| adenine and pyridine nucleotides in forms of verticillium dahliae kleb. differing in pathogenicity. | | 1975 | 1214 |
| pentaketide metabolites of verticillium dahliae. ii. accumulation of naphthol derivatives by the aberrant-melanin mutant brm-2. | | 1977 | 557166 |
| ultrastructural and chemical distinction of melanins formed by verticillium dahliae from (+)-scytalone, 1,8-dihydroxynaphthalene, catechol, and l-3,4-dihydroxyphenylalanine. | microsclerotia of three melanin-deficient mutants of verticillium dahliae formed malanin from (+)-scytalone, 1,8-dihydroxynaphthalene, catechol, and l-3,4-dihydroxyphenylalanine. the melanins formed from (+)-scytalone or 1,8-dihydroxynaphthalene resembled wild-type melanin chemically and ultrastructurally, whereas the melanins formed from catechol and l-3,4-dihydroxyphenlalanine were different. this suggests that scytalone and 1,8-dihydroxynaphthalene but no catechol or l-3,4-dihydroxyphenylalan ... | 1978 | 565670 |
| osmotic effects on radial growth rate and specific growth rate of three soil fungi. | the radial growth rate on osmotically adjusted agar medium and the relative specific growth rate in osmotically adjusted liquid medium were determined for rhizoctonia solani, pythium ultimum, and verticillium dahliae. on basal medium, an isolate of p. ultimum and r. solani had similar radial growth rates of 0.52 and 0.47 mm/h, respectively, whereas v. dahliae grew at a rate of 0.08 mm/h. radial growth rate was reduced 50% at osmotic potentials of -16, -27, and -32 bars for p. ultimum, r. solani, ... | 1978 | 570448 |
| [multiple forms of pectin trans-eliminase from verticillium dahliae klebahn -- cotton wilt agent]. | from the culture liquid filtrate of verticillium dahliae--cotton wilt agent--pectin trans-eliminase (ec) was isolated. the enzyme was isolated and examined, using ultrafiltration, gel filtration, ion exchange chromatography, isoelectrofocusing, and electrophoresis. the fungus was found capable to produce several forms of pectin trans-eliminase that differed in their molecular weight, charge, synthesis and release regulation, substrate action (position of bonding breakdowns in the pectin polymer ... | 1978 | 571122 |
| ultrastructure of melanin formation in verticillium dahliae with (+)-scytalone as a biosynthetic intermediate. | transmission and scanning electron microscopy showed that melanin of wild-type verticillium dahliae occurred as granules in microsclerotial cell walls and in a fibrillar network encapsulating the walls. an albino microsclerotial mutant and a brown microsclerotial mutant of v. dahliae did not form melanin granules. when albino microsclerotia were treated with (+)-scytalone (a metabolite that the brown mutant accumulates), they formed melanin granules and turned black. these granules were similar ... | 1976 | 945119 |
| use of mutants to establish (+)-scytalone as an intermediate in melanin biosynthesis by verticillium dahliae. | melanin biosynthesis in verticillium dahliae kleb, was studied with mutants deficient for normal black melanin or for production of microsclerotia. seven genetically different mutants had apparent blocks in melanin biosynthesis. four mutants (brm-1 to -4) produced brown microsclerotia and extruded pigments into media; three (alm-1 to -3) produced albino microsclerotia. other mutants produced no microsclerotia (nms) or had greatly reduced numbers of microsclerotia (rms). mutation alm-1 was due to ... | 1976 | 945120 |
| pentaketide metabolites of verticillium dahliae. 3. identification of (-)-3,4-dihydro-3,8-dihydroxy-1(2h)-naphtalenone((-)-vermelone) as a precursor to melanin. | | 1976 | 945335 |
| the effect of volatile and gaseous metabolites of swelling seeds on germination of fungal spores. | effects of volatile and gaseous metabolites of swelling seeds of pea, bean, wheat, corn cucumber, tomato, lentil, carrot, red papper and lettuce on germination of spores of five genera of fungi were found to depend rather on the fungal than on the plant genus. germination of spores of botrytis cinerea, mucor racemosus and trichoderma viride was most severely inhibited. spores of verticillium dahliae were less sensitive and germination of spores of fusarium oxysporum was inhibited only in two cas ... | 1975 | 1176038 |
| expression and regulation of casein kinase 2 during heat shock in verticillium dahliae. | a homogeneous preparation of casein kinase 2 has been isolated from the phytopathogenic fungus verticillium dahliae (the parasite of cotton). the enzyme consists of three subunits with molecular masses of 53, 41, and 38 kda. highly specific immune serum against casein kinase 2 has been obtained. by means of immunoblotting, enzyme-linked immunosorbent assay (elisa), and immunochemical isolation on protein a-sepharose, it is shown that the amount of casein kinase 2 increases under heat shock condi ... | 1992 | 1562326 |
| production of xylanase by verticillium dahliae. | xylan induced the production of xylanase by verticillium dahliae. other cellulolytic enzymes such as glucanase and beta-glucosidase were synthesized in smaller quantities. the process of degradation indicated that xylanase behaved like a typical endo-enzyme causing first production of high mol, wt. products, and indicated that v. dahliae produced at least three enzymes which degrade xylan. | 1991 | 1775094 |
| [research of antifungal substances secreted by higher fungi in culture]. | a screening in vitro of antifungic activity of 24 strains of basidiomycetes was realized with their culture filtrate. lycoperdon perlatum pers. = pers., oudemansiella platyphylla (pers. ex fr.) mos., agrocybe dura (bolt) singer have shown an activity against candida albicans, candida tropicalis and aspergillus fumigatus; pholiota spumosa (fr.) singer towards botrytis cinerea and lycoperdon perlatum pers. = pers. towards alternaria solani, botrytis cinerea and verticillium dahliae. more extensive ... | 1990 | 2082797 |
| [organization of genes of ribosomal rna from the mushroom verticillium dahliae]. | using yeast probe, a complete ribosomal dna unit from a plant pathogenic fungus, verticillium dahliae, was cloned into a plasmid vector ptz19r. partial dna sequence of the clones, when compared to the yeast ribosomal dna sequence, allowed to establish the physical map of the fungal rdna. the overall organization was shown to be similar to other fungal rdnas previously known. | 1990 | 2094815 |
| [a comparative analysis of chromatographic, kinetic and antigenic properties of casein kinase type 2 from bovine liver and the phytopathogenic mushroom verticillium dahliae]. | homogeneous preparations of casein kinases 2 have been isolated from bovine liver and the phytopathogenic fungus verticillium dahliae. the isolation procedure was similar in both cases and included consecutive chromatography on heparin-sepharose, phosphocellulose pii and monoq. the bovine liver enzyme consists of two subunits with molecular masses of 38 kda and 27 kda, while the fungus kinase has three subunits with mr of 53 kda, 41 kda and 38 kda. self-phosphorylation of casein kinase 2 in vitr ... | 1990 | 2096946 |
