bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Nutrition on Deformation Disease inGypsophila paniculataMother Plants
Authors: Ben-Yephet Y. , Reuven M. , Zviebil A. , Szmulewich Y. , Lavkovits I. , Markovits T. , Soriano S. , Bar-Yosef B.
Published: May 2007

Deformation disease of Gypsophila paniculata mother plants reduces cutting yields as much as 50% but does not kill the mother plants. In preliminary experiments, fertigation of G. paniculata mother plants with a 20:20:20 compound fertilizer (N, P, K, plus microelements) at an N concentration of 720 mg/liter reduced the expression of deformation disease compared with the conventional N concentration of 360 mg/liter. The current study determined which component of the compound fertilizer reduced the disease. Experiments were carried out in 10-liter buckets packed with naturally infested 0- to 8-mm black tuff (Scoria = crushed volcanic stones). Irrigation was applied once a day at 0.5 liter per bucket with the nutrient under test being added at 1.0 liter per bucket via the water once a week. Treatments included: (i) four levels of 20:20:20 fertilizer; (ii) four levels of each of N, P, K, and Fe-Zn-Mn mixture (ME) corresponding to their concentrations in the compound fertilizer; (iii) different N sources (20:20:20, (NH4)2SO4, KNO3, NH4NO3, urea); and (iv) three pHs of the irrigation water at each of three NO3and NH4application levels. Increasing the 20:20:20 fertilizer concentration reduced the disease level from 82 to 96% with N at 180 mg/liter to 6 to 10% with N at 720 mg/liter. When either P, K, or ME was the sole variable, increasing concentrations had no significant effect on the disease, but elevating a mixed source nitrogen concentration from 0 to 180, 360, and 720 mg/liter (as 50% urea, 40% NH4NO3, and 10% KNO3) significantly reduced deformation similar to that observed with increasing concentrations of the 20:20:20 fertilizer. mong tested N sources, NH4was the most effective in reducing the disease (almost to zero at an N concentration of 360 g/liter). Low disease incidence (0 to 10%) was always associated with effluent pH of 6 or lower. Irrigating with acidified water pH 5.5) in the presence of N, as NH4at 180 mg/liter, additionally reduced disease from 56% under tap water (pH 7.8) rrigation to 11%. Similar acidification in the presence of NO-3N at 180 mg/liter was ineffective in reducing the disease, but ater basification in the presence of NO-3N reduced disease incidence from 93 to 38% 90 days after planting. The N, P, K, Fe, and Zn concentrations in gypsophila cuttings were similar under the three tested levels of NH4, NO3, and 20:20:20, whereas the concentration of Mn increased with increasing N. The Mn concentration in cuttings was inversely correlated with disease and is probably an important factor to understanding the physiological background of the deformation disease.
Insect-Transmitted Urediniospores of the Rust Puccinia punctiformis Cause Systemic Infections in Established Cirsium arvense Plants
Authors: Wandeler H. , Bacher S.
Published: May 2007

The rust fungus Puccinia punctiformis has potential as a biological control agent for creeping thistle Cirsium arvense, because systemically infected shoots usually die before flowering. The mechanism of rust transfer as well as the spore type responsible for systemic infections have been a source of controversy. One possibility of successful transmission is the use of the weevil Ceratapion onopordi as a vector. Our results from a garden experiment show that urediniospores transmitted by the weevil are able to induce systemic infections in established thistle clones. Furthermore, the weevil origin and the date of rust treatment significantly influenced the number of rust-infected shoots. The earlier a shoot was treated, the higher the probability of rust transmission. These results challenge the current belief that teliospores passing through the soil and infecting root buds are the major cause of systemic infections in the field. Further research on biological control of creeping thistle should therefore concentrate on the application of urediniospores to enhance systemic rust infections.
An Indigenous Virulent Strain of Erwinia amylovora Lacking the Ubiquitous Plasmid pEA29
Authors: Llop Pablo , Donat Victoria , Rodríguez Margarita , Cabrefiga Jordi , Ruz Lídia , Palomo José Luis , Montesinos Emilio , López María M.
Published: May 2007

An atypical strain of Erwinia amylovora was isolated near an outbreak of fire blight at a nursery in Spain in 1996. It was obtained from a Crataegus plant showing typical symptoms and was identified as E. amy-lovora by biochemical tests and enrichment-enzyme-linked immuno-sorbent assay, but not by polymerase chain reaction using primers based on the pEA29 sequence. Nevertheless, with primers from chromosomal regions, the isolate gave the expected amplification band. This strain carries one plasmid of ≈70 kb, with no homology with the 29-kb plasmid common to all pathogenic strains, or with a large plasmid present in some E. amylovora strains. Growth of the strain in minimal medium without thiamine was slower compared with cultures in the same medium with thiamine, a characteristic typical of strains cured of the 29-kb plasmid. Nevertheless, aggressiveness assays on pear, apple, and Pyracantha plants and in immature pear fruit showed that this strain exhibited a virulence level similar to other strains containing pEA29. To the best of our knowledge, this is the first report of the isolation from naturally infected plant material of a pathogenic strain of E. amylovora without pEA29, but with a plasmid of ≈70 kb not previously described.
Beta-Cyanoalanine Synthase as a Molecular Marker for Induced Resistance by Fungal Glycoprotein Elicitor and Commercial Plant Activators
Authors: Takahashi Hideki , Ishihara Takeaki , Hase Shu , Chiba Ayaka , Nakaho Kazuhiro , Arie Tsutomu , Teraoka Tohru , Iwata Michiaki , Tugane Taneaki , Shibata Daisuke , Takenaka Shigehito
Published: May 2007

