Oxytropis

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Roger D. Bull - One of the best experts on this subject based on the ideXlab platform.

Zeki Kaya - One of the best experts on this subject based on the ideXlab platform.

  • phylogenetic relationships among native Oxytropis species in turkey using the trnl intron trnl f igs and trnv intron cpdna regions
    Turkish Journal of Botany, 2016
    Co-Authors: Ayten Tekpinar, Seher Karaman Erkul, Zeki Aytaç, Zeki Kaya
    Abstract:

    We tested the phylogenetic utility of three chloroplast DNA loci, i.e. the trnL(UAA) intron, trnL(UAA)-F(GAA) intergenic spacer (IGS), and trnV(UAC) intron, across thirteen native Oxytropis species. Our objective was to determine whether any of these chloroplast DNA markers could be beneficial to figure out phylogenetic relationships among Oxytropis species. To increase the interspecific sampling, nine sequences of the trnL intron and trnL-F regions were retrieved from GenBank and included in the analyses. No sequence of the trnV intron region was available in the database and so only sequences of the native species were used for the analyses. Phylogenies derived from maximum likelihood and maximum parsimony analyses indicated that the trnL intron and trnV intron regions provided better resolution for relationships among species with respect to the trnL-F region. The highest variable and parsimony informative sites were observed in the trnL intron region, while the lowest sites were seen in the trnV intron. Less variable sites for the trnV intron region were expected since no foreign sequences could be included in the analysis. Oxytropis lazica was phylogenetically separated from native species and clustered with foreign ones when the trnL intron and trnL-F regions were analyzed. The result obtained from the trnV intron region proved that Oxytropis engizekensis Duman and Vural may be a synonym of O. persica Boiss. A previous study that used morphological characters arrived at a similar conclusion.

Daniel Cook - One of the best experts on this subject based on the ideXlab platform.

  • evidence for nonpathogenic relationships of alternaria section undifilum endophytes within three host locoweed plant species
    Botany, 2018
    Co-Authors: Aziza Ibrahim Noor, Peter H Cooke, Daniel Cook, Amy Nava, Rebecca Creamer
    Abstract:

    Locoweeds are legumes of the Astragalus and Oxytropis genera that live symbiotically with the fungal endosymbionts belonging to Alternaria section Undifilum. These endophytes produce the toxin swainsonine, which causes a neurological syndrome (locosim) when the plant is ingested by grazing animals. Here we characterize the growth of the endophytes within plant tissues using confocal and scanning electron microscopy (SEM) to define the physical relationship. Microscopy results revealed the endophyte distribution to be densely networked, with the mycelia aligned parallel to the long axis of the plant stem and with no pathology to plant cell walls, xylem, or phloem. The autofluorescent mycelia were located intercellulary in the pith of stems. Stereofluoroscope observations of Alternaria Oxytropis, A. cinerea, and A. fulva in phytoagar showed that mycelial growth occurred at the tip of hyphae. The growth rate of A. cinerea was significantly faster than for A. Oxytropis or A. fulva. All three species of endoph...

  • Detection of swainsonine and isolation of the endophyte Undifilum from the major locoweeds in Inner Mongolia
    Biochemical Systematics and Ecology, 2012
    Co-Authors: Xinlei Gao, Dale R Gardner, Daniel Cook, Kip E. Panter, Stephen T. Lee, Ling Yan, Bing Han, Meng-li Zhao
    Abstract:

    Abstract Locoweeds are Astragalus and Oxytropis species that contain the toxic alkaloid swainsonine, causing widespread poisoning of livestock in Inner Mongolia. Taxa ( Astragalus , Oxytropis , Sphaerophysa , and Sophora species) suspected of causing locoism and/or poisoning in Inner Mongolia were surveyed for swainsonine and Undifilum , the fungal endophyte responsible for the production of swainsonine. Swainsonine was detected at concentrations greater than 0.01% in Astragalus variabilis and Oxytropis glabra . The endophyte Undifilum was detected by culturing and PCR in samples containing swainsonine concentrations greater than 0.01%. In some specimens of A. variabilis and O. glabra swainsonine was not detected or concentrations were less than 0.01%. In these samples the endophyte could not be cultured, but was detected by PCR. Additionally, contrary to previous reports the quinolizidine alkaloids, thermopsine, anagyrine, and lupanine, were not detected in O. glabra and Oxytropis ochrocephala , however the quinolizidine alkaloids, sophoridine, sophocarpine, and sophoramine were detected in Sophora alopecuroides as previously reported.

