bluuebunny/crossref_metadata_2025 huggingface dataset:
A One-Step, Immunochromatographic Lateral Flow Device Specific toRhizoctonia solaniand Certain Related Species, and Its Use to Detect and QuantifyR. solaniin Soil
Authors: Thornton Christopher R. , Groenhof Andrew C. , Forrest Robert , Lamotte Richard
Published: May 2007

A murine hybridoma cell line GD2 secreting an immunoglobulin (Ig)M monoclonal antibody (MAb) was produced against surface antigens from an anastomosis group (AG) 4 isolate of Rhizoctonia solani (teleomorph: Thanatephorus cucumeris). Ascites were produced in mice using GD2 hybridoma cells and used to develop a rapid immunochromatographic lateral flow device (LFD) for the detection of antigens from R. solani and certain related Rhizoctonia spp. The LFD was tested for specificity against surface antigens from related and unrelated soil fungi. Antigens from representative isolates of R. solani AGs 1, 2-1, 2-3, 2-t, 3, 4, 5, 6, 7, 8, 9, 10, 11, and BI gave a positive response in LFD tests, as did antigens from Thanatephorus orchidicola, T. praticola, R. fragariae (teleomorph: Ceratorhiza fragariae), Ceratorhiza goodyerae-repentis, Ceratobasidium cornigerum, and binucleate AGE. Antigens from R. solani AGs 2-2, 2-2IIIB, and 2-2IV and from the related fungi R. carotae, R. cerealis (teleomorph: Ceratobasium cereale), R. crocorum (teleomorph: Helicobasidium brebissonii), R. oryzae (teleomorph Waitea circinata), and R. zeae gave negative responses, as did antigens from a range of unrelated fungi and oomycetes including Fusarium, Gliocladium, Trichoderma, Pythium, and Phytophthora spp. The usefulness of the LFD to detect R. solani was demonstrated in soils naturally infested with R. solani AG3. There was close agreement between results of LFD tests and conventional plate enrichment tests employing selective medium. The specificity of the technique was confirmed by polymerase chain reaction PCR using R. solani AG3-specific primers and by analyses based on sequences of the internal transcribed spacer (ITS)1-5.8S-ITS2 rRNA-encoding regions of unrelated fungi recovered from soil samples. The LFD was used to quantify R. solani AG4 in artificially infested soil samples (chopped potato soil inoculum). Estimates of CFU per gram of soil were derived using a most-probable number technique, which was based on the presence or absence of a detectable signal in the LFD. Estimates of CFU obtained in LFD tests and those obtained in a plate-trapped antigen enzyme-linked immunosorbent assay incorporating MAb GD2 were identical (449 CFU g-1of soil).
The Am Gene Controlling Resistance to Alfalfa mosaic virus in Tomato Is Located in the Cluster of Dominant Resistance Genes on Chromosome 6
Authors: Parrella Giuseppe , Moretti André , Gognalons Patrick , Lesage Marie-Laure , Marchoux George , Gebre-Selassie Kashay , Caranta Carole
Published: May 2007

The dominant gene Am from Lycopersicon hirsutum f. sp. glabratum PI134417 confers resistance to most strains of Alfalfa mosaic virus, including the recently identified necrotic strains. The phenotypic response includes a lack of symptom development following mechanical inoculation of leaves. To study the resistance mechanism controlled by Am, biological (back-inoculation to susceptible hosts), serological (double-antibody sandwich, enzyme-linked immunosorbent assay), and molecular (reverse transcription-polymerase chain reaction and hybridization with specific riboprobes) methods of virus detection have been conducted on mechanically inoculated PI134417 leaves. The virus was never recovered, indicating that Am acts by an inhibition of viral accumulation during the early events of the virus life cycle. Am has been mapped genetically to the short arm of tomato chromosome 6 in the resistance hotspot, which includes the R-genes Mi and Cf-2/Cf-5 and the quantitative resistance factors Ty-1, Ol-1, and Bw-5.
Quantitative Trait Loci (QTL) Analysis Reveals Both Broad-Spectrum and Isolate-Specific QTL for Scab Resistance in an Apple Progeny Challenged with Eight Isolates of Venturia inaequalis
Authors: Calenge F. , Faure A. , Goerre M. , Gebhardt C. , Van de Weg W. E. , Parisi L. , Durel C.-E.
Published: May 2007

The major scab resistance gene Vf, extensively used in apple breeding programs, was recently overcome by the new races 6 and 7 of the fungal pathogen Venturia inaequalis. New, more durable, scab resistance genes are needed in apple breeding programs. F1 progeny derived from the cross between partially resistant apple cv. Discovery and apple hybrid ‘TN10-8’ were inoculated in the greenhouse with eight isolates of V. inaequalis, including isolates able to overcome Vf. One major resistance gene, Vg, and seven quantitative trait loci (QTL) were identified for resistance to these isolates. Three QTL on linkage group (LG)12, LG13, and LG15 were clearly isolate-specific. Another QTL on LG5 was detected with two isolates. Three QTL on LG1, LG2, and LG17 were identified with most isolates tested, but not with every isolate. The QTL on LG2 displayed alleles conferring different specificities. This QTL co-localized with the major scab resistance genes Vr and Vh8, whereas the QTL on LG1 colocalized with Vf. These results contribute to a better understanding of the genetic basis of the V. inaequalis-Malus × domestica interaction.
Survival of Phytophthora infestans in Surface Water
Authors: Porter Lyndon D. , Johnson Dennis A.
Published: May 2007