The biocontrol agent Pythium oligandrum produces glycoprotein elicitor in the cell wall fraction, designated CWP, and induces resistance to a broad range of pathogens. To understand the mechanism of CWP-induced resistance to pathogens, gene expression at the early stage of CWP treatment in tomato roots was analyzed using a cDNA array. At 4 h after CWP treatment, 144 genes were up-regulated and 99 genes were down-regulated. In the 144 up-regulated genes, nine genes exhibited about eightfold increased expression. Analysis of the response of these nine genes to three commercial plant activators indicated that a high level of one gene, beta-cyanoalanine synthase gene (LeCAS) encoding hydrogen cyanide (HCN) detoxification enzyme, was stably induced in tomato roots by such treatment. However, expression of LeCAS was not significantly induced in tomato roots at 4 h by abiotic stresses, whereas only a very low level of induction of such expression by cold stress was observed. This LeCAS expression was also induced after exogenous treatment with a low level of 1-amino-cyclopropane-1-carboxylate as the precursor of ethylene, but not with either salicylic acid or methyl jas-monate. The induction of LeCAS expression in CWP-treated and plant activator-treated roots is likely to be caused by the detoxification of HCN during ethylene production. Transient activation of LeCAS expression caused by ethylene production in tomato roots may be a general phenomenon in fungal elicitor-induced and synthetic plant activator-induced resistance. LeCAS seems to be useful for screening possible novel plant activators for plant protection against pathogens.
A New Spore Precipitator with Polarized Dielectric Insulators for Physical Control of Tomato Powdery Mildew
Authors: Matsuda Yoshinori , Ikeda Hiroki , Moriura Nobuyuki , Tanaka Norio , Shimizu Kunihiko , Oichi Wataru , Nonomura Teruo , Kakutani Koji , Kusakari Shin-ichi , Higashi Katsuhide , Toyoda Hideyoshi
Published: May 2007

In an attempt to physically protect greenhouse tomato plants from the powdery mildew fungus Oidium neolycopersici, we developed a new electrostatic spore precipitator in which a copper wire conductor is linked to an electrostatic generator and covered with a transparent acrylic cylinder (insulator). The conductor was negatively charged by the generator, and the electrostatic field created by the conductor was used to dielectrically polarize the insulator cylinder. The dielectrically polarized cylinder also produced an electrostatic force without a spark discharge. This force was directly proportional to the potential applied to the conductor and was used to attract conidia of the pathogen. The efficacy of this spore precipitator in protecting hydroponically cultured tomato plants from powdery mildew was evaluated in the greenhouse. The hydroponic culture troughs were covered with a cubic frame installed with the spore precipitator, and the disease progress on precipitator-guarded and unguarded seedlings was traced after the conidia were disseminated mechanically from inoculum on tomato plants. Seedlings in the guarded troughs remained uninfected during the entire experiment, in spite of rapid spread of the disease to all leaves of the unguarded seedlings.
Clarification of the Etiology of Glomerella Leaf Spot and Bitter Rot of Apple Caused by Colletotrichum spp. Based on Morphology and Genetic, Molecular, and Pathogenicity Tests
Authors: González Eugenia , Sutton Turner B. , Correll James C.
Published: May 2007

Morphological characteristics and vegetative compatibility groups (VCGs) of 486 isolates of Glomerella cingulata, Colletotrichum gloeosporioides, and C. acutatum collected from apple leaves with Glomerella leaf spot (GLS) symptoms and fruit with bitter rot symptoms in the United States and Brazil were studied. From this collection, 155 isolates of G. cingulata (93 from fruit, 61 from leaves, and 1 from buds), 42 isolates of C. gloeosporioides from fruit, and 14 isolates of C. acutatum (10 from fruit and 4 from leaves) were studied using mitochondrial (mt)DNA restriction fragment length polymorphism (RFLP) haplotypes. A subset of 24 isolates was studied by examining the sequence of a 200-bp intron of the glyceraldehyde 3-phosphate dehydrogenase (GDPH) nuclear gene. In addition, 98 isolates were tested for pathogenicity on leaves of cvs. Gala and Golden Delicious in the greenhouse, and 24 isolates were tested for pathogenicity on fruit of cv. Gala in growth chambers. In total, 238 and 225 isolates of G. cingulata were separated into four distinct morphological types and six VCGs, respectively. Five morphological types and six VCGs were identified among 74 and 36 isolates of C. gloeosporioides, respectively. Three morphological types and four VCGs were identified among 74 and 23 isolates of C. acutatum, respectively. Seven different mtDNA RFLP haplotypes were observed within isolates of G. cingulata, two within isolates of C. gloeosporioides, and two within isolates of C. acutatum. Phylogenetic trees, inferred based on maximum likelihood and maximum parsimony methods using the intron sequence, produced similar topologies. Each species was separated into distinct groups. All isolates tested were pathogenic on fruit, though only isolates with specific VCGs and haplotypes were pathogenic to leaves. Vegetative compatibility was a better tool than molecular characters for distinguishing isolates of G. cingulata pathogenic on both leaves and fruit from the ones pathogenic only on fruit. Isolates of G. cingulata capable of causing both GLS and bitter rot were included in haplotypes and groups based on the sequence analysis of the 200-bp intron that also included isolates capable of causing bitter rot only. Additionally, isolates of G. cingulata from the United States and Brazil which cause GLS were included in different haplotypes and sequence analysis groups. Therefore, one hypothesis is that isolates of G. cingulata from the United States capable of causing both GLS on foliage and bitter rot on fruit may have arisen independently of Brazilian isolates of G. cingulata capable of causing both GLS and bitter rot, and the two groups of isolates may represent distinct populations.
Alteration of Photosynthesis in Grapevines Affected by Esca
Authors: Petit A.-N. , Vaillant N. , Boulay M. , Clément C. , Fontaine F.
Published: May 2007

