bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification and Characterization of Nip, Necrosis-Inducing Virulence Protein of Erwinia carotovora subsp. carotovora
Authors: Mattinen Laura , Tshuikina Marina , Mäe Andres , Pirhonen Minna
Published: May 2007

Erwinia carotovora subsp. carotovora is a gram-negative bacterium that causes soft rot disease of many cultivated crops. When a collection of E. carotovora subsp. carotovora isolates was analyzed on a Southern blot using the harpin-encoding gene hrpN as probe, several harpinless isolates were found. Regulation of virulence determinants in one of these, strain SCC3193, has been characterized extensively. It is fully virulent on potato and in Arabidopsis thaliana. An RpoS (SigmaS) mutant of SCC3193, producing elevated levels of secreted proteins, was found to cause lesions resembling the hypersensitive response when infiltrated into tobacco leaf tissue. This phenotype was evident only when bacterial cells had been cultivated on solid minimal medium at low pH and temperature. The protein causing the cell death was purified and sequenced, and the corresponding gene was cloned. The deduced sequence of the necrosis-inducing protein (Nip) showed homology to necrosis- and ethylene-inducing elicitors of fungi and oomycetes. A mutant strain of E. carotovora subsp. carotovora lacking the nip gene showed reduced virulence in potato tuber assay but was unaffected in virulence in potato stem or on other tested host plants.
Fungal Elicitation of Signal Transduction-Related Plant Genes Precedes Mycorrhiza Establishment and Requires the dmi3 Gene in Medicago truncatula
Authors: Weidmann Stéphanie , Sanchez Lisa , Descombin Julie , Chatagnier Odile , Gianinazzi Silvio , Gianinazzi-Pearson Vivienne
Published: May 2007

Suppressive subtractive hybridization and expressed sequence tag sequencing identified 29 plant genes which are upregulated during the appressorium stage of mycorrhiza establishment between Medicago truncatula J5 (Myc+) and Glomus mosseae. Eleven genes coding plant proteins with predicted functions in signal transduction, transcription, and translation were investigated in more detail for their relation to early events of symbiotic interactions. Expression profiling showed that the genes are activated not only from the appressorium stage up to the fully established symbiosis in the Myc+ genotype of M. truncatula, but also when the symbionts are not in direct cell contact, suggesting that diffusible fungal molecules (Myc factors) play a role in the induction of a signal-transduction pathway. Transcript accumulation in roots of a mycorrhiza-defective Myc- dmi3 mutant of M. truncatula is not modified by appressorium formation or diffusible fungal molecules, indicating that the signal transduction pathway is required for a successful G. mosseae-M. truncatula interaction leading to symbiosis development. The symbiotic nodulating bacterium Sinorhizobium meliloti does not activate the 11 genes, which supposes early discrimination by plant roots between the microbial symbionts.
Relationships Among Endo-Polygalacturonase, Oxalate, pH, and Plant Polygalacturonase-Inhibiting Protein (PGIP) in the Interaction Between Sclerotinia sclerotiorum and Soybean
Authors: Favaron Francesco , Sella Luca , D'Ovidio Renato
Published: May 2007

The necrotrophic fungal pathogen Sclerotinia sclerotiorum secretes oxalic acid and endo-polygalacturonase (endo-PG) in host plants. Oxalic acid acidifies the plant tissue to values more suitable to endo-PG activity. However, we observed that the infected soybean seedlings possessed a pH of 3.8, which is below that optimal for endo-PG activity (4.5 to 5.0). We investigated, therefore, the effects of pH (from 5.0 to 3.6) and oxalate (5 to 20 mM) on the activity of the major basic endo-PG (PGb) and towards an acidic endo-PG (PGa) secreted by S. sclerotiorum during soybean infection. We verified that only PGb activity is stimulated by oxalate, while at the lowest pH levels, PGa escapes the inhibition of a soybean polygalacturonase-inhibiting protein (PGIP). These results, performed on polygalacturonic acid, were apparently consistent with data obtained from studies with soybean hypocotyl segments, in which PGb activity was increased by oxalate and PGa maintained its activity also at pH 3.6, possibly because at this pH the PGIP contained in the plant tissue is inactive. Reverse transcription-polymerase chain reaction analysis showed that, during soybean infection, the expression of the putative pga gene is delayed in comparison to the basic one. The different temporal expressions of the two endo-PGs and their differing responses to pH, oxalate, and PGIP seem to be consistent with a possible maximization of the fungal PG activity in the host tissue.
Harpin Inactivates Mitochondria in Arabidopsis Suspension Cells
Authors: Krause Maren , Durner Jörg
Published: May 2007

Harpin is a well-known proteinaceous bacterial elicitor that can induce an oxidative burst and programmed cell death in various host plants. Given the demonstrated roles of mitochondria in animal apoptosis, we investigated the effect of harpin from Pseudomonas syringae on mitochondrial functions in Arabidopsis suspension cells in detail. Fluorescence microscopy in conjunction with double-staining for reactive oxygen species (ROS) and mitochondria suggested co-localization of mitochondria and ROS generation. Plant defense responses or cell death after pathogen attack have been suggested to be regulated by the concerted action of ROS and nitric oxide (NO). However, although Arabidopsis cells respond to harpin treatment with NO generation, time course analyses suggest that NO generation is not involved in initial responses but, rather, is a consequence of cellular decay. Among the fast responses we observed was a decrease of the mitochondrial membrane potential Δψm and, possibly as a direct consequence, of ATP production. Furthermore, treatment of Arabidopsis cells with harpin protein induced a rapid cytochrome C release from mitochondria into the cytosol, which is regarded as a hallmark of programmed cell death or apoptosis. Northern and DNA array analyses showed strong induction of protecting or scavenging systems such as alternative oxidase and small heat shock proteins, components that are known to be associated with cellular stress responses. In sum, the presented data suggest that harpin inactivates mitochondria in Arabidopsis cells.
Importance of opgHXcv of Xanthomonas campestris pv. vesicatoria in Host-Parasite Interactions
Authors: Minsavage G. V. , Mudgett M. B. , Stall R. E. , Jones J. B.
Published: May 2007

