bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Fusicoccin on the Early Infection Process of Legume Roots by Rhizobium spp
Authors: Donaire Juan Pedro , Rodríguez-Rosales María Pilar , Soto María José , Sanjuan Juan , Olivares José
Published: May 2007

Nod factors, the first detectable signals produced by Rhizobium spp., were reported to induce cytosolic pH changes and plasma membrane depolarization in root hairs as specific responses. In this study, it has been found that fusicoccin inhibits nodulation of alfalfa roots. This inhibition was only observed when fusicoccin was applied in the earlier steps of the bacteria-plant interaction. The observed effect is similar to that caused by the undissociated permeant acetic acid, which decreases the cytoplasmic pH and, like fusicoccin, significantly stimulates net H+ efflux. These results suggest that the fusicoccin-induced plasma membrane H+-ATPase activity and membrane hyperpolarization could be responsible for the nodulation inhibition observed. Moreover, it was found that nodulation was inhibited by removing external calcium with EGTA. When fusicoccin is present, a lower concentration of EGTA is necessary to inhibit nodulation. Furthermore, the addition of Ca2+ ionophore A23187 was found to inhibit H+ efflux by roots. These observations support the idea that the Nod factor-triggered calcium signal modulates the activity of the proton pump.
Identification of Pathogenicity Mutants of the Rice Blast Fungus Magnaporthe grisea by Insertional Mutagenesis
Authors: Balhadère Pascale V. , Foster Andrew J. , Talbot Nicholas J.
Published: May 2007

Restriction enzyme-mediated DNA integration (REMI) mutagenesis was used to identify mutants of Magnaporthe grisea impaired in pathogenicity. Three REMI protocols were evaluated and the frequency of REMIs determined. An REMI library of 3,527 M. grisea transformants was generated in three genetic backgrounds, and 1,150 transformants were screened for defects in pathogenicity with a barley cut leaf assay. Five mutants were identified and characterized. Two mutants (2029 and 2050) were impaired in appressorium function. Two other mutants, 125 and 130, were altered in conidial morphology, conidiogenesis, and appressorium function. Mutant 130 was also a methionine auxotroph and methionine auxotrophy co-segregated with the reduction in pathogenicity. An additional mutant, 80, showed reduced pathogenicity on blast-susceptible rice cultivars but was fully pathogenic on barley. The reduction of pathogenicity in mutant 80 was associated with a delay in conidial germination and appressorium development. Genetic analysis suggested single-gene segregation for each mutant, but only two of the mutations co-segregated with the hygromycin resistance marker. The genetic loci in mutants 2029, 2050, 125, 130, and 80 were termed PDE1, PDE2, IGD1, MET1, and GDE1, respectively. pde1 and pde2 were non-allelic to cpkA, a mutation in the catalytic subunit of cyclic AMP (cAMP)-dependent protein kinase A with a very similar phenotype. The results indicate the utility of REMI for studying fungal pathogenicity, but also highlight the requirement for rigorous genetic and phenotypic analysis.
Isolation of theLEMMI9Gene and Promoter Analysis During a Compatible Plant-Nematode Interaction
Authors: Escobar Carolina , De Meutter Jan , Aristizábal Fabio A. , Sanz-Alférez Soledad , del Campo Francisca F. , Barthels Nathalie , Van der Eycken Walter , Seurinck Jef , van Montagu Marc , Gheysen Godelieve , Fenoll Carmen
Published: May 2007

Plant-endoparasitic root-knot nematodes feed on specialized giant cells that they induce in the vascular cylinder of susceptible plants. Although it has been established that a number of plant genes change their expression pattern during giant cell differentiation, virtually no data are available about the mechanisms involved in that change. One possibility is differential promoter recognition by the transcription factor(s) responsible for the expression of specific genes. We have isolated and characterized a genomic clone from tomato containing the promoter region of LEMMI9, one of the few plant genes that have been reported to be highly expressed in galls (predominantly in giant cells). The analysis of transgenic potato plants carrying a LEMMI9 promoter-β glucuronidase (GUS) fusion has demonstrated that the tomato promoter was activated in Meloidogyne incognita-induced galls in a heterologous system. We have located putative regulatory sequences in the promoter and have found that nuclear proteins from the galls formed specific DNA-protein complexes with the proximal region of the LEMMI9 promoter. The nuclear protein-binding sequence mapped to a region of 111 bp immediately upstream from the TATA box. This region contains a 12-bp repeat possibly involved in the formation of DNA-protein complexes, which might be related to the LEMMI9 transcriptional activation in the giant cells.
An hrcU-Homologous Gene Mutant of Xanthomonas campestris pv. glycines 8ra That Lost Pathogenicity on the Host Plant but Was Able to Elicit the Hypersensitive Response on Nonhosts
Authors: Oh Changsik , Heu Sunggi , Yoo Jae-Yong , Cho Yongsup
Published: May 2007

Transposon mutagenesis was used to isolate nonpathogenic mutants of Xanthomonas campestris pv. glycines 8ra, which causes bacterial pustule disease in soybean. A 6.1-kb DNA region in which a mutation gave loss of pathogenicity was isolated and found to carry six open reading frames (ORFs). Four ORFs had homology with hrcU, hrcV, hrcR, and hrcS genes of Ralstonia solanacearum and X. campestris pv. vesicatoria. One nonpathogenic mutant, X. campestris pv. glycines H80, lost pathogenicity on soybean but was able to elicit the hypersensitive response (HR) on nonhost pepper and tomato plants. This mutant still multiplied as well as the wild type in the leaves or cotyledons of soybean. Although the DNA and amino acid sequences showed high homology with known hrp genes, the hrcU-homolog ORF is not required for HR induction on nonhost plants, pepper and tomato, or for the multiplication of bacteria in the host plant. This gene was only required for the pathogenic symptoms of X. campestris pv. glycines 8ra on soybean.
Carbon Metabolism in Developing Soybean Root Nodules: The Role of Carbonic Anhydrase
Authors: Kavroulakis Nektarios , Flemetakis Emanouil , Aivalakis Georgios , Katinakis Panagiotis
Published: May 2007