To further understand the development of esca disease in grapevine, its physiological impact on plants grown in the vineyard was characterized, focusing mainly on photosynthesis. For this purpose, the state of the photosynthetic apparatus was evaluated in symptomatic and asymptomatic leaves in esca-infected grape plants, and wood carbohydrates stored in annual canes were assayed. In symptomatic leaves, esca greatly affected grapevine physiology. Foliar symptoms were associated with both stomatal closure and alteration of the photosynthetic apparatus as revealed by (i) a decrease in CO2 assimilation, transpiration, and a significant increase in intercellular CO2 concentration; (ii) a strong drop in both the maximum fluorescence yield and the effective Photosystem II quantum yield; and (iii) a reduction of total chlorophyll but a stable carotenoid content. On symptomatic canes, all these variables were affected more on leaves with symptoms than without symptoms, suggesting a gradation in photosynthesis disruptions in the plant according to the degree of symptom severity. In addition, canes of symptomatic plants had reduced carbohydrate reserves during the winter rest, whether they exhibit symptoms or not. The following year, the lower pool of reserves may contribute to a significant decrease in plant development, as well as a global loss in plant vigor.
High Variation in Pathogenicity of Genetically Closely Related Strains ofXanthomonas albilineans, the Sugarcane Leaf Scald Pathogen, in Guadeloupe
Authors: Champoiseau P. , Daugrois J.-H. , Pieretti I. , Cociancich S. , Royer M. , Rott P.
Published: May 2007

Pathogenicity of 75 strains of Xanthomonas albilineans from Guadeloupe was assessed by inoculation of sugarcane cv. B69566, which is susceptible to leaf scald, and 19 of the strains were selected as representative of the variation in pathogenicity observed based on stalk colonization. In vitro production of albicidin varied among these 19 strains, but the restriction fragment length polymorphism pattern of their albicidin biosynthesis genes was identical. Similarly, no genomic variation was found among strains by pulsed-field gel electrophoresis. Some variation among strains was found by amplified fragment length polymorphism, but no relationship between this genetic variation and variation in pathogenicity was found. Only 3 (pilB, rpfA, and xpsE) of 40 genes involved in pathogenicity of bacterial species closely related to X. albilineans could be amplified by polymerase chain reaction from total genomic DNA of all nine strains tested of X. albilineans differing in pathogenicity in Guadeloupe. Nucleotide sequences of these genes were 100% identical among strains, and a phylogenetic study with these genes and housekeeping genes efp and ihfA suggested that X. albilineans is on an evolutionary road between the X. campestris group and Xylella fastidiosa, another vascular plant pathogen. Sequencing of the complete genome of Xanthomonas albilineans could be the next step in deciphering molecular mechanisms involved in pathogenicity of X. albilineans.
Genetic Variability in the Potato Pathogen Colletotrichum coccodes as Determined by Amplified Fragment Length Polymorphism and Vegetative Compatibility Group Analyses
Authors: Heilmann Larry J. , Nitzan Nadav , Johnson Dennis A. , Pasche Julie S. , Doetkott Curt , Gudmestad Neil C.
Published: May 2007

Amplified fragment length polymorphism (AFLP) using three primer sets was used to characterize 211 Colletotrichum coccodes isolates from North America, 112 of which were assigned to six vegetative compatibility groups (VCGs) using nitrate nonutilizing (nit) mutants. These isolates clustered into five corresponding groups by unweighted pairgroup method with arithmetic means-based cluster analysis of AFLP banding patterns. Isolates of C. coccodes belonging to NA-VCG1 and NA-VCG3 were closely related, as were isolates belonging to NA-VCG2 and NA-VCG5. Based on bootstrap analysis of AFLP data, the two isolates originally assigned to NA-VCG4 clustered with isolates belonging to NA-VCG2 and NA-VCG5. C. coccodes isolates that clustered with two isolates belonging to NA-VCG6 were the most diverged from other groups, including seven isolates collected from hosts other than potato. As opposed to the bootstrap analysis, a quadratic discriminant analysis (QDA) of AFLP data correctly categorized the two isolates of NA-VCG4. Furthermore, in isolates where VCG determinations had been made, this model correctly classified isolates of all VCGs. QDA classifications were identical to those made by the bootstrap analysis, with the exception of VCG4. Overall, classifications made by the QDA model were strongly correlated (r = 0.970, P < 0.001) to the VCGs assigned by traditional methods. All 99 C. coccodes isolates evaluated only by AFLP also were subjected to QDA, leading to the assignment of a presumptive VCG for each isolate. No isolates of VCG4 or VCG6 were identified by QDA within this population. Symptoms of black dot developed in plants inoculated with isolates collected from both potato and non-potato hosts. However, total yield was not significantly reduced by infection with non-potato isolates. The lack of any additional groups identified by AFLP analysis may be an indicator of a limited level of genetic variation among North American C. coccodes isolates. AFLP is a much more efficient technique for subspecific characterization in C. coccodes than VCG analysis utilizing nit mutants and will provide an effective means by which the population biology of this pathogen can be further investigated worldwide.
Differential Selection on Rhynchosporium secalis During Parasitic and Saprophytic Phases in the Barley Scald Disease Cycle
Authors: Abang Mathew M. , Baum Michael , Ceccarelli Salvatore , Grando Stefania , Linde Celeste C. , Yahyaoui Amor , Zhan Jiasui , McDonald Bruce A.
Published: May 2007