  • detection and localization of the endophyte undifilum Oxytropis in locoweed tissues
    Botany, 2012
    Co-Authors: Roxanna Reyna, Daniel Cook, Daniel Grum, Peter H Cooke, Rebecca Creamer
    Abstract:

    Poisoning of livestock owing to grazing on locoweeds results in significant economic losses in the western United States. Some Oxytropis spp. locoweeds contain a seed-transmitted endophytic fungus, Undifilum Oxytropis, which produces the toxic alkaloid swainsonine. We sought to localize and characterize growth patterns of the fungus within leaves and petioles of Oxytropis lambertii Pursh and Oxytropis sericea Nutt. to help define the types of interactions between the fungus and its hosts. Vegetative hyphae were observed within locoweed tissues using integrated imaging. Topographical images from scanning electron microscopy revealed the presence of the endophyte in the pith tissue of petioles. The fungus was identified between plant cells but did not appear to penetrate host cells. Transmission electron microscopy images of thin sections revealed that hyphae were closely associated with host cell walls. Oxytropis sericea was innoculated with green fluorescent protein-transformed U. Oxytropis and observed by confocal microscopy, confirming the presence of the endophyte hyphae in leaves and petioles. The fungus was identified only in the pith of petioles using fluorescence and in the vascular bundle throughout extracellular spaces in leaves. These results revealed no signs of a pathogenic interaction between plant and fungus and support the hypothesis of a mutualistic or commensal relationship. Resume : Le locoisme, empoisonnement du betail qui broute les Oxytropis, occasionne d'importantes pertes economiques dans l'ouest des Etats-Unis. Certains Oxytropis contiennent un champignon endophyte transmis par la graines, l'Undifilum Oxytropis, produisant l'alcaloide toxique swainsonine. Les auteurs ont cherche alocaliser et acaracteriser le patron de croissance du champignon dans les feuilles et les petioles des Oxytropis lambertii Pursh et Oxytropis sericea Nutt. afin de pouvoir definir les types d'interactions entre le champignon et ses hotes. Ils ont observe les hyphes vegetatifs dans les tissus des Oxytropis al'aide de l'imagerie integree. Les images topographiques obtenues en microscopie electronique par balayage revelent la presence de l'endophyte dans les tissus de la moelle des petioles. Ils ont identifie le champignon entre les cellules de la plante, mais il ne semble pas penetrer les cellules. La microscopie electronique par balayage de sections minces montre une etroite association entre les hyphes et les parois cellulaires des cellule hotes. Les auteurs ont utilise les proteines transformees fluorescentes de l'U. Oxytropis pour inoculer l'O. sericea; l'observation en microscopie confocale confirme la presence des hyphes des endophytes dans les feuilles et les petioles. Les auteurs ont identifie le champignon seulement dans la moelle des petioles en utilisant la fluorescence, ainsi que dans les faisceaux vasculaires de l'ensemble des espaces extracellulaires des feuilles. Les resultats ne montrent aucun signe d'interaction pathogene entre la plante et le champignon et supportent l'hypothese d'une relation mutualiste ou commensaliste. Mots-cles : Undifilum, locoisme, microscopie electronique, microscopie confocale. (Traduit par la Redaction)

  • Influence of endophyte genotype on swainsonine concentrations in Oxytropis sericea.
    Toxicon, 2012
    Co-Authors: Daniel Cook, Daniel Grum, Dale R Gardner, Kevin D Welch, James A Pfister
    Abstract:

    Abstract Locoism is a toxic syndrome of livestock caused by the ingestion of a subset of legumes belonging to the Astragalus and Oxytropis genera known as “locoweeds”. Locoweeds contain the toxic indolizidine alkaloid swainsonine, which is produced by the endophytic fungi Undifilum species. Previously we reported that swainsonine concentrations differ between populations of Oxytropis sericea. We hypothesized that the genotype of the plant, endophyte, or an interaction of the two may be responsible for the differences in swainsonine concentration between populations of O. sericea. To test this hypothesis, plants derived from seeds collected at each location were grown in a common garden, Undifilum Oxytropis isolates from each location were cultured and grown in a common environment, and a plant genotype by endophyte cross inoculation was performed. Here we show that the genotype of the endophyte is responsible for the differences in swainsonine concentrations observed in the two populations of O. sericea.

