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Hans D. Vanetten - One of the best experts on this subject based on the ideXlab platform.
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The supernumerary chromosome of Nectria haematococca that carries pea-pathogenicity-related genes also carries a trait for pea rhizosphere competitiveness.
Applied and environmental microbiology, 2008Co-Authors: M. Rodriguez-carres, Gerard J. White, D. Tsuchiya, M. Taga, Hans D. VanettenAbstract:Fungi are found in a wide range of environments, and the ecological and host diversity of the fungus Nectria haematococca has been shown to be due in part to unique genes on different supernumerary chromosomes. These chromosomes have been called "conditionally dispensable" (CD) since they are not needed for axenic growth but are important for expanding the host range of individual isolates. From a biological perspective, the CD chromosomes can be compared to bacterial plasmids that carry unique genes that can define the habits of these microorganisms. The current study establishes that the N. haematococca PDA1-CD chromosome, which contains the genes for pea pathogenicity (PEP cluster) on pea roots, also carries a gene(s) for the utilization of homoserine, a compound found in large amounts in pea root exudates. Competition studies demonstrate that an isolate that lacks the PEP cluster but carries a portion of the CD chromosome which includes the homoserine utilization (HUT) gene(s) is more competitive in the pea rhizosphere than an isolate without the CD chromosome.
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Tissue-specific localization of pea root infection by Nectria haematococca. Mechanisms and consequences.
Plant physiology, 2005Co-Authors: Uvini Gunawardena, Hans D. Vanetten, Marianela Rodriguez, David C. Straney, John T. Romeo, Martha C. HawesAbstract:Root infection in susceptible host species is initiated predominantly in the zone of elongation, whereas the remainder of the root is resistant. Nectria haematococca infection of pea (Pisum sativum) was used as a model to explore possible mechanisms influencing the localization of root infection. The failure to infect the root tip was not due to a failure to induce spore germination at this site, suppression of pathogenicity genes in the fungus, or increased expression of plant defense genes. Instead, exudates from the root tip induce rapid spore germination by a pathway that is independent of nutrient-induced germination. Subsequently, a factor produced during fungal infection and death of border cells at the root apex appears to selectively suppress fungal growth and prevent sporulation. Host-specific mantle formation in response to border cells appears to represent a previously unrecognized form of host-parasite relationship common to diverse species. The dynamics of signal exchange leading to mantle development may play a key role in fostering plant health, by protecting root meristems from pathogenic invasion.
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pisatin demethylase genes are on dispensable chromosomes while genes for pathogenicity on carrot and ripe tomato are on other chromosomes in Nectria haematococca
Molecular Plant-microbe Interactions, 2002Co-Authors: Deanna L. Funnell, Hans D. VanettenAbstract:Studies on the wide-host-range fungus Nectria haematococca MP VI have shown a linkage between virulence on pea and five of nine PDA genes that encode the ability to detoxify the pea phytoalexin, pisatin. Most of the PDA genes are on chromosomes of approximately 1.6 megabases (Mb) and two of these genes, PDA1-2 and PDA6-1, have been demonstrated to reside on approximately 1.6-Mb chromosomes that can be lost during meiosis. Prior studies also have shown that the dispensable chromosome carrying PDA6-1 contains a gene (MAK1) necessary for maximum virulence on chickpea. The present study evaluated whether the other approximately 1.6-Mb chromosomes that carry PDA genes also are dispensable, their relationship to each other, and whether they contain genes for pathogenicity on hosts other than pea or chickpea. DNA from the PDA1-1 chromosome (associated with virulence on pea) and the PDA6-1 chromosome (associated with virulence on chickpea) were used to probe blots of contour-clamped homogeneous electric field (CH...
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Distribution of the pea pathogenicity ( PEP) genes in the fungus Nectria haematococca mating population VI.
