cenocepacia strains.
The Sensitivity of the Epidemic Growth Rate to Weather Variables, with an Application to Yellow Rust on Wheat
Authors: Papastamati Konstantina , van den Bosch Frank
Published: May 2007

We first show how to estimate the exponential epidemic growth rate, r, for different combinations of three weather variables. Then we derive a method to quantify the sensitivity of r to a weather variable as a function of the pathogen life cycle variables of latent period, basic reproductive number, and the mean and standard deviation of the sporulation curve. The method can be used to identify the most important weather variable and pathogen life cycle component in terms of epidemic progress. The method is applied to yellow rust, caused by Puccinia striiformis, on winter wheat. We conclude that the most important weather variable for the progress of yellow rust is temperature, followed by dew period and light quantity. By far, the most important pathogen life cycle component is the basic reproductive number, especially at low and high temperatures. This disagrees with the general view that latent period is the most important variable at low temperatures. We discuss explanations of this.
Pseudomonas Biocontrol Agents of Soilborne Pathogens: Looking Back Over 30 Years
Authors: Weller David M.
Published: May 2007

Pseudomonas spp. are ubiquitous bacteria in agricultural soils and have many traits that make them well suited as biocontrol agents of soilborne pathogens. Tremendous progress has been made in characterizing the process of root colonization by pseudomonads, the biotic and abiotic factors affecting colonization, bacterial traits and genes contributing to rhizosphere competence, and the mechanisms of pathogen suppression. This review looks back over the last 30 years of Pseudomonas biocontrol research and highlights key studies, strains, and findings that have had significant impact on shaping our current understanding of biological control by bacteria and the direction of future research.
Artificial Infection of Rosellinia necatrix with Purified Viral Particles of a Member of the Genus Mycoreovirus Reveals Its Uneven Distribution in Single Colonies
Authors: Sasaki Atsuko , Kanematsu Satoko , Onoue Mari , Oikawa Yuri , Nakamura Hitoshi , Yoshida Kouji
Published: May 2007

Rosellinia necatrix mycoreovirus 3 (W370) (RnMYRV-3/W370, described as RnMYRV-3 in this paper), a member of the newly established genus Mycoreovirus within the family Reoviridae, is the hypovirulence factor of the white root rot fungus, Rosellinia necatrix. Two virus-free fungal isolates (W37 and W97) that were somatically incompatible with the virus-harboring field isolate (W370) were transfected with purified RnMYRV-3 particles. Virus infection was confirmed by electrophoresis and northern hybridization of viral double-stranded RNA. RnMYRV-3 was transmissible from transfected strains to their respective, virus-free counterparts via hyphal anastomosis. Virus-transfected strains produced smaller lesions on apple fruits than did their virus-free counterparts. Virus-cured strains were indistinguishable from wild-type strains in culture morphology and displayed approximately the same virulence level on apples. Virus-transfected strains had “mosaic” colony portions consisting of thin, fast-growing and dense, slow-growing mycelia, and grew more slowly as a whole than their virus-free, parental strains. The level of virus accumulation varied among virus-transfected subcultures and within its single colonies. Virus-transfected strains were occasionally cured, as was W370. Such a phenomenon may be ascribed to uneven viral distribution in single colonies and the difficulty in viral transmission to virus-free hyphae.
Mutations Associated with Resistance-Breaking Isolates of Beet necrotic yellow vein virus and Their Allelic Discrimination Using TaqMan Technology
Authors: Acosta-Leal R. , Rush C. M.
Published: May 2007

Genetic resistance in sugar beet (Beta vulgaris) to Beet necrotic yellow vein virus (BNYVV), which causes the disease rhizomania, is conferred by the single dominant gene Rz1. However, since 2002, Rz1 cultivars grown in the Imperial Valley of California have been increasingly damaged by a new strain of BNYVV. Viral RNA 3 was extracted from asymptomatic and symptomatic sugar beets and, after amplification and sequencing of a region including the p25 cistron, two polymorphic sites, A67V and D135E, associated with the capability of the virus to overcome resistance were identified. Using the real-time reverse transcription-polymerase chain reaction allelic discrimination technique, TaqMan probes designed to detect the responsible nucleotide substitutions permitted the differentiation between wild type (WT) and resistance-breaking (RB) isolates. This method also allowed easy detection of mixed infections by giving a heterozygous call, which was verified by DNA sequencing of individual clones. The capability of this technology to typify numerous isolates facilitated the analysis of the spatial distribution of virus haplotypes in the field. Thus, RB variants were mostly baited from yellow strips with high incidence of rhizomania, whereas WT variants predominated in the surrounding green areas. Mixed infections were found mainly in green areas and transitional zones. The predominance of the RB isolates in yellow strips suggests that they have gained fitness in Rz1 cultivars and will eventually become the dominant haplotype.
Seasonal Dynamics of Taphrina deformans Inoculum in Peach Orchards
Authors: Rossi V. , Bolognesi M. , Giosuè S.
Published: May 2007