A full-length cDNA clone encoding carbonic anhydrase (CA) was isolated from a soybean nodule cDNA library. In situ hybridization and immunolocalization were performed in order to assess the location of CA transcripts and protein in developing soybean nodules. CA transcripts and protein were present at high levels in all cell types of young nodules, whereas in mature nodules they were absent from the central tissue and were concentrated in cortical cells. The results suggested that, in the earlier stages of nodule development, CA might facilitate the recycling of CO2 while at later stages it may facilitate the diffusion of CO2 out of the nodule system. In parallel, sucrose metabolism was investigated by examination of the temporal and spatial transcript accumulation of sucrose synthase (SS) and phosphoenolpyruvate carboxylase (PEPC) genes, with in situ hybridization. In young nodules, high levels of SS gene transcripts were found in the central tissue as well as in the parenchymateous cells and the vascular bundles, while in mature nodules the levels of SS gene transcripts were much lower, with the majority of the transcripts located in the parenchyma and the pericycle cells of the vascular bundles. High levels of expression of PEPC gene transcripts were found in mature nodules, in almost all cell types, while in young nodules lower levels of transcripts were detected, with the majority of them located in parenchymateous cells as well as in the vascular bundles. These data suggest that breakdown of sucrose may take place in different sites during nodule development.
Bean dwarf mosaic virus BV1 Protein Is a Determinant of the Hypersensitive Response and Avirulence in Phaseolus vulgaris
Authors: Garrido-Ramirez E. R. , Sudarshana M. R. , Lucas W. J. , Gilbertson R. L.
Published: May 2007

The capacities of the begomoviruses Bean dwarf mosaic virus (BDMV) and Bean golden yellow mosaic virus (BGYMV) to differentially infect certain common bean (Phaseolus vulgaris) cultivars were used to identify viral determinants of the hypersensitive response (HR) and avirulence (avr) in BDMV. A series of hybrid DNA-B components, containing BDMV and BGYMV sequences, was constructed and coinoculated with BDMV DNA-A (BDMV-A) or BDMVA-green florescent protein into seedlings of cv. Topcrop (susceptible to BDMV and BGYMV) and the BDMV-resistant cvs. Othello and Black Turtle Soup T-39 (BTS). The BDMV avr determinant, in bean hypocotyl tissue, was mapped to the BDMV BV1 open reading frame and, most likely, to the BV1 protein. The BV1 also was identified as the determinant of the HR in Othello. However, the HR was not required for resistance in Othello nor was it associated with BDMV resistance in BTS. BDMV BV1, a nuclear shuttle protein that mediates viral DNA export from the nucleus, represents a new class of viral avr determinant. These results are discussed in terms of the relationship between the HR and resistance.
Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-BreakingLettuce mosaic virusIsolate in Susceptible and Resistant Lettuce Cultivars
Authors: German-Retana Sylvie , Candresse Thierry , Alias Emmanuel , Delbos René-Pierre , Le Gall Olivier
Published: May 2007

The RNA genome of a resistance-breaking isolate of Lettuce mosaic virus (LMV-E) was engineered to express the jellyfish green fluorescent protein (GFP) or β-glucuronidase (GUS) fused to the helper-component proteinase (HC-Pro) to study LMV invasion and spread in susceptible and resistant lettuce cultivars. Virus accumulation and movement were monitored by either histochemical GUS assays or detection of GFP fluorescence under UV light. The GFP- and GUS-tagged viruses spread systemically in the susceptible lettuce cultivars Trocadero and Vanguard, where they induced attenuated symptoms, compared with the wild-type virus. Accumulation of the GFP-tagged virus was reduced but less affected than in the case of the GUS-tagged virus. Systemic movement of both recombinant viruses was very severely affected in Vanguard 75, a lettuce cultivar nearly isogenic to Vanguard but carrying the resistance gene mo12. Accumulation of the recombinant viruses in systemically infected leaves was either undetectable (GUS-tag) or erratic, strongly delayed, and inhibited by as much as 90% (GFP-tag). As a consequence, and contrary to the parental virus, the recombinant viruses were not able to overcome the protection afforded by the mo12gene. Taken together, these results indicate that GUS or GFP tagging of the HC-Pro of LMV has significant negative effects on the biology of the virus, abolishing its resistance-breaking properties and reducing its pathogenicity in susceptible cultivars.
Mesorhizobium loti Increases Root-Specific Expression of a Calcium-Binding Protein Homologue Identified by Promoter Tagging in Lotus japonicus
Authors: Webb K. Judith , Skøt Leif , Nicholson Margaret N. , Jørgensen Bodil , Mizen Sue
Published: May 2007

