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At least 199 records · Page 11Linked to original sources

Tuberculina-Thanatophytum/Rhizoctonia crocorum-Helicobasidium: a unique mycoparasitic-phytoparasitic life strategy.

Tuberculina species are mitosporic parasites of rust fungi. Phylogenetically they belong to the Urediniomycetidae, therefore being closely related to their rust fungal hosts. We reveal by means of molecular analyses, ultrastructural and morphological features, observations in the field, and infection experiments that species of the genus Tuberculina and the violet root rot (Helicobasidium/Rhizoctonia crocorum) are stages of the life-cycle of one holomorph. This opens up new perspectives on parasitic life strategies as the resulting life-cycle is based on interkingdom host jumping between rusts and spermatophytes. In addition, we point at the consequences for any practical application dealing with Helicobasidium as an economically important plant pathogen and Tuberculina as a biological agent in rust control.

Basidiomycota↗

Identification and characterization of Armillaria tabescens from the southeastern United States.

In the southeastern USA, Armillaria root rot disease on peach (Prunus persica) is caused by Armillaria tabescens, and to a lesser degree by A. mellea. Recent attempts to genetically characterize A. tabescens isolates using rDNA indicated the existence of heterozygosity in diploid isolates. In order to clarify this heterozygosity, DNA from stipe and single spore cultures of A. tabescens isolate SC.MF-1.01 was characterized using RFLP and sequence analysis. Direct sequencing of rDNA amplicons from diploid stipe tissue indicated heterozygosity in the IGS1 and ITS2 regions. IGS1 copies A and B, and ITS copies A and B segregated in a 1:1 ratio in single spore progeny, whereby IGS1 copy A always segregated with ITS copy A and IGS1 copy B always segregated with ITS copy B. The results indicate the existence of divergent haplotypes in the two nuclei of SC.MF-1.01 diploid mycelium and a single locus for rDNA tandem repeats. Additional IGS1 copies, designated IGS1 copy C, D, and E, with variable AluI restriction sites were cloned from SC.MF-1.01 diploid mycelium indicating polymorphism within ribosomal tandem repeats in A. tabescens. IGS1 copies C, D, and E were found once each in 100 clones analyzed, and therefore seemed to be rare compared to copies A (44 clones) and B (53 clones). RFLP and sequence analysis of the ribosomal IGS1 and ITS1 regions in North American A. tabescens isolates indicated that heterozygosity is common in A. tabescens diploid mycelium. A PCR-based molecular technique was developed to distinguish A. tabescens from many other North American Armillaria species, including A. mellea.

Agaricales↗

Identification of 1,8-dihydroxynaphthalene melanin in Monosporascus cannonballus and the analysis of hexaketide and pentaketide compounds produced by wild-type and pigmented isolates of the fungus.

Monosporascus cannonballus causes root rot and vine decline in muskmelons and watermelons. Wild types of this fungus often undergo degenerative changes that have been associated with yellow to brown pigmentation, hypovirulence, dsRNA infection, and decreased production of perithecia. In this study, degenerate isolates that produced yellow to brown pigments and no perithecia were obtained from wild-type cultures that had been stored for extended periods of time. Cultures of the degenerate isolates were found to accumulate five related hexaketides when grown on potato-dextrose agar (PDA). In contrast, these hexaketides were present only in minute amounts in wild-type cultures unless grown on NaCl-amended PDA. 1,8-Dihydroxynaphthalene melanin was established to be present in wild-type M. cannonballus and absent in the degenerate isolates. Various melanin-related metabolites, however, were produced by the variants. Tricyclazole in PDA cultures blocked melanin biosynthesis by the wild types but had little effect on hexaketide production by the degenerate isolates.

Melanins↗

Antifungal activity of camptothecin, trifolin, and hyperoside isolated from Camptotheca acuminata.

Leaf spots and root rots are major fungal diseases in Camptotheca acuminata that limit cultivation of the plant for camptothecin (CPT), a promising anticancer and antiviral alkaloid. Bioassays showed that pure CPT and flavonoids (trifolin and hyperoside) isolated from Camptotheca effectively control fungal pathogens in vitro, including Alternaria alternata, Epicoccum nigrum, Pestalotia guepinii, Drechslera sp., and Fusarium avenaceum, although antifungal activity of these compounds in the plant is limited. CPT inhibited mycelial growth by approximately 50% (EC50) at 10-30 microg/mL and fully inhibited growth at 75-125 microg/mL. The flavonoids were less effective than CPT at 50 microg/mL, particularly within 20 days after treatment, but more effective at 100 or 150 microg/mL. CPT, trifolin, and hyperoside may serve as leads for the development of fungicides.