Coverless petri dishes with water suspensions of sporangia and zoospores of Phytophthora infestans were embedded in sandy soil in eastern Washington in July and October 2001 and July 2002 to quantify longevity of spores in water under natural conditions. Effects of solar radiation intensity, presence of soil in petri dishes (15 g per dish), and a 2-h chill period on survival of isolates of clonal lineages US-8 and US-11 were investigated. Spores in water suspensions survived 0 to 16 days under nonshaded conditions and 2 to 20 days under shaded conditions. Mean spore survival significantly increased from 1.7 to 5.8 days when soil was added to the water. Maximum survival time of spores in water without soil exposed to direct sunlight was 2 to 3 days in July and 6 to 8 days in October. Mean duration of survival did not differ significantly between chilled and nonchilled sporangia, but significantly fewer chilled spores survived for extended periods than that of nonchilled spores. Spores of US-11 and US-8 isolates did not differ in mean duration of survival, but significantly greater numbers of sporangia of US-8 survived than did sporangia of US-11 in one of three trials.
Barrage Zone Formation Between Vegetatively Incompatible Fusarium graminearum (Gibberella zeae) Isolates
Authors: McCallum B. D. , Tekauz A. , Gilbert J.
Published: May 2007

Vegetative compatibility has been used to assess the population biology of many fungal plant pathogens. However, for many species, including Fusarium graminearum, this has meant making auxotrophic mutants to force heterokaryon formation. A method was developed to observe barrage zones of thick, raised mycelium at the junctions of vegetatively incompatible F. graminearum isolates. The appearance of the barrage zones was influenced by the growth medium and the light. Barrage zones on V8 agar were thicker and better defined than those on potato dextrose agar, Spezieller Nahrstoffarmer agar, and water agar. The addition of ground wheat kernels to V8 agar enhanced barrage zone formation. Incubating the cultures under constant light at 2,150 lx produced more distinct barrage zones than constant light at 3,400 lx, constant darkness, or ambient room light. Forty-three F. graminearum isolates from 34 vegetative compatibility groups, determined previously using nit auxotrophic mutants, were paired in all combinations using these optimized conditions. Isolates in different vegetative compatibility groups typically formed distinct, thick barrage zones at their junctions. Pairs of isolates in the same vegetative compatibility group had a very slight or no visible reaction, or rarely, a distinct “line gap” of sparse mycelium. Subcultures from the same isolate typically had no visible reaction at their colony junctions; however, subcultures from some isolates had thin, slight barrage zones. This method was used to identify the proportion of each of four F. graminearum isolates from infected barley spikes in the field, inoculated previously with a mixture of conidia from these four isolates. Barrage zone formation represents a rapid method to screen vegetative compatibility groups in F. graminearum and may be useful for other Fusarium species.
Genetic and Pathogenic Analyses of Colletotrichum gloeosporioides Isolates from Strawberry and Noncultivated Hosts
Authors: Xiao C. L. , MacKenzie S. J. , Legard D. E.
Published: May 2007

Colletotrichum crown rot of strawberry in Florida is caused primarily by Colletotrichum gloeosporioides. To determine potential inoculum sources, isolates of Colletotrichum spp. from strawberry and various noncultivated plants growing in the areas adjacent to strawberry fields were collected from different sites. Species-specific internal transcribed spacer primers for C. gloeosporioides and C. acutatum were used to identify isolates to species. Random amplified polymorphic DNA (RAPD) markers were used to determine genetic relationships among isolates recovered from noncultivated hosts and diseased strawberry plants. Selected isolates also were tested for pathogenicity on strawberry plants in the greenhouse. In all, 39 C. gloeosporioides and 3 C. acutatum isolates were recovered from diseased strawberry crowns, and 52 C. gloeosporioides and 1 C. acutatum isolate were recovered from noncultivated hosts. In crown inoculation tests, 18 of the 52 C. gloeosporioides isolates recovered from noncultivated hosts were pathogenic to strawberry. Phylogenetic analysis using RAPD marker data divided isolates of C. gloeosporioides from noncultivated hosts into two separate clusters. One cluster contained 50 of the 52 isolates and a second cluster contained 2 isolates that were homothallic in culture. Isolates from strawberry were interspersed within the cluster containing the 50 isolates that were recovered from noncultivated hosts. The results are not inconsistent with the hypothesis that C. gloeosporioides isolates obtained from strawberry and noncultivated hosts adjacent to strawberry fields are from the same population and that noncultivated hosts can serve as potential inoculum sources for Colletotrichum crown rot of strawberry.
Genetic Constitution and Pathogenicity of Lolium Isolates of Magnaporthe oryzae in Comparison with Host Species-Specific Pathotypes of the Blast Fungus
Authors: Tosa Y. , Hirata K. , Tamba H. , Nakagawa S. , Chuma I. , Isobe C. , Osue J. , Urashima A. S. , Don L. D. , Kusaba M. , Nakayashiki H. , Tanaka A. , Tani T. , Mori N. , Mayama S.
Published: May 2007