  • Influence of Phenological Stage on Swainsonine and Endophyte Concentrations in Oxytropis sericea
    Journal of Chemical Ecology, 2012
    Co-Authors: Daniel Cook, Daniel Grum, Dale R Gardner, James A Pfister, Kevin D Welch
    Abstract:

    Locoweeds are defined as Astragalus and Oxytropis species that cause intoxication due to the alkaloid swainsonine. Swainsonine concentrations in Oxytropis sericea were influenced by location, plant part, and the developmental stage of the plant. Concentrations followed similar trends at each location, generally increasing over the growing season in above-ground parts until the plant reaches maturity with no change in concentration in the crowns. At the onset of senescence, swainsonine decreased in floral parts to less than half of the peak concentration. Similar to swainsonine concentrations, endophyte amounts were influenced by location, plant part, and the developmental stage of the plant. Likewise, endophyte amounts generally increased over the growing season in above ground parts and remained static in the crowns at all four locations. Swainsonine in Oxytropis sericea was positively associated with the endophyte Undifilum , which is responsible for swainsonine biosynthesis.

Rebecca Creamer - One of the best experts on this subject based on the ideXlab platform.

  • evidence for nonpathogenic relationships of alternaria section undifilum endophytes within three host locoweed plant species
    Botany, 2018
    Co-Authors: Aziza Ibrahim Noor, Peter H Cooke, Daniel Cook, Amy Nava, Rebecca Creamer
    Abstract:

    Locoweeds are legumes of the Astragalus and Oxytropis genera that live symbiotically with the fungal endosymbionts belonging to Alternaria section Undifilum. These endophytes produce the toxin swainsonine, which causes a neurological syndrome (locosim) when the plant is ingested by grazing animals. Here we characterize the growth of the endophytes within plant tissues using confocal and scanning electron microscopy (SEM) to define the physical relationship. Microscopy results revealed the endophyte distribution to be densely networked, with the mycelia aligned parallel to the long axis of the plant stem and with no pathology to plant cell walls, xylem, or phloem. The autofluorescent mycelia were located intercellulary in the pith of stems. Stereofluoroscope observations of Alternaria Oxytropis, A. cinerea, and A. fulva in phytoagar showed that mycelial growth occurred at the tip of hyphae. The growth rate of A. cinerea was significantly faster than for A. Oxytropis or A. fulva. All three species of endoph...

  • detection and localization of the endophyte undifilum Oxytropis in locoweed tissues
    Botany, 2012
    Co-Authors: Roxanna Reyna, Daniel Cook, Daniel Grum, Peter H Cooke, Rebecca Creamer
    Abstract:

    Poisoning of livestock owing to grazing on locoweeds results in significant economic losses in the western United States. Some Oxytropis spp. locoweeds contain a seed-transmitted endophytic fungus, Undifilum Oxytropis, which produces the toxic alkaloid swainsonine. We sought to localize and characterize growth patterns of the fungus within leaves and petioles of Oxytropis lambertii Pursh and Oxytropis sericea Nutt. to help define the types of interactions between the fungus and its hosts. Vegetative hyphae were observed within locoweed tissues using integrated imaging. Topographical images from scanning electron microscopy revealed the presence of the endophyte in the pith tissue of petioles. The fungus was identified between plant cells but did not appear to penetrate host cells. Transmission electron microscopy images of thin sections revealed that hyphae were closely associated with host cell walls. Oxytropis sericea was innoculated with green fluorescent protein-transformed U. Oxytropis and observed by confocal microscopy, confirming the presence of the endophyte hyphae in leaves and petioles. The fungus was identified only in the pith of petioles using fluorescence and in the vascular bundle throughout extracellular spaces in leaves. These results revealed no signs of a pathogenic interaction between plant and fungus and support the hypothesis of a mutualistic or commensal relationship. Resume : Le locoisme, empoisonnement du betail qui broute les Oxytropis, occasionne d'importantes pertes economiques dans l'ouest des Etats-Unis. Certains Oxytropis contiennent un champignon endophyte transmis par la graines, l'Undifilum Oxytropis, produisant l'alcaloide toxique swainsonine. Les auteurs ont cherche alocaliser et acaracteriser le patron de croissance du champignon dans les feuilles et les petioles des Oxytropis lambertii Pursh et Oxytropis sericea Nutt. afin de pouvoir definir les types d'interactions entre le champignon et ses hotes. Ils ont observe les hyphes vegetatifs dans les tissus des Oxytropis al'aide de l'imagerie integree. Les images topographiques obtenues en microscopie electronique par balayage revelent la presence de l'endophyte dans les tissus de la moelle des petioles. Ils ont identifie le champignon entre les cellules de la plante, mais il ne semble pas penetrer les cellules. La microscopie electronique par balayage de sections minces montre une etroite association entre les hyphes et les parois cellulaires des cellule hotes. Les auteurs ont utilise les proteines transformees fluorescentes de l'U. Oxytropis pour inoculer l'O. sericea; l'observation en microscopie confocale confirme la presence des hyphes des endophytes dans les feuilles et les petioles. Les auteurs ont identifie le champignon seulement dans la moelle des petioles en utilisant la fluorescence, ainsi que dans les faisceaux vasculaires de l'ensemble des espaces extracellulaires des feuilles. Les resultats ne montrent aucun signe d'interaction pathogene entre la plante et le champignon et supportent l'hypothese d'une relation mutualiste ou commensaliste. Mots-cles : Undifilum, locoisme, microscopie electronique, microscopie confocale. (Traduit par la Redaction)

  • Potential role for saccharopine reductase in swainsonine metabolism in endophytic fungus, Undifilum Oxytropis
    Fungal biology, 2012
    Co-Authors: Suman Mukherjee, Angus L. Dawe, Rebecca Creamer
    Abstract:

    Locoweed plants in the southwestern United States often harbour a slow-growing endophytic fungus, Undifilum Oxytropis (Phylum: Ascomycota; Order: Pleosporales), which produces a toxic alkaloid, swainsonine. Consumption of U. Oxytropis by grazing animals induces a neurological disorder called locoism for which the toxic alkaloid swainsonine has been reported to be the causal agent. Little is known about the biosynthetic pathway of swainsonine in endophytic fungi, but previous studies on non-endophytic ascomycetous fungi indicate that pipecolic acid and saccharopine are key intermediates. We have used degenerate primers, Rapid amplification of cDNA ends (RACE)-PCR and inverse PCR to identify the gene sequence of U. Oxytropis saccharopine reductase. To investigate the role of this gene product in swainsonine metabolism, we have developed a gene deletion system for this slow-growing endophyte based on our recently established transformation protocol. A strain of U. Oxytropis lacking saccharopine reductase had decreased levels of saccharopine and lysine along with increased accumulation of pipecolic acid and swainsonine. Thus, saccharopine reductase influences the accumulation of swainsonine and its precursor, pipecolic acid, in U. Oxytropis.

  • Proteomic analysis of the endophytic fungus Undifilum Oxytropis
    AFRICAN JOURNAL OF BIOTECHNOLOGY, 2012
    Co-Authors: Omar Holguin, Jianhua Wang, Guoxia Geng, Jianna Wang, Tanner Schaub, Caiju Hao, Rebecca Creamer
    Abstract:

    The filamentous Ascomycete fungus Undifilum Oxytropis is an endophyte of locoweed plants of the genera Oxytropis that produces a toxic alkaloid swainsonine. Swainsonine, an alpha-mannosidase inhibitor causes a general toxicosis and neurological problems (locoism) when consumed by grazing animals. Swainsonine is also being assessed for its anti-cancer properties. While the ecology of U. Oxytropis has been studied, little is known about the genetics and proteomics of any swainsonine-producing fungus. To help understand the proteins in U. Oxytropis , the proteome of U. Oxytropis was analyzed using 2-dimensional electrophoresis (2-DE). Proteins from U. Oxytropis mycelia were extracted and separated by in-gel isoelectric focusing (IEF). The entire immobilized pH gradient (IPG) strip was cut into a set of gel sections and each gel section was digested with trypsin and then identified using liquid chromatography tandem mass spectrometry (LC-MS/MS). 2-DE maps were also developed for U. Oxytropis to define its proteome. In the isoelectric point (pI) range of 3-11 and 10-250 kDa ranges, more than 450 spots were detected in 2-DE silver-stained gels, and 52 proteins were identified by LC-MS/MS. Most of the identified proteins were involved in energy production, oxidoreductase activity, carbohydrate metabolic process, amino acid and cellular ketone metabolic process. A large group of identified proteins were related to stress proteins and heat shock proteins. This work presents the first two-dimensional reference map of this alkaloid-producing fungus. Details of the proteome serve as a baseline for further study of this swainsonine-producing fungus and are essential for a reverse genetic analysis of the fungus. Keywords: Undifilum Oxytropis fungus, two-dimensional gel electrophoresis, proteome reference map, liquid chromatography tandem mass spectrometry, swainsonine

  • Seasonal Changes in Undifilum Colonization and Swainsonine Content of Locoweeds
    Journal of Chemical Ecology, 2012
    Co-Authors: Jorge Achata Böttger, Rebecca Creamer, Dale Gardner
    Abstract:

    Locoweeds ( Astragalus and Oxytropis ) are leguminous plants that are toxic due to a symbiotic association with the endophytic fungus Undifilum Oxytropis . The fungus produces the alkaloid swainsonine, an α-mannosidase-inhibitor that causes serious damage to mammals when consumed. A real-time PCR technique was developed to quantify the colonization extent of Undifilum in locoweeds and to compare it to the swainsonine concentration in the plants. Amplification of the endophyte nuclear ITS region allowed reliable quantification of Undifilum DNA from field plants and in vitro cultures. Swainsonine concentration was highly correlated ( ρ  = 0.972, P  

Dale R Gardner - One of the best experts on this subject based on the ideXlab platform.

  • Detection of swainsonine and isolation of the endophyte Undifilum from the major locoweeds in Inner Mongolia
    Biochemical Systematics and Ecology, 2012
    Co-Authors: Xinlei Gao, Dale R Gardner, Daniel Cook, Kip E. Panter, Stephen T. Lee, Ling Yan, Bing Han, Meng-li Zhao
    Abstract:

    Abstract Locoweeds are Astragalus and Oxytropis species that contain the toxic alkaloid swainsonine, causing widespread poisoning of livestock in Inner Mongolia. Taxa ( Astragalus , Oxytropis , Sphaerophysa , and Sophora species) suspected of causing locoism and/or poisoning in Inner Mongolia were surveyed for swainsonine and Undifilum , the fungal endophyte responsible for the production of swainsonine. Swainsonine was detected at concentrations greater than 0.01% in Astragalus variabilis and Oxytropis glabra . The endophyte Undifilum was detected by culturing and PCR in samples containing swainsonine concentrations greater than 0.01%. In some specimens of A. variabilis and O. glabra swainsonine was not detected or concentrations were less than 0.01%. In these samples the endophyte could not be cultured, but was detected by PCR. Additionally, contrary to previous reports the quinolizidine alkaloids, thermopsine, anagyrine, and lupanine, were not detected in O. glabra and Oxytropis ochrocephala , however the quinolizidine alkaloids, sophoridine, sophocarpine, and sophoramine were detected in Sophora alopecuroides as previously reported.

  • Influence of endophyte genotype on swainsonine concentrations in Oxytropis sericea.
    Toxicon, 2012
    Co-Authors: Daniel Cook, Daniel Grum, Dale R Gardner, Kevin D Welch, James A Pfister
    Abstract:

    Abstract Locoism is a toxic syndrome of livestock caused by the ingestion of a subset of legumes belonging to the Astragalus and Oxytropis genera known as “locoweeds”. Locoweeds contain the toxic indolizidine alkaloid swainsonine, which is produced by the endophytic fungi Undifilum species. Previously we reported that swainsonine concentrations differ between populations of Oxytropis sericea. We hypothesized that the genotype of the plant, endophyte, or an interaction of the two may be responsible for the differences in swainsonine concentration between populations of O. sericea. To test this hypothesis, plants derived from seeds collected at each location were grown in a common garden, Undifilum Oxytropis isolates from each location were cultured and grown in a common environment, and a plant genotype by endophyte cross inoculation was performed. Here we show that the genotype of the endophyte is responsible for the differences in swainsonine concentrations observed in the two populations of O. sericea.