Alternaria↗

Mechanisms of natural soil suppressiveness to soilborne diseases.

Suppressive soils are characterized by a very low level of disease development even though a virulent pathogen and susceptible host are present. Biotic and abiotic elements of the soil environment contribute to suppressiveness, however most defined systems have identified biological elements as primary factors in disease suppression. Many soils possess similarities with regard to microorganisms involved in disease suppression, while other attributes are unique to specific pathogen-suppressive soil systems. The organisms operative in pathogen suppression do so via diverse mechanisms including competition for nutrients, antibiosis and induction of host resistance. Non-pathogenic Fusarium spp. and fluorescent Pseudomonas spp. play a critical role in naturally occurring soils that are suppressive to Fusarium wilt. Suppression of take-all of wheat, caused by Gaeumannomyces graminis var. tritici, is induced in soil after continuous wheat monoculture and is attributed, in part, to selection of fluorescent pseudomonads with capacity to produce the antibiotic 2,4-diacetylphloroglucinol. Cultivation of orchard soils with specific wheat varieties induces suppressiveness to Rhizoctonia root rot of apple caused by Rhizoctonia solani AG 5. Wheat cultivars that stimulate disease suppression enhance populations of specific fluorescent pseudomonad genotypes with antagonistic activity toward this pathogen. Methods that transform resident microbial communities in a manner which induces natural soil suppressiveness have potential as components of environmentally sustainable systems for management of soilborne plant pathogens.

Antibiosis↗

Cloning and characterization of a totivirus double-stranded RNA from the plant pathogenic fungus, Helicobasidium mompa Tanaka.

Virus-like particles (VLPs, named HmTV1-17), about 40 nm in diameter were found in the violet root rot fungus Helicobasidium mompa Tanaka strain No. 17, which had been isolated from an apple tree. Purified preparations of HmTV1-17 contained two species of double-stranded RNA (dsRNA), designated 17L and 17S. cDNAs were constructed from HmTV1-17 genomic dsRNAs purified using CF-11 cellulose column chromatography. The sequences of 17L and 17S cDNA comprised 5,207 and 2,096 bp, respectively. Although 17S has no large open reading flame (ORF) on either strand, 17L has two large overlapping ORFs. The 5' located ORF1 encodes the coat protein (CP, 788 amino acids), whereas the gene product of ORF2, which is in the -1 frame relative to ORF1, shows the typical features of a RNA dependent RNA polymerase (RDRP, 845 amino acids). Phylogenetic analysis based on RDRP showed that HmTV1-17 is closely related to Sphaeropsis sapinea SsRV1, a member of the genus Totivirus from filamentous fungus S. sapinea.

Amino Acid Sequence↗

Can synthetic pesticides be replaced with biologically-based alternatives?--an industry perspective.

Agricultural chemical companies have invested in the discovery and development of biological pesticides to complement synthetic pesticides for the control of insects, diseases, and weeds on agronomic and horticultural crops. For plant disease control, companies envisage biological fungicides entering markets where they have the best chance of performing and which are most receptive to using biological control methods. Fewer regulatory requirements can mean faster registration for a biological than a synthetic pesticide. However, industry's requirements for competitive performance, effective formulations, and economic production can mean significant investments in time and money for a biological pesticide, although total investment may be less than for a synthetic pesticide. One biocontrol project in which industry has invested is baculoviruses for insect control. Insect baculoviruses, genetically modified to kill insects faster than wild-type viruses, are attractive biocontrol agents because their selectivity to insect pests and safety to beneficial insects and mammals enable them to compete with synthetic insecticides. Industry is looking for similar biocontrol opportunities in disease control. Biocontrol agents for seedling disease, root rot, and postharvest disease control have been registered by the EPA and are trying to compete with synthetic fungicides for market share. To date, effective biocontrol agents have not been identified for the control of serious foliar diseases, such as grape downy mildew, potato late blight, wheat powdery mildew, and apple scab. Farmers must rely on synthetic fungicides and agronomic methods to control these diseases for the foreseeable future.

Animals↗

Characterization and in vitro expression patterns of an exopolygalacturonase encoding gene from Fusarium oxysporum f.sp. radicis lycopersici.