Tn5 insertion mutants of Xanthomonas campestris pv. vesicatoria were inoculated into tomato and screened for reduced virulence. One mutant exhibited reduced aggressiveness and attenuated growth in planta. Southern blot analyses indicated that the mutant carried a single Tn5 insertion not associated with previously cloned pathogenicity-related genes of X. campestris pv. vesicatoria. The wild-type phenotype of this mutant was restored by one recombinant plasmid (pOPG361) selected from a genomic library of X. campestris pv. vesicatoria 91–118. Tn3-gus insertion mutagenesis and sequence analyses of a sub-clone of pOPG361 identified a 1,929-bp open reading frame (ORF) essential for complementation of the mutants. The predicted protein encoded by this ORF was highly homologous to the previously reported pathogenicity-related HrpM protein of Pseudomonas syringae pv. syringae and OpgH of Erwinia chrysanthemi. Based on homology, the new locus was designated opgHXcv. Manipulation of the osmotic potential in the intercellular spaces of tomato leaves by addition of mannitol at low concentrations (25 to 50 mM) compensates for the opgHXcv mutation.
Identification of a Novel Pseudomonas syringae Psy61 Effector with Virulence and Avirulence Functions by a HrpL-Dependent Promoter-Trap Assay
Authors: Losada L. , Sussan T. , Pak K. , Zeyad S. , Rozenbaum I. , Hutcheson S. W.
Published: May 2007

The hrp pathogenicity island of Pseudomonas syringae encodes a type III secretion system (TTSS) that translocates effectors into plant cells. Most genes encoding effectors are dispersed in the P. syringae genome. Regardless of location, all are regulated coordinately by the alternative sigma factor HrpL. An HrpL-dependent promoter-trap assay was developed to screen genomic libraries of P. syringae strains for promoters whose activity in Escherichia coli is dependent on an inducible hrpL construct. Twenty-two HrpL-dependent promoter fragments were isolated from P. syringae Psy61 that included promoters for known HrpL-dependent genes. One fragment also was isolated that shared no similarity with known genes but retained a near consensus HrpL-dependent promoter. The sequence of the region revealed a 375-amino acid open reading frame encoding a 40.5-kDa product that was designated HopPsyL. HopPsyL was structurally similar to other secreted effectors and carried a putative chloroplast-targeting signal and two predicted transmembrane domains. HopPsyL′:′ AvrRpt2 fusions were translocated into host cells via the P. syringae pv. tomato DC3000 hrp TTSS. A hopPsyL∷kan mutant of Psy61 exhibited strongly reduced virulence in Phaseolus vulgaris cv. Kentucky Wonder, but did not appear to act as a defense response suppressor. The ectopically expressed gene reduced the virulence of Pseudomonas syringae DC3000 transformants in Arabidopsis thaliana Col-0. The gene was shown to be conserved in 6 of 10 P. syringae pv. syringae strains but was not detected in 35 strains of other pathovars. HopPsyL appears to be a novel TTSS-dependent effector that functions as a host-species-specific virulence factor in Psy61.
The White Barley Mutant Albostrians Shows Enhanced Resistance to the Biotroph Blumeria graminis f. sp. hordei
Authors: Jain Sanjay Kumar , Langen Gregor , Hess Wolfgang , Börner Thomas , Hückelhoven Ralph , Kogel Karl-Heinz
Published: May 2007

We performed cytological and molecular analyses of the interaction between the biotrophic barley powdery mildew fungus Blumeria graminis f. sp. hordei and white and green leaves of the barley albostrians mutant. The leaves have the same nuclear genotype but differ from each other in respect to plastid differentiation. White leaves showed enhanced penetration resistance to B. graminis f. sp. hordei, associated with higher epidermal H2O2 accumulation beneath the appressorial germ tubes and protein cross-linking in papillae. Very low basal salicylic acid content was found in white leaves, which further confirmed that H2O2 accumulation and penetration resistance in barley are independent of salicylic acid. Expression analysis of stress and defense-related genes, including such being involved in reactive oxygen species production and cell death regulation, revealed stronger constitutive or pathogen-induced transcript accumulation in white leaves. We discuss the data on the basis of the finding that white albostrians leaves exhibit a supersusceptible interaction phenotype with the hemibiotrophic fungus Bipolaris sorokiniana.
The Avr1b Locus of Phytophthora sojae Encodes an Elicitor and a Regulator Required for Avirulence on Soybean Plants Carrying Resistance Gene Rps1b
Authors: Shan Weixing , Cao Minh , Leung Dan , Tyler Brett M.
Published: May 2007