The dynamics of the inoculum of Taphrina deformans were studied during a 4-year period by (i) inspecting curled leaves for the presence of asci, (ii) placing deposition spore samplers within the tree canopies, and (iii) exposing potted peach plants (trap plants). These three approaches produced consistent results. Four main periods characterized the dynamics of the inoculum: the first period coincides with the parasitic stage of the pathogen's life cycle and the other periods with the saprophytic stage. Mid- to late spring (first period) was characterized by the presence of asci on infected leaves which produced and ejected large quantities of ascospores in 96% of the samplings. Rainfall was not necessary for ascospore dispersal, which was favored by air temperature <20°C and relative humidity ≥80% or wetness duration >8 h. In summer and autumn (second period), blastospores were trapped in 54 and 24% of samplings, respectively, with low spore numbers. In the winter (third period), blasto-spores were trapped in the lowest numbers and in only 6% of samplings. In late winter to early spring (fourth period), blastospores were found in 56% of samples, with increasing numbers. Rainfall significantly influenced blastospore dispersal and temperature was correlated with the seasonality found during the saprophytic stage.
Phosphite Inhibits Development of the Nematodes Heterodera avenae and Meloidogyne marylandi in Cereals
Authors: Oka Yuji , Tkachi Nadia , Mor Mishael
Published: May 2007

Phosphonic acid (H3PO3) solutions were applied to wheat or to bristle oat as soil drenches before inoculation with juveniles of the sedentary, endoparasitic nematodes Heterodera avenae or Meloidogyne marylandi. All the solutions, which were pH adjusted and added at levels as low as 0.63 mg of phosphite (HPO32-) per plant, reduced the numbers of H. avenae females and M. marylandi egg masses. Phosphate (PO43-), applied as potassium phosphate at the same concentrations, did not reduce the number of female nematodes on the wheat. Addition of phosphate to the phosphite solutions did not change the inhibitory effect of phosphite on H. avenae, but it reduced phosphite's effect on M. marylandi. Phosphite also reduced the number of H. avenae females when applied as many as 20 days after addition of nematodes. The phosphite treatment did not prevent M. marylandi juveniles from penetrating wheat roots or inducing giant cells. However, phosphite inhibited giant cell development: 14 days after inoculation, the giant cells in the phosphite-treated wheat were almost completely vacuolated, whereas those in untreated wheat contained dense cytoplasm.
Ageratum yellow vein China virus Is a Distinct Begomovirus Species Associated with a DNAβ Molecule
Authors: Xiong Qing , Fan Sanwei , Wu Jianxiang , Zhou Xueping
Published: May 2007

Ageratum conyzoides plants exhibiting yellow vein symptoms, collected near Haikou, Hainan Province, China, contained begomoviral DNA-A-like molecules. The complete sequences of the molecules from two samples, Hn2 and Hn2-19, were shown to consist of 2,768 and 2,748 nucelotides (nt), respectively. These sequences have more than 97% nucleotide sequence identity, but less than 86% identity with other reported begomovirus sequences. In line with the taxonomic convention for begomoviruses, Hn2 and Hn2-19 are therefore considered to represent isolates of a distinct begomovirus species, for which the name Ageratum yellow vein China virus (AYVCNV) is proposed. Sequence alignment shows AYVCNV has arisen by recombination among viruses related to Ageratum yellow vein virus, Papaya leaf curl China virus, and an unidentified begomovirus. Southern blot analyses revealed that all plants sampled contained molecules resembling DNAβ. DNAβ molecules from three samples were 1,323 or 1,324 nt long and had >98% sequence identity but <81% identity with previously reported DNAβ sequences. Infectious clones of Hn2 and its associated DNAβ were constructed and agroinoculated to plants. Hn2 alone caused sporadic asymptomatic systemic infection of Nicotiana benthamiana, N. glutinosa, Lycopersicon esculentum, Petunia hybrida, and A. conyzoides but its accumulation was much enhanced in plants co-inoculated with DNAβ. The co-inoculated N. benthamiana, N. glutinosa, P. hybrida, and L. esculentum plants developed leaf curling or leaf crinkling symptom; those in A. conyzoides were typical of ageratum yellow vein disease. When the DNAβ molecules associated with four other Chinese begomoviruses were coinoculated with Hn2 to N. benthamiana and N. glutinosa, the DNAβ molecules were replicated, and the plants developed systemic symptoms of types that were specific for each DNAβ. This illustrates that there is less specific interaction between monopartite begomovirus and DNAβ than between the DNA-A and DNA-B of begomoviruses with bipartite genomes.
Systemic Modulation of Gene Expression in Tomato by Trichoderma hamatum 382
Authors: Alfano G. , Ivey M. L. Lewis , Cakir C. , Bos J. I. B. , Miller S. A. , Madden L. V. , Kamoun S. , Hoitink H. A. J.
Published: May 2007