AIMS: In this work, we report the isolation, characterization and expression pattern in in vitro cultures of an EXOPG encoding gene (pgx2), a novel EXOPG encoding gene of Fusarium oxysporum f.sp. radicis lycopersici, responsible for foot crown and root rot disease in tomato plants. The gene was compared with other fungal polygalacturonases (PGs) previously reported. METHODS AND RESULTS: Partial sequences of the purified EXOPG native protein were used to design primers that amplified a genomic fragment by PCR. The amplified genomic fragment was used as a probe to screen a genomic library. One isolated clone was analysed. The complete genomic, cDNA and the deduced amino acid sequences were compared with other fungal EXOPGs and ENDOPGs. Regulation of pgx2 expression was analysed by Northern blot in in vitro cultures supplemented with different carbon sources. CONCLUSIONS: Pgx2 was present as single copy in the haploid genome of several Fusarium species. PGX2 showed the conserved amino acid motifs typical of PGs and those reported for fungal EXOPGs. Pgx2 was regulated at transcriptional level showing similar expression pattern to other EXOPG encoding gene (pgx1) when the fungus was cultured on different carbon sources suggesting a coordinate expression of both genes. This similarity would be supported by the presence of common putative regulatory motifs in the upstream regions of both genes. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the analysis of a novel EXOPG gene of the tomato pathogen F. oxysporum f.sp. radicis lycopersici, a contribution to the understanding of the role of cell-wall-degrading enzymes produced by fungi during pathogenesis.

Amino Acid Sequence↗

Expression of a Phytophthora sojae necrosis-inducing protein occurs during transition from biotrophy to necrotrophy.

Phytophthora sojae is an oomycete that causes stem and root rot on soybean plants. To discover pathogen factors that produce disease symptoms or activate plant defense responses, we identified putative secretory proteins from expressed sequence tags (ESTs) and tested selected candidates using a heterologous expression assay. From an analysis of 3035 ESTs originating from mycelium, zoospore, and infected soybean tissues, we identified 176 putative secreted proteins. A total of 16 different cDNAs predicted to encode secreted proteins ranging in size from 6 to 26 kDa were selected for expression analysis in Nicotiana benthamiana using an Agrobacterium tumefaciens binary potato virus X (PVX) vector. This resulted in the identification of a 25.6-kDa necrosis-inducing protein that is similar in sequence to other proteins from eukaryotic and prokaryotic species. The genomic region encoding the P. sojae necrosis-inducing protein was isolated and the expression pattern of the corresponding gene determined by RNA blot hybridization and by RT-PCR. The activity of this P. sojae protein was compared to proteins of similar sequence from Fusarium oxysporum, Bacillus halodurans, and Streptomyces coelicolor by PVX-based expression in N. benthamiana and by transient expression via particle bombardment in soybean tissues. The P. sojae protein was a powerful inducer of necrosis and cell death in both assays, whereas related proteins from other species varied in their activity. This study suggests that the P. sojae necrosis-inducing protein facilitates the colonization of host tissues during the necrotrophic phase of growth.

Agrobacterium tumefaciens↗

Chitinolytic activity of an endophytic strain of Bacillus cereus.

Bacillus cereus strain 65, previously isolated as an endophyte of Sinapis, was shown to produce and excrete a chitinase with an apparent molecular mass of 36 kDa. The enzyme was classified as a chitobiosidase because it was able to cleave diacetylchitobiose (GlcNAc)2 from the non-reducing end of trimeric chitin derivatives. The chitinase exhibited activity over the pH range 4.5-7.5 and was stable between pH 4.0 and 8.5. The enzyme had an isoelectric point of 6.4. Application of B. cereus 65 directly to soil significantly protected cotton seedlings from root rot disease caused by Rhizoctonia solani.

Antifungal Agents↗

Isolation of a superfamily of candidate disease-resistance genes in soybean based on a conserved nucleotide-binding site.

The tobacco N and Arabidopsis RPS2 genes, among several recently cloned disease-resistance genes, share highly conserved structure, a nucleotide-binding site (NBS). Using degenerate oligonucleotide primers for the NBS region of N and RPS2, we have amplified and cloned the NBS sequences from soybean. Each of these PCR-derived NBS clones detected low-or moderate-copy soybean DNA sequences and belongs to 1 of 11 different classes. Sequence analysis showed that all PCR clones encode three motifs (P-loop, kinase-2, and kinase-3a) of NBS nearly identical to those in N and RPS2. The intervening region between P-loop and kinase-3a of the 11 classes has high (26% average) amino acid sequence similarity to the N gene although not as high (19% average) to RPS2. These 11 classes represent a superfamily of NBS-containing soybean genes that are homologous to N and RPS2. Each class or subfamily was assessed for its positional association with known soybean disease-resistance genes through near-isogenic line assays, followed by linkage analysis in F2 populations using restriction fragment length polymorphisms. Five of the 11 subfamilies have thus far been mapped to the vicinity of known soybean genes for resistance to potyviruses (Rsv1 and Rpv), Phytophthora root rot (Rps1, Rps2, and Rps3), and powdery mildew (rmd). The conserved N- or RPS2-homologous NBS sequences and their positional associations with mapped soybean-resistance genes suggest that a number of the soybean disease-resistance genes may belong to this superfamily. The candidate subfamilies of NBS-containing genes identified by genetic mapping should greatly facilitate the molecular cloning of disease-resistance genes.