Competition among eight Rhynchosporium secalis isolates was assessed during parasitic and saprophytic phases of the disease cycle in field experiments conducted at two locations and over two growing seasons. The eight isolates were inoculated onto six barley populations exhibiting varying degrees of resistance. Microsatellite analysis of 2,866 isolates recovered from the field experiments showed significant, and sometimes opposite, changes in the frequencies of R. secalis genotypes during the growing season (parasitic phase) and between growing seasons (saprophytic phase). Isolates that showed the most complex virulence in greenhouse seedling assays had the lowest fitness in the field experiment. Significant differences in isolate fitness were found on different host populations and in different environments. Selection coefficients were large, indicating that evolution can occur rapidly in field populations. Although inoculated isolates had the lowest overall fitness on the moderately resistant landrace cv. Arabi Aswad, some isolates were more virulent and consistently increased in frequency on this landrace, suggesting a risk of directional selection and possible erosion of the resistance following its widespread deployment in monoculture. These results provide the first direct evidence that R. secalis pathogen genotypes differ in their saprophytic ability and parasitic fitness under field conditions.
A Pectate Lyase Homolog, xagP, in Xanthomonas axonopodis pv. glycines Is Associated with Hypersensitive Response Induction on Tobacco
Authors: Kaewnum S. , Prathuangwong S. , Burr T. J.
Published: May 2007

Xanthomonas axonopodis pv. glycines is the causal agent of bacterial pustule disease of soybeans. A transposon insertional mutant (KU-P-M670) of X. axonopodis pv. glycines derived from wild-type strain KU-P-34017 lost the ability to induce the hypersensitive response (HR) on tobacco and pepper but retained its HR induction capacity on cucumber, sesame, and tomato. The mutation also resulted in loss of ability to cause a potato soft rot and express pectolytic activity at pH 6.5. An approximate 1.4-kb DNA fragment carrying the transposon insertion contained a single open reading frame that showed high homology with PSTRU-3, a pectate lyase gene in X. axonopodis pv. malvacearum. Complemented KU-P-M670 regained HR induction on tobacco and also pectolytic activity. Treatment of plants with inhibitors of eukaryotic metabolism blocked HR induction by wild-type strains and by complemented KU-P-M670. The presence of the pectate lyase homolog, which we designated xagP, in 26 X. axonopodis pv. glycines strains was highly correlated with their ability to induce an HR on tobacco. To our knowledge, this is the first study indicating a role for a functional pectate lyase in induction of a plant HR.
A Multiplex Real-Time Polymerase Chain Reaction (TaqMan) Assay for the Simultaneous Detection of Meloidogyne chitwoodi and M. fallax
Authors: Zijlstra C. , Van Hoof R. A.
Published: May 2007

This study describes a multiplex real-time polymerase chain reaction (PCR) approach for the simultaneous detection of Meloidogyne chitwoodi and M. fallax in a single assay. The approach uses three fluorogenic minor groove binding (MGB) TaqMan probes: one FAM-labeled to detect M. chitwoodi, one VIC-labeled to detect M. fallax, and one NED-labeled to detect the internal amplification control (IAC) to monitor false negative results. One common primer set is used for the amplification of part of the internal transcribed spacer (ITS) region of M. chitwoodi and M. fallax and one primer set for the amplification of the IAC. The test enabled detection of M. chitwoodi and/or M. fallax in DNA samples extracted from batches of juveniles, from single juveniles, and from infected plant material. Compared with current assays to detect M. chitwoodi and M. fallax, the multiplex real-time PCR offers the following advantages: it is faster because the test can simultaneously detect both quarantine species without the need for post-PCR processing; and it is at least 10 times more sensitive than a comparable regular PCR also targeting the ITS sequence. Inclusion of the IAC facilitates the interpretation of the FAM and VIC cycle threshold (Ct) values and can prevent the scoring of false negative results when FAM, VIC, and NED Ct values are high. The test allows precise quantification when only one of the two species is present in the sample. However, experiments with mixtures of genomic DNA of M. chitwoodi and M. fallax revealed that the ability of the multiplex real-time PCR assay to detect small quantities of DNA of one species is reduced when large quantities of DNA of the other species are present.
Identification of Sources of Resistance to Phoma medicaginis Isolates in Medicago truncatula SARDI Core Collection Accessions, and Multigene Differentiation of Isolates
Authors: Ellwood Simon R. , Kamphuis Lars G. , Oliver Richard P.
Published: May 2007