Fungal isolates from gray leaf spot on perennial ryegrass (prg isolates) were characterized by DNA analyses, mating tests, and pathogenicity assays. All of the prg isolates were interfertile with Triticum isolates and clustered into the crop isolate group (CC group) on a dendrogram constructed from rDNA-internal transcribed spacer 2 sequences. Since the CC group corresponded to a newly proposed species, Magnaporthe oryzae, all of the prg isolates were designated M. oryzae. However, DNA fingerprinting with MGR586, MGR583, and Pot2 showed that the prg isolates are divided into two distinct populations, i.e., TALF isolates and WK isolates. The TALF isolates were virulent only on Lolium species, whereas the WK isolates were less specific, suggesting that gray leaf spot can be caused not only by Lolium-specific isolates but also by less specific isolates. We designated the TALF isolates as Lolium pathotype. The TALF isolates showed diverse karyotypes in spite of being uniform in DNA fingerprints, suggesting that theyare unstable in genome organization.
The Low-Temperature-Induced Viable-But-Nonculturable State Affects the Virulence of Ralstonia solanacearum Biovar 2
Authors: van Overbeek Leo S. , Bergervoet Jan H. W. , Jacobs Frans H. H. , van Elsas Jan D.
Published: May 2007

The physiology and virulence of Ralstonia solanacearum biovar 2 strain 1609, kept in water at 4 and 20°C, were studied. At 20°C, total cell and plate count (colony forming units; CFU) numbers were similar, between log 5.03 and log 5.55 CFU, and log 5.03 and log 5.51 cells per ml, at days 0 and 132, respectively. However, CFU in the cultures kept at 4°C dropped from log 6.78 CFU/ml at day 0 to below detection after 84 days. The presence of catalase in the agar resulted in higher CFU, and at day 84, log 1.95 CFU/ml still was detectable. No colonies were observed at day 125. The presence of viable-but-nonculturable (VBNC) cells in the 4°C cultures was confirmed using SYTO9 viability staining. Viable cell numbers were log 1.77 higher than CFU on plates with catalase. At day 84 and after 125 days, log 3.70 viable cells per ml still were present. Shifts in subpopulations differing in viability were found by flow cytometric sorting of 4°C-treated cells stained with SYTO9 (healthy) and propidium iodide (PI; compromised). The SYTO9-stained cell fractions dropped from 99 to 39%, and the PI-stained fractions increased from 0.7 to 33.3% between days 0 and 125. At 20°C, the SYTO9-stained fraction remained stable at 99% until day 132. SYTO9-stained cells sorted from 4°C cultures at day 100 were injected into tomato plants. Upon incubation for 30 days, these plants did not show wilting. However, more than log 4.19 CFU and log 8.17 cells were recovered from these plants. Cells from colonies isolated from the nonwilted plants did not regain their virulence as demonstrated by subsequent injection into several new sets of tomato plants. Cells from 4°C cultures injected at day 125 were not able to cause wilting of, or proliferate in, tomato plants. The threat posed by VBNC R. solanacearum cells upon incubation at 4°C was thus ephemeral because cells lost their capacity to cause disease after 125 days.
Partial Resistance to Septoria Tritici Blotch (Mycosphaerella graminicola) in Wheat Cultivars Arina and Riband
Authors: Chartrain L. , Brading P. A. , Widdowson J. P. , Brown J. K. M.
Published: May 2007

Partial resistance to Septoria tritici blotch (STB) and its inheritance were investigated in a doubled-haploid population of a cross between cvs. Arina and Riband. The former has good partial resistance whereas the latter is susceptible. In adult plant trials in polytunnels, STB disease scores were negatively correlated with heading date. Resistance was not specific to any of the three fungal isolates used in these tests. A quantitative trait locus (QTL) for partial resistance to STB was identified in Riband on chromosome 6B and is named QStb.psr-6B-1. No QTL controlling a major part of the Arina resistance was identified, suggesting that its resistance may be dispersed and polygenic. There was no correlation between the lines' mean disease scores at the seedling and adult stages, implying that partial resistance to STB is developmentally regulated. Seedling resistance to the isolate IPO323 was isolate-specific and controlled by a single gene in Arina, probably allelic with the Stb6 gene in cv. Flame that confers resistance to the same isolate. The implications of these results for wheat breeding programs are discussed.
Morphology, Life Cycle Biology, and DNA Sequence Analysis of Rust Fungi on Garlic and Chives from California
Authors: Anikster Y. , Szabo L. J. , Eilam T. , Manisterski J. , Koike S. T. , Bushnell W. R.
Published: May 2007

In the late 1990s, commercial garlic fields in California (CA) were devastated by an outbreak of rust caused by Puccinia allii. We compared collections of the pathogen from garlic (Allium sativum) and chives (A. schoenoprasum) in central CA and Oregon (OR) to collections from garlic and leek (A. porrum and A. ampeloprasum) in the Middle East. Teliospores from the CA and OR collections were smaller in length, width, and projected cross-sectional area compared with collections from the Middle East. CA and OR collections had a shortened life cycle, in which pycnia and aecia were not formed. Germinating teliospores produced a two-celled promycelium, resulting in two basidiospores, each initially with two nuclei, indicating that this rust was homothallic. In addition, the morphology of the substomatal vesicles was different between the CA-OR (fusiform) and the Middle Eastern (bulbous) collections. DNA sequence analysis of the nuclear ribosomal internal transcribed spacer region showed that the CA and OR rust collections formed a well-supported cluster distinct from the Middle Eastern and European samples. These results suggest that the rust on garlic and chives in CA and OR is a different species than the rust fungus on garlic and leek in the Middle East.
A Rapid Diagnostic Test to Distinguish Between American and European Populations of Phytophthora ramorum
Authors: Kroon Laurens P. N. M. , Verstappen Els C. P. , Kox Linda F. F. , Flier Wilbert G. , Bonants Peter J. M.
Published: May 2007