| glucose oxidase as the antifungal principle of talaron from talaromyces flavus. | analysis of an authentic sample of the antifungal antibiotic talaron from the biocontrol fungus talaromyces flavus indicated that approximately 40% of the solid sample was glucose oxidase. high-performance liquid chromatography elution profiles of the antimicrobial activity of talaron coeluted with those of glucose oxidase. fluorescence emission and excitation wavelength maxima for talaron were similar to those of glucose oxidase from aspergillus niger. the molecular weight of talaron was 152,00 ... | 1990 | 2279238 |
| biosynthetic arginine decarboxylase in phytopathogenic fungi. | it has been reported that while bacteria and higher plants possess two different pathways for the biosynthesis of putrescine, via ornithine decarboxylase (odc) and arginine decarboxylase (adc); the fungi, like animals, only use the former pathway. we found that contrary to the earlier reports, two of the phytopathogenic fungi (ceratocystis minor and verticillium dahliae) contain significant levels of adc activity with very little odc. the adc in these fungi has high ph optimum (8.4) and low km ( ... | 1989 | 2497290 |
| elicitor stimulation of the defense response in cultured plant cells monitored by fluorescent dyes. | addition of fungal elicitors to plant cells in suspension is known to stimulate biochemical changes in the plant cell leading to production of defense compounds. in this paper we demonstrate that introduction of elicitors from the pathogenic fungus verticillium dahliae to cultured cotton, tobacco, or soybean cells leads to a rapid, dramatic change in the fluorescence of several membrane-associated potentiometric or ph-sensitive dyes. the fluorescence transitions occur abruptly following a brief ... | 1986 | 3753013 |
| melanin biosynthesis and the metabolism of flaviolin and 2-hydroxyjuglone in wangiella dermatitidis. | melanin biosynthesis in the human pathogen wangiella dermatitidis was inhibited by tricyclazole, causing pentaketide melanin metabolites to accumulate in the cultures. one of these metabolites, scytalone, was racemic and thus different than the (+)-enantiomer from verticillium dahliae. an albino mutant of w. dermatitidis metabolized scytalone to a pigment ultrastructurally identical to wild-type melanin. cell-free homogenates of the wild type carried out typical reductive and dehydrative reactio ... | 1985 | 4037982 |
| the development of lignitubers in roots after infection by verticillium dahliae kleb. | | 1971 | 4101737 |
| electrophoretic protein and enzyme patterns and antigenic structure in verticillium dahliae and v. albo-atrum. | | 1974 | 4374661 |
| [strain differences in the structure of verticillium dahliae dna]. | | 1973 | 4795946 |
| a quantitative bioassay for determining low numbers of microsclerotia of verticillium dahliae in field soils. | | 1974 | 4822783 |
| [effect of vitamins on the growth rate of the germ tubes and germinating microsclerotia and the subsequent morphogenesis of verticillium dahliae kleb]. | | 1974 | 4857618 |
| [effect of the antifungal antibiotic 18-80 on nucleic acids of verticillium dahliae]. | | 1972 | 5065190 |
| the fine structure of verticillium dahliae kleb. colonizing cellophane. | | 1971 | 5103059 |
| [electron microscopic study of the structure of verticillium dahliae]. | | 1971 | 5104437 |
| metabolites of verticillium dahliae: the isolation and structure elucidation of some disaccharides and a trisaccharide. | | 1971 | 5165992 |
| interactions between heterodera rostochiensis woll. and verticillium dahliae kleb. on potatoes and the effect of ccc on both. | | 1971 | 5167334 |
| [requirement of specific nutrient substances of the cotton wilt pathogen--verticillium dahliae kleb]. | | 1965 | 5893626 |
| [the role of pectolytic enzymes of the verticillium dahliae fungus in the development of cotton wilt]. | | 1965 | 5895993 |
| [weeds as host-plants for verticillium albo-atrum rke. et berth. including verticillium dahliae kleb]. | | 1966 | 5953031 |
| pentaketide metabolites of melanin synthesis in the dematiaceous fungus wangiella dermatitidis. | melanin synthesis in the dematiaceous, polymorphic hyphomycete wangiella dermatitidis, a human pathogen, was investigated by biochemical and physiological techniques. mutants with a decrease or loss in melanin synthesis were induced and isolated. melanin precursors were obtained from the mutants, purified, and then identified by comparison with authentic compounds from verticillium dahliae . isolation of scytalone , vermelone , flaviolin , and 1,8- dihydroxynaphthalene from the mutants of wangie ... | 1984 | 6539583 |
| volatile and gaseous metabolites released by germinating seeds of lentil and maize cultivars with different susceptibilities to fusariosis and smut. | the effect of volatile and gaseous metabolites released by germinating seeds of lentil cultivars more and less susceptible to fusariosis on the germination of spores of mucor racemosus, trichoderma viride, verticillium dahliae and botrytis cinerea was found to depend rather on the fungal genus than on the lentil cultivar. however, spores of fusarium oxysporum reacted more sensitively during germination to the presence of exudates of both cultivars, when the more susceptible lentil displayed a st ... | 1980 | 7189738 |
| effect of foliar application of antibiotics and gibberellic acid on the rhizosphre microflora of pea, infected with verticillium dahliae. | a study was made of the effects of foliar spray of bacitracin, chloramphenicol and gibberellic acid on the rhizosphere microflora of pea seedlings (pisum sativum l.) infected with verticillium dahliae. the antibiotics increased fungus and actinomycete counts and reduced the bacterial populations in the rhizosphere. gibberellic acid at 10 ppm concentration reduced all three groups of microorganisms while at 100 ppm fungi and actinomycetes increased slightly. invariably the rhizosphere effect was ... | 1980 | 7191387 |
| molecular analysis of japanese isolates of verticillium dahliae and v. albo-atrum. | sixteen isolates of different pathogenicity groups of the plant pathogen verticillium dahliae and four isolates of v. albo-atrum from japan were analysed by means of an rapd (random amplified polymorphic dna) method using a pcr (polymerase chain reaction). verticillium dahliae and v. albo-atrum could be distinguished by rapd analysis. four pathogenicity groups of v. dahliae could also be classified to a certain extent by this method. similarities and differences in banding patterns obtained by r ... | 1995 | 7639996 |