  • Seasonal Changes in Undifilum Colonization and Swainsonine Content of Locoweeds
    Journal of chemical ecology, 2012
    Co-Authors: Jorge Achata Böttger, Rebecca Creamer, Dale R Gardner
    Abstract:

    Locoweeds (Astragalus and Oxytropis) are leguminous plants that are toxic due to a symbiotic association with the endophytic fungus Undifilum Oxytropis. The fungus produces the alkaloid swainsonine, an α-mannosidase-inhibitor that causes serious damage to mammals when consumed. A real-time PCR technique was developed to quantify the colonization extent of Undifilum in locoweeds and to compare it to the swainsonine concentration in the plants. Amplification of the endophyte nuclear ITS region allowed reliable quantification of Undifilum DNA from field plants and in vitro cultures. Swainsonine concentration was highly correlated (ρ = 0.972, P < 0.001) with the proportion of Undifilum DNA during the first 4 weeks of in vitro culture growth. Species of Astragalus and Oxytropis were sampled seasonally in New Mexico and Colorado for two years. High swainsonine concentration in plant samples was associated with high levels of endophyte DNA, except in plant reproductive tissues.

  • Influence of Phenological Stage on Swainsonine and Endophyte Concentrations in Oxytropis sericea
    Journal of Chemical Ecology, 2012
    Co-Authors: Daniel Cook, Daniel Grum, Dale R Gardner, James A Pfister, Kevin D Welch
    Abstract:

    Locoweeds are defined as Astragalus and Oxytropis species that cause intoxication due to the alkaloid swainsonine. Swainsonine concentrations in Oxytropis sericea were influenced by location, plant part, and the developmental stage of the plant. Concentrations followed similar trends at each location, generally increasing over the growing season in above-ground parts until the plant reaches maturity with no change in concentration in the crowns. At the onset of senescence, swainsonine decreased in floral parts to less than half of the peak concentration. Similar to swainsonine concentrations, endophyte amounts were influenced by location, plant part, and the developmental stage of the plant. Likewise, endophyte amounts generally increased over the growing season in above ground parts and remained static in the crowns at all four locations. Swainsonine in Oxytropis sericea was positively associated with the endophyte Undifilum , which is responsible for swainsonine biosynthesis.

  • swainsonine and endophyte relationships in astragalus mollissimus and astragalus lentiginosus
    Journal of Agricultural and Food Chemistry, 2011
    Co-Authors: Daniel Cook, Daniel Grum, Dale R Gardner, Kevin D Welch, James A Pfister, Benedict T. Green
    Abstract:

    Locoweeds are defined as Astragalus and Oxytropis species that induce locoism due to the toxic alkaloid swainsonine. Swainsonine was detected in all parts of Astragalus lentiginosus and Astragalus mollissimus , with greater concentrations found in the aboveground parts. Undifilum Oxytropis , a fungal endophyte responsible for the synthesis of swainsonine, was detected in all plant parts of A. lentiginosus and A. mollissimus. The amount of endophyte within a plant part does not always correspond to the concentration of swainsonine in the same part. Plants of A. mollissimus and A. lentiginosus can be divided into two chemotypes: those that contain swainsonine (>0.1%; chemotype 1) and those that contain little or no detectable swainsonine (<0.01%; chemotype 2). Chemotype 1 plants in both species had quantitatively higher amounts of endophyte compared to chemotype 2 plants. Swainsonine and endophyte amounts were not uniformly distributed within stalks of the same plant. For that reason, repeated sampling of stalks from the same plant during one growing season may provide misleading results. Sequence variants of U. Oxytropis exist within populations of A. mollissimus, A. lentiginosus, and Oxytropis sericea and do not correlate with chemotype. These findings suggest several possible reasons for differential concentrations of swainsonine that will be tested in future work.

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