Current genetics, 2002Co-Authors: Esteban D. Temporini, Hans D. VanettenAbstract:Previous studies identified a cluster of six genes that are expressed in the fungus Nectria haematococca mating population VI during infection of pea. Four of these genes were shown to contribute to pathogenicity on pea and were called PEP genes for pea pathogenicity. The cluster is located on a "conditionally dispensable" (CD) chromosome and has features similar to bacterial pathogenicity islands. In this study, the occurrence and location of members of the PEP cluster were analyzed in laboratory strains and nine pea pathogenic and 16 non-pea pathogenic field isolates of N. haematococca. Our results indicate that all pea-pathogenic isolates have homologues for all six genes present in the PEP cluster and the homologues appear to be clustered. PEP homologues are also present in isolates that are not pathogenic on pea, although none of these isolates have homologues of all six genes. In addition, PEP homologues are found in CD chromosomes and in other chromosomes. Isolates without PEP homologues are virulent on ripe tomato fruits and carrot roots, indicating that PEP genes are not required for pathogenicity on these hosts.
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Genes determining pathogenicity to pea are clustered on a supernumerary chromosome in the fungal plant pathogen Nectria haematococca
The Plant journal : for cell and molecular biology, 2001Co-Authors: Yinong Han, Ulla K Benny, H. Corby Kistler, Xiaoguang Liu, Hans D. VanettenAbstract:Three genes that contribute to the ability of the fungus Nectria haematococca to cause disease on pea plants have been identified. These pea pathogenicity (PEP) genes are within 25 kb of each other and are located on a supernumerary chromosome. Altogether, the PEP gene cluster contains six transcriptional units that are expressed during infection of pea tissue. The biochemical function of only one of the genes is known with certainty. This gene, PDA1, encodes a specific cytochrome P450 that confers resistance to pisatin, an antibiotic produced by pea plants. The three new PEP genes, in addition to PDA1, can independently increase the ability of the fungus to cause lesions on pea when added to an isolate lacking the supernumerary chromosome. Based on predicted amino acid sequences, functions for two of these three genes are hypothesized. The deduced amino acid sequence of another transcribed portion of the PEP cluster, as well as four other open reading frames in the cluster, have a high degree of similarity to known fungal transposases. Several of the features of the PEP cluster -- a cluster of pathogenicity genes, the presence of transposable elements, and differences in codon usage and GC content from other portions of the genome -- are shared by pathogenicity islands in pathogenic bacteria of plants and animals.
Kerry Odonnell - One of the best experts on this subject based on the ideXlab platform.
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molecular phylogeny of the Nectria haematococca fusarium solani species complex
Mycologia, 2000Co-Authors: Kerry OdonnellAbstract:AbstractPhylogenetic relationships and biogeography of the phytopathogenic Nectria haematococca-Fusarium solani species complex, section Martiella of Fusarium, were inferred from sequence data from...
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molecular phylogeny of the Nectria haematococca fusarium solani species complex
Mycologia, 2000Co-Authors: Kerry OdonnellAbstract:Phylogenetic relationships and biogeography of the phytopathogenic Nectria haematococca-Fusarium solani species complex, section Martiella of Fusarium, were inferred from sequence data from the nuc...
Masatoki Taga - One of the best experts on this subject based on the ideXlab platform.
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Meiotic inheritance of a fungal supernumerary chromosome and its effect on sexual fertility in Nectria haematococca.
Fungal biology, 2015Co-Authors: Hamid S. Garmaroodi, Masatoki TagaAbstract:PDA1-conditionally dispensable chromosome (CDC) of Nectria haematococca MP VI has long served as a model of supernumerary chromosomes in plant pathogenic fungi because of pathogenicity-related genes located on it. In our previous study, we showed the dosage effects of PDA1-CDC on pathogenicity and homoserine utilization by exploiting tagged PDA1-CDC with a marker gene. CDC content of mating partners and progenies analyzed by PCR, PFGE combined with Southern analysis and chromosome painting via FISH. In this study, we analyzed mode of meiotic inheritance of PDA1-CDC in several mating patterns with regard to CDC content and found a correlation between CDC content of parental strains with fertility of crosses. The results showed non-Mendelian inheritance of this chromosome followed by duplication or loss of the CDC in haploid genome through meiosis that probably were due to premature centromere division, not by nondisjunction as reported for the supernumerary chromosomes in other species. Correlation of CDC with fertility is the first time to be examined in fungi in this study.