| pcr primers that allow intergeneric differentiation of ascomycetes and their application to verticillium spp. | a pair of conserved pcr primers, designated nms1 and nms2, that amplify a region in the mitochondrial small rrna gene region were designed for fungi belonging to the class ascomycetes. these primers were tested with members of eight fungal genera (aspergillus, fusarium, magnaporthe, mycospharella, neurospora, saccharomyces, sclerotinia, verticillium) and 10 verticillium species (verticillium albo-atrum, verticillium chlamydosporium, verticillium cinnebarium, verticillium dahliae, verticillium fu ... | 1994 | 7811072 |
| phenylalanine ammonia-lyase from cotton (gossypium hirsutum) hypocotyls: properties of the enzyme induced by a verticillium dahliae phytotoxin. | phenylalanine ammonia-lyase (ec 4.3.1.5), induced by a verticillium dahliae phytotoxin, has been purified to electrophoretic homogeneity from cotton hypocotyls by differential ammonium sulfate fractionation and hydrophobic interaction chromatography, with a yield of 52%. the enzyme is a tetramer with a molecular weight of 332,000 to 337,000. the isoelectric point is 4.6, and no isoforms were observed. the subunits of the enzyme are unstable and breaks down to fragments with m(r)'s of 69,000 and ... | 1994 | 8043606 |
| toxicity of cotton phytoalexins to zoopathogenic fungi. | the sesquiterpenoid phytoalexins desoxyhemigossypol, desoxymethoxyhemigossypol, and hemigossypolone formed in cotton (gossypium hirsutum and g. barbadense) stem xylem infected with verticillium dahliae were shown to be highly toxic to zoopathogenic fungi. this appears to be the first study of the toxicity of terpenoid phytoalexins to zoopathogenic fungi. the toxicities of the phytoalexins expressed as mic (micrograms ml-1) values were 8 to 128 against four isolates of candida albicans and one is ... | 1993 | 8167949 |
| differential enhancement of benzophenanthridine alkaloid content in cell suspension cultures of sanguinaria canadensis under conditions of combined hormonal deprivation and fungal elicitation. | an elicitation protocol, resulting in the accumulation of sanguinarine in suspension cultures of papaver bracteatum, was assessed for induction of the same alkaloid in sanguinaria canadensis. although only a trace constituent of p. bracteatum plants, sanguinarine is a major alkaloid (1-3% dry wt) of s. canadensis rhizomes. by combining hormonal deprivation for various intervals and a 3-day fungal (verticillium dahliae) elicitation, benzophenanthridine alkaloid accumulation was induced in s. cana ... | 1993 | 8229008 |
| high-affinity binding of a protein-lipopolysaccharide phytotoxin from verticillium dahliae to cotton membranes. | the presence of a specific binding site for a protein-lipopolysaccharide (plp) phytotoxin isolated from culture filtrates of verticillium dahliae has been demonstrated in plasma membranes from cotton seedlings. the 125i-labelled plp was used as a ligand in binding assays with plasma membrane enriched fractions. root tissue exhibited the highest amount of binding activity compared to hypocotyl and cotyledon tissue. binding of the [125i]plp was saturable, reversible, and with an affinity (kd = 1.4 ... | 1993 | 8253197 |
| direct dna extraction for pcr-mediated assays of soil organisms. | by using the rdna of a plant wilt pathogen (verticillium dahliae) as the target sequence, a direct method for the extraction of dna from soil samples which can be used for pcr-mediated diagnostics without a need for further dna purification has been developed. the soil organisms are disrupted by grinding in liquid nitrogen with the natural abrasives in soil, and losses due to degradation and adsorption are largely eliminated by the addition of skim milk powder. the dna from disrupted cells is ex ... | 1995 | 8526511 |
| cloning, expression, and characterization of (+)-delta-cadinene synthase: a catalyst for cotton phytoalexin biosynthesis. | in cotton, sesquiterpene phytoalexins are elicited in response to bacterial or fungal infection. a gossypium arboreum cell suspension culture which produces the sesquiterpene phytoalexin gossypol showed a time-dependent 10-fold increase in a 1.9-kb mrna in response to a challenge by a preparation from verticillium dahliae. the mrna prepared from these elicited cultures was used to isolated two cdna clones that contain open frames coding for proteins of 554 amino acids with m(r) 64,096 and 64,118 ... | 1995 | 8554317 |
| differential induction of 3-hydroxy-3-methylglutaryl coa reductase in two cotton species following inoculation with verticillium. | gossypium barbadense cottons are typically more resistant to wilt pathogens than are gossypium hirsutum cultivars. both species make terpenoid phytoalexins in response to infection, implicating isoprenoid biosynthesis as a factor in resistance. conserved regions in plant 3-hydroxy-3-methylglutaryl coenzyme a reductase (hmgr), the first enzyme in the terpene biosynthesis pathway, were used to design polymerase chain reaction primers for cloning a fragment of a cotton hmgr gene. the clone was used ... | 1995 | 8664497 |
| in vitro analysis of the role of glucose oxidase from talaromyces flavus in biocontrol of the plant pathogen verticillium dahliae. | culture filtrates from talaromyces flavus grown on glucose contained high levels of glucose oxidase activity, while culture filtrates from t. flavus grown on xylan contained negligible glucose oxidase activity. culture filtrates from t-flavus grown on both media contained complex protein profiles. however, only culture filtrates from t. flavus grown on glucose inhibited germination of microsclerotia of verticillium dahliae in in vitro inhibition assays. a polyclonal antiserum preparation, pabgo- ... | 1996 | 8795208 |
| cloning and heterologous expression of a second (+)-delta-cadinene synthase from gossypium arboreum. | screening of a gossypium arboreum l. cv. nanking cdna library resulted in the identification and cloning of a second (+)-delta-cadinene synthase. a probe for the screens was prepared by pcr using primers based on conserved sequences in farnesyl diphosphate cyclases and genomic dna as a template. this second cdna clone encodes a protein that is 80% identical to the recently described (+)-delta-cadinene synthases cad1-c1 and c14 from g. arboreum and maintains a significant degree of homology to th ... | 1996 | 8904844 |
| [cloning of dna probes for detection and identification of pathogenic fungi verticillium dahliae i verticillium tricorpus]. | | 1996 | 9026717 |