A new devastating disease in the United States, commonly known as Sudden Oak Death, is caused by Phytophthora ramorum. This pathogen, which previously was described attacking species of Rhododendron and Viburnum in Germany and the Netherlands, has established itself in forests on the central coast of California and is killing scores of native oak trees (Lithocarpus densiflora, Quercus agrifolia, Q. kelloggii, and Q. parvula var. shrevei). The phytosanitary authorities in the European Union consider non-European isolates of P. ramorum as a threat to forest trees in Europe. To date, almost all European isolates are mating type A1 while those from California and Oregon are type A2. The occurrence of both mating types in the same region could lead to a population capable of sexual recombination, which could generate a new source of diversity. To prevent contact between these two populations, a rapid, reliable, and discriminating diagnostic test was developed to easily distinguish the two populations. Based on a DNA sequence difference in the mitochondrial Cytochrome c oxidase subunit 1 (Cox1) gene, we developed a single-nucleotide polymorphism (SNP) protocol to distinguish between isolates of P. ramorum originating in Europe and those originating in the United States. A total of 83 isolates of P. ramorum from Europe and 51 isolates from the United States were screened and all isolates could be consistently and correctly allocated to either the European or the U.S. populations using the SNP protocol.
Research Priorities for Rice Pest Management in Tropical Asia: A Simulation Analysis of Yield Losses and Management Efficiencies
Authors: Willocquet Laetitia , Elazegui Francisco A. , Castilla Nancy , Fernandez Luzviminda , Fischer Kenneth S. , Peng ShaoBing , Teng Paul S. , Srivastava R. K. , Singh H. M. , Zhu Defeng , Savary Serge
Published: May 2007

A simulation study was conducted to assess the current and prospective efficiency of rice pest management and develop research priorities for lowland production situations in tropical Asia. Simulation modeling with the RICEPEST model provided the flexibility required to address varying production situations and diverse pest profiles (bacterial leaf blight, sheath blight, brown spot, leaf blast, neck blast, sheath rot, white heads, dead hearts, brown plant-hoppers, insect defoliators, and weeds). Operational definitions for management efficacy (injury reduction) and management efficiency (yield gain) were developed. This approach enabled the modeling of scenarios pertaining to different pest management strategies within the agroecological contexts of rice production and their associated pest injuries. Rice pests could be classified into two broad research priority-setting categories with respect to simulated yield losses and management efficiencies. One group, including weeds, sheath blight, and brown spot, consists of pests for which effective pest management tools need to be developed. The second group consists of leaf blast, neck blast, bacterial leaf blight, and brown plant-hoppers, for which the efficiency of current management methods is to be maintained. Simulated yield losses in future production situations indicated that a new type of rice plant with high-harvest index and high-biomass production (“New Plant Type”) was more vulnerable to pests than hybrid rice. Simulations also indicated that the impact of deployment of host resistance (e.g., through genetic engineering) was much larger when targeted against sheath blight than when targeted against stem borers. Simulated yield losses for combinations of production situations and injury profiles that dominate current lowland rice production in tropical Asia ranged from 140 to 230 g m-2. For these combinations, the simulated efficiency of current pest management methods, expressed in terms of relative yield gains, ranged from 0.38 to 0.74. Overall, the analyses indicated that 120 to 200 × 106tons of grain yield are lost yearly to pests over the 87 × 106ha of lowland rice in tropical Asia. This also amounts to the potential gain that future pest management strategies could achieve, if deployed.
An Antibody Against the SecA Membrane Protein of One Phytoplasma Reacts with Those of Phylogenetically Different Phytoplasmas
Authors: Wei Wei , Kakizawa Shigeyuki , Jung Hee-Young , Suzuki Shiho , Tanaka Minoru , Nishigawa Hisashi , Miyata Shin-ichi , Oshima Kenro , Ugaki Masashi , Hibi Tadaaki , Namba Shigetou
Published: May 2007

Antisera raised against phloem-limited phytoplasmas generally react only with the phytoplasma strain used to produce the antigen. There is a need for an antiserum that reacts with a variety of phytoplasmas. Here, we show that an antiserum raised against the SecA membrane protein of onion yellows phytoplasma, which belongs to the aster yellows 16S-group, detected eight phytoplasma strains from four distinct 16S-groups (aster yellows, western X, rice yellow dwarf, and elm yellows). In immunoblots,≈96-kDa SecA protein was detected in plants infected with each of the eight phytoplasmas. Immunohistochemical staining of thin sections prepared from infected plants was localized in phloem tissues. This antiserum should be useful in the detection and histopathological analysis of a wide range of phytoplasmas.
Potential of the Mycoparasite, Verticillium lecanii, to Protect Citrus Fruit Against Penicillium digitatum, the Causal Agent of Green Mold: A Comparison with the Effect of Chitosan
Authors: Benhamou Nicole
Published: May 2007