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Cytological karyotyping and characterization of a 410 kb minichromosome in Nectria haematococca MPI.
Mycologia, 2012Co-Authors: Ahmed M. A. Mahmoud, Masatoki TagaAbstract:Karyotypes of the cucurbit pathogen Nectria haematococca MPI (anamorph Fusarium solani f. sp. cucurbitae race 1) was studied using the two standard strains ATCC18098 and ATCC18099. Complete separation of all chromosomes was difficult with pulsed field gel electrophoresis due to both the large size and co-migration of chromosomes. In contrast, cytological karyotyping was done successfully with fluorescence microscopy combined with the germ tube burst method for sample preparation to visualize mitotic metaphase chromosomes. For each strain the basic chromosome number (CN) was nine, which revises previous chromosome estimates of n = 4. Chromosomes were morphologically characterized by their sizes, intensely fluorescing segments, and protrusion of rDNA. In addition to the basic chromosome complement, ATCC18098 had a mini-chromosome of ~410 kb present as a single copy in somatic nuclei. Chromosome fluorescence in situ hybridization indicated that this mini-chromosome is not a derivative from the other chromosomes in the genome. In addition, crossing experiments suggested that it was transmitted in a Mendelian manner to the ascospore progeny.
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The genome of Nectria haematococca: contribution of supernumerary chromosomes to gene expansion
PLoS Genetics, 2009Co-Authors: Jeffrey J. Coleman, Catherine C. Wasmann, Masatoki Taga, Gerard J. White, Steve D. Rounsley, Marianela Rodriguez-carres, Alan Kuo, Jane Grimwood, Jeremy Schmutz, Shiguo ZhouAbstract:The ascomycetous fungus Nectria haematococca, (asexual name Fusarium solani), is a member of a group of >50 species known as the "Fusarium solani species complex". Members of this complex have diverse biological properties including the ability to cause disease on >100 genera of plants and opportunistic infections in humans. The current research analyzed the most extensively studied member of this complex, N. haematococca mating population VI (MPVI). Several genes controlling the ability of individual isolates of this species to colonize specific habitats are located on supernumerary chromosomes. Optical mapping revealed that the sequenced isolate has 17 chromosomes ranging from 530 kb to 6.52 Mb and that the physical size of the genome, 54.43 Mb, and the number of predicted genes, 15,707, are among the largest reported for ascomycetes. Two classes of genes have contributed to gene expansion: specific genes that are not found in other fungi including its closest sequenced relative, Fusarium graminearum; and genes that commonly occur as single copies in other fungi but are present as multiple copies in N. haematococca MPVI. Some of these additional genes appear to have resulted from gene duplication events, while others may have been acquired through horizontal gene transfer. The supernumerary nature of three chromosomes, 14, 15, and 17, was confirmed by their absence in pulsed field gel electrophoresis experiments of some isolates and by demonstrating that these isolates lacked chromosome-specific sequences found on the ends of these chromosomes. These supernumerary chromosomes contain more repeat sequences, are enriched in unique and duplicated genes, and have a lower G+C content in comparison to the other chromosomes. Although the origin(s) of the extra genes and the supernumerary chromosomes is not known, the gene expansion and its large genome size are consistent with this species' diverse range of habitats. Furthermore, the presence of unique genes on supernumerary chromosomes might account for individual isolates having different environmental niches
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Duplication of a Conditionally Dispensable Chromosome Carrying Pea Pathogenicity (PEP) Gene Clusters in Nectria haematococca