| production of an extracellular trypsin-like protease by the fungal plant pathogen verticillium dahliae. | the plant pathogenic fungus verticillium dahliae produced extracellular alkaline protease activity when grown in liquid medium supplemented with a protein source. a serine protease was purified 80-fold in a single step, using cation-exchange chromatography, from the filtrate of cultures grown with skim milk as a protein source. n-terminal amino acid sequence analysis of the 30-kda protein (vdp30) that copurified with the serine protease activity suggested that vdp30 is a trypsin-like protein. th ... | 1997 | 9090111 |
| two isoforms of np24: a thaumatin-like protein in tomato fruit. | two isoforms (i and ii) of the thaumatin-like protein np24 have been discovered in extracts of tomato fruit. np24 i increased ca 8-fold during ripening of tomato fruit, whereas np24 ii increased slightly as the fruit began to change colour and them remained constant. both proteins were purified to homogeneity and one of them, np24 i, was obtained in crystalline form. the proteins are similar except for differences in binding to cation exchangers and in two amino acids of the n-terminal amino aci ... | 1997 | 9115696 |
| adaptation of proteases and carbohydrates of saprophytic, phytopathogenic and entomopathogenic fungi to the requirements of their ecological niches. | the abilities of isolates of saprophytes (neurospora crassa, aspergillus nidulans), an opportunistic human pathogen (aspergillus fumigatus), an opportunistic insect pathogen (aspergillus flavus), plant pathogens (verticillium albo-atrum, verticillium dahliae, nectria haematococca), a mushroom pathogen (verticillium fungicola) and entomopathogens (verticillium lecanii, beauveria bassiana, metarhizium anisopliae) to utilize plant cell walls and insect cuticle components in different nutrient media ... | 1997 | 9202474 |
| isolation of the glucose oxidase gene from talaromyces flavus and characterisation of its role in the biocontrol of verticillium dahliae. | the glucose oxidase gene from the biocontrol fungus talaromyces flavus has been isolated and shown to be only 64% identical at the amino-acid sequence level to the similar enzyme from aspergillus niger. a transformation system has been developed for both t. flavus and the related t. macrosporus and has been used to create talaromyces spp. which either over-express or are deficient in glucose oxidase. in vitro inhibition experiments on verticillium dahliae using culture filtrates from these trans ... | 1997 | 9371889 |
| interlaboratory comparison of methods to quantify microsclerotia of verticillium dahliae in soil | in a comparison of different methods for estimating verticillium dahliae in soil, 14 soil samples were analyzed in a blinded fashion by 13 research groups in seven countries, using their preferred methods. one group analyzed only four samples. twelve soil samples were naturally infested, and two had known numbers of microsclerotia of v. dahliae added to them. in addition, a control was included to determine whether transport had an effect on the results. results differed considerably among the r ... | 1998 | 9758809 |
| identification of disease response genes expressed in gossypium hirsutum upon infection with the wilt pathogen verticillium dahliae. | verticillium wilt is a vascular disease of cotton (gossypium spp.) caused by the fungal pathogen verticillium dahliae. to begin to understand the molecular mechanisms of the disease response in cotton cultivars that display superior wilt tolerance, such as gossypium hirsutum cv. sicala v-1, a cdna library was constructed with mrna isolated from root tissue of sicala v-1, 24 h after inoculation with v. dahliae. the library was screened by a differential screening technique which was successful in ... | 1999 | 10412907 |
| induction of delta-cadinene synthase and sesquiterpenoid phytoalexins in cotton by verticillium dahliae. | phytoalexin biosynthesis occurred earlier in the resistant cotton cultivar seabrook sea island 12b2 (sbsi) (gossypium barbadense) than in the susceptible cotton cultivar rowden (g. hirsutum) after inoculation with a defoliating isolate of the pathogen verticillium dahliae. this was demonstrated by significantly higher levels of phytoalexins in sbsi 12 h after inoculation. furthermore, by 48 h after inoculation of sbsi, the phytoalexins hemigossypol and desoxyhemigossypol achieved levels (23.9 an ... | 1999 | 10552825 |
| purification and characterization of s-adenosyl-l-methionine: desoxyhemigossypol-6-o-methyltransferase from cotton plants. an enzyme capable of methylating the defense terpenoids of cotton. | cotton contains a unique group of terpenoids including desoxyhemigossypol, hemigossypol, gossypol, hemigossypolone, and the heliocides that are part of the plant's defense system against pathogenic fungi and insects. desoxyhemigossypol is a key intermediate in the biosynthesis of these compounds. we have isolated, purified, and characterized from cotton stele tissue infected with verticillium dahliae a methyltransferase (s-adenosyl-l-met: desoxyhemigossypol-6-o-methyltransferase) that specifical ... | 1999 | 10557251 |
| expression pattern of genes encoding farnesyl diphosphate synthase and sesquiterpene cyclase in cotton suspension-cultured cells treated with fungal elicitors. | cotton plants accumulate sesquiterpene aldehydes in pigment glands. the two enzymes farnesyl diphosphate synthase (fps) and (+)-delta-cadinene synthase (cad), a sesquiterpene cyclase, are involved in the biosynthesis of these secondary metabolites. a full-length cdna (garfps) encoding fps was isolated from gossypium arboreum and identified by in vitro enzymatic assay of the garfps protein heterologously expressed in escherichia coli. treatment of g. arboreum suspension-cultured cells with an eli ... | 1999 | 10624017 |
| transgenic expression of a gene encoding a synthetic antimicrobial peptide results in inhibition of fungal growth in vitro and in planta. | transgenic tobacco plants producing the synthetic antimicrobial peptide d4e1, encoded by a gene under the control of an enhanced cauliflower mosaic virus 35s rna promoter, were obtained by agrobacterium-mediated transformation. successful transformation was demonstrated by pcr and southern hybridization analysis of tobacco dnas. expression of the synthetic d4e1 gene was shown by rt-pcr of tobacco mrna. crude protein extracts from leaf tissue of transformed plants significantly reduced the number ... | 2000 | 10729616 |
| the complete dna sequence of the nuclear ribosomal rna gene complex of verticillium dahliae: intraspecific heterogeneity within the intergenic spacer region. | the complete dna sequence of the nuclear ribosomal rna gene complex of verticillium dahliae: intraspecific heterogeneity within the intergenic spacer region. fungal genetics and biology 29, 19-27. the complete sequence of the nuclear ribosomal dna gene complex of the phytopathogenic fungus verticillium dahliae has been determined. the tandemly repeated unit was 7216 bp long and appears to be the shortest rdna cluster described so far among filamentous fungi. primer pairs were designed for amplif ... | 2000 | 10779396 |