The potential of the mycoparasite, Verticillium lecanii, at protecting citrus fruits against green mold was explored at the cellular level. Treatmentthe fruit with V. lecanii or chitosan prior to inoculation with the causal agent of green mold, Penicillium digitatum, markedly reduced disease development compared with that of nontreated control citrus fruits in which symptoms were visible by 3 days after inoculation with the pathogen. Scanning electron microscope investigations of citrus samples, collected 5 days after inoculation with the pathogen, revealed striking differences in the extent of cell surface colonization between treated and nontreated fruits. Pathogen hyphae, which sporulated abundantly at the surface of control fruits, were collapsed and severely damaged in V. lecanii and chitosan-treated fruits. Histological observations of citrus samples confirmed that restriction of pathogen colonization at the cell surface correlated with a pronounced disorganization of the pathogen hyphae. In addition, host cell changes, mainly characterized by the deposition of a new material in the exocarp cells and the thickening of cell walls, were observed. Ultrastructural investigations of citrus samples revealed that the pathogen multiplied abundantly through much of the mesocarp and exocarp tissues in V. lecanii-free citrus fruits, whereas in V. lecanii-treated citrus, pathogen growth was restricted. Penicillium hyphae that penetrated the mesocarp tissue were markedly altered. Labeling with the wheat germ agglutinin/ovomucoid-gold complex for the localization of chitin resulted in an irregular labeling of Penicillium cell walls, even at a time when in an irregular labeling of Penicillium cell walls, even at a time when these were markedly altered. Cytochemical investigations revealed that callose and lignin-like compounds accumulated at sites of pathogen colonization in the exocarp tissue. Evidence is provided in this study that V. lecanii as well as chitosan are equally capable of inducing a striking response in P. digitatum-infected citrus fruits. The marked differences observed in the rate and extent of colonization as well as in pathogen cell viability between control and treated citrus fruits demonstrate that both treatments have the ability to induce the transcriptional activation of defense genes leading to the accumulation of structural and biochemical compounds at strategic sites.
Spinach curly top virus: A Newly Described Curtovirus Species from Southwest Texas with Incongruent Gene Phylogenies
Authors: Baliji Surendranath , Black Mark C. , French Roy , Stenger Drake C. , Sunter Garry
Published: May 2007

A curtovirus associated with a disease of spinach was isolated in southwest Texas during 1996. Disease symptoms included severe stunting and chlorosis, with younger leaves curled, distorted, and dwarfed. Viral DNA was purified and an infectious clone obtained. Agroinoculation using a construct bearing full-length tandem repeats of the cloned viral genome resulted in systemic infection of species in six of seven plant families tested, indicating that the virus has a wide host range. Symptoms produced in spinach agroinoculated with cloned viral DNA were similar to those observed in the field. Viral single-stranded and double-stranded DNA forms typical of curtovirus infection were detected in host plants by Southern blot hybridization. The complete sequence of the infectious clone comprised 2,925 nucleotides, with seven open reading frames encoding proteins homologous to those of other curtoviruses. Complete genome comparisons revealed that the spinach curtovirus shared 64.2 to 83.9% nucleotide sequence identity relative to four previously characterized curtovirus species: Beet curly top virus, Beet severe curly top virus, Beet mild curly top virus, and Horseradish curly top virus. Phylogenetic analysis of individual open reading frames indicated that the evolutionary history of the three virion-sense genes was different from that of the four complementary-sense genes, suggesting that recombination among curtoviruses may have occurred. Collectively, these results indicate that the spinach curtovirus characterized here represents a newly described species of the genus Curtovirus, for which we propose the name Spinach curly top virus.
Cerebroside Elicitor Confers Resistance to Fusarium Disease in Various Plant Species
Authors: Umemura Kenji , Tanino Shigeki , Nagatsuka Tadako , Koga Jinichiro , Iwata Michiaki , Nagashima Kenji , Amemiya Yoshimiki
Published: May 2007

In the rice blast fungus pathosystem, cerebroside, a compound categorized as a sphingolipid, was found in our previous study to be a non-racespecific elicitor, which elicits defense responses in rice. Here we describe that cerebroside C is produced in diverse strains of Fusarium oxysporum, a common soilborne agent of wilt disease affecting a wide range of plant species. In addition, some type of cerebroside elicitor involving cerebroside A, B, or C was detected in other soilborne phytopathogens, such as Pythium and Botrytis. Treatment of lettuce (Lactuca sativa), tomato (Lycopersicon esculentum), melon (Cucumis melo), and sweet potato (Ipomoea batatas) with cerebroside B resulted in resistance to infection by each pathogenic strain of F. oxysporum. Induction of pathogenesis-related genes and H2O2 production by treatment with cerebroside B were observed in tomato root tissues. The cerebroside elicitor showed no antifungal activity against F. oxysporum in vitro, indicating that the cerebroside elicitor activates defense mechanisms to confer resistance to Fusarium disease. These results suggest that cerebroside functions as a non-race-specific elicitor in a wide range of plant-phytopathogenic fungus interactions. Additionally, cerebroside elicitor serves as a potential biologically derived control agent.
Mediation of Partial Resistance to Rice Blast Through Anaerobic Induction of Ethylene
Authors: Singh Manjul P. , Lee Fleet N. , Counce Paul A. , Gibbons Julia H.
Published: May 2007