Phoma medicaginis is a necrotrophic fungal pathogen, commonly found infecting the annual medic Medicago truncatula. To differentiate eight P. medicaginis isolates, five gene regions were examined: actin, β-tubulin, calmodulin, translation elongation factor 1-α (EF-1α), and the internal transcribed spacer ribosomal DNA. Sequence comparisons showed that specimens isolated from M. truncatula in Western Australia formed a group that was consistently different from, but allied to, a P. medicaginis var. medicaginis type specimen. EF-1α contained a hyper-variable 55-bp repeat unit, which forms the basis of a rapid polymerase chain reaction-based method of reliably distinguishing isolates. Characterization of three isolates showed that all exhibited a narrow host range, causing disease only in M. sativa and M. truncatula among eight commonly cultivated legume species sampled. Infection of 86 M. truncatula single-seeded accessions showed a continuous distribution in disease phenotypes, with the majority of accessions susceptible. On a 1-to-5 disease reaction scale increasing in severity, individual fungal isolates showed means of 2.6 to 3.2, and scores ranged from 1 to 4.8 among accessions. The results presented here suggest that M. truncatula harbors specific and diverse sources of resistance to individual P. medicaginis genotypes.
Induction of Soil Suppressiveness Against Rhizoctonia solani by Incorporation of Dried Plant Residues into Soil
Authors: Kasuya Masahiro , Olivier Andriantsoa R. , Ota Yoko , Tojo Motoaki , Honjo Hitoshi , Fukui Ryo
Published: May 2007

Suppressive effects of soil amendment with residues of 12 cultivars of Brassica rapa on damping-off of sugar beet were evaluated in soils infested with Rhizoctonia solani. Residues of clover and peanut were tested as noncruciferous controls. The incidence of damping-off was significantly and consistently suppressed in the soils amended with residues of clover, peanut, and B. rapa subsp. rapifera ‘Saori’, but only the volatile substance produced from water-imbibed residue of cv. Saori exhibited a distinct inhibitory effect on mycelial growth of R. solani. Nonetheless, disease suppression in such residue-amended soils was diminished or nullified when antibacterial antibiotics were applied to the soils, suggesting that proliferation of antagonistic bacteria resident to the soils were responsible for disease suppression. When the seed (pericarps) colonized by R. solani in the infested soil without residues were replanted into the soils amended with such residues, damping-off was suppressed in all cases. In contrast, when seed that had been colonized by microorganisms in the soils containing the residues were replanted into the infested soil, damping-off was not suppressed. The evidence indicates that the laimosphere, but not the spermosphere, is the site for the antagonistic microbial interaction, which is the chief principle of soil suppressiveness against Rhizoctonia damping-off.
Selection for Fungicide Resistance Within a Growing Season in Field Populations of Phytophthora infestans at the Center of Origin
Authors: Grünwald Niklaus J. , Sturbaum Anne K. , Montes Gaspar Romero , Serrano Edith Garay , Lozoya-Saldaña Hector , Fry William E.
Published: May 2007

The central highlands of Mexico should provide an optimal testing ground for evaluating the potential threat of selection for resistance to fungicides in the population of Phytophthora infestans. We evaluated the hypotheses that exposure to the fungicides azoxystrobin, cymoxanil, dimethomorph, fluazinam, mancozeb, metalaxyl, and propamocarb hydrochloride would lead to (i) a shift in the sensitivity distributions (i.e., selection) and (ii) a lower genotypic diversity of the population. We compared populations from unsprayed plots with populations that had been exposed to several applications of each of the fungicides within a single field season. This study provides novel baseline data and shows that the Toluca valley P. infestans population has a wide range of sensitivities to the fungicides fluazinam, cymoxanil, dimethomorph, metalaxyl, and propamocarb. Directional selection toward resistance combined with a reduction in genetic diversity of the P. infestans population was observed only for the fungicide metalaxyl. The results obtained provide direct experimental support for continuing vigilance regarding further introductions of exotic strains of P. infestans into the United States.
Macroarray Detection of Plant RNA Viruses Using Randomly Primed and Amplified Complementary DNAs from Infected Plants
Authors: Agindotan Bright , Perry Keith L.
Published: May 2007

Membrane-based macroarrays provide a relatively inexpensive technology with the potential to detect hundreds of pathogens in a single assay. For the simultaneous detection of a large number of pathogens, it is necessary to obtain sufficient nucleic acids for labeling, and any amplification reactions need to be performed using unbiased, pathogen-non-specific primers. A nonradioactive macroarray system is described to test for plant RNA viruses using 70-mer oligonucleotide probes immobilized on nylon membranes. Starting with a total plant RNA extract, complementary DNA (cDNA) and second-strand syntheses were carried out using an anchor primer sequence with random pentamers coupled at the 3′ end. Subsequent synthesis by polymerase chain reaction using the anchor primer alone resulted in a relatively unbiased amplification of plant and viral RNAs. These cDNAs were chemically labeled and the product used as a target in hybridization analyses. The system was validated using RNA extracts from plants infected with Cucumber mosaic virus, Potato virus Y, and Potato leaf roll virus (PLRV). Despite the relative excess of host-derived nonviral sequences, viral RNAs were amplified between 100- and 1,000-fold and were detected in single and mixed infections. The macroarray sensitivity was comparable to that of double-antibody sandwich enzyme-linked immunosorbent assay, with PLRV being detected in sap dilutions of 1:100. The potential for the development of a relatively inexpensive multipathogen detection system is discussed.
Sequence Tagged Site Markers to Rsp1, Rsp2, and Rsp3 Genes for Resistance to Septoria Speckled Leaf Blotch in Barley
Authors: Lee S. H. , Neate S. M.
Published: May 2007