| expression pattern of (+)-delta-cadinene synthase genes and biosynthesis of sesquiterpene aldehydes in plants of gossypium arboreum l. | the cotton (+)-delta-cadinene synthase, a sesquiterpene cyclase, is encoded by a complex gene family which, based on homology, can be divided into two subfamilies: cad1-a and cad1-c. southern blots revealed several members of the cad1-c subfamily, and a single member of the cad1-a subfamily, in the diploid gossypium arboreum genome. one of the cad1-c genes, cad1-c3, was isolated from this species. according to reverse transcriptase-polymerase chain reaction, transcripts of both cad1-c and cad1-a ... | 2000 | 10787059 |
| terpenoids and flavonoids from artemisia species. | a phytochemical reinvestigation of the aerial parts of artemisia sieversiana gave a new guaianolide and two known flavones (chrysosplenetin and 5-hydroxy-3',4',6,7-tetramethoxyflavone). antifungal fractions derived from the chloroform extract of a. annua afforded two cadinane derivatives (arteannuin b and artemisinin), oleanolic acid, beta-sitosterol, stigmasterol, and the four flavones artemetin, bonanzin, eupalitin and chrysosplenetin. their structures were elucidated by spectral methods. all ... | 2000 | 10865468 |
| the complete dna sequence of the nuclear ribosomal rna gene complex of verticillium dahliae: intraspecific heterogeneity within the intergenic spacer region. | the complete sequence of the nuclear ribosomal dna gene complex of the phytopathogenic fungus verticillium dahliae has been determined. the tandemly repeated unit was 7216 bp long and appears to be the shortest rdna cluster described so far among filamentous fungi. primer pairs were designed for amplification of the region spanning half of the 28s subunit, the intergenic spacer (igs), and the 5' end of 18s subunit of a number of verticillium strains, isolated from various hosts and geographic or ... | 2000 | 10919381 |
| establishment of introduced antagonistic bacteria in the rhizosphere of transgenic potatoes and their effect on the bacterial community. | in a field release experiment, rifampicin resistant mutants of two antagonistic plant-associated bacteria were used for seed tuber inoculation of transgenic t4 lysozyme expressing potatoes, transgenic control potatoes and non-transgenic parental potatoes. the t4 lysozyme tolerant pseudomonas putida qc14-3-8 was originally isolated from the tuber surface (geocaulosphere) of t4 lysozyme producing plants and showed in vitro antibacterial activity to the bacterial pathogen erwinia carotovora ssp. at ... | 2000 | 10922502 |
| fungal pathogen protection in potato by expression of a plant defensin peptide. | defensins are small cysteine-rich peptides with antimicrobial activity. we demonstrate that the alfalfa antifungal peptide (alfafp) defensin isolated from seeds of medicago sativa displays strong activity against the agronomically important fungal pathogen verticillium dahliae. expression of the alfafp peptide in transgenic potato plants provides robust resistance in the greenhouse. importantly, this resistance is maintained under field conditions. there have been no previous demonstrations of a ... | 2000 | 11101813 |
| successful strategy for the selection of new strawberry-associated rhizobacteria antagonistic to verticillium wilt. | in order to isolate and characterize new strawberry-associated bacteria antagonistic to the soil-borne pathogenic fungus verticillium dahliae kleb., rhizobacterial populations from two different strawberry species, greenish strawberry (fragaria viridis) and garden strawberry (f. x ananassa) obtained after plating onto king's b and glycerol-arginine agar, were screened for in vitro antagonism toward v. dahliae. the proportion of isolates with antifungal activity determined in in vitro assay again ... | 2000 | 11142403 |
| identification of group-i introns at three different positions within the 28s rdna gene of the entomopathogenic fungus metarhizium anisopliae var. anisopliae. | using a set of heterologous primers designed from the 3'-end of the 28s rrna gene of verticillium dahliae the corresponding gene region of 30 isolates of the entomopathogenic fungus metarhizium anisopliae var. anisopliae was amplified. the polymerase chain reaction products obtained could be classified into four groups varying in size from 1.0 to 2.2 kb. sequence analyses of representative pcr products revealed the presence of five distinct introns, positioned in three different insertion sites. ... | 2000 | 11170737 |
| inhibition of polygalacturonase from verticillium dahliae by a polygalacturonase inhibiting protein from cotton. | an extracellular endo-polygalacturonase (pgase) [e.c. 3.2.1.15] was isolated from 18-day-old culture filtrates of verticillium dahliae and partially purified using gel permeation chromatography. the band responsible for pgase activity was electrophoretically characterized as having a molecular mass of approximately 29 500 and an isoelectric point of 5.4. kinetic studies indicate a km of 3.3 mg ml(-1) and vmax of 0.85 micromol reducing units min(-1) ml(-1) with polygalacturonic acid as substrate. ... | 2001 | 11382229 |
| broad-spectrum antimicrobial activity in vitro of the synthetic peptide d4e1. | broad-spectrum antimicrobial activity of a synthetic peptide, d4e1, is documented in this paper. d4e1 inhibited the growth of several fungal phytopathogens belonging to four classes-ascomycetes, basidiomycetes, deuteromycetes, and oomycetes, and two bacterial pathogens, pseudomonas syringae pv. tabaci and xanthomonas campestris pv. malvacearum race 18. the minimum inhibitory concentration (mic) of d4e1 required to completely inhibit the growth of all fungi studied ranged from 4.67 to 25 microm. ... | 2001 | 11409968 |
| the complete dna sequence of the nuclear ribosomal rna gene complex of verticillium dahliae: intraspecific heterogeneity within the intergenic spacer region. | the complete sequence of the nuclear ribosomal dna gene complex of the phytopathogenic fungus verticillium dahliae has been determined. the tandemly repeated unit was 7216 bp long and appears to be the shortest rdna cluster described so far among filamentous fungi. primer pairs were designed for amplification of the region spanning half of the 28s subunit, the intergenic spacer (igs), and the 5' end of 18s subunit of a number of verticillium strains, isolated from various hosts and geographic or ... | 2000 | 11419483 |