The correlation between anaerobic soil conditions and increased resistance to rice blast disease has long been observed without benefit of an adequate explanation. We researched flood depth, dissolved oxygen (DO), and ethylene relative to expression of partial blast resistance in cvs. M-201, Newbonnet, LaGrue, Mars, and Cypress. Cultivar blast index (BI) and flood DO decreased with increasing flood depth. BIs were positively correlated with DO. Total leaf blast lesions were 3.4 and 3.2 times greater in cvs. M-201 and LaGrue growing in a 5.0-μl liter-1 DO nutrient solution than when growing in a 0.1-μl liter-1 DO solution. Treatment with 0.25 mM ethephon, which releases ethylene, lowered BIs of Newbonnet, LaGrue, and Cypress growing upland when applied drench, foliar, or foliar-drench. If flooded, BIs of ethephon-treated cultivars were decreased by drench and foliar-drench applications only. BIs of upland plants were unchanged, whereas BIs of analogous flooded plants increased following treatment with 0.31 mM aminoethoxyvinylglycine (AVG), an ethylene biosynthesis inhibitor. We hypothesize that varying anaerobic conditions mediate production of phytohormones, particularly ethylene, which modify expression of inherent partial blast resistance in these rice cultivars.
Quantification of Root and Stem Colonization of Watermelon byFusarium oxysporumf. sp.niveumand Its Use in Evaluating Resistance
Authors: Zhou X. G. , Everts K. L.
Published: May 2007

Colonization of watermelon root and stem tissues by Fusarium oxysporum f. sp. niveum race 1 and its relationship to the apparent resistance to Fusarium wilt was investigated. In each of 2 years, 17 differentially susceptible watermelon cultivars and one accession were tested in the greenhouse, and 7 cultivars also were tested in the field. Colonization by a chlorate-resistant marked isolate of the fungus was assayed by plating homogenized tissue samples on a selective medium. Six days after inoculation, seedlings of highly resistant, moderately resistant, and susceptible cultivars had F. oxysporum f. sp. niveum race 1 CFU counts in the lower stems of 102, 103, and 104CFU/g of fresh tissue, respectively. Percent wilt (Y) of the seedlings was positively correlated with colonization (X) by F. oxysporum f. sp. niveum race 1 in roots (Y = 21.2 ln [X + 1] - 140.7, R2= 0.85) or lower stems (Y = 17.3 ln [X + 1] - 78.6, R2= 0.86). Percent wilt (Y) also was correlated with the ratio (Xr, 0 to 1 values) of lower stem to root colonization (Y = 34 ln Xr+ 112, R2= 0.36). Field evaluations confirmed these relationships, and a link between cultivar resistance and a reduced rate of spread of the fungus in primary stems during a season was observed. Fruit yield decreased with increased tissue colonization at linear rates of 9.9 to 12.7 t/ha per ln (CFU/g + 1) (R2≥ 0.58). The greenhouse seedling stem colonization assay described may be utilized as a collaborative method to quantify Fusarium wilt resistance in watermelon.
Molecular and Symptom Analyses of Phytoplasma Strains from Lettuce Reveal a Diverse Population
Authors: Zhang Jianhua , Hogenhout Saskia A. , Nault Lowell R. , Hoy Casey W. , Miller Sally A.
Published: May 2007

Epidemics of aster yellows in lettuce in Ohio are caused by at least seven distinct phytoplasma strains in the aster yellows (AY) group. Five of the strains are newly reported: AY-BW, AY-WB, AY-BD3, AY-SS, and AY-SG. All seven strains were characterized based on symptoms in aster and lettuce, and by polymerase chain reaction (PCR). Strain AY-BD2 (formerly ‘Bolt’) causes yellowing and leaf distortion in lettuce and bolting in aster, whereas strain AY-S (formerly ‘Severe’) causes stunting, leaf clustering, and phyllody. Strain AY-WB causes yellowing and wilting in lettuce and witches'-broom in aster. Strain AY-SG induces horizontal growth in lettuce and aster plants. Strain AY-BW causes chlorosis of emerging leaves and abnormally upright growth of leaf petioles. AY-SS causes symptoms similar to those caused by AY-S but has a different PCR-restriction fragment length polymorphism (RFLP) banding pattern. Strains AY-BD2 and AY-BD-3 cause mild leaf and stem distortion in lettuce but are differentiated by PCR-RFLP. All phytoplasma strains collected from lettuce in Ohio belong to the 16SrI group. AY-WB belongs to the 16SrI-A subgroup and the other six belong to the 16SrI-B subgroup. Five of the seven strains were distinguished from each other by primer typing. The results of phylogenetic analyses of sequences of the 16S rRNA genes were basically consistent with the classification based on PCR-RFLP, in which AY-WB clustered with phytoplasmas of the 16rIA subgroup and the other Ohio lettuce strains clustered with phytoplasmas in the 16SrI-B subgroup.
Overview of Selected Multivariate Statistical Methods and Their Use in Phytopathological Research
Authors: Sanogo S. , Yang X. B.
Published: May 2007

To disentangle the nature of a pathosystem or a component of the system such as disease epidemics for descriptive or predictive purposes, mensuration is conducted on several variables of the physical and chemical environment, pathogenic populations, and host plants. For instance, it may be desired to (i) distinguish pathogenic variation among several isolates of a pathogen based on disease severity; (ii) identify the most important variables that characterize the structure of an epidemic; and (iii) assess the potential of developing regional scale versus site-specific postmanagement schemes using weather and site variation. In all these cases, a simultaneous handling of several variables is required, and entails the use of multivariate statistics such as discriminant analysis, multivariate analysis of variance, correspondence analysis, and canonical correlation analysis. These tools have been used to varying degree in the phytopathological literature. A succinct overview of these tools is presented with cited examples.
Meta-Analysis in Plant Pathology: Synthesizing Research Results
Authors: Rosenberg M. S. , Garrett K. A. , Su Z. , Bowden R. L.
Published: May 2007