A promoter tagging program in the legume Lotus japonicus was initiated to identify plant genes involved in the nitrogen-fixing symbiosis between legumes and rhizobia. Seven transformed plant lines expressing the promoterless reporter gene uidA (β-glucuronidase; GUS) specifically in roots and/or nodules were identified. Four of these expressed GUS in the roots only after inoculation with nodule-forming Mesorhizobium loti. In one line (T90), GUS activity was found in the root epidermis, including root hairs. During seedling growth, GUS expression gradually became focused in developing nodules and disappeared from root tissue. No GUS activity was detected when a non-nodulating mutant of M. loti was used to inoculate the plants. The T-DNA insertion in this plant line was located 1.3 kb upstream of a putative coding sequence with strong homology to calcium-binding proteins. Four motifs were identified, which were very similar to the “EF hands” in calmodulin-related proteins, each binding one Ca2+. We have named the gene LjCbp1 (calcium-binding protein). Northern (RNA) analyses showed that this gene is expressed specifically in roots of L. japonicus. Expression was reduced in roots inoculated with non-nodulating M. loti mutants and in progeny homozygous for the T-DNA insertion, suggesting a link between the T-DNA insertion and this gene.
Determination of the T-DNA Transfer and the T-DNA Integration Frequencies upon Cocultivation of Arabidopsis thaliana Root Explants
Authors: De Buck Sylvie , De Wilde Chris , Van Montagu Marc , Depicker Ann
Published: May 2007

Using the Cre/lox recombination system, we analyzed the extent to which T-DNA transfer to the plant cell and T-DNA integration into the plant genome determine the transformation and cotransformation frequencies of Arabidopsis root cells. Without selection for transformation competence, the stable transformation frequency of shoots obtained after cocultivation and regeneration on nonselective medium is below 0.5%. T-DNA transfer and expression occur in 5% of the shoots, indicating that the T-DNA integrates in less than 10% of the transiently expressing plant cells. A limited fraction of root cells, predominantly located at the wounded sites and in the pericycle, are competent for interaction with agrobacteria and the uptake of a T-DNA, as demonstrated by histochemical GUS staining. When selection for transformation competence is applied, the picture is completely different. Then, approximately 50% of the transformants show transient expression of a second, nonselected T-DNA and almost 50% of these cotransferred T-DNAs are integrated into the plant genome. Our results indicate that both T-DNA transfer and T-DNA integration limit the transformation and cotransformation frequencies and that plant cell competence for transformation is based on these two factors.
Rhizobium etli CE3 Carries vir Gene Homologs on a Self-Transmissible Plasmid
Authors: Bittinger M. A. , Gross J. A. , Widom J. , Clardy J. , Handelsman J.
Published: May 2007

RosR is a transcriptional regulator important for determining cell-surface characteristics and nodulation competitiveness in Rhizobium etli CE3. We identified a 15-kb region that contains genes with similarity to members of the virB, virC, virG, and virE operons of Agrobacterium tumefaciens and demonstrated that RosR directly regulates one operon in this region. These genes were located on plasmid pa of R. etli CE3, which is self-transmissible between R. etli and A. tumefaciens.
Apple Contains Receptor-like Genes Homologous to the Cladosporium fulvum Resistance Gene Family of Tomato with a Cluster of Genes Cosegregating with Vf Apple Scab Resistance
Authors: Vinatzer Boris A. , Patocchi Andrea , Gianfranceschi Luca , Tartarini Stefano , Zhang Hong-Bin , Gessler Cesare , Sansavini Silviero
Published: May 2007

Scab caused by the fungal pathogen Venturia inaequalis is the most common disease of cultivated apple (Malus × domestica Borkh.). Monogenic resistance against scab is found in some small-fruited wild Malus species and has been used in apple breeding for scab resistance. Vf resistance of Malus floribunda 821 is the most widely used scab resistance source. Because breeding a high-quality cultivar in perennial fruit trees takes dozens of years, cloning disease resistance genes and using them in the transformation of high-quality apple varieties would be advantageous. We report the identification of a cluster of receptor-like genes with homology to the Cladosporium fulvum (Cf) resistance gene family of tomato on bacterial artificial chromosome clones derived from the Vf scab resistance locus. Three members of the cluster were sequenced completely. Similar to the Cf gene family of tomato, the deduced amino acid sequences coded by these genes contain an extracellular leucine-rich repeat domain and a transmembrane domain. The transcription of three members of the cluster was determined by reverse transcription-polymerase chain reaction to be constitutive, and the transcription and translation start of one member was verified by 5′ rapid amplification of cDNA ends. We discuss the parallels between Cf resistance of tomato and Vf resistance of apple and the possibility that one of the members of the gene cluster is the Vf gene. Cf homologs from other regions of the apple genome also were identified and are likely to present other scab resistance genes.
Rsv1-Mediated Resistance Against Soybean mosaic virus-N Is Hypersensitive Response-Independent at Inoculation Site, but Has the Potential to Initiate a Hypersensitive Response-like Mechanism
Authors: Hajimorad M. R. , Hill J. H.
Published: May 2007