| bulk and rhizosphere soil bacterial communities studied by denaturing gradient gel electrophoresis: plant-dependent enrichment and seasonal shifts revealed. | the bacterial rhizosphere communities of three host plants of the pathogenic fungus verticillium dahliae, field-grown strawberry (fragaria ananassa duch.), oilseed rape (brassica napus l.), and potato (solanum tuberosum l.), were analyzed. we aimed to determine the degree to which the rhizosphere effect is plant dependent and whether this effect would be increased by growing the same crops in two consecutive years. rhizosphere or soil samples were taken five times over the vegetation periods. to ... | 2001 | 11571180 |
| molecular cloning and functional identification of (+)-delta-cadinene-8-hydroxylase, a cytochrome p450 mono-oxygenase (cyp706b1) of cotton sesquiterpene biosynthesis. | in cotton, gossypol and related sesquiterpene aldehydes are present in the glands of aerial tissues and in epidermal cells of roots. a cytochrome p450 was found to be expressed in aerial tissues of glanded cotton cultivars, but not or at an extremely low level in the aerial tissues of a glandless cultivar. its cdna was then isolated from gossypium arboreum l. after expression in saccharomyces cerevisiae, the p450 was found to catalyse the hydroxylation of (+)-delta-cadinene, forming 8-hydroxy-(+ ... | 2001 | 11696190 |
| expression of an antimicrobial peptide via the chloroplast genome to control phytopathogenic bacteria and fungi. | the antimicrobial peptide msi-99, an analog of magainin 2, was expressed via the chloroplast genome to obtain high levels of expression in transgenic tobacco (nicotiana tabacum var. petit havana) plants. polymerase chain reaction products and southern blots confirmed integration of msi-99 into the chloroplast genome and achievement of homoplasmy, whereas northern blots confirmed transcription. contrary to previous predictions, accumulation of msi-99 in transgenic chloroplasts did not affect norm ... | 2001 | 11706168 |
| sesquiterpenoids in root exudates of solanum aethiopicum. | five known sesquiterpenoids, solavetivone, lubimin, lubiminoic acid, aethione and lubiminol were isolated from the root exudates recovered from solanum aethiopicum by a newly proposed method using charcoal. quantitative analysis of the sesquiterpenoids in the root exudates of s. aethiopicum and s. melongena suggested that relatively large amounts of the sesquiterpenoids were exuded from the roots. antifungal activity of the sesquiterpenoids against fusarium oxysporum and verticillium dahliae was ... | 2001 | 11724373 |
| elemental sulfur and thiol accumulation in tomato and defense against a fungal vascular pathogen. | the occurrence of fungicidal, elemental s is well documented in certain specialized prokaryotes, but has rarely been detected in eukaryotes. elemental s was first identified in this laboratory as a novel phytoalexin in the xylem of resistant genotypes of theobroma cacao, after infection by the vascular, fungal pathogen verticillium dahliae. in the current work, this phenomenon is demonstrated in a resistant line of tomato, lycopersicon esculentum, in response to v. dahliae. a novel gas chromatog ... | 2002 | 11788760 |
| evaluation of potential biocontrol rhizobacteria from different host plants of verticillium dahliae kleb. | a screening approach was developed to assess the potential of rhizobacterial strains to control verticillium wilt caused by verticillium dahliae kleb. | 2001 | 11851803 |
| isozyme variation in verticillium dahliae isolates from crete. | fifteen isolates of verticillium dahliae (eight of race 1, seven of race 2; most from the island of crete, greece) were examined for isozyme and molecular variation. among the isozyme banding patterns (zymograms) of six enzymes that were "activity-stained" after electrophoresis in 9% polyacrylamide gels, differences were observed in diaphorase, alpha-esterase, peroxidase and superoxide dismutase; 2, 2, 3 and 5 different types of zymograms were recorded, respectively. the zymograms could not be c ... | 2002 | 12058396 |
| plant-dependent genotypic and phenotypic diversity of antagonistic rhizobacteria isolated from different verticillium host plants. | to study the effect of plant species on the abundance and diversity of bacterial antagonists, the abundance, the phenotypic diversity, and the genotypic diversity of rhizobacteria isolated from potato, oilseed rape, and strawberry and from bulk soil which showed antagonistic activity towards the soilborne pathogen verticillium dahliae kleb. were analyzed. rhizosphere and soil samples were taken five times over two growing seasons in 1998 and 1999 from a randomized field trial. bacterial isolates ... | 2002 | 12089011 |
| activation of phospholipase a by plant defense elicitors. | participation of phospholipase a (plase a) in plant signal transduction has been documented for auxin stimulation of growth but not for elicitation of any plant defense response. in this paper, we report two independent assays for monitoring plase a induction in plant cells and have used these assays to evaluate whether transduction of defense-related signals might require plase a activation. oligogalacturonic acid, a potent elicitor of the soybean (glycine max) h2o2 burst, was unable to stimula ... | 1996 | 12226235 |
| antagonistic effects of several bacteria on verticillium dahliae the causal agent of cotton wilt. | experiments were carried out with 89 bacterial isolates that were collected from cotton rhizosphere in gorgan province. the antagonistic effects of bacterial isolates on verticillium dahliae klebahn were studied using dual culture test. five highly effective isolates were selected from these antagonists for subsequent studies. according to the biochemical, physiological and morphological tests, isolates 2020 and 3 were identified as pseudomonas fluorescens and isolate 204, 202 and 309 as bacillu ... | 2001 | 12425025 |
| potato-associated bacteria and their antagonistic potential towards plant-pathogenic fungi and the plant-parasitic nematode meloidogyne incognita (kofoid & white) chitwood. | to study the effect of microenvironments on potato-associated bacteria, the abundance and diversity of bacteria isolated from the rhizosphere, phyllosphere, endorhiza, and endosphere of field grown potato was analyzed. culturable bacteria were obtained after plating on r2a medium. the endophytic populations averaged 10(3) and 10(5) cfu/g (fresh wt.) for the endosphere and endorhiza. respectively, which were lower than those for the ectophytic microenvironments, with 10(5) and 10(7) cfu/g (fresh ... | 2002 | 12455609 |
| sequence tag analysis of gene expression during pathogenic growth and microsclerotia development in the vascular wilt pathogen verticillium dahliae. | two cdna libraries were constructed from cultures of the vascular wilt fungus verticillium dahliae, grown either in simulated xylem fluid medium (sxm) or under conditions that induce near-synchronous development of microsclerotia. expressed sequence tags (ests) were obtained for over 1000 clones from each library. most sequences in the two est collections were unique; nearly 55% of the translated ests had strong similarity to protein sequences in the ncbi nonredundant database. ests correspondin ... | 2003 | 12553936 |