A light sphagnum peat mix inoculated with Trichoderma hamatum 382 consistently provided a significant (P = 0.05) degree of protection against bacterial spot of tomato and its pathogen Xanthomonas euvesicatoria 110c compared with the control peat mix, even though this biocontrol agent did not colonize aboveground plant parts. To gain insight into the mechanism by which T. hamatum 382 induced resistance in tomato, high-density oligonucleotide microarrays were used to determine its effect on the expression pattern of 15,925 genes in leaves just before they were inoculated with the pathogen. T. hamatum 382 consistently modulated the expression of genes in tomato leaves. We identified 45 genes to be differentially expressed across the replicated treatments, and 41 of these genes could be assigned to at least one of seven functional categories. T. hamatum 382-induced genes have functions associated with biotic or abiotic stress, as well as RNA, DNA, and protein metabolism. Four extensin and extensin-like proteins were induced. However, besides pathogenesis-related protein 5, the main markers of systemic acquired resistance were not significantly induced. This work showed that T. hamatum 382 actively induces systemic changes in plant physiology and disease resistance through systemic modulation of the expression of stress and metabolism genes.
Identification of a Novel Fusarium Head Blight Resistance Quantitative Trait Locus on Chromosome 7A in Tetraploid Wheat
Authors: Kumar S. , Stack R. W. , Friesen T. L. , Faris J. D.
Published: May 2007

Fusarium head blight (FHB) caused by Fusarium graminearum is one of the most destructive diseases of durum (Triticum turgidum sp. durum) and common wheat (T. aestivum). Promising sources of FHB resistance have been identified among common (hexaploid) wheats, but the same is not true for durum (tetraploid) wheats. A previous study indicated that chromosome 7A from T. turgidum sp. dicoccoides accession PI478742 contributed significant levels of resistance to FHB. The objectives of this research were to develop a genetic linkage map of chromosome 7A in a population of 118 recombinant inbred lines derived from a cross between the durum cv. Langdon (LDN) and a disomic LDN-T. turgidum sp. dicoccoides PI478742 chromosome 7A substitution line [LDN-DIC 7A(742)], and identify a putative FHB resistance quantitative trait locus (QTL) on chromosome 7A derived from LDN-DIC 7A(742). The population was evaluated for type II FHB resistance in three greenhouse environments. Interval regression analysis indicated that a single QTL designated Qfhs.fcu-7AL explained 19% of the phenotypic variation and spanned an interval of 39.6 cM. Comparisons between the genetic map and a previously constructed physical map of chromosome 7A indicated that Qfhs.fcu-7AL is located in the proximal region of the long arm. This is only the second FHB QTL to be identified in a tetraploid source, and it may be useful to combine it with the QTL Qfhs.ndsu-3AS in order to develop durum wheat germ plasm and cultivars with higher levels of FHB resistance.
Asexual Genetic Exchange in the Barley Pathogen Rhynchosporium secalis
Authors: Forgan Angus H. , Knogge Wolfgang , Anderson Peter A.
Published: May 2007

The causal agent of barley scald, Rhynchosporium secalis, is a haploid anamorphic ascomycete with no known sexual stage. Nevertheless, a high degree of genetic variation has been observed in fungal populations on commercial barley cultivars and parasexuality has been suggested to contribute to this variation. In order to test whether asexual genetic exchange can occur, isolates of R. secalis were transformed to hygromycin B resistance or phleomycin resistance. Mixtures of transformants were co-inoculated either on agar or in planta and screened for the occurrence of dual-antibiotic-resistant colonies. No dual-antibiotic-resistant colonies resulted from mixing transformants of different fungal isolates. In contrast, with transformants originating from the same fungal isolate, asexual exchange of markers was demonstrated on agar plates and in planta. This is the first definitive evidence of asexual genetic exchange in R. secalis.
Production and Characterization of a Monoclonal Antibody Raised Against Surface Antigens from Mycelium of Gaeumannomyces graminis var. tritici: Evidence for an Extracellular Polyphenol Oxidase
Authors: Thornton Christopher R. , Dewey Frances M. , Gilligan Christopher A.
Published: May 2007