Rsv1, a single dominant gene in soybean PI 96983, confers resistance to most strains of Soybean mosaic virus (SMV), including strain G2. The phenotypic response includes the lack of symptoms and virus recovery from mechanically inoculated leaves. To study the resistance mechanism, SMV-N (an isolate of strain G2) was introduced into PI 96983 by grafting. Hypersensitive response (HR)-like lesions occurred on the stems, petioles, and leaf veins, and virus was recovered from these lesions. The response demonstrated the cytological and histological characteristics of HR as well as elevated transcription of a soybean salicylic acid-inducible, pathogenesis-related (PR-1) protein gene. Mechanical inoculation of PI 96983 primary leaves with a high level of SMV-N virions caused no symptoms or up regulation of the PR-1 protein gene transcript. Furthermore, inoculation with infectious viral RNA did not alter the resistance phenotype. The data suggest that interaction of SMV-N with Rsv1 has the potential to induce an HR-like defense reaction. Rsv1-mediated resistance in the inoculated leaf, however, is HR-independent and operates after virion disassembly.
No Evidence for Binding Between Resistance Gene Product Cf-9 of Tomato and Avirulence Gene Product AVR9 of Cladosporium fulvum
Authors: Luderer Rianne , Rivas Susana , Nürnberger Thorsten , Mattei Benedetta , Van den Hooven Henno W. , Van der Hoorn Renier A. L. , Romeis Tina , Wehrfritz Josa-M. , Blume Beatrix , Nennstiel Dirk , Zuidema Douwe , Vervoort Jacques , De Lorenzo Giulia , Jones Jonathan D. G. , De Wit Pierre J. G. M. , Joosten Matthieu H. A. J.
Published: May 2007

The gene-for-gene model postulates that for every gene determining resistance in the host plant, there is a corresponding gene conditioning avirulence in the pathogen. On the basis of this relationship, products of resistance (R) genes and matching avirulence (Avr) genes are predicted to interact. Here, we report on binding studies between the R gene product Cf-9 of tomato and the Avr gene product AVR9 of the pathogenic fungus Cladosporium fulvum. Because a high-affinity binding site (HABS) for AVR9 is present in tomato lines, with or without the Cf-9 resistance gene, as well as in other solanaceous plants, the Cf-9 protein was produced in COS and insect cells in order to perform binding studies in the absence of the HABS. Binding studies with radio-labeled AVR9 were performed with Cf-9-producing COS and insect cells and with membrane preparations of such cells. Furthermore, the Cf-9 gene was introduced in tobacco, which is known to be able to produce a functional Cf-9 protein. Binding of AVR9 to Cf-9 protein produced in tobacco was studied employing surface plasmon resonance and surface-enhanced laser desorption and ionization. Specific binding between Cf-9 and AVR9 was not detected with any of the procedures. The implications of this observation are discussed.
Overexpression of the Tomato Asc-1 Gene Mediates High Insensitivity to AAL Toxins and Fumonisin B1 in Tomato Hairy Roots and Confers Resistance to Alternaria alternata f. sp. lycopersici in Nicotiana umbratica Plants
Authors: Brandwagt Bas F. , Kneppers Tarcies J. A. , Nijkamp H. John J. , Hille Jacques
Published: May 2007

The sphinganine-analog mycotoxins (SAMs) fumonisin B1 and AAL toxins are inhibitors of eukaryotic sphinganine N-acyltransferase in vitro. Treatment of eukaryotes with SAMs generally results in an accumulation of sphingoid base precursors and a depletion of complex sphingolipids. The asc,asc genotypes of tomato (Lycopersicon esculentum) and Nicotiana umbratica are sensitive to SAMs and host of the AAL toxin-producing fungus Alternaria alternata f. sp. lycopersici. Codominant insensitivity to SAMs in tomato is mediated by the Asc-1 gene, and sensitivity is associated with a frame-shift mutation present in asc-1. We investigated the function of Asc-1 in mediating insensitivity to SAMs and resistance to the fungus by overexpression of asc-1 and Asc-1. In this study, it is shown that overexpression of these genes did not lead to visual symptoms in tomato hairy roots and N. umbratica plants. Overexpression of asc-1 did not influence the (in)sensitivity to SAMs. Overexpression of Asc-1 in SAM-sensitive hairy roots and N. umbratica plants, however, mediated a high insensitivity to SAMs and resistance to plant infection by Alternaria alternata f. sp. lycopersici.
Pseudomonas viridiflava and P. syringae—Natural Pathogens of Arabidopsis thaliana
Authors: Jakob Katrin , Goss Erica M. , Araki Hitoshi , Van Tam , Kreitman Martin , Bergelson Joy
Published: May 2007

We report the isolation and identification of two natural pathogens of Arabidopsis thaliana, Pseudomonas viridiflava and Pseudomonas syringae, in the midwestern United States. P. viridiflava was found in six of seven surveyed Arabidopsis thaliana populations. We confirmed the presence in the isolates of the critical pathogenicity genes hrpS and hrpL. The pathogenicity of these isolates was verified by estimating in planta bacterial growth rates and by testing for disease symptoms and hypersensitive responses to A. thaliana. Infection of 21 A. thaliana ecotypes with six locally collected P. viridiflava isolates and with one P. syringae isolate showed both compatible (disease) and incompatible (resistance) responses. Significant variation in response to infection was evident among Arabidopsis ecotypes, both in terms of symptom development and in planta bacterial growth. The ability to grow and cause disease symptoms on particular ecotypes also varied for some P. viridiflava isolates. We believe that these pathogens will provide a powerful system for exploring coevolution in natural plant-pathogen interactions.
Patterns of Pectin Methylesterase Transcripts in Developing Stem Nodules of Sesbania rostrata
Authors: Lievens Sam , Goormachtig Sofie , Herman Sylvia , Holsters Marcelle
Published: May 2007

