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This article in CS

  1. Vol. 37 No. 4, p. 1265-1269
     
    Received: June 11, 1996
    Published: July, 1997


    * Corresponding author(s): ehiatt@uga.cc.uga.edu
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doi:10.2135/cropsci1997.0011183X003700040040x

Monoclonal Antibodies for Detection of Neotyphodium coenophialum

  1. E. E. Hiatt ,
  2. N. S. Hill,
  3. J. H. Bouton and
  4. C. W. Mims
  1. Dep. of Plant Pathology, Univ. of Georgia, Athens, GA 30602

Abstract

Abstract

A simple endophyte detection system is vital for breeding efforts designed to produce low alkaloid, endophyte-infected tall rescue (Festuca arundinacea Schreb.) cultivars. The objectives of this research were to generate monoclonal antibodies specific to endophytes for immunochcmical detection purposes. Protein extracts from Neotyphodium coenophialum (Morgan-Jones and Gains) Glenn, Bacon, and Hanlin comb. nov. isolate EDN11w ere injected into Balb/c mice to elicit an immuner esponse. Hybridomac ell lines were screened for antibody production to the Neotyphodium extract and for cross recognition of protein extracts from N. uncinatum Glen, Bacon, and Hanlin and N. starrii Glen, Bacon, and Hanlin, as well as other fungi in the genera Ciadosporlum, Penicilllum, Fusarium, and Aspergillus. Three hybridomace ll lines produceda ntibodies that reacted to Neotyphodium spp. proteins, but not with proteins isolated from the other fungi. Monoclonal antibody 5C7 was immunochemically tested for specificity to endophyte in leaf sheaths of plant genotype DN11. Fluorescent staining suggested the antibody preferentially bound to the cell wall of the endophyte. Antibodies were tested for affinity to protein extracts from 14 endophyte-infected and endophyte-free tall fescue genotypes by an enzyme-linked immunosorbenta ssay (ELISA) method. Mean absorbance values for the protein extracts from the endophyte-infected genotypcs differed and ranged from 0.317 to 0.583. Mean absorbance values for the endophyte-free forms did not differ (P ≥ 0.05). All infected plants had higher absorbance values than non-infected plants regardless of antibody tested. Monoclonal antibodies can be used in an ELISA to detect Neotyphodium spp. fungal mycelia present in tall fescue.

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