A murine monoclonal antibody (MAb) of immunoglobulin class M (IgM) was raised against surface antigens from Gaeumannomyces graminis var. tritici and, by enzyme-linked immunosorbent assay, recognized isolates of G. graminis var. tritici, G. graminis var. avenae and G. graminis var. graminis. Characterization of the antigen by heat and protease treatments showed that the epitope recognized by the MAb was a protein. Antigen production was detected only in live mycelia. Immunofluorescence studies showed that the antigen was associated with both the broad melanized macrohyphae and hyaline mycelia of G. graminis var. tritici. Secretion of antigen into an aqueous minimal medium was promoted only by exposure of live mycelia to certain phenolic substrates, including monophenols ortho-, para-, and meta-cresol; 3,4,5-trihydroxybenzoic acid (gallic acid); and phenolic amino acid L-3-(3,4-dihydroxyphenyl) alanine (L-DOPA). Antigen secretion was not promoted by 3-(4-hydroxyphenyl) alanine (L-tyrosine). The MAb reacted strongly with purified enzyme laccase (polyphenol oxidase, EC 1.10.3.2) but did not recognize purified tyrosinase (monophenol oxidase, EC 1.14.18.1). Moreover, chemicals that bind to copper and inhibit copper-containing enzymes such as laccase completely inhibited antigen secretion in response to L-DOPA. The MAb was tested for specificity against a wide range of fungi, common yeast species, and gram positive and negative bacteria. It did not recognize antigens from a broad range of unrelated fungi, including Gliocladium roseum, Fusarium sp., Phoma exigua, Phialophora fastigiata, Penicillium crustosum, Pythium ultimum, Rhizopus stolonifer, Rhizoctonia carotae, R. oryzae, R. tuliparum, and Trichoderma viride, nor did it recognize surface antigens from yeasts or bacteria. The MAb cross-reacted with antigens from Botrytis spp., Chaetomium globosum, R. cerealis, and R. solani. However, secretion of antigen by R. solani and R. cerealis was not promoted by L-DOPA, and secretion by C. globosum in response to the phenolic amino acid was significantly less compared to G. graminis var. tritici.
Detection and Description of Spatial Patterns of Bacterial Brown Spot of Snap Beans Using Cyclic Samples
Authors: Hudelson B. D. , Clayton M. K. , Smith K. P. , Upper C. D.
Published: May 2007

Snap bean plants within seven-row segments that ranged from 65 to 147 m were sampled, using a cyclic sampling plan. In the cyclic sampling plan, only 6 of every 31 plants were sampled, but sampled plants were spaced such that pairs of plants that were 1, 2, 3, 4,…, 1,525 plants apart could be identified within each sample. Every leaflet on every sampled plant was assessed for bacterial brown spot, and the proportion of disease leaflets per plant was determined. Arcsine square-root-transformed disease incidence values were analyzed for spatial patterns by autocorrelation and spectral analyses. Disease patterns were detected at several different scales within a single snap bean row, at distances that ranged from ˜20 to ˜100 m. Approximately 23 to 53% of the disease variability in the samples could be described by sine and cosine curves, indicating a substantial component of regularity in the disease patterns. Possible origins for these regular patterns, including cultural practices and seed infestation, are discussed.
Population Structure and Disease Development ofCryphonectria parasiticain European Chestnut Forests in the Presence of Natural Hypovirulence
Authors: Bissegger Martin , Rigling Daniel , Heiniger Ursula
Published: May 2007

The Cryphonectria parasitica populations in two 6-year-old European chestnut (Castanea sativa) coppices were investigated in southern Switzerland over a period of 4 years. Occurrence of white isolates indicating an infection with Cryphonectria hypovirus, vegetative compatibility groups (VCGs), hypovirulence conversion capacity, and mating types were used to characterize the populations. Sampling of randomly chosen cankers in the first year yielded 59% white isolates in one and 40% in the other population. The distribution of the VCGs and mating types was similar among white and orange isolates, indicating a homogeneous infection of the two populations by the hypovirus. Fourteen VCGs were found in the first population, 16 VCGs in the second. Altogether, 21 VCGs were determined. The same three VCGs dominated in both populations, comprising more than 60% of all isolates. Several VCGs were represented only by white isolates. Five of the six most common VCGs were clustered in two hypovirulence conversion groups, with almost 100% hypovirus transmission within each cluster. Repeated sampling of the same cankers in 1990, 1992, and 1994 did not reveal an increase of white isolates. The portion of blighted stems rose from 37% to about 60% in both plots within 4 years. In this time, chestnut blight killed 15% and competition an additional 21% of the sprouts. Predominantly, sprouts with low diameters at breast height were killed. The growth rate of new cankers was high in their first year and decreased gradually in the following years. A role of hypovirulence in the decline of disease severity was evident since (i) cankers yielding white isolates grew slower and killed considerably fewer sprouts than cankers with orange isolates; and (ii) the majority of the cankers yielded white isolates at least once during the 4-year observation period.