Differential display was applied to the early stages of the interaction between the tropical legume Sesbania rostrata and its microsymbiont Azorhizobium caulinodans ORS571. An upregulated clone that is similar to pectin methylesterase-encoding genes was isolated (Srpme1). The full-length sequence of Srpme1 was used to localize PME transcripts in situ during S. rostrata stem-nodule development. Several expression patterns were distinguished, hinting at general roles in vascular tissue development and cell division or expansion and at symbiosis-specific functions, such as uninfected cell differentiation.
Autoregulation of Root Nodule Formation: Signals of Both Symbiotic Partners Studied in a Split-Root System of Vicia sativa subsp. nigra
Authors: van Brussel Anton A. N. , Tak Teun , Boot Kees J. M. , Kijne Jan W.
Published: May 2007

Inhibition of root nodule formation on leguminous plants by already induced or existing root nodules is called autoregulation of root nodule formation (AUT). Optimal conditions for AUT were determined using a split-root technique newly developed for Vicia sativa subsp. nigra. Infection of a root A with nodulating Rhizobium leguminosarum bv. viciae bacteria systemically inhibited nodulation of a spatially separated root B inoculated 2 days later with the same bacteria. This treatment gives complete AUT (total absence of nodules on root B). Only partial AUT of root B was obtained by incubation of root A with mitogenic nodulation (Nod) factors or with a noninfective strain producing normal mitogenic Nod factors. Nonmitogenic Nod factors did not evoke AUT. We identified two systemic plant signals induced by Rhizobium bacteria. Signal 1 (at weak buffering) was correlated with sink formation in root A and induced acidification of B-root medium. This signal is induced by treatment of root A with (i) nodulating rhizobia, (ii) mitogenic Nod factors, (iii) nonmitogenic Nod factors, or (iv) the cytokinin zeatin. Signal 2 (at strong buffering) could only be evoked by treatment with nodulating rhizobia or with mitogenic Nod factors. Most probably, this signal represents the specific AUT signal. Induction of complete AUT appears to require actively dividing nodule cells in nodule primordia, nodule meristems, or both of root A.
Virus-Induced Gene Silencing in Transgenic Plants Expressing the Minor Capsid Protein of Beet western yellows virus
Authors: Brault V. , Pfeffer S. , Erdinger M. , Mutterer J. , Ziegler-Graff V.
Published: May 2007

Transgenic Nicotiana benthamiana expressing the minor coat protein P74 of the phloem-limited Beet western yellows virus (BWYV) exhibited an unusual spatial pattern of post-transcriptional gene silencing (PTGS) when infected with BWYV or related viruses. Following infection, transgenic P74 and its mRNA accumulated to only low levels, 21 to 23 nucleotide RNAs homologous to the transgene appeared, and the transgene DNA underwent methylation. The infecting viral RNA, however, was not subject to significant silencing but multiplied readily and produced P74 in the phloem tissues, although the P74 encoded by the transgene disappeared from the phloem as well as the nonvascular tissues.
Natural Resistance to Clover yellow vein virus in Beans Controlled by a Single Recessive Locus
Authors: Sato Masanao , Masuta Chikara , Uyeda Ichiro
Published: May 2007

We characterized the resistance of the common bean cv. Jolanda to Clover yellow vein virus no. 30 (ClYVV). After inoculation, the virus was detected in neither inoculated nor upper leaves, suggesting that the resistance operates at either the viral replication or cell-to-cell movement level. To analyze the mechanism of resistance, we developed a green fluorescent protein (GFP)-tagged ClYVV, and monitored GFP fluorescence at sites of infection on ClYVV-inoculated leaves. No GFP fluorescence was detected in Jolanda, whereas its expression in single cells and spread on inoculated leaves were observed clearly in susceptible cultivars. ClYVV-introduced Jolanda cells were found to be still viable; therefore, it is unlikely that the restriction of multiplication was due to rapid cell death. Genetic analysis indicated that a single recessive locus controlled the resistant phenotype of Jolanda. We designated this locus desc (determinant of susceptibility to ClYVV). Meanwhile, a spontaneous mutant virus that overcomes the resistance (ClYVV-Br) was isolated. Inoculation assays using chimeric viruses suggested that a viral ge-nome-linked protein (VPg) might be the avirulence determinant. The resistance mechanism may be associated with the role of VPg in the viral infection cycle.
Meloidogyne javanicaChorismate Mutase 1 Alters Plant Cell Development
Authors: Doyle Elizabeth A. , Lambert Kris N.
Published: May 2007

Root-knot nematodes are obligate plant parasites that alter plant cell growth and development by inducing the formation of giant cells for feeding. Nematodes inject secretions from their esophageal glands through their stylet and into plant cells to induce giant cell formation. Meloidogyne javanica chorismate mutase 1 (MjCM-1) is one such esophageal gland protein likely to be secreted from the nematode as giant cells form. MjCM-1 has two domains, an N-terminal chorismate mutase (CM) domain and a C-terminal region of unknown function. It is the N-terminal CM domain of the protein that is the predominant form produced in root-knot nematodes. Transgenic expression of MjCM-1 in soybean hairy roots results in a phenotype of reduced and aborted lateral roots. Histological studies demonstrate the absence of vascular tissue in hairy roots expressing MjCM-1. The phenotype of MjCM-1 expressed at low levels can be rescued by the addition of indole-3-acetic acid (IAA), indicating MjCM-1 overexpression reduces IAA biosynthesis. We propose MjCM-1 lowers IAA by causing a competition for chorismate, resulting in an alteration of chorismate-derived metabolites and, ultimately, in plant cell development. Therefore, we hypothesize that MjCM-1 is involved in allowing nematodes to establish a parasitic relationship with the host plant.
A Global Analysis of Protein Expression Profiles in Sinorhizobium meliloti: Discovery of New Genes for Nodule Occupancy and Stress Adaptation
Authors: Djordjevic Michael A. , Chen Han Cai , Natera Siria , Van Noorden Giel , Menzel Christian , Taylor Scott , Renard Clotilde , Geiger Otto , Weiller Georg F.
Published: May 2007