We have used map-based approaches to clone a locus containing two genes, Avr1b-1 and Avr1b-2, required for avirulence of the oomycete pathogen Phytophthora sojae (Kaufmann & Gerdemann) on soybean plants carrying resistance gene Rps1b. Avr1b-1 was localized to a single 60-kb bacterial artificial chromosome (BAC) clone by fine-structure genetic mapping. Avr1b-1 was localized within the 60-kb region by identification of an mRNA that is expressed in a race-specific and infection-specific manner and that encodes a small secreted protein. When the Avr1b-1 protein was synthesized in the yeast Pichia pastoris and the secreted protein infiltrated into soybean leaves, it triggered a hypersensitive response specifically in host plants carrying the Rps1b resistance gene. This response eventually spread to the entire inoculated plant. In some isolates of P. sojae virulent on Rps1b-containing cultivars, such as P7081 (race 25) and P7076 (race 19), the Avr1b-1 gene had numerous substitution mutations indicative of strong divergent selection. In other isolates, such as P6497 (race 2) and P9073 (race 25), there were no substitutions in Avr1b-1, but Avr1b-1 mRNA did not accumulate. Genetic complementation experiments with P6497 revealed the presence of a second gene, Avr1b-2, required for the accumulation of Avr1b-1 mRNA. Avr1b-2 was genetically mapped to the same BAC contig as Avr1b-1, using a cross between P7064 (race 7) and P6497. The Avr1k gene, required for avirulence on soybean cultivars containing Rps1k, was mapped to the same interval as Avr1b-1.
Identification and Characterization of Victorin Sensitivity in Arabidopsis thaliana
Authors: Lorang Jennifer M. , Carkaci-Salli Nurgul , Wolpert Thomas J.
Published: May 2007

Cochliobolus victoriae is a necrotrophic fungus that produces a host-selective toxin called victorin. Victorin is considered to be host selective because it has been known to affect only certain allohexaploid oat cultivars containing the dominant Vb gene. Oat cultivars containing Vb are also the only genotypes susceptible to C. victoriae. Assays were developed to screen the “nonhost” plant of C. victoriae, Arabidopsis thaliana, for victorin sensitivity. Sensitivity to victorin was identified in six of 433 bulk populations of Arabidopsis. In crosses of Col-4 (victorin-insensitive) × victorin-sensitive Arabidopsis ecotypes, victorin sensitivity segregated as a single dominant locus, as it does in oats. This Arabidopsis locus was designated LOV, for locus orchestrating victorin effects. Allelism tests indicate that LOV loci are allelic or closely linked in all six victorin-sensitive ecotypes identified. LOV was localized to the north arm of Arabidopsis thaliana chromosome I. The victorin-sensitive Arabidopsis line LOV1 but not the victorin-insensitive line Col-4 was susceptible to C. victoriae infection. Consequently, the LOV gene appears to be a genetically dominant, disease susceptibility gene.
Gene Expression Profile Changes in Cotton Root and Hypocotyl Tissues in Response to Infection with Fusarium oxysporum f. sp. vasinfectum
Authors: Dowd Caitriona , Wilson Iain W. , McFadden Helen
Published: May 2007

Microarray analysis of large-scale temporal and tissue-specific plant gene expression changes occurring during a susceptible plant-pathogen interaction revealed different gene expression profile changes in cotton root and hypocotyl tissues. In hypocotyl tissues infected with Fusarium oxysporum f. sp. vasinfectum, increased expression of defense-related genes was observed, whereas few changes in the expression levels of defense-related genes were found in infected root tissues. In infected roots, more plant genes were repressed than were induced, especially at the earlier stages of infection. Although many known cotton defense responses were identified, including induction of pathogenesis-related genes and gossypol biosynthesis genes, potential new defense responses also were identified, such as the biosynthesis of lignans. Many of the stress-related gene responses were common to both tissues. The repression of drought-responsive proteins such as aquaporins in both roots and hypocotyls represents a previously unreported response of a host to pathogen attack that may be specific to vascular wilt diseases. Gene expression results implicated the phytohormones ethylene and auxin in the disease process. Biochemical analysis of hormone level changes supported this observation.
RCH1, a Locus inArabidopsisThat Confers Resistance to the Hemibiotrophic Fungal PathogenColletotrichum higginsianum
Authors: Narusaka Yoshihiro , Narusaka Mari , Park Pyoyun , Kubo Yasuyuki , Hirayama Takashi , Seki Motoaki , Shiraishi Tomonori , Ishida Junko , Nakashima Maiko , Enju Akiko , Sakurai Tetsuya , Satou Masakazu , Kobayashi Masatomo , Shinozaki Kazuo
Published: May 2007

When challenged with the crucifer pathogen Colletotrichum higginsianum, Arabidopsis thaliana ecotype Columbia (Col-0) was colonized by the fungus within 2 to 3 days, developing brown necrotic lesions surrounded by a yellow halo. Lesions spread from the inoculation site within 3 to 4 days, and subsequently continued to expand until they covered the entire leaf. Electron microscopy confirmed that C. higginsianum is a hemibiotroph on Arabidopsis, feeding initially on living cells as a biotroph before switching to a necrotrophic mode of growth. A collection of 37 ecotypes of Arabidopsis varied in their responses to infection by C. higginsianum. The ecotype Eil-0 was highly resistant, with symptoms limited to necrotic flecking and with only very limited fungal colonization. Analyses suggested that the hypersensitive response and reactive oxygen species may be important in this defense response. Expression analyses with cDNA microarrays indicated that the defense reaction depends primarily on the jasmonic acid- and ethylene-dependent signaling pathways and, to a lesser extent, on the salicylate-dependent pathway. Crosses between the Eil-0 and Col-0 ecotypes suggested that the resistance in Eil-0 was dominant and was conferred by a single locus, which we named RCH1. RCH1 is the first resistance locus to be identified from Arabidopsis against the hemibiotrophic fungus genus Colletotrichum.
Inorganic Cations Mediate Plant PR5 Protein Antifungal Activity through Fungal Mnn1- and Mnn4-Regulated Cell Surface Glycans
Authors: Salzman Ron A. , Koiwa Hisashi , Ibeas José Ignacio , Pardo José M. , Hasegawa P. M. , Bressan Ray A.
Published: May 2007

