Localization of endophytic Undifilum fungi in locoweed seed and influence of environmental parameters on a locoweed in vitro culture system

Oldrup, Erik; McLain-Romero, Jennifer; Padilla, Anna; Moya, Andrew; Gardner, Dale; Creamer, Rebecca
May 2010
Botany;May2010, Vol. 88 Issue 5, p512
Academic Journal
Endophytic Undifilum oxytropis (Q. Wang, Nagao & Kakish) Pryor, Creamer, Shoemaker, McLain-Romero, & Hambleton found within toxic locoweeds (Astragalus and Oxytropis spp.) produces the alkaloid swainsonine, which is responsible for locoism in grazing animals. We sought to determine the location of U. oxytropis within locoweed seed, develop endophyte free plants, and assess the influence of environmental stresses on locoweed and endophyte cultures. Undifilum was identified within the parenchyma layers in seeds of Astragalus lentiginosus M.E. Jones using light microscopy and polymerase chain reaction. Astragalus lentiginosus and Oxytropis sericea Nutt. seedlings produced in embryo culture without seed coats did not contain swainsonine or fungus. Plants produced from whole seed contained U. oxytropis in both foliage and root tissues. When the in-vitro cultured plants of O. sericea and U. oxytropis cultures were subjected to environmental stresses including high temperature, low and high pH media, nutrient deficient media, and polyethylene glycol (PEG) amended media to simulate water deficit, both dry mass and swainsonine levels were affected. Swainsonine levels were greatest for O. sericea and Undifilum cultures in PEG or hydrochloric acid amended media. Plants grown in PEG-amended media had significantly greater dry mass, while Undifilum grown in PEG-amended media had lower dry mass than other treatments. L'endophyte Undifilum oxytropis (Q. Wang. Nagao & Kakish) Pryor, Creamer, Shoemaker, McLain-Romero & Hambleton, que l'on retrouve chez les plantes locogènes (Astragalus et Oxytropis spp.), produit l'alcaloïde de Swainson, responsable du locoïsme chez des animaux herbivores. Les auteurs ont cherché à déterminer la localisation de la substance responsable dans la semence, ont développé des plantes sans endophytes, et ont évalué l'influence des stress environnementaux sur les plantes locogènes et les cultures d'endophytes. Ils ont identifié l'Undifilum dans les couches parenchymatiques de l'Astragalus lentiginosus M.E. Jones, à l'aide de la microscopie photonique et de la réaction de polymérisation en chaîne. Les plantules de l'A. lentiginosus et de l'Oxytropis sericea Nutt., produites à partir de cultures d'embryons sans téguments, ne contiennent ni le champignon ni la swainsonine. Par contre, les plantes produites à partir de graines entières contiennent l'U. oxytropis dans leurs tissus foliaires et racinaires. On affecte les poids secs des plantes et leurs teneurs en swainsonine, lorsqu'on soumet des plants de l'O. sericea et l'U. oxytropis cultivés in vitro à des stress environnementaux, incluant une température élevée, des milieux à haut et faible pH, des déficiences en nutriments, ainsi qu'un apport en polyéthylène glycol (PEG) pour simuler un déficit en eau. On obtient des teneurs en swainsonine plus importantes chez l'O. sericea et l'U. oxytropis lorsqu'on apporte dans leurs milieux de culture du PEG ou de l'acide chlorhydrique. Les plantes cultivées en présence de PEG montrent des poids secs plus élevés, alors que l'U. oxytropis montre des poids secs plus faibles en présence du même traitement.


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