A proteomic examination of Sinorhizobium meliloti strain 1021 was undertaken using a combination of 2-D gel electrophoresis, peptide mass fingerprinting, and bioinformatics. Our goal was to identify (i) putative symbiosis- or nutrientstress-specific proteins, (ii) the biochemical pathways active under different conditions, (iii) potential new genes, and (iv) the extent of posttranslational modifications of S. meliloti proteins. In total, we identified the protein products of 810 genes (13.1% of the genome's coding capacity). The 810 genes generated 1,180 gene products, with chromosomal genes accounting for 78% of the gene products identified (18.8% of the chromosome's coding capacity). The activity of 53 metabolic pathways was inferred from bioinformatic analysis of proteins with assigned Enzyme Commission numbers. Of the remaining proteins that did not encode enzymes, ABC-type transporters composed 12.7% and regulatory proteins 3.4% of the total. Proteins with up to seven transmembrane domains were identified in membrane preparations. A total of 27 putative nodulespecific proteins and 35 nutrient-stress-specific proteins were identified and used as a basis to define genes and describe processes occurring in S. meliloti cells in nodules and under stress. Several nodule proteins from the plant host were present in the nodule bacteria preparations. We also identified seven potentially novel proteins not predicted from the DNA sequence. Post-translational modifications such as N-terminal processing could be inferred from the data. The posttranslational addition of UMP to the key regulator of nitrogen metabolism, PII, was demonstrated. This work demonstrates the utility of combining mass spectrometry with protein arraying or separation techniques to identify candidate genes involved in important biological processes and niche occupations that may be intransigent to other methods of gene expression profiling.
Targeted Activation Tagging of the Arabidopsis NBS-LRR gene, ADR1, Conveys Resistance to Virulent Pathogens
Authors: Grant John J. , Chini Andrea , Basu Debrabata , Loake Gary J.
Published: May 2007

A transgenic Arabidopsis line containing a chimeric PR-1∷luciferase (LUC) reporter gene was subjected to mutagenesis with activation tags. Screening of lines via high-throughput LUC imaging identified a number of dominant Arabidopsis mutants that exhibited enhanced PR-1 gene expression. Here, we report the characterization of one of these mutants, designated activated disease resistance (adr) 1. This line showed constitutive expression of a number of key defense marker genes and accumulated salicylic acid but not ethylene or jasmonic acid. Furthermore, adr1 plants exhibited resistance against the biotrophic pathogens Peronospora parasitica and Erysiphe cichoracearum but not the necrotrophic fungus Botrytis cinerea. Analysis of a series of adr1 double mutants suggested that adr1-mediated resistance against P. parasitica was salicylic acid (SA)-dependent, while resistance against E. cichoracearum was both SA-dependent and partially NPR1-dependent. The ADR1 gene encoded a protein possessing a number of key features, including homology to subdomains of protein kinases, a nucleotide binding domain, and leucine-rich repeats. The controlled, transient expression of ADR1 conveyed striking disease resistance in the absence of yield penalty, highlighting the potential utility of this gene in crop protection.
Characterization of the Xanthomonas AvrXv4 Effector, a SUMO Protease Translocated into Plant Cells
Authors: Roden Julie , Eardley Leah , Hotson Andrew , Cao Yajuan , Mudgett Mary Beth
Published: May 2007

Homologs of the Yersinia virulence factor YopJ are found in both animal and plant bacterial pathogens, as well as in plant symbionts. The conservation of this effector family indicates that several pathogens may use YopJ-like proteins to regulate bacteria-host interactions during infection. YopJ and YopJ-like proteins share structural homology with cysteine proteases and are hypothesized to functionally mimic small ubiquitin-like modifier (SUMO) proteases in eukaryotic cells. Strains of the phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria are known to possess four YopJ-like proteins, AvrXv4, AvrBsT, AvrRxv, and XopJ. In this work, we have characterized AvrXv4 to determine if AvrXv4 functions like a SUMO protease in planta during Xanthomonas-plant interactions. We provide evidence that X. campestris pv. vesicatoria secretes and translocates the AvrXv4 protein into plant cells during infection in a type III-dependent manner. Once inside the plant cell, AvrXv4 is localized to the plant cytoplasm. By performing AvrXv4 deletion and mutational analysis, we have identified amino acids required for type III delivery and for host recognition. We show that AvrXv4 recognition by resistant plants requires a functional protease catalytic core, the domain that is conserved in all of the putative YopJ-like cysteine proteases. We also show that AvrXv4 expression in planta leads to a reduction in SUMO-modified proteins, demonstrating that AvrXv4 possesses SUMO isopeptidase activity. Overall, our studies reveal that the YopJ-like effector AvrXv4 encodes a type III SUMO protease effector that is active in the cytoplasmic compartment of plant cells.
The Erwinia chrysanthemi EC16 hrp/hrc Gene Cluster Encodes an Active Hrp Type III Secretion System That Is Flanked by Virulence Genes Functionally Unrelated to the Hrp System
Authors: Rojas Clemencia M. , Ham Jong Hyun , Schechter Lisa M. , Kim Jihyun F. , Beer Steven V. , Collmer Alan
Published: May 2007