Molecular plant-microbe interactions : MPMI, 2007Co-Authors: Hamid S. Garmaroodi, Masatoki TagaAbstract:A supernumerary chromosome called a conditionally dispensable chromosome (CDC) is essential for pathogenicity of Nectria haematococca on pea. Among several CDCs discovered in N. haematococca, the PDA1 CDC that harbors the pisatin demethylation gene PDA1 is one of the best-studied CDCs and serves as a model for plant-pathogenic fungi. Although the presence of multiple copies is usual for supernumerary chromosomes in other eukaryotes, this possibility has not been examined well for any CDCs in N. haematococca. In this study, we produced strains with multiple copies of the PDA1 CDC by protoplast fusion and analyzed dosage effects of this chromosome. Using multiple methods, including cytological chromosome counting and fluorescence in situ hybridization, the fusion products between two transformants derived from the same strain that bears a single PDA1 CDC were shown to contain two PDA1 CDCs from both transformants and estimated to be haploid resulting from the deletion of an extra set or sets of A chromosomes in the fused nuclei. In phenotype assays, dosage effects of PDA1 CDC in the fusion products were evident as increased virulence and homoserine-utilizing ability compared with the parents. In a separate fusion experiment, PDA1 CDC accumulated up to four copies in a haploid genome.
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Visualization of a conditionally dispensable chromosome in the filamentous ascomycete Nectria haematococca by fluorescence in situ hybridization.
Fungal genetics and biology : FG & B, 1999Co-Authors: Masatoki Taga, Minoru Murata, Hans D. VanettenAbstract:Abstract Supernumerary chromosomes, termed “conditionally dispensable” (CD) chromosomes, are known in Nectria haematococca. Because these CD chromosomes had been revealed solely by pulsed-field gel electrophoresis, their morphological properties were unknown. In this study, we visualized a 1.6-Mb CD chromosome of this fungus by three different types of fluorescence in situ hybridization. The CD chromosome at mitotic metaphase was similar in its appearance to the other chromosomes in the genome. Heterochromatic condensation was not distinct in the CD chromosome, suggesting that it is primarily euchromatic. It was also evident that the CD chromosome is unique and not a duplicate of other chromosomes in the genome. At interphase and prophase, the CD chromosome was not dispersed throughout the nucleus, but occupied a limited domain. Occasionally, occurrence of two distinct unattached copies of the CD chromosome were observed during interphase and metaphase.
Sarah F. Covert - One of the best experts on this subject based on the ideXlab platform.
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Nht2, a copia LTR retrotransposon from a conditionally dispensable chromosome in Nectria haematococca.
Current genetics, 2002Co-Authors: April M. Shiflett, Jürg Enkerli, Sarah F. CovertAbstract:In the plant pathogenic ascomycete Nectria haematococca mating population (MP) VI, the conditionally dispensable chromosomes are unstable during sexual reproduction. During mapping of such a chromosome, three dispersed repeats were identified. Nht2, one of these repeated elements, is a long terminal repeat (LTR) retrotransposon that is 5.9 kb in length. Its deduced amino acid sequence is homologous to the four enzymatic domains characteristic of copia retrotransposons, but it contains multiple stop codons and probably is no longer able to transpose autonomously. Nht2's LTRs differ at ten positions and the characteristics of these differences resemble the changes induced by repeat-induced point mutation (RIP) in Neurospora crassa. The likelihood that Nectria haematococca MP VI has a RIP-like process, however, is reduced by the fact that a multi-copy transposon cloned from the same ascospore isolate as Nht2 encodes an intact open reading frame. Nht2 is broadly distributed among isolates collected from a variety of host plants. A limited survey of three field isolates suggests that Nht2 is on only one or a few chromosomes in every genome. Nht2's degeneracy and its widespread distribution within the species both suggest that it is an ancient element within N. haematococca MP VI.
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Physical map of a conditionally dispensable chromosome in Nectria haematococca mating population VI and location of chromosome breakpoints.