Antimicrobial activities of many defense proteins are profoundly altered by inorganic cations, thereby controlling disease pathologies in a number of mammalian systems, such as cystic fibrosis in humans. Protein-based active defense systems in plants also are influenced by cations; however, little is known of how these cation effects are mediated. Cytotoxicity of the pathogenesis-related protein osmotin against the model fungus Saccharomyces cerevisiae was progressively abolished by K+. By the use of S. cerevisiae mannosylation mutants, this effect was shown to require mannosephosphate residues in the cell wall. However, osmotin activity was not suppressed by even high concentrations of Ca2+. Rather, submillimolar levels of Ca2+ specifically facilitated osmotin's activity, as well as its binding to the cell surface. This effect also was dependent on mannosephosphate groups on the cell surface, and appeared to require negative charge on a portion of the osmotin protein. Results suggest that Ca2+ modulates osmotin action by facilitating its binding to the fungal cell surface, but that K+ blocks this interaction by competing for binding to mannosephosphate groups. Therefore, we have identified glycan interaction as a mechanism for antimicrobial protein activity modulation by cations, a pattern that may apply to diverse innate defense responses.
Basal Defenses Induced in Pepper by Lipopolysaccharides Are Suppressed by Xanthomonas campestris pv. vesicatoria
Authors: Keshavarzi Mansureh , Soylu Soner , Brown Ian , Bonas Ulla , Nicole Michel , Rossiter John , Mansfield John
Published: May 2007

The nonpathogenic hrcC mutant of Xanthomonas campestris pv. vesicatoria 85-10∷hrpA22 multiplied in pepper leaves if it was mixed with pathogenic strains of X. campestris pv. vesicatoria. Reactions to the mutant alone included localized deposition of phenolics and callose in papillae, and alterations to the plant cell wall leading to increased electron density. Electron microscopy showed that the localized responses were suppressed in the presence of wild-type bacteria but other wall changes occurred at some sites, involving cellulose-rich ingrowth of the wall. Multiplication of the hrp mutant in mixed inocula was confirmed by tagging 85-10∷hrpA22 using immunocytochemical location of AvrBs3 expressed from the plasmid pD36. Elicitors of callose deposition and other wall changes were isolated from the hrcC mutant. Activity in extracts of bacteria was attributed to the presence of high molecular weight lipopolysaccharides (LPS). Wild-type X. campestris pv. vesicatoria suppressed induction of structural changes caused by purified LPS. Results obtained suggest that effector proteins produced by phytopathogenic bacteria and delivered by the type III secretion system may have a key role in suppressing the basal defense responses activated by bacterial LPS, which lead to restricted multiplication of nonpathogens such as hrp mutants.
Aberrant mRNA Processing of the Maize Rp1-D Rust Resistance Gene in Wheat and Barley
Authors: Ayliffe Michael A. , Steinau Martin , Park Robert F. , Rooke Lee , Pacheco Maria G. , Hulbert Scot H. , Trick Harold N. , Pryor Anthony J.
Published: May 2007

The maize Rp1-D gene confers race-specific resistance against Puccinia sorghi (common leaf rust) isolates containing a corresponding avrRp1-D avirulence gene. An Rp1-D genomic clone and a similar Rp1-D transgene regulated by the maize ubiquitin promoter were transformed independently into susceptible maize lines and shown to confer Rp1-D resistance, demonstrating that this resistance can be transferred as a single gene. Transfer of these functional transgenes into wheat and barley did not result in novel resistances when these plants were challenged with isolates of wheat stem rust (P. graminis), wheat leaf rust (P. triticina), or barley leaf rust (P. hordei). Regardless of the promoter employed, low levels of gene expression were observed. When constitutive promoters were used for transgene expression, a majority of Rp1-D transcripts were truncated in the nucleotide binding site-encoding region by premature polyadenylation. This aberrant mRNA processing was unrelated to gene function because an inactive version of the gene also generated such transcripts. These data demonstrate that resistance gene transfer between species may not be limited only by divergence of signaling effector molecules and pathogen avirulence ligands, but potentially also by more fundamental gene expression and transcript processing limitations.
The Transcriptome of Rhizobacteria-Induced Systemic Resistance in Arabidopsis
Authors: Verhagen Bas W. M. , Glazebrook Jane , Zhu Tong , Chang Hur-Song , van Loon L. C. , Pieterse Corné M. J.
Published: May 2007

Plants develop an enhanced defensive capacity against a broad spectrum of plant pathogens after colonization of the roots by selected strains of nonpathogenic, fluorescent Pseudomonas spp. In Arabidopsis thaliana, this rhizobacteria-induced systemic resistance (ISR) functions independently of salicylic acid but requires responsiveness to the plant hormones jasmonic acid and ethylene. In contrast to pathogen-induced systemic acquired resistance, rhizobacteria-mediated ISR is not associated with changes in the expression of genes encoding pathogenesis-related proteins. To identify ISR-related genes, we surveyed the transcriptional response of over 8,000 Arabidopsis genes during rhizobacteria-mediated ISR. Locally in the roots, ISR-inducing Pseudomonas fluorescens WCS417r bacteria elicited a substantial change in the expression of 97 genes. However, systemically in the leaves, none of the approximately 8,000 genes tested showed a consistent change in expression in response to effective colonization of the roots by WCS417r, indicating that the onset of ISR in the leaves is not associated with detectable changes in gene expression. After challenge inoculation of WCS417r-induced plants with the bacterial leaf pathogen P. syringae pv. tomato DC3000, 81 genes showed an augmented expression pattern in ISR-expressing leaves, suggesting that these genes were primed to respond faster or more strongly upon pathogen attack. The majority of the primed genes was predicted to be regulated by jasmonic acid or ethylene signaling. Priming of pathogen-induced genes allows the plant to react more effectively to the invader encountered, which might explain the broad-spectrum action of rhizobacteria-mediated ISR.
Cytological, Genetic, and Molecular Analysis to Characterize Compatible and Incompatible Interactions Between Medicago truncatula and Colletotrichum trifolii
Authors: Torregrosa Carine , Cluzet Stéphanie , Fournier Joëlle , Huguet Thierry , Gamas Pascal , Prospéri Jean-Marie , Esquerré-Tugayé Marie-Thérèse , Dumas Bernard , Jacquet Christophe
Published: May 2007

