Comparison of endophyte detection in fescue and ryegrass seeds using an immunoblot assay and a microscopic method
Neotyphodium coenophialum is an endophytic fungus that grows symbiotically in the intercellular spaces between seed or plant cells of some cool-season grasses. N. coenophialum can contribute to insect and drought tolerance, although the presence