Erwinia chrysanthemi is a host-promiscuous plant pathogen that possesses a type III secretion system (TTSS) similar to that of the host-specific pathogens E. amylovora and Pseudomonas syringae. The regions flanking the TTSS-encoding hrp/hrc gene clusters in the latter pathogens encode various TTSS-secreted proteins. DNA sequencing of the complete E. chrysanthemi hrp/hrc gene cluster and approximately 12 kb of the flanking regions (beyond the previously characterized hecA adhesin gene in the left flank) revealed that the E. chrysanthemi TTSS genes were syntenic and similar (>50% amino-acid identity) with their E. amylovora orthologs. However, the hrp/hrc cluster was interrupted by a cluster of four genes, only one of which, a homolog of lytic transglycosylases, is implicated in TTSS functions. Furthermore, the regions flanking the hrp/hrc cluster lacked genes that were likely to encode TTSS substrates. Instead, some of the genes in these regions predict ABC transporters and methyl-accepting chemotaxis proteins that could have alternative roles in virulence. Mutations affecting all of the genes in the regions flanking or interrupting the hrp/hrc cluster were constructed in E. chrysanthemi CUCPB5047, a mutant whose reduced pectolytic capacity can enhance the phenotype of minor virulence factors. Mutants were screened in witloof chicory leaves and then in potato tubers and Nicotiana clevelandii seedlings. Mu dII1734 insertion in one gene, designated virA, resulted in strongly reduced virulence in all three tests. virA is immediately downstream of hecA, has an unusually low G+C content of 38%, and predicts an unknown protein of 111 amino acids. The E. chrysanthemi TTSS was shown to be active by its ability to translocate AvrPto-Cya (a P. syringae TTSS effector fused to an adenylate cyclase reporter that is active in the presence of eukaryote calmodulin) into N. benthamiana However, VirA(1–61)¯ Cya was not translocated into plant cells, and virA expression was not affected by mutations in E. chrysanthemi Hrp regulator genes hrpL and hrpS. Thus, the 44-kb region of the E. chrysanthemi EC16 genome that is centered on the hrp/hrc cluster encodes a potpourri of virulence factors, but none of these appear to be a TTSS effector.
First Report of Aberia caffra and Quercus cerris as Hosts of Inonotus rickii
Authors: Annesi T. , Coppola R. , D'Amico L. , Motta E.
Published: May 2007

Inonotus rickii (Pat.) D. Reid (Hymenochaetaceae), widespread in tropical and subtropical countries, produces cankers and decay in several hardwoods. In Italy, it was recorded for the first time approximately 20 years ago on Parkinsonia sp. and Schinus molle L. in Sicily (2) and more recently on Acer negundo L. and Celtis australis L. in Rome (1). During May 2004, a survey was performed in Sicily in public and botanical gardens where the fungus had been detected in the past. Although the previously attacked plants had been removed, more trees were showing decline symptoms, with sparse foliage and some dieback. Typical brown, powdery masses of chlamydospores of I. rickii were observed near wounds or pruning cuts on Quercus cerris L. in Palermo and Sambucus nigra L. and Aberia caffra Hook. F. & Harv. in Catania. From each fungal specimen, fragments were isolated and cultured on malt agar. Cultures produced acute, thick-walled, brown setae (as much as 250 µm long) and abundant chlamydospores that were yellowish brown, thick walled, irregularly shaped, and 10 to 12 × 8 to 12 µm. Each isolate conformed to I. rickii (3). Cultures and voucher specimens have been deposited in the ISPaVe collection (PF40, PF41, and PF42) and the ROPV herbarium (772, 773, and 774). To our knowledge, this is the first report of I. rickii in living Q. cerris and A. caffra. S. nigra is a new host in Italy. This record shows that the pathogen has fully established itself in southern Europe where it may expand its host range.

References: (1) T. Annesi et al. For. Pathol. 33:405, 2003. (2) M. Intini. Micol. Ital. 20:49, 1988. (3) J. A. Stalpers. Karstenia 40:167, 2000.
An Experimental Inoculation System to Study Citrus-Xylella fastidiosa Interactions
Authors: Lopes S. A. , Teixeira D. C. , Fernandes N. G. , Ayres A. J. , Torres S. C. Z. , Barbosa J. C. , Li W. B.
Published: May 2007

Difficulties in reproducing the citrus variegated chlorosis (CVC) disease symptoms in experimental plants have delayed implementation of studies to better understand the essential aspects of this important disease. In an extensive study, cultivars of sweet orange (Citrus sinensis) were inoculated with Xylella fastidiosa using procedures that included root immersion, and stem absorption, pricking, or infiltration of the inoculum into plants of different ages. Inoculum consisted of 5-day-old cultures or cell suspensions of CVC strain 9a5c diluted in phosphate-buffered saline. Inoculated plants and controls were grown, or transferred just after inoculation, to 5-liter pots or 72-cell foam trays. Approximately 4, 5, 9, and 12 months after inoculation, leaves were collected and processed for polymerase chain reaction analysis or X. fastidiosa isolation on BCYE agar medium. Root immersion and stem inoculation of 4- and 6-month-old plants resulted in low percentages of symptomatic (0 to 7%) and plants positive by isolation (0 to 9%). Pinpricked or injected stems of 1-month-old seedlings resulted in high percentages of plants symptomatic (29 and 90% in Pera Rio, 75, 59, and 83% in Valencia, and 77% in Natal) or positive by isolation (26 and 93% in Pera Rio, 98, 96, and 83% in Valencia, and 77% in Natal). In foam trays, the seedlings grew less, the incubation period was shorter, and disease severity was higher than in pots. This system allows testing of higher numbers of plants in a reduced space with a more precise reproduction of the experimental conditions.
Inoculum Sources and Survival of Xanthomonas axonopodis pv. allii in Colorado
Authors: Gent David H. , Lang Jillian M. , Bartolo Michael E. , Schwartz Howard F.
Published: May 2007