Five random amplified polymorphic DNA markers, two in coupling (OPAH5545C, and OPBA12314C) and three in repulsion phase (UBC285158R, OPC2441R, and OPB17451R), closely linked to Rsp genes conferring resistance to Septoria speckled leaf blotch (SSLB), were identified using bulked segregant analysis in three F2 populations, each containing a Rsp gene. These markers were converted into the sequence tagged site (STS) markers SUBC285, SOPC2, SOPAH5, and SOPBA12. Another STS marker (MWG938) linked to Rsp2 in coupling phase was also identified in an F2 population from the cross Robust/CIho 4780. The STS markers were tested on a set of 42 resistant and susceptible barley germplasm lines and 98 landraces. The expected sizes of marker fragments associated with each allele at Rsp loci were present in resistant or susceptible accessions. Efficiency of marker-assisted selection (MAS) for Rsp1, Rsp2, and Rsp3 using STS markers were evaluated in three F2–3 populations in the greenhouse and the field. Results of testing F2–3 progeny demonstrated that the accuracy of MAS was, with one exception, greater than 97% in the greenhouse and in two field locations (90% in the Osnabrock, ND trial for Rsp2). The STS markers closely linked to Rsp genes also identified the SSLB resistance corresponding to Rsp1, Rsp2, or Rsp3 in gene pyramiding F2 populations. The STS markers tightly linked to Rsp genes may be useful for M and for pyramiding with other genes in barley breeding for SSLB resistance.
Diversity and Ecology of Biocontrol Pseudomonas spp. in Agricultural Systems
Authors: McSpadden Gardener Brian B.
Published: May 2007

Diverse Pseudomonas spp. may act as biological controls of plant pathogens, but the ecology of those natural populations is not well understood. And, while biocontrol potential has been identified in multiple pseudomonad strains, the linkages between genotype and phenotype have yet to be fully delineated. However, intensive studies of one class of biocontrol strains, i.e., those that can produce 2,4-diacetylphloroglucionl (DAPG), have provided new insights into the diversity, distribution, and interactions of biocontrol pseudomonads. Those studies also laid the foundation for future research and development of pseudomonad-based biocontrol strategies. Over the past several years, numerous studies have also revealed that biocontrol pseudomonads are widely distributed in agricultural soils, and that multiple crop and soil factors can affect their abundance and activities. Recent work has shown that a variety of farm management practices that reduce soilborne disease pressure can also alter the rhizosphere abundance of DAPG producers in complex ways. Such studies provide support for the hypothesis of an ecological feedback mechanism whereby a native biocontrol population increase and subsequently reduce root disease severity following infection. It is well established that complex biological interactions can take place among bio-control pseudomonads, plant pathogens, their hosts, and other members of the microbial community. The net result of such interactions likely dilutes biocontrol efficacy at the field scale. Nonetheless, inoculation can be effective, and several successful applications of biocontrol pseudomonads have been developed. Future applications of microbial ecology research will hopefully improve the consistency and efficacy of bio-control mediated by Pseudomonas spp. Current applications and future opportunities for improving pseudomonad-based biological control are discussed.
Roles of Diffusible Signals in Communication Among Plant-Associated Bacteria
Authors: Pierson Leland S. , Pierson Elizabeth A.
Published: May 2007

In nature, Pseudomonas species compete and co-exist in mixed communities with a diversity of prokaryotic and eukaryotic micro- and macroorganisms. Many bacteria produce various signals that control gene expression and thus contribute to specific bacterial behaviors and coordinate essential functions with other members of the community. The best-studied signaling compounds are N-acyl-homoserine lactones (AHLs), which are involved in quorum sensing (QS) regulation and are produced by a diverse range of bacterial taxa. To date, research on QS has focused on how signals control gene expression in the bacterial cell and the role of these signals in positive and negative communication among different groups of organisms. Additionally, mechanisms for AHL decay and AHL utilization as sole carbon/energy sources have been identified. Some host organisms produce compounds that can mimic AHLs, and some bacterial signals can influence host gene expression. Thus, interkingdom communication may be more widespread than previously believed. Our current understanding of individual, community and bacterial-host interactions is still in its infancy and there are many exciting discoveries yet to be made.
Characterization and Expression of β-1,3-Glucanase Genes in Jujube Fruit Induced by the Microbial Biocontrol Agent Cryptococcus laurentii
Authors: Tian Shi P. , Yao Hong J. , Deng Xin , Xu Xiang B. , Qin Guo Z. , Chan Zhu L.
Published: May 2007

Two β-1,3-glucanase genes were cloned from jujube (Ziziphus jujuba Mill) fruit and designated Glu-1 and Glu-2 (GenBank accession numbers DQ012940 and DQ093571), respectively. The expression of Glu-1 and Glu-2 in jujube fruit in response to wounding and microbial biocontrol agent was evaluated by semi-quantitative reverse-transcription polymerase chain reaction analysis. Wounding and treatment with Cryptococcus laurentii stimulated an increase in β-1,3-glucanase (EC 3.2.1.39) activity in jujube fruit. Analysis of gene expression proved that Glu-1 was highly induced both by wounding and C. laurentii, whereas Glu-2 was broadly not responsive to the yeast. The expression of Glu-1 was noticeably enhanced with increased concentrations of C. laurentii, suggesting that Glu-1 may play a role in defense responses to fungal pathogens. The results hold true at the levels of gene activation and enzyme accumulation in jujube fruit treated by both stimuli, resulting in a significant decrease in disease incidence and lesion diameter, thus providing evidence that changes in β-1,3-glucanase activity are related to expression of the genes. Taken together, these findings suggest the possible use of β-1,3-glucanase activity as a biochemical marker for screening jujube fruit against fungal pathogens, and also provide a mechanistic framework for the functions of β-1,3-glucanase in defense responses.
Fruit Exocarp Phenols in Relation to Quiescence and Development ofMonilinia fructicolaInfections inPrunusspp.: A Role for Cellular Redox?
Authors: Lee Miin-Huey , Bostock Richard M.
Published: May 2007