Genetics, 2000Co-Authors: Jürg Enkerli, Garima Bhatt, Heather Reed, Angela Briley, Sarah F. CovertAbstract:Certain isolates of the plant pathogenic fungus Nectria haematococca mating population (MP) VI contain a 1.6-Mb conditionally dispensable (CD) chromosome carrying the phytoalexin detoxification genes MAK1 and PDA6-1. This chromosome is structurally unstable during sexual reproduction. As a first step in our analysis of the mechanisms underlying this chromosomal instability, hybridization between overlapping cosmid clones was used to construct a map of the MAK1 PDA6-1 chromosome. The map consists of 33 probes that are linked by 199 cosmid clones. The polymerase chain reaction and Southern analysis of N. haematococca MP VI DNA digested with infrequently cutting restriction enzymes were used to close gaps and order the hybridization-derived contigs. Hybridization to a probe extended from telomeric repeats was used to anchor the ends of the map to the actual chromosome ends. The resulting map is estimated to cover 95% of the MAK1 PDA6-1 chromosome and is composed of two ordered contigs. Thirty-eight percent of the clones in the minimal map are known to contain repeated DNA sequences. Three dispersed repeats were cloned during map construction; each is present in five to seven copies on the chromosome. The cosmid clones representing the map were probed with deleted forms of the CD chromosome and the results were integrated into the map. This allowed the identification of chromosome breakpoints and deletions.
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Inducing the loss of conditionally dispensable chromosomes in Nectria haematococca during vegetative growth.
Current genetics, 1998Co-Authors: Hans D. Vanetten, Jürg Enkerli, Sam Jorgensen, Sarah F. CovertAbstract:A procedure for inducing and detecting the loss of conditionally dispensable (CD) chromosomes in filamentous fungi during vegetative growth was developed using Nectria haematococca mating population VI as a model. CD chromosomes in two different isolates of N. haematococca were tagged via integrative transformation with a gene conferring resistance to hygromycin B. In each case the transformation vector included chromosome-specific DNA in order to direct its homologous recombination with the desired chromosome. Chromosome loss was induced by exposing tagged isolates to inhibitory concentrations of benomyl either for protracted periods of time on solid medium or for short periods of time in liquid medium. After exposure to benomyl, isolates that lost the tagged chromosome were identified by their loss of resistance to hygromycin B. Electrophoretic karyotyping was used to verify that isolates which failed to grow on hygromycin B lacked an intact CD chromosome. Ten other chemicals known to interfere with mitotic events or cell development in other organisms did not induce CD chromosome loss in N. haematococca.
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maackiain detoxification contributes to the virulence of Nectria haematococca mp vi on chickpea
Molecular Plant-microbe Interactions, 1998Co-Authors: Jürg Enkerli, Garima Bhatt, Sarah F. CovertAbstract:Nectria haematococca mating population (MP) VI isolates that contain the MAK1 gene are able to degrade maackiain, a chickpea (Cicer arietinum) phytoalexin, to a less toxic compound. To test the contribution of MAK1 to the virulence of N. haematococca MP VI on chickpea, the MAK1 gene was disrupted in a highly virulent Mak+ isolate or added to a weakly virulent Mak- isolate via transformation. The disruption of MAK1 decreased virulence to a moderate level, while addition of multiple copies of MAK1 increased virulence to either a moderate or a high level. These data demonstrate that maackiain detoxification is a determinant, but not the only determinant, of virulence in N. haematococca MP VI isolates capable of causing disease on chickpea. MAK1 is located on a 1.6-Mb conditionally dispensable chromosome. To ascertain if there are additional genes influencing virulence toward chickpea stems on the MAK1 chromosome, the loss of this chromosome was chemically induced in an isolate containing the disrupted MAK1 g...