Meta-analysis is a set of statistical procedures for synthesizing research results from a number of different studies. An estimate of a statistical effect, such as the difference in disease severity for plants with or without a management treatment, is collected from each study along with a measure of the variance of the estimate of the effect. Combining results from different studies will generally result in increased statistical power so that it is easier to detect small effects. Combining results from different studies may also make it possible to compare the size of the effect as a function of other predictor variables such as geographic region or pathogen species. We present a review of the basic methodology for meta-analysis. We also present an example of meta-analysis of the relationship between disease severity and yield loss for foliar wheat diseases, based on data collected from a decade of fungicide and nematicide test results.
Nonparametric Tests in Plant Disease Epidemiology: Characterizing Disease Associations
Authors: Turechek W. W.
Published: May 2007

Nonparametric tests are suited to many statistical applications, including experimental design, regression, and time series analysis, for example. Often these tests are thought of as alternatives to their parametric counterparts when certain assumptions about the underlying population are questionable. Although suited for this scenario, there are a number of nonparametric tests that fill unique niches in the analysis of data, for example, characterizing interspecific associations. Quantifying the degree of association between two or more pathogens or diseases at a defined spatial scale is essential to gain a thorough understanding of disease dynamics, generate testable hypothesis behind the mechanisms that cause association, and is often necessary in modeling applications. In this paper, nonparametric approaches to characterizing interspecific associations will be covered. Specifically, I will address the use of rank correlation coefficients and the development of a randomization procedure for testing the Jaccard index of association against a null model.
Bayesian Analysis in Plant Pathology
Authors: Mila A. L. , Carriquiry A. L.
Published: May 2007

Bayesian methods are currently much discussed and applied in several disciplines from molecular biology to engineering. Bayesian inference is the process of fitting a probability model to a set of data and summarizing the results via probability distributions on the parameters of the model and unobserved quantities such as predictions for new observations. In this paper, after a short introduction of Bayesian inference, we present the basic features of Bayesian methodology using examples from sequencing genomic fragments and analyzing microarray gene-expressing levels, reconstructing disease maps, and designing experiments.
Biological Control of Botrytis cinerea by Volatiles of ‘Isabella’ Grapes
Authors: Kulakiotu Eleni K. , Thanassoulopoulos Constantine C. , Sfakiotakis Evangelos M.
Published: May 2007

The effect of volatiles from cv. Isabella (Vitis labrusca) on the growth of Botrytis cinerea was tested in vitro and in situ, in the latter case on ‘Roditis’ grapes (V. vinifera), at various temperatures. The goal of the research was to determine whether the volatiles emitted by Isabella grapes could be effective biocontrol agents of Botrytis cinerea. The closed Mariotte system was used as a bioassay method to analyze quantitatively the biological action of these volatiles on fungal growth and disease development. The in vitro experiments revealed the inhibitory action of the Isabella volatiles on the sporulation and sclerotia formation of the fungus, as well as the stimulating action of the Roditis volatiles on the sporulation of the fungus. The in situ study confirmed the antifungal action of the Isabella volatiles as they reduced the inoculum and pathogenicity of B. cinerea. The antibiotic action was more pronounced at 21°C. The study indicates that Isabella volatiles act as biocontrol agents of B. cinerea.
Identification of Phytophthora Isolates to Species Level Using Restriction Fragment Length Polymorphism Analysis of a Polymerase Chain Reaction-Amplified Region of Mitochondrial DNA
Authors: Martin Frank N. , Tooley Paul W.
Published: May 2007

Polymerase chain reaction primers spanning the mitochondrially encoded coxI and II genes have been identified that were capable of amplifying target DNA from all 152 isolates of 31 species in the genus Phytophthora that were tested. Digestion of the amplicons with restriction enzymes generated species-specific restriction fragment length polymorphism banding profiles that were effective for isolate classification to a species level. Of the 24 species in which multiple isolates were examined, intraspecific polymorphisms were not observed for 16 species, while 5 species exhibited limited intraspecific polymorphism that could be explained by the addition/loss of a single restriction site. Intraspecific polymorphisms were observed for P. megakarya, P. megasperma, and P. syringae; however, these differences may be a reflection of the variation that exists in these species as reported in the literature. Although digestion with AluI alone could differentiate most species tested, single digests with a total of four restriction enzymes were used in this investigation to enhance the accuracy of the technique and minimize the effect of intraspecific variability on correct isolate identification. The use of the computer program BioNumerics simplified data analysis and identification of isolates. Successful template amplification was obtained with DNA recovered from hyphae using a boiling miniprep procedure, thereby reducing the time and materials needed for conducting this analysis.
Evolutionarily conserved genes preferentially accumulate introns
Authors: Carmel Liran , Rogozin Igor B. , Wolf Yuri I. , Koonin Eugene V.
Published: May 2007