Monilinia fructicola causes brown rot of Prunus species and usually remains quiescent on immature fruit but reactivates when fruit are mature. The dihydroxycinnamates caffeic acid and its quinate ester, chlorogenic acid, abundant in the exocarp of peach fruit, had no effect on fungal growth but markedly inhibited the production of the cell wall degrading enzymes polygalacturonase and cutinase in M. fructicola cultures. This inhibition was related to changes in the electrochemical redox potentials of the cultures, as measured with a redox electrode. Fungal culture filtrates had lower electrochemical redox potentials when the growth medium contained caffeic acid than in caffeic acid-free medium. Levels of total intracellular glutathione, the reduced form of which serves as a major cellular antioxidant, increased significantly in M. fructicola cells in response to external caffeic acid. The presence of caffeic acid, chlorogenic acid, or reduced glutathione in conidial suspensions of M. fructicola did not inhibit germination on flower petals and fruit, but inhibited appressorium formation from germinated conidia and subsequent brown rot lesion development. These results suggest that intracellular antioxidant levels in the pathogen can be influenced by phenols present in host tissue and that changes in the redox environment may influence gene expression and differentiation of structures associated with infection by the pathogen. The possible relationship of host phenols to quiescence and subsequent development of M. fructicola infections is discussed.
Calcium Restores Prepenetration Morphogenesis Abolished by Methylglyoxal-Bis-Guanyl Hydrazone inCochliobolus miyabeanusInfecting Rice
Authors: Ahn Il-Pyung , Suh Seok-Cheol
Published: May 2007

Cochliobolus miyabeanus forms a specialized infection structure, an appressorium, to infect its host rice plants. Curtailment of prepenetration development by spermidine and spermine was more evident in appressorium development and germination remained unaffected, whereas putrescine and methylglyoxal-bis-guanyl hydrazone (MGBG) impaired both morphogenetic events. Exogenous calcium nullified the inhibitory effect of MGBG on the prepenetration development in vitro and in vivo and the disease progression. High levels of polyamines were detected in freshly collected conidia, but the amounts were reduced during germination and appressorium formation. MGBG fortified the decrease of polyamines within conidia under development and calcium amendment did not affect the reduction. Hard-surface contact augmented messenger RNA synthesis of calmodulin gene (CmCaM) and protein kinase C (PKC) activity in germinating or appressorium-forming conidia. Calcium restored transcription of CmCaM and upregulation of PKC activity suppressed by MGBG. Taken together, fine-tuning of intracellular polyamine transition is indispensable for the conidial germination and appressorium formation in C. miyabeanus. Biochemical and molecular analyses revealed that the MGBG-acting site or sites are upstream of Ca2+-dependent signaling pathways regulating prepenetration morphogenesis of C. miyabeanus causing rice brown leaf spot.
Adaptation ofPhytophthora infestansto Partial Resistance in Potato: Evidence from French and Moroccan Populations
Authors: Andrivon Didier , Pilet Fabian , Montarry Josselin , Hafidi Majida , Corbière Roselyne , Achbani El Hassan , Pellé Roland , Ellissèche Daniel
Published: May 2007

The use of partially resistant cultivars should become an essential component of a sustainable management strategy of potato late blight, caused by Phytophthora infestans. It is therefore important to determine to what extent P. infestans populations can be selected for increased aggressiveness by potato cultivars with different levels of partial resistance. To this end, we sampled P. infestans populations from France and Morocco, chosen as locations where late blight occurs regularly but which differ in the distribution of potato cultivars. Cross-inoculation experiments were used to determine the aggressiveness of all populations to potato cvs. Bintje (prevalent in France but not grown in Morocco) and Désirée (popular in Morocco but cultivated to a very small extent in France). French populations were more aggressive on cv. Bintje than on cv. Désirée, irrespective of the site they were sampled from. Their aggressiveness increased between early and late samplings, suggesting that both cultivars selected for increased aggressiveness during epidemics. By contrast, Moroccan populations were more aggressive on Désirée, regarded as partially resistant in Europe, than on Bintje, highly susceptible under European conditions. These data indicate that P. infestans populations adapt to locally dominant cultivars, irrespective of their resistance levels, and can therefore overcome polygenic, partial resistance. This adaptive pattern may render partial resistance nondurable if not properly managed.
Colonization of Fiber Cells by Colletotrichum graminicola in Wounded Maize Stalks
Authors: Venard C. , Vaillancourt L.
Published: May 2007