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Maackiain Detoxification Contributes to the Virulence of Nectria haematococca MP VI on Chickpea
Molecular Plant-Microbe Interactions®, 1998Co-Authors: Jürg Enkerli, Garima Bhatt, Sarah F. CovertAbstract:Nectria haematococca mating population (MP) VI isolates that contain the MAK1 gene are able to degrade maackiain, a chickpea (Cicer arietinum) phytoalexin, to a less toxic compound. To test the contribution of MAK1 to the virulence of N. haematococca MP VI on chickpea, the MAK1 gene was disrupted in a highly virulent Mak+ isolate or added to a weakly virulent Mak- isolate via transformation. The disruption of MAK1 decreased virulence to a moderate level, while addition of multiple copies of MAK1 increased virulence to either a moderate or a high level. These data demonstrate that maackiain detoxification is a determinant, but not the only determinant, of virulence in N. haematococca MP VI isolates capable of causing disease on chickpea. MAK1 is located on a 1.6-Mb conditionally dispensable chromosome. To ascertain if there are additional genes influencing virulence toward chickpea stems on the MAK1 chromosome, the loss of this chromosome was chemically induced in an isolate containing the disrupted MAK1 gene. Loss of the MAK1 chromosome did not reduce virulence toward chickpea stems further, thus indicating that no additional genes for virulence on this part of the host plant are located on the MAK1 chromosome.
David C. Straney - One of the best experts on this subject based on the ideXlab platform.
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Tissue-specific localization of pea root infection by Nectria haematococca. Mechanisms and consequences.
Plant physiology, 2005Co-Authors: Uvini Gunawardena, Hans D. Vanetten, Marianela Rodriguez, David C. Straney, John T. Romeo, Martha C. HawesAbstract:Root infection in susceptible host species is initiated predominantly in the zone of elongation, whereas the remainder of the root is resistant. Nectria haematococca infection of pea (Pisum sativum) was used as a model to explore possible mechanisms influencing the localization of root infection. The failure to infect the root tip was not due to a failure to induce spore germination at this site, suppression of pathogenicity genes in the fungus, or increased expression of plant defense genes. Instead, exudates from the root tip induce rapid spore germination by a pathway that is independent of nutrient-induced germination. Subsequently, a factor produced during fungal infection and death of border cells at the root apex appears to selectively suppress fungal growth and prevent sporulation. Host-specific mantle formation in response to border cells appears to represent a previously unrecognized form of host-parasite relationship common to diverse species. The dynamics of signal exchange leading to mantle development may play a key role in fostering plant health, by protecting root meristems from pathogenic invasion.
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Modulation of cAMP and phosphodiesterase activity by flavonoids which induce spore germination of Nectria haematococca MP VI (Fusarium solani).
Physiological and Molecular Plant Pathology, 2000Co-Authors: Savita Bagga, David C. StraneyAbstract:Flavonoids exuded from legume roots stimulate spore germination of a number of soilborne fungi which interact with these plants, and so may act as host cues to initiate interaction. Macroconidia of Nectria haematococca MPVI (anamorph Fusarium solani), a pathogen of pea (Pisum sativum), germinate in response to the same flavonoids which induce nod gene expression in pea-specific rhizobia. Pisatin, the isoflavonoid phytoalexin of pea, also induces germination. The present study found that cAMP levels in macroconidia were transiently induced by flavonoid treatment. Combined with previous pharmacological studies, this indicates that flavonoid signalling utilizes the cAMP pathway. A hypothesis that flavonoids modulate cAMP levels through direct inhibition of N. haematococca cAMP phosphodiesterase was tested. A low KMcAMP phosphodiesterase activity from macroconidia was tested for inhibition by 10 flavonoids. Naringenin, a strong inducer of germination, was a strong inhibitor of phosphodiesterase (apparent Ki33 μm). There was a general correlation between strength of induction and inhibition of phosphodiesterase. Pisatin, structurally distinct from the others, appeared to be an exception to this trend. The results suggest that the ability of specific flavones and flavanones to inhibit cAMP phosphodiesterase is a potential mechanism through which they can induce cAMP levels and so promote germination.