Introns that interrupt eukaryotic protein-coding sequences are generally thought to be nonfunctional. However, for reasons still poorly understood, positions of many introns are highly conserved in evolution. Previous reconstructions of intron gain and loss events during eukaryotic evolution used a variety of simplified evolutionary models that yielded contradicting conclusions and are not suited to reveal some of the key underlying processes. We combine a comprehensive probabilistic model and an extended data set, including 391 conserved genes from 19 eukaryotes, to uncover previously unnoticed aspects of intron evolution—in particular, to assign intron gain and loss rates to individual genes. The rates of intron gain and loss in a gene show moderate positive correlation. A gene’s intron gain rate shows a highly significant negative correlation with the coding-sequence evolution rate; intron loss rate also significantly, but positively, correlates with the sequence evolution rate. Correlations of the opposite signs, albeit less significant ones, are observed between intron gain and loss rates and gene expression level. It is proposed that intron evolution includes a neutral component, which is manifest in the positive correlation between the gain and loss rates and a selection-driven component as reflected in the links between intron gain and loss and sequence evolution. The increased intron gain and decreased intron loss in evolutionarily conserved genes indicate that intron insertion often might be adaptive, whereas some of the intron losses might be deleterious. This apparent functional importance of introns is likely to be due, at least in part, to their multiple effects on gene expression.
Characterization of the opossum immune genome provides insights into the evolution of the mammalian immune system
Authors: Belov Katherine , Sanderson Claire E. , Deakin Janine E. , Wong Emily S.W. , Assange Daniel , McColl Kaighin A. , Gout Alex , de Bono Bernard , Barrow Alexander D. , Speed Terence P. , Trowsdale John , Papenfuss Anthony T.
Published: May 2007

The availability of the first marsupial genome sequence has allowed us to characterize the immunome of the gray short-tailed opossum (Monodelphis domestica). Here we report the identification of key immune genes, including the highly divergent chemokines, defensins, cathelicidins, and Natural Killer cell receptors. It appears that the increase in complexity of the mammalian immune system occurred prior to the divergence of the marsupial and eutherian lineages ∼180 million years ago. Genomes of ancestral mammals most likely contained all of the key mammalian immune gene families, with evolution on different continents, in the presence of different pathogens leading to lineage specific expansions and contractions, resulting in some minor differences in gene number and composition between different mammalian lineages. Gene expansion and extensive heterogeneity in opossum antimicrobial peptide genes may have evolved as a consequence of the newborn young needing to survive without an adaptive immune system in a pathogen laden environment. Given the similarities in the genomic architecture of the marsupial and eutherian immune systems, we propose that marsupials are ideal model organisms for the study of developmental immunology.
Age‐ and gender‐related differences in blood amphetamine concentrations in apprehended drivers: lack of association with clinical evidence of impairment
Authors: Jones Alan Wayne
Published: May 2007

Background  New legislation aimed at combating driving under the influence of drugs (DUID) in Sweden stipulated zero‐concentrations in blood for scheduled substances. DUID suspects (n = 300), with amphetamine as the only psychoactive drug identified in blood, were investigated in relation to age and gender. In a smaller retrospective sample (n = 70) the relationship between clinical tests of impairment were compared with the concentration of amphetamine in blood.

Measurements  All forensic blood samples were subjected to a broad toxicological screening analysis by immunoassay methods [enzyme multiplied immunoassay technique/cloned enzyme donor immunoassay (EMIT/CEDIA)] and positive results were verified by gas chromatography‐mass spectrometry (GC‐MS). The limit of quantitation (LOQ) for determination of amphetamine in blood was 0.03 mg/l. People suspected of being under the influence of drugs were examined by a physician who asked various questions about state of health and use of drugs and also administered simple psychomotor and cognitive tests of impairment. After conducting these tests the physician concluded whether the suspects were not impaired, slightly, moderately or highly impaired by drugs other than alcohol.

Findings  Among 300 DUID suspects with amphetamine in blood there were 246 men (82%) and 54 women (18%). Mean age (± SD) of the men was 37.1 ± 8.7 years compared with 35.5 ± 7.1 years for the women (P > 0.05). The frequency distribution of blood amphetamine concentration was positively skewed with mean, median and highest values of 1.0 mg/l, 0.9 mg/l and 7.1 mg/l, respectively. The mean concentrations were slightly higher in the women 1.11 mg/l (median 1.0 mg/l) compared with 0.97 mg/l (median 0.8 mg/l) in the men (P > 0.05). There was a weak but statistically significant correlation between the person's age and the concentration of amphetamine in blood (r = 0.18, P < 0.05). The results of clinical tests of impairment showed no relationship with the concentration of amphetamine in blood according to analysis of variance (P > 0.05).

Conclusions  The lack of association between degree of drug influence and the concentration of amphetamine in blood speaks against the notion of introducing concentration per se limits or graded penalties depending on the blood‐concentration of this stimulant. Zero‐concentration limits or LOQ‐limits are a much more pragmatic way to enforce DUID legislation.
Rituximab for the treatment of autoimmune cytopenias in children with immune deficiency
Authors: Kim J. J. , Thrasher A. J. , Jones A. M. , Davies E. G. , Cale C. M.
Published: May 2007

Autoimmune cytopenias are well recognised in patients with primary immune deficiency, but treatment may be difficult. We report eight children with autoimmune cytopenias (autoimmune haemolytic anaemia, autoimmune thrombocytopenia, autoimmune neutropenia) complicating immune deficiency states (common variable immunodeficiency, Wiskott‐Aldrich Syndrome, autoimmune lymphoproliferative syndrome, combined immunodeficiency) treated with between 1 and 3 courses of rituximab (anti‐CD20). Responses occurred for 90% of treatments but relapse rates (after a median of 53 weeks) were high (78%). We conclude that rituximab is an effective treatment for autoimmune cytopenias in children with immune deficiencies, but repeated courses of treatment may be needed.