In this study, a new pathosystem was established using the model plant Medicago truncatula and Colletotrichum trifolii, the causal agent of anthracnose on Medicago sativa. Screening of a few M. truncatula lines identified Jemalong and F83005.5 as resistant and susceptible to Colletotrichum trifolii race 1, respectively. Symptom analysis and cytological studies indicated that resistance of Jemalong was associated with a hypersensitive response of the plant. The two selected lines were crossed, and inoculations with C. trifolii were performed on the resulting F1 and F2 progenies. Examination of the disease phenotypes indicated that resistance was dominant and was probably due to a major resistance gene. Molecular components of the resistance were analyzed through macroarray experiments. Expression profiling of 126 expressed sequence tags corresponding to 92 genes, which were selected for their putative functions in plant defense or signal transduction, were compared in Jemalong and F83005.5 lines. A strong correlation was observed between the number of up-regulated genes and the resistance phenotype. Large differences appeared at 48 h postinoculation; more than 40% of the tested genes were up-regulated in the Jemalong line compared with only 10% in the susceptible line. Interestingly, some nodulin genes were also induced in the resistant line upon inoculation with C. trifolii.
Proteomic Analysis of Resistance Mediated by Rcm 2.0 and Rcm 5.1, Two Loci Controlling Resistance to Bacterial Canker of Tomato
Authors: Coaker Gitta L. , Willard Belinda , Kinter Michael , Stockinger Eric J. , Francis David M.
Published: May 2007

Two quantitative trait loci from Lycopersicon hirsutum, Rcm 2.0 and Rcm 5.1, control resistance to Clavibacter michiganensis subsp. michiganensis, the causal agent of bacterial canker of tomato. Lines containing Rcm 2.0 and Rcm 5.1 and a susceptible control line were compared at 72 and 144 h postinoculation, using 2-dimensional gel electrophoresis to identify proteins regulated in response to C. michiganensis subsp. michiganensis infection. A total of 47 proteins were subjected to tandem mass spectrometry. Database queries with resulting spectra identified tomato genes for 26 proteins. The remaining 21 proteins were either identified in other species or possessed no homology to known proteins. Spectra were interpreted to deduce peptide amino acid sequences that were then used to query publicly available data. This approach identified tomato genes or expressed sequence tags for 44 of the proteins analyzed. Three superoxide dismutase (SOD) enzymes were differentially regulated among genotypes, and patterns of hydrogen peroxide accumulation were genotype- and tissue-specific, indicating a role for oxidative stress in response to C. michiganensis subsp. michiganensis. Steady-state mRNA and protein levels for SOD, thioredoxin M-type, S-adenosylhomocysteine hydrolase, and pathogenesis-related proteins demonstrated similar patterns of differential regulation. Lines containing Rcm 2.0 and Rcm 5.1 accumulate different proteins and steady-state mRNAs in response to inoculation, suggesting that the two loci may confer resistance through distinct mechanisms.
Rice cDNA Microarray-Based Gene Expression Profiling of the Response to Flagellin Perception in Cultured Rice Cells
Authors: Fujiwara Satsuki , Tanaka Noriko , Kaneda Takashi , Takayama Seiji , Isogai Akira , Che Fang-Sik
Published: May 2007

Incompatible strains of Acidovorax avenae elicit an immune response in cultured rice cells, with immunity specifically induced by the flagellin of the incompatible strain. To identify genes regulated by flagellin perception signaling in cultured rice cells, gene expression patterns were analyzed with rice cDNA microarrays, including 3,353 independent rice cDNA clones. In all, 131 genes were differentially expressed between incompatible and compatible interactions. K-means clustering showed that 94 genes were upregulated and 32 genes were downregulated during incompatible interactions, whereas only 5 genes were upregulated during compatible interactions. Among the 126 genes that were up- or downregulated during incompatible interactions, expression of 46 genes was decreased when cultured rice cells were inoculated with a flagellin-deficient incompatible strain (Δfla1141-2), indicating that approximately 37% of the 126 genes were directly controlled by flagellin perception. Real-time reverse-transcription polymerase chain reaction analysis using flagellins purified from incompatible or compatible strains was performed to confirm flagellin-regulated expression of candidate genes selected by microarray analysis. Results showed that induction of some genes involved in the immune response is regulated not only by the flagellin perception pathway, but also by another recognition molecule-perception pathway.
Verticillium Wilt in Spinach Seed Production
Authors: du Toit Lindsey J. , Derie Mike L. , Hernandez-Perez Pablo
Published: May 2007

There are no previous reports of Verticillium wilt in fresh and processing spinach (Spinacia oleracea) crops in the United States. In 2002, a hybrid spinach seed crop in the Pacific Northwest developed late-season wilt symptoms. Assays of the harvested seed and stock seed of the male and female parents revealed 59.5, 44.0, and 1.5%, respectively, were infected with Verticillium dahliae. Assays of 13 stock or commercial seed lots grown in 2002 and 62 commercial lots harvested in 2003 in Denmark, Holland, New Zealand, and the United States revealed the prevalence of Verticillium spp. in commercial spinach seed. Sixty-eight lots (89%) were infected with Verticillium spp. at incidences ranging from 0.3 to 84.8%. Five spinach seed isolates of V. dahliae were pathogenic on each of three spinach cultivars by root-dip inoculation. V. dahliae was detected on 26.4% of the seed from 7 of 11 inoculated plants but on none of the seed from 6 control plants, demonstrating systemic movement of V. dahliae. Seed-to-seed transmission was also demonstrated by planting naturally infected seed lots. This is the first report of Verticillium wilt of spinach in the primary region of spinach seed production in the United States.
Infection Structures of Host-Specialized Isolates of Uromyces viciae-fabae and of Other Species of Uromyces Infecting Leguminous Crops
Authors: Emeran A. A. , Sillero J. C. , Niks R. E. , Rubiales D.
Published: May 2007