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Regulatory signals influencing expression of the PDA1 gene of Nectria haematococca MPVI in culture and during pathogenesis of pea
Molecular Plant-Microbe Interactions®, 1999Co-Authors: Rana Khan, David C. StraneyAbstract:The PDA1 gene of the filamentous fungus Nectria haematococca MPVI (anamorph: Fusarium solani) encodes a cytochrome P450 monooxygenase that detoxifies pisatin, the isoflavonoid phytoalexin produced by its host, garden pea (Pisum sativum). PDA1 is regulated by several signals in culture that may control its expression during pathogenesis of pea. It is induced by pisatin and repressed by glucose and amino acids. Deletion analysis was performed on the PDA1 promoter to define regulatory regions, using a β-glucuronidase (GUS) reporter gene fusion. The results identified a region between -287 and -429, relative to the start of transcription, that mediated repression by either glucose or amino acids in culture, independent from pisatin induction. Transformants bearing PDA1 promoter constructs displaying altered regulation in response to the different signals were used to infect pea epicotyls in order to correlate regulation in culture with that observed during pathogenesis of the host. Removal of the nutritional ...
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Identification of elements in the PDA1 promoter of Nectria haematococca necessary for a high level of transcription in vitro.
Molecular & general genetics : MGG, 1996Co-Authors: Yijun Ruan, David C. StraneyAbstract:Expression of the PDA1 gene in the ascomycete Nectria haematococca MPVI (anamorph: Fusarium solani) is induced by exposure of mycelium to pisatin, an isoflavonoid phytoalexin produced by its host plant, garden pea. The PDA1 gene encodes a cytochrome P-450 monooxygenase which detoxifies pisatin. Regulatory elements controlling transcription from the PDA1 promoter were identified using a homologous Nectria in vitro transcription system through analysis of 5' deletions, specific oligonucleotide competition, and fusion of upstream segments to a heterologous promoter. A promoter-distal element which provided transcriptional activation was localized to a 35-bp region positioned -514 to -483 upstream of the transcriptional start site. This 35-bp region binds a previously characterized pisatin-responsive DNA-binding factor (PRF) and thus may provide pisatin-responsive control of transcription. A second promoter-proximal positive-acting region was found to be necessary for promoter transcription in both homologous and heterologous extracts, and so is likely to bind less genespecific transcription activator(s). A negative-acting element located between these two positive regions may act to make the positive-acting elements interdependent. The identification of an activator responding to pisatin provides a model for the control of a number of genes and processes controlled by host-specific signals, particularly the flavonoids.
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Analysis of determinants of binding and transcriptional activation of the pisatin-responsive DNA binding factor of Nectria haematococca.
Molecular plant-microbe interactions : MPMI, 1996Co-Authors: He J, Ruan Y, David C. StraneyAbstract:Pisatin is a fungistatic isoflavonoid produced by garden pea. Field isolates of the ascomycete Nectria haematococca mating population VI (anamorph: Fusarium solani) that are highly virulent on pea have been found to possess the PDA1 gene encoding a pisatin detoxifying activity. Expression of PDA1 is specifically and highly induced by exposure of mycelia to pisatin. A pisatin-responsive DNA-binding activity has previously been identified with properties suggestive of a transcriptional regulator of PDA1. In this study, the sequence determinants for binding this pisatin-responsive factor (PRF) were localized to a 14-bp region through analysis of sequence alterations that reduced PRF binding. Using a homologous in vitro transcription system, a transcriptional activator of PDA1 was shown to be present in mycelial extracts that shared the sequence specificity characteristic of the PRF, indicating function of the DNA-binding protein in transcriptional control. A 70-kDa protein was shown to be a DNA-binding component of PRF by three independent assays for DNA-binding proteins: Southwestern (DNA-protein) blotting, UV-crosslinking, and binding to immobilized DNA. These results characterize a transcriptional activator acting on the PDA1 promoter that is responsive to a host-specific compound and provide insight into the regulation of fungal genes in response to plant flavonoids.