Amino Acid Sequence↗

A novel tetratricopeptide repeat-containing J-protein localized in a plasma membrane-bound protein complex of the phytopathogenic oomycete Phytophthora megasperma.

Phytoalexins originating from plant tissues may cause within cells of fungi or oomycetes a change in the localization of actin, tubulin and chaperones. To test the hypothesis in a filamentously growing oomycete, we compared the distribution of cellular markers in the presence and absence of hydroxystilbene phytoalexins. Using cDNA from the phytopathogenic organism Phytophthora megasperma, the causal agent of root rot on soybean and many other plants, and including probes for Hsp70 and Hsp40, we cloned a DnaJ-protein (Jcp) with the capacity of interacting with both a particular Hsp70 isoform via its J-domain and with other proteins via its tetratricopeptide repeat (TPR) domain. Antisera raised against the bacterially expressed protein Jcp allowed the analysis of its intracellular localization during hyphal growth. Following the subfractionation of cell homogenates, we detected virtually all immunoreactive Jcp in the plasma membrane-enriched fraction and as constituent of a membrane-associated protein complex. In agreement with the biochemical findings, immunocytochemical stains of hyphae showed Jcp as part of cortical patches positioned along the plasma membrane similar to the distribution of actin patches. Confocal microscopy, however, revealed that the Jcp-containing patches did not generally co-localize with the patches visualized by the actin stain. The 59-kDa Jcp, characterized by a large 8-fold TPR domain at the N-terminal region and a J-domain close to the C-terminus, is a good candidate for bridging the gap between Hsp70 and Hsp90 by protein-protein interactions. By administration of plant-derived phytoalexins it was shown that the presence of resveratrol or piceatannol significantly reduces the amount of the Jcp-containing patches, but does not lead to a relocalization of intracellular Jcp.

Amino Acid Sequence↗

Two pathogenesis-related peroxidases in greengram (Vigna radiata (L.) wilczek) leaves and cultured cells induced by Macrophomina phaseolina (Tassi) Goid. and its elicitor.

An elicitor has been isolated from Macrophomina phaseolina, the root rot and leaf blight pathogen of greengram. Suspension-cultured cells of greengram were established which responded to the fungal elicitor. When greengram leaves were inoculated with M. phaseolina two new peroxidases appeared. Similarly, two new peroxidases could be detected in suspension-cultured greengram cells when treated with the fungal elicitor. These peroxidases were purified by column chromatography and their molecular masses were 27 and 38 kDa. The new peroxidases detected in both leaves and cultured cells appear to be similar with the same molecular weights.

Basidiomycota↗

Molecular characterization of Macrophomina phaseolina and Fusarium species by a single primer RAPD technique.

Charcoal root rot and wilt, are two economically important diseases of many crop plants in North and South America, Asia and Africa and some parts of Europe. Genetic variation in 43 isolates of Macrophomina phaseolina and 22 isolates of Fusarium species, collected from geographically distinct regions over a range of hosts, was studied using random amplified polymorphic DNA (RAPD) markers. Initially, 210 arbitrary nucleotide (10-mer) primers were tested for amplification of genomic DNA of one M. phaseolina isolate, 70 primers amplified the genomic DNA of M. phaseolina. One primer OPA-13 (5'-CAGCACCCAC-3') produced fingerprint profiles, which clearly distinguished between the different isolates of M. phaseolina. UPGMA analysis classified these isolates into five major groups. By primer OPA-13, 22 isolates of pathogenic and non-pathogenic Fusarium species of different formae-speciales and races, were also distinguished from M. phaseolina. This marker is useful for distinguishing between these two important plant pathogens irrespective of hosts, virulence spectrum and races. This is the first report of reliable diagnosis of two soilborne pathogens (root/collar rot and wilt causing pathogens) at the level of isolates, formae-speciales and races by a single primer RAPD procedure with uniform PCR conditions.

Basidiomycota↗

Mitochondrial genetics in a natural population of the plant pathogen armillaria.