A study was made of the morphology of urediniospores and primary infection structures of 12 isolates of six legume-infecting species of Uromyces. Infection structures were sufficient to distinguish among species. Isolates of Uromyces viciae-fabae proved to be specialized with respect to host, because each isolate infected only cultivars of the species from which it was collected. Host-specialized isolates of U. viciae-fabae also were morphologically distinct, differing in both spore dimensions and infection structure morphology. In particular, the shape and dimensions of the substomatal vesicle were distinctive. These results support the view that U. viciae-fabae sensu lato is a species complex.
Susceptibility of Oregon Forest Trees and Shrubs to Phytophthora ramorum: A Comparison of Artificial Inoculation and Natural Infection
Authors: Hansen E. M. , Parke J. L. , Sutton W.
Published: May 2007

Phytophthora ramorum is an invasive pathogen in some mixed-hardwood forests in California and southwestern Oregon, where it causes sudden oak death (SOD) on some members of Fagaceae, ramorum shoot dieback on some members of Ericaceae and conifers, and ramorum leaf blight on diverse hosts. We compared symptoms of P. ramorum infection resulting from four different artificial inoculation techniques with the symptoms of natural infection on 49 western forest trees and shrubs; 80% proved susceptible to one degree or another. No single inoculation method predicted the full range of symptoms observed in the field, but whole plant dip came closest. Detached-leaf-dip inoculation provided a rapid assay and permitted a reasonable assessment of susceptibility to leaf blight. Both leaf age and inoculum dose affected detached-leaf assays. SOD and dieback hosts often developed limited leaf symptoms, although the pattern of midrib and petiole necrosis was distinctive. Stem-wound inoculation of seedlings correlated with field symptoms for several hosts. The results suggested that additional conifer species may be damaged in the field. Log inoculation provided a realistic test of susceptibility to SOD, but was cumbersome and subject to seasonal variability. Pacific rhododendron, salmonberry, cascara, and poison oak were confirmed as hosts by completing Koch's postulates. Douglas-fir was most susceptible to shoot dieback shortly after budburst, with infection occurring at the bud.
First Report of Citrus Blight in Costa Rica
Authors: Villalobos W. , Moreira L. , Derrick K. S. , Beretta M. J. G. , Rivera C.
Published: May 2007

Citrus blight (CB), causing a chronic decline of citrus, has been an important disease in Florida for over 100 years. CB was first reported in Brazil in the 1980s and is now responsible for the removal of nearly 10% of the trees from production annually. No causal agent has been identified, but CB has been root-graft transmitted to healthy trees, suggesting that the causal agent is infectious (3). Since 1997, CB symptoms were observed in several groves in northern Costa Rica, the most important citrus area of approximately 25,000 ha. Symptoms observed include a general decline and wilt of the tree canopy, off-color leaves, leaf drop, twig dieback, small fruit, delayed blossom, poor growth, and death. A survey near Guanacaste revealed CB symptoms in 7-yr-old Valencia and Pineapple orange trees (Citrus sinensis (L.) Osbeck) grafted on Carrizo citrange (C. sinensis (L.) Osbeck × Poncirus trifoliata (L.) Raf.) rootstock. Since 1997, 6% of the trees in this area have been replanted annually because of CB symptoms. Similar situations were observed in other groves in the northern citrus area. Dot immunobinding assays (DIBA) (1) were used to detect the P12 protein associated with CB with 20 of 22 trees showing CB-like symptoms giving a positive test. Zinc (Zn) accumulation in trunk wood and water uptake tests were done according to Roistacher (2) in 8 healthy and 20 symptomatic trees which were positive for CB using DIBA. The average Zn concentration of 16 declining trees was 4.6 ± 1.9, whereas the average concentration for 8 healthy trees was 2.0 ± 0.9. The average water uptake in 1 min was 14 ml for healthy trees, and virtually zero for the 20 symptomatic trees. These diagnostic tests confirm the presence of CB in the northern citrus area of Costa Rica, and the surveys indicate the disease is beginning to spread and become economically important. To our knowledge, this is the first report of CB in commercial citrus in Costa Rica.

References: (1) K. S. Derrick et al. Plant Dis. 74:168, 1990. (2) C. N. Roistacher. Pages 57–66 in: Graft-Transmissible Diseases of Citrus. Handbook for Detection and Diagnosis. C.N. Roistacher, ed. Food and Agriculture Organization, Rome, 1991. (3) D. P. H. Tucker et al. Plant Dis. 68:979, 1984.
Alternaria yaliinficiens sp. nov. on Ya Li Pear Fruit: From Interception to Identification
Authors: Roberts Rodney G.
Published: May 2007