| regioselective glucosylation of pyridoxine by microorganisms. | microorganisms from culture collections and isolates from nature were screened for the ability to catalyze the regioselective glucosylation of pyridoxine (pn) to produce pyridoxine 5'-alpha-d-glucoside (pn-5'-alpha-g) or pyridoxine 4'-alpha-d-glucoside (pn-4'-alpha-g). transglucosylation activity specific to 5'-position of pn was found in fungi belonging to genera such as coriolus and verticillium, and activity at the 4'-position of pn was found in bacteria belonging to genera such as bacillus a ... | 2003 | 12723596 |
| improvement in 5'-position-selective glucosylation of pyridoxine by verticillium dahliae tpu 4900. | optimization of culture and reaction conditions for 5'-position-selective transglucosylation to pyridoxine by verticillium dahliae tpu 4900 was investigated. v. dahliae tpu 4900 had high transglucosylation activity when grown with soluble starch as a carbon source and organic nitrogens such as esusan meat as a nitrogen source at 15-20 degrees c. both the yield of pyridoxine 5'-alpha-d-glucoside (pn-5'-alpha-g) and the 5'-position-selectivity reached a maximum when an intact-cell reaction was don ... | 2003 | 12723597 |
| design and development of a dna array for rapid detection and identification of multiple tomato vascular wilt pathogens. | fusarium wilt, caused by fusarium oxysporum f. sp. lycopersici, and verticillium wilt, caused by either verticillium albo-atrum or verticillium dahliae, are devastating diseases of tomato (lycopersicon esculentum) found worldwide. monitoring is the cornerstone of integrated pest management of any disease. the lack of rapid, accurate, and reliable means by which plant pathogens can be detected and identified is one of the main limitations in integrated disease management. in this paper, we descri ... | 2003 | 12799009 |
| synthesis and fungicidal activity of lipophylic n- and o-acyl derivatives of beta-hydroxy dl-alpha-amino acids. | synthesis of n- and o-acyl derivatives of dl-serine and threo-dl-phenylserine was accomplished by a regioselective acylation of the corresponding amino acid. the residues introduced into amino acid structure contain hydrophobic long chain or aromatic, namely lauroyl, myristoyl and phenylacetyl moieties. the fungicidal activity against six strains of fungi was studied. several compounds were found to be effective against growth of fungi, and o-myristoyl-dl-serine 2 and n-phenylacetyl-threo-dl-phe ... | 2003 | 12872931 |
| cynodontin: a fungal metabolite with antifungal properties. | a red pigment that accumulates in cultures of a drechslera avenae pathotype with specificity for avena sterilis was isolated and identified as the anthraquinone cynodontin (3-methyl-1,4,5,8-tetrahydroxyanthraquinone). satisfactory yield of the compound was obtained with 20-60 day incubations at temperatures between 20 and 27 degrees c. cynodontin was tested in vitro for fungitoxicity and was found to be a potent inhibitor of the growth of sclerotinia minor, sclerotinia sclerotiorum, and botrytis ... | 2003 | 12903946 |
| cloning of two contrasting high-affinity sulfate transporters from tomato induced by low sulfate and infection by the vascular pathogen verticillium dahliae. | two cdnas, lest1-1 (af347613) and lest1-2 (af347614), encoding sulfate transporters have been cloned from tomato (lycopersicon esculentum mill.) by reverse transcription-polymerase chain reaction and their expression characterised. sharing 76% identity at the amino acid level, the transporters are phylogenetically associated with the group-1, high-affinity plant sulfate transporters. both were shown to have high affinity for sulfate by uptake kinetic analysis using a yeast (saccharomyces cerevis ... | 2003 | 12937983 |
| identification of a locus controlling verticillium disease symptom response in arabidopsis thaliana. | verticillium dahliae klebahn is a soil-borne fungal pathogen causing vascular diseases. the pathogen penetrates the host through the roots, spreads through the xylem, and systemically colonizes both resistant and susceptible genotypes. to elucidate the genetic and molecular bases of plant-verticillium interactions, we have developed a pathosystem utilizing arabidopsis thaliana and an isolate of v. dahliae pathogenic to both cruciferous and non-cruciferous crops. relative tolerance (based on symp ... | 2003 | 12940951 |
| morphology of verticillium dahliae and v. tricorpus on semi-selective media used for the detection of v. dahliae in soil. | the morphology of two soil-borne verticillium species, v. dahliae and v. tricorpus, was studied on two semi-selective agar media, in the absence and presence of soil. morphology of the fungi differed considerably between the media, with respect to presence and shape of microsclerotia, dark hyphae (i.e. short melanised hyphae attached to the microsclerotia) and dark mycelium (i.e. melanised mycelium throughout the colony). on modified soil extract agar (msea), a pectate based agar, v. dahliae alw ... | 2003 | 12967209 |
| mining data from potato pedigrees: tracking the origin of susceptibility and resistance to verticillium dahliae in north american cultivars through molecular marker analysis. | potato ( solanum tuberosum l.) cultivated in north america is an autotetraploid species with a narrow genetic base. most of the popular commercial cultivars are susceptible to verticillium dahliae, a fungal pathogen causing verticillium wilt disease, though some cultivars with relatively high resistance also exist. we have used the available pedigree information to track the origin of susceptibility and resistance to verticillium wilt present in cultivated potatoes. one hundred thirty-nine potat ... | 2004 | 14523519 |
| linkage disequilibrium mapping of a verticillium dahliae resistance quantitative trait locus in tetraploid potato ( solanum tuberosum) through a candidate gene approach. | we have used the linkage disequilibrium mapping method to test for an association between a candidate gene marker and resistance to verticillium dahliae in tetraploid potato. a probe derived from the tomato verticillium resistance gene ( ve1) identified homologous sequences ( stve1) in potato, which in a diploid population map to chromosome 9, in a position analogous to that of the tomato resistance gene. when a molecular marker closely linked (1.5 cm) to the homologues was used as a candidate g ... | 2004 | 14523522 |