Transmission and propagation of mitochondrial genotypes in fungi have not been previously investigated in the field. This study examined the distribution of nuclear and mitochondrial genotypes in a natural, local population of the fungal (Basidiomycetes) root-rot pathogen, Armillaria. Six vegetative clones, ranging in size up to 635 m, were identified on the basis of mating-type alleles. Mitochondrial DNA (mtDNA) restriction fragment patterns indicated that each vegetative clone has one, unique mtDNA type. However, as in other basidiomycetous fungi, biparental transmission of mitochondria following laboratory matings of sexually compatible haploid isolates of Armillaria resulted in a uniformly diploid mycelium that was a mosaic for both parental mitochondrial types. Therefore, either matings between monosporous, haploid isolates are uncommon in nature, or when mating does occur, cytoplasmic markers of one partner predominate during subsequent vegetative growth.

Basidiomycota↗

Organization, expression and evolution of a disease resistance gene cluster in soybean.

PCR amplification was previously used to identify a cluster of resistance gene analogues (RGAs) on soybean linkage group J. Resistance to powdery mildew (Rmd-c), Phytophthora stem and root rot (Rps2), and an ineffective nodulation gene (Rj2) map within this cluster. BAC fingerprinting and RGA-specific primers were used to develop a contig of BAC clones spanning this region in cultivar "Williams 82" [rps2, Rmd (adult onset), rj2]. Two cDNAs with homology to the TIR/NBD/LRR family of R-genes have also been mapped to opposite ends of a BAC in the contig Gm_Isb001_091F11 (BAC 91F11). Sequence analyses of BAC 91F11 identified 16 different resistance-like gene (RLG) sequences with homology to the TIR/NBD/LRR family of disease resistance genes. Four of these RLGs represent two potentially novel classes of disease resistance genes: TIR/NBD domains fused inframe to a putative defense-related protein (NtPRp27-like) and TIR domains fused inframe to soybean calmodulin Ca(2+)-binding domains. RT-PCR analyses using gene-specific primers allowed us to monitor the expression of individual genes in different tissues and developmental stages. Three genes appeared to be constitutively expressed, while three were differentially expressed. Analyses of the R-genes within this BAC suggest that R-gene evolution in soybean is a complex and dynamic process.

Amino Acid Sequence↗

Anatomical-based defense responses of Scots pine (Pinus sylvestris) stems to two fungal pathogens.

We investigated the cellular responses of stem tissues of mature Scots pine (Pinus sylvestris L.) trees to inoculations with two fungal pathogens. The bark beetle vectored fungus, Leptographium wingfieldii Morelet, induced longer lesions in the bark, stronger swelling of polyphenolic parenchyma cells, more polyphenol accumulation and increased ray parenchyma activity compared with the root rot fungus, Heterobasidion annosum (Fr.) Bref., or mechanical wounding. Axial resin ducts in the xylem are a general feature of the preformed defenses of Scots pine, but there was no clear induction of additional traumatic axial resin ducts in response to wounding or fungal infection. The anatomical responses of Scots pine to pathogen infection were localized to the infection site and were attenuated away from bark lesions. The responses observed in Scots pine were compared with published studies on Norway spruce (Picea abies (L.) Karst.) for which anatomically based defense responses have been well characterized.

Ascomycota↗

Changes in host chitinase isoforms in relation to wounding and colonization by Heterobasidion annosum: early and strong defense response in 33-year-old resistant Norway spruce clone.

We studied the defense reactions of 33-year-old susceptible and resistant clones of Norway spruce (Picea abies (L.) Karst.) to the major root-rot fungus Heterobasidion annosum (Fr.) Bref. and determined if tissue cultures can be used as a model system for studying defense responses of mature trees at the molecular level. Quantitative PCR analysis of genomic DNA obtained from samples taken at different times along the lesion length in living bark indicated that the fungus was present in higher amounts and extended further into the host tissue in the susceptible clone than in the resistant clone. In protein extracts from the same lesion samples, there were differences in temporal and spatial changes in host chitinase isoform profiles between the resistant and susceptible clones. Host chitinase isoforms with pI values approximately 4.8, 4.4 and 3.7 increased more during the first 7 days after wounding and inoculation and extended further along the lesion length in the resistant clone than in the susceptible clone. These results suggest that the time from wounding and infection to induction of defense-related expression is shorter in the resistant clone indicating a more efficient host defense response than in the susceptible clone. Tissue cultures from the same clones were not resistant to H. annosum and showed no difference in the timing of the increase in chitinase isoforms in response to the pathogen. However, tissue cultures from both clones showed an increase in chitinase isoforms within 6 to 24 h past inoculation, indicating that increased chitinase expression in response to the pathogen is part of a general defense response common to both mature clones and tissue cultures.

Basidiomycota↗