In 2001, numerous Ya Li pear fruit from China with a disease apparently caused by an Alternaria sp. were intercepted at U.S. ports, raising concern because the disease was unknown in the United States. To identify the etiologic agent of the disease, single-spore cultures were established from intercepted fruit and were characterized by pathogenicity to wounded fruit, morphology, and sporulation pattern on three agar media, random amplified polymorphic DNA fragment pattern analysis, and species-specific polymerase chain reaction (PCR). These data indicated that the disease likely was caused by more than one species of Alternaria, and that none of the isolates from intercepted Ya Li fruit was A. alternata or A. gaisen. A species-specific PCR assay for A. gaisen demonstrated that no isolate from Ya Li was A. gaisen. A PCR assay for the AMT gene demonstrated that no isolate from Ya Li was A. ‘mali’. Because of these data and the absence of a description to accommodate at least one of the taxa involved, A. yaliinficiens R. G. Roberts sp. nov. is described, and the disease caused by this and other species of Alternaria on Ya Li pear from China is referred to as “chocolate spot of Ya Li pear” to distinguish it from black spot of Japanese pear.
Characterization of Phytophthora capsici Isolates from Processing Pumpkin in Illinois
Authors: Islam S. Z. , Babadoost M. , Lambert K. N. , Ndeme A. , Fouly H. M.
Published: May 2007

This study was conducted to investigate pathogenic, morphologic, and genetic variations among Phytophthora capsici isolates from processing pumpkin (Cucurbita moschata) fields in Illinois. Random amplified polymorphic DNA (RAPD) markers were employed to assess genetic variation among 24 isolates of P. capsici from 10 individual fields at six locations. Unweighted mean pair group analysis clustered isolates into six groups. The genetic distances ranged from 0.03 to 0.45. Inoculation of pumpkin seedlings in the greenhouse revealed that the isolates belonged to six distinct genetic groups differing significantly (P = 0.05) in virulence. Isolates tested exhibited four growth patterns in culture: cottony, rosaceous, petaloid, and stellate. P. capsici isolates, including an ATCC isolate (ATCC-15427), with cottony growth pattern did not grow at 36°C. The mean oospore diameter of A1 mating type isolates was greater than that of A2 mating type isolates. Nine of 24 isolates tested produced chlamydospores in V8-CaCO3 liquid medium.
Use of a Rainfall Frequency Threshold to Adjust a Degree-Day Model of Ascospore Maturity of Venturia inaequalis
Authors: Stensvand Arne , Eikemo Håvard , Gadoury David M. , Seem Robert C.
Published: May 2007

Estimates of ascospore maturity generated by a model developed previously in New Hampshire, United States, were compared with the cumulative release of ascospores in southern Norway as monitored by volumetric spore traps at one site for 12 years, and at two additional sites for 2 years. In locations and years with frequent rain events, model-estimated ascospore maturity closely approximated observed ascospore release. However, in years with protracted dry periods of 1 to 3 weeks with no or little rain, not only was spore release delayed, but release continued to lag behind predicted maturity even after several rain events subsequent to the dry interval. By halting degree-day (base = 0°C) accumulation if 7 consecutive days without rain occurred, accuracy of the model during “dry” years was greatly improved, without substantially affecting accuracy in “wet” seasons. With minimal additional effort on the part of the user, this simple modification increases the accuracy of model-derived estimates of ascospore maturity when lack of rain slows ascospore maturation.
First Report of Southern blight Incited by Sclerotium rolfsii on Dichondra repens in Italy
Authors: Garibaldi A. , Minuto A. , Gullino M. L.
Published: May 2007

Kidney weed (Dichondra repens) is increasingly used for low maintenance turf in Italy, particularly for gardens and parks in areas characterized by mild climate. During September 2003, on the D. repens turf of a private garden located near Imperia (northern Italy), yellow, circular areas as much as 60 cm in diameter appeared with the grass becoming chlorotic and thin. A ring of the patch at its periphery exhibited a reddish brown color and eventually died. An area of green grass remained in the center of the patch. Rings of dead grass enlarged rapidly during hot, humid weather. In the presence of abundant moisture, a white mycelium occurred on the dying grass at the periphery of the ring. White or light-to-dark brown sclerotia (1 to 3 mm in diameter) developed from mycelium on the dead grass. The diseased tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar amended with 100 mg/l of streptomycin sulphate. Sclerotium rolfsii was consistently isolated from infected plants. Pathogenicity of three isolates obtained from infected plants was confirmed by inoculating healthy D. repens turf (1 m2 plots and three replicates). Inoculum that consisted of wheat kernels infested with mycelium and sclerotia of each isolate was placed on the soil surface. Noninoculated plots served as controls. The inoculation trial was repeated once. Turf was covered with a plastic film for 7 days, kept at temperatures ranging between 22 and 25°C, and watered as needed. Inoculated plants developed symptoms of leaf yellowing within 11 days, soon followed by the appearance of white mycelium and sclerotia, and then eventually wilted. Control plants remained symptomless. Sclerotium rolfsii was re-isolated from inoculated plants. To our knowledge, this is the first report of S. rolfsii on D. repens in Italy. This disease has been reported on kidney weed in several countries such as the United States (3), Brazil (1), and India (2).

References: (1) M. Menezes and J. A. A. Lima. Fitossanidade 1:18, 1974. (2) K. Ranganathan and N. Shanmugam. Indian Phytopathol, 27:113, 1974. (3) J. D. Smith et al. Fungal Diseases of Amenity Turf Grasses. E & F.N. Spon, London, 1989.
Root Rot of Black Spruce Caused by Cylindrocladium canadense in Eastern North America
Authors: Vujanovic V. , St-Arnaud M.
Published: May 2007

During October 2002, symptoms of root rot of black spruce, Picea mariana (Mill.) B.S.P., were observed in the St-Modeste (47°46′N, 69°36′W) conifer nursery (400 km northeast of Montreal, Quebec, Canada). Disease severity was low in the greenhouse-produced mother plants and 1-year-old seedlings and moderate in field-grown 2- and 3-year-old seedlings. A species of Cylindrocladium was isolated on potato dextrose agar from 12 symptomatic seedlings from the greenhouse and 12 from the field. The isolates produced chestnut-colored colonies and chlamydospores, both of which were typical of C. canadense Kang, Crous & Schoch (2). DNA was extracted from representative isolates (MTF 101, MTF 102), and the internal transcribed spacer (ITS) of the rDNA gene was amplified and sequenced (GenBank Accession Nos. AY705980 and AY705981). There was a 99% match with a sequence of C. canadense (GenBank Accession No. AF348256). However, there was approximately 10% divergence with the ITS sequence of C. floridanum (GenBank Accession No AF307343). MTF101 and MTF102 were pathogenic on black spruce seedlings when fungal suspension (106 CFU/ml) was added to germinating seeds in petri plates or infiltrated into roots of 2-week-old seedlings growing in sterilized, moist, sandy soil in the greenhouse. Within 3 weeks, inoculated seedlings exhibited typical root necrosis, while control seedlings were symptomless. C. canadense was reisolated only from symptomatic seedlings. The occurrence of C. canadense in eastern North America has significant implications for forestry regeneration. Previously, only C. floridanum had been reported as pathogenic in the St-Modeste nursery and in eastern North America(1).

References: (1) R. C. Hamelin et al. Appl. Environ. Microbiol. 62:4026, 1996. (2) J. C. Kang et al. Syst. Appl. Microbiol. 24:206, 2001.
Nicotiana occidentalis: A New Herbaceous Host for Blueberry scorch virus
Authors: Lowery D. T. , French C. J. , Bernardy M.
Published: May 2007

Blueberry scorch virus(BlScV) is a carlavirus that causes a serious disease of blueberries (Vaccinium corymbosum L.) in North America (2). In aphid-transmission studies of BlScV using blueberry as host and test species, we found the rate of transmission to be low, and a lengthy incubation period was required before BlScV could be detected. For sequencing studies, RNA extraction from blueberry using standard methods was unreliable and inefficient. These problems prompted a search for alternate hosts. Of 12 herbaceous hosts screened for BlScV transmission using the blueberry aphid, Ericaphis fimbriata Richards, with mechanical transmission, only Nicotiana occidentalis (Wheeler) became infected. After 3 to 4 weeks, infection of N. occidentalis with BlScV resulted in mild symptoms that included pronounced leaf twisting and swollen leaf veins. Infection with BlScV was confirmed using a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) kit (Agdia Inc., Elkhart, IN), polyclonal antibodies to BlScV from the antiserum collection at the Pacific Agri-Food Research Center, Summerland, British Columbia, Canada, and reverse transcription-polymerase chain reaction (RT-PCR). Forward (5′-NTAAACACTCCCGAATATAC-3′) and reverse (5′-CAGATTGCTTATCCGGCTTC-3′) primers were designed with the published sequence of BlScV isolate NJ-02 (GenBank Accession No. NC003499). An amplicon of the expected size was generated and sequenced. BLAST analysis indicated that the nucleotide sequence of the amplified fragment was 87% identical to the corresponding sequence in NJ-02 (1). N. occidentalis was readily infected with BlScV following aphid or mechanical inoculations from blueberry. With E. fimbriata as the aphid vector, the transmission rate from blueberry to N. occidentalis was approximately 26%, compared with 70% for mechanical inoculations. Mechanical transfer of BlScV between infected N. occidentalisplants resulted in a 100% transmission rate. Recently, with N. occidentalis, we have completely sequenced two strains of BlScV from British Columbia, Canada and identified several aphid vector species. The identification of N. occidentalis as an herbaceous host of BlScV greatly facilitates future studies on the virus.

References: (1) T. D. Cavileer et al. J. Gen. Virol. 75: 711, 1994. (2) R. R. Martin and P. R. Bristow. Phytopathology 78:1636, 1988.
First Report of Soybean Rust Caused by Phakopsora pachyrhizi on Dry Beans in South Africa
Authors: du Preez E. D. , van Rij N. C. , Lawrance K. F. , Miles M. R. , Frederick R. D.
Published: May 2007

During April 2004, a 150-m2 dry bean (Phaseolus vulgaris) plot growing adjacent to rust-infected soybean (Glycine max) at Cedara Agricultural Research Farm (29°32′S 30°16′E) was observed to be infected with two distinct rust types. Common bean rust (caused by Uromyces appendiculatus) with reddish brown uredinia and black telia was readily identified. A second rust with smaller sporulating uredinia (1.0 to 1.5 mm2), which were gray in appearance, was also found. Visual rust severity on the dry bean plants, which were in mid pod-fill, was high (approximately 30 to 40% disease incidence). Twenty plants were examined and observed to be infected with both rusts. With microscopic examination of no fewer than 20 leaves per plant, the urediniospores from the smaller lesions were determined to be morphologically similar to Phakopsora pachyrhizi (3). Real-time fluorescent polymerase chain reaction assays on six leaves and sequence analysis of the nuclear ribosomal internal transcribed spacer region 2 (1) verified the identity of the urediniospores as P. pachyrhizi. Although P. vulgaris is a known host of P. pachyrhizi, to our knowledge this is the first time since the arrival of soybean rust in 2001 that P. pachyrhizi has been observed on an alternate host plant in South Africa (2). Since dry beans are grown all year in frost-free areas, the implications are that dry beans may serve as an important overwintering host and source of inoculum for seasonal soybean rust outbreaks.

References: (1) R. D. Frederick et al. Phytopathology 92:217, 2002. (2) Z. A. Pretorius et al. Plant Dis. 85:1288, 2001. (3) J. B. Sinclair and G. L. Hartman. Soybean Rust. Pages 25–26 in: Compendium of Soybean Diseases, 4th ed. G. L. Hartman et al. eds. The American Phytopathological Society, St. Paul, MN, 1999.