Colonization of wounded maize stalks by a wild-type strain of Colletotrichum graminicola was compared with colonization by a C. graminicola mutant that is avirulent on maize leaves, and by a wild-type strain of C. sublineolum that is normally a pathogen of sorghum but not maize. Local infection by all strains at the wound site resulted in formation of primary lesions consisting of disintegrated parenchyma cells beneath an intact rind and epidermis. However, subsequent rapid longitudinal expansion of the primary lesion occurred only in infections with the wild-type C. graminicola strain, and proceeded specifically through the fiber cells associated with the vascular bundles and the rind. Hyphae emerged from the fiber cells to produce discontinuous secondary lesions. There was no evidence that C. graminicola is a vascular wilt pathogen. Resistance of wounded cv. Jubilee maize stalks to the mutant strain of C. graminicola and to C. sublineolum was associated with restriction of colonization and spread of the pathogen through the fibers, as well as with the limitation of localized destruction of parenchyma cells at the wound site.
Instability of Propiconazole Resistance and Fitness in Monilinia fructicola
Authors: Cox K. D. , Bryson P. K. , Schnabel G.
Published: May 2007

The fitness and the dynamics of demethylation inhibitor fungicide (DMI) sensitivity in isolates of Monilinia fructicola sensitive (no growth at 0.3 mg/liter propiconazole) and resistant (≥50% relative growth at 0.3 mg/liter propiconazole) to propiconazole were investigated. Overall, there was no considerable compromise in the fitness of resistant isolates compared to sensitive isolates of M. fructicola at the time of collection. Resistant and sensitive isolates differed in their sensitivity to propiconazole (P < 0.001) and incubation period (P = 0.044), but not in latent period, growth rate, spore production, and spore germination frequency (P > 0.05). Consecutive transferring on potato dextrose agar had an impact on conidia production, conidial germination, and growth rate (P < 0.0001). Consecutive transferring also had an impact on propiconazole sensitivity in resistant isolates. In the resistant isolates, sensitivity to propiconazole increased (R2 = 0.960, P = 0.0034) within the first eight transfers. Similarly, sensitivity to propiconazole increased by 273% over the course of 34 months in cold storage in propiconazole-resistant isolates. Our results show that propiconazole resistance is unstable in vitro and that standard subculturing and cold storage procedures impact propiconazole sensitivity of resistant isolates. The instability of propiconazole resistance in M. fructicola may have important implications for disease management in that a reversion to propiconazole sensitivity could potentially occur in the absence of DMI fungicide pressure in the field.
Characterization of a Regional Population ofFusarium oxysporumf. sp.niveumby Race, Cross Pathogenicity, and Vegetative Compatibility
Authors: Zhou X. G. , Everts K. L.
Published: May 2007

Eighty-eight isolates of Fusarium oxysporum f. sp. niveum, collected from wilted watermelon plants and infested soil in Maryland and Dela-ware, were characterized by cross pathogenicity to muskmelon, race, and vegetative compatibility. Four isolates (4.5%) were moderately pathogenic to ≥2 of 18 muskmelon cultivars in a greenhouse test, and one representative isolate also was slightly pathogenic in field microplots. The four isolates all were designated as race 2, and were in vegetative compatibility group (VCG) 0082. Of the 74 isolates to which a VCG could be assigned, 41 were in VCG 0080, the VCG distributed most widely; 27 were in VCG 0082, and were distributed in half of the 20 watermelon fields surveyed; and 6 were in the newly described VCG 0083, and were restricted to three fields. Among the isolates in VCG 0080, 8 were designated as race 0, 21 as race 1, and 12 as race 2. Of the isolates in VCG 0082, 6 were designated as race 0, 11 as race 1, and 10 as race 2. All isolates in VCG 0083 were designated as race 2. Isolates from more than one race within the same VCG or isolates from more than one VCG were recovered from single plants and fields. No differences in aggressiveness on differential watermelon cultivars were observed among isolates from different VCGs of the same race. A diverse association between virulence and VCG throughout the Mid-Atlantic region suggests that the pathotypes of F. oxysporum f. sp. niveum may be of local origin or at least long existent in the region.
Relationship of Sphinganine Analog Mycotoxin Contamination in Maize Silage to Seasonal Weather Conditions and to Agronomic and Ensiling Practices
Authors: Mansfield Michele A. , Archibald Douglas D. , Jones A. Daniel , Kuldau Gretchen A.
Published: May 2007

Sphinganine analog mycotoxins (SAMs) are reported in maize and maize based feeds. Our objectives were to detect and quantify fumonisins B1 and B2 and Alternaria toxins (AAL toxins) AAL-TA and AAL-TB and determine how agronomic practices, weather conditions, and ensiling affected the occurrence and levels in maize silage. Silage was collected at harvest and after ensiling in 2001 and 2002 from 30 to 40 dairies, representing four regions in Pennsylvania. SAMs were quantified using high pressure liquid chromatography (HPLC) with fluorescence detection and high pressure liquid chromatography-mass spectrometry HPLC-MS. The average concentrations and ranges were as follows: fumonisin B1 2.02 μg/g (0.20 to 10.10), fumonisin B2 0.98 μg/g (0.20 to 20.30), AAL-TA 0.17 μg/g (0.20 to 2.01), and AAL-TB 0.05 μg/g (0.03 to 0.90). Fumonisin B1 was the most frequently detected toxin (92%) in all samples, followed by fumonisin B2 (55%), AAL-TA (23%), and -TB (13%). Temperature during maize development was positively correlated with fumonisin occurrence and levels and negatively with AAL-TA, while moisture events were negatively correlated with fumonisins and positively with AAL-TA. Fumonisin levels were higher in silage harvested at later developmental stages (dough through physiological maturity). Ensiling did not affect toxin concentration nor did agronomic practices (tillage system, inoculant use, or silo type) or silage characteristics (dry matter, pH, or organic acid concentration). This is the first report of AAL-TB in silage and on factors that affect SAM frequency and levels in maize silage.