Xanthomonas leaf blight, caused by the bacterium Xanthomonas axonopodis pv. allii, is an emerging disease of onion in the western United States and worldwide, but few management strategies have been developed because little is known about disease epidemiology and pathogen survival. Therefore, we sought to identify and quantify primary inoculum sources of the pathogen in Colorado. Growth chamber and field studies evaluated survival and dissemination of X. axonopodis pv. allii in association with weed, alternate host, and volunteer onion plants, irrigation water, and crop debris. Epiphytic X. axonopodis pv. allii was recovered from the foliage of nine asymptomatic weed species and Medicago sativa, but the bacterium was not recovered from plants in locations where an epidemic of Xanthomonas leaf blight did not occur the prior year. The bacterium also was isolated from volunteer onion with characteristic Xanthomonas leaf blight symptoms. A rifampicin mutant of X. axonopodis pv. allii strain O177 was recovered consistently from the irrigation tail water of onion fields inoculated with the bacterium; populations as large as 3.02 × 104 CFU/ml were recovered. X. axonopodis pv. allii was recovered from infested onion leaves 9 months after they were placed on the soil surface or buried to a depth of 25 cm, but culturable populations of the pathogen decreased 104 to 106 more in buried leaves. Cultural practices that avoid or eliminate X. axonopodis pv. allii inoculum sources should reduce Xanthomonas leaf blight losses to onion.
First Report of Gray Mold in Tomato Caused by Botrytis cinerea in Baja California, Mexico
Authors: Holguín-Peña R. J. , Arcos F. G.
Published: May 2007

San Quintin Valley, a 60-mile-long coastal plain (30°30′N, 116°W) in the Baja California Peninsula, is one of the major fresh tomato (Lycopersicon esculentum Mill.) production areas in Mexico with more than 8,000 ha. During the last 10 years, the valley's tomato production has declined because of gray mold and stem canker diseases. Flower rot, reddish brown margins on the leaves and stems, and fruit with a gray mold were observed on field-grown tomato plants (Roma type cv. Tequila) in the autumn of 2003. Severity ranging from 55 to 60% was observed at harvest. Infected tissues were sampled and disinfested by immersion in 1% NaOCl for 1 min, rinsed in sterile water, and placed on malt extract agar at 22°C. Fungal conidia were then transferred to 2% potato dextrose agar (PDA). The resulting fungal colonies were definitively identified as Botrytis cinerea Pers.:Fr. The colonies of B. cinerea were first hyaline and white and became dark gray after 96 h. Mycelia were septate with dark branched conidiophores. Conidia were unicellular, ellipsoid, and ranged from 5 to 8 × 8 to 14 μm. Profuse black sclerotia developed in 7-day-old cultures. Infection site analyses in diseased flowers at different stages during the bloom were done with scanning electron microscopy. Fungal hyphae were located predominantly on the receptacle areas, whereas conidia were located in the ovaries as described previously (3). The identity of B. cinerea was confirmed by a restriction digest with ApoI of the 413-kb polymerase chain reaction amplification product obtained with BA2f/BA1r primers (1) and random amplified polymorphic DNA banding patterns (2). Pathogenicity tests were done by spray inoculation of 1-ml aqueous conidial suspension (106 CFU/ml) on 20 healthy plants during the blossom stage. An equal number of plants sprayed with sterile water was used as the control. Plants were incubated at 20 ± 2°C for 5 days. The fungus was reisolated from diseased flowers and peduncles after surface disinfestation (2.5% NaOCl) and plating on PDA. No symptoms were observed in the noninoculated controls. To our knowledge, this is the first report of B. cinerea causing gray mold disease on tomato in Baja California.

References: (1) K. Nielsen et al. Plant Dis. 86:682, 2002. (2) S. Rigotti et al. FEMS Microbiol. Lett. 209:169, 2002. (3) O. Viret et al. Phytopathology 94:850, 2004.
Quantitative Trait Loci Conditioning Resistance to Phaeosphaeria Leaf Spot of Maize Caused by Phaeosphaeria maydis
Authors: Carson M.L. , Stuber C.W. , Senior M.L.
Published: May 2007

Phaeosphaeria leaf spot (PLS) is a potentially important disease of maize (Zea mays) that has appeared in winter breeding nurseries in southern Florida. Inbred lines related to B73 are particularly susceptible to Phaeosphaeria leaf spot, whereas inbreds related to Mo17 are highly resistant. A previous study of the inheritance of resistance to Phaeosphaeria leaf spot in the cross B73 × Mo17 found that resistance is highly heritable and controlled by mostly additive gene action at three or four loci. In this study, we used 158 recombinant inbred (RI) lines derived from the cross B73 × Mo17 to map quantitative trait loci (QTL) governing resistance. The RI lines along with the parent inbred lines and the F1 were evaluated for PLS resistance in replicated trials over two winter growing seasons in southern Florida. Using the composite interval mapping (CIM) function of PLABQTL software, five QTL on four different chromosomes were found to control PLS resistance in Mo17. In addition, the × additive interaction between two of these QTL was found to be significant. Our results are in close agreement with the previous study, where generation mean analysis was used to study the inheritance of resistance to PLS.