| cloning and targeted disruption, via agrobacterium tumefaciens-mediated transformation, of a trypsin protease gene from the vascular wilt fungus verticillium dahliae. | a gene encoding a trypsin protease was isolated from a tomato isolate of verticillium dahliae. the gene, designated vtp1, contains two introns and is predicted to encode a protein of 256 amino acids. the gene is present in v. dahliae isolates from different host plants and in v. albo-atrum; weakly hybridizing sequences are present in v. tricorpus. vtp1 cdna sequences were identified in a sequence tag analysis of genes expressed under growth conditions that promote microsclerotia development. rep ... | 2004 | 14618375 |
| use of borate to control the 5'-position-selective microbial glucosylation of pyridoxine. | nearly 100% 5'-position selectivity of transglucosylation from maltodextrin to pyridoxine (pn) by cells of verticillium dahliae tpu 4900 was observed when the reaction was carried out with borate. the same effect of borate was observed not only during synthesis of pyridoxine 5'-alpha-d-glucoside by partially purified enzyme of this strain but also during synthesis of this compound by other microorganisms and with other enzymes (alpha-glucosidase and cyclomaltodextrin glucanotransferase). the eff ... | 2003 | 14660349 |
| molecular cloning of a potential verticillium dahliae resistance gene slve1 with multi-site polyadenylation from solanum licopersicoides. | caused by verticillium spp. pathogens, verticillium wilt is a common detrimental disease damaging yield production of many important crops. isolation of verticillium wilt resistance genes and their transgenic application is a fundamental way to control this disease. here we report the cloning and sequence characterization of a potential verticillium dahliae kleb. resistance gene (ve) from solanum lycopersicoides dun. (designated as slve1). the nucleotide sequence of slve1 is 3400 bp with an orf ... | 2003 | 14756424 |
| the role of the jasmonate response in plant susceptibility to diverse pathogens with a range of lifestyles. | plants defend themselves against attack from insects and pathogens with various resistance strategies. the jasmonate and salicylate signaling pathways are two induced responses that protect plants against these attackers. knowledge of the range of organisms that are affected by each response is important for understanding how plants coordinate their defenses against multiple attackers and the generality of effect of different resistance mechanisms. the jasmonate response is known to protect plan ... | 2004 | 15133157 |
| phylogenetic analysis of verticillium species based on nuclear and mitochondrial sequences. | three different genes were sequenced from isolates of five plant-pathogenic verticillium species, verticillium albo-atrum, verticillium dahliae, verticillium longisporum, verticillium nigrescens, and verticillium tricorpus. the sequences covered parts of the mitochondrial cytochrome b gene ( cob), the mitochondrial small subunit rrna gene ( rns) and the nuclear its2 region. when the sequences were combined, the five species clustered in five monophyletic groups, with v. nigrescens distantly rela ... | 2004 | 15138663 |
| study on interaction between root-knot nematode meloidogyne javanica and wilt fungus verticillium dahliae on olive seedlings in greenhouse. | verticillium dahliae has been reported as a limiting factor in cotton, olive, potato and tomato fields from several countries in the world. root-knot nematodes meloidogyne javanica causes considerable damage to olive groves in olive growing areas. since the presence of these two pathogens in olive trees and seedlings were confirmed in golestan province, this study was proposed to find the mode of their action and interaction with olive seedlings in greenhouse. the non-defoliant strain of the fun ... | 2003 | 15149103 |
| elemental sulphur as an induced antifungal substance in plant defence. | man's oldest fungicide has probably long functioned in this role in plants, as a natural component of induced antifungal defence. elemental sulphur (s(0)) is the only inorganic phytoalexin and the only phytoalexin produced by so many different taxa. s(0) (detected by gc-ms as (32)s(8)) is produced in representative species of sterculiaceae (cocoa), solanaceae (tomato, tobacco), malvaceae (cotton), and leguminosae (french bean) in response to xylem-invading fungal and bacterial pathogens. product ... | 2004 | 15181110 |
| endophytic bacterial communities of field-grown potato plants and their plant-growth-promoting and antagonistic abilities. | to study the effect of plant growth on potato-associated bacteria, the composition and properties of bacteria colonizing the endosphere of field-grown potato were analyzed by a multiphasic approach. the occurrence and diversity of potato-associated bacteria were monitored by a cultivation-independent approach, using terminal restriction fragment length polymorphism analysis of 16s rdna. the patterns obtained revealed a high heterogeneity of community composition and suggested the existence of pl ... | 2004 | 15213748 |
| vdnep, an elicitor from verticillium dahliae, induces cotton plant wilting. | verticillium wilt is a vascular disease of cotton. the causal fungus, verticillium dahliae, secretes elicitors in culture. we have generated approximately 1,000 5'-terminal expressed sequence tags (ests) from a cultured mycelium of v. dahliae. a number of ests were found to encode proteins harboring putative signal peptides for secretion, and their cdnas were isolated. heterologous expression led to the identification of a protein with elicitor activities. this protein, named v. dahliae necrosis ... | 2004 | 15294839 |
| identification and quantification of gossypol in cotton by using packed micro-tips columns in combination with hplc. | self-packed micro-tip columns containing a c18-bonded silica stationary phase, based on the same principles as solid-phase extraction methods, were used to obtain gossypol and related sesquiterpenoid aldehyde-enriched fractions. the enriched metabolite fractions were then analyzed by optimized high-performance liquid chromatography (hplc) with a c18 column (4.6 mm x 25 cm) eluted with the binary mobile phase acetonitrile-0.1% aqueous tfa solution (80:20). this method has proven to be highly repr ... | 2004 | 15372136 |
| colonization and persistence of a plant growth-promoting bacterium pseudomonas fluorescens strain cs85, on roots of cotton seedlings. | pseudomonas fluorescens cs85, which was previously isolated from the rhizosphere of cotton seedlings, acts as both a plant growth-promoting bacterium and a biocontrol agent against cotton pathogens, including rhizoctonia solani, colletotrichum gossypii, fusarium oxysporum f sp. vasinfectum, and verticillium dahliae. strain cs85 was labeled separately with luxab and gusa. the labeled strains were stably maintained and had high levels of expression of the marker genes, luxab and gusa, after succes ... | 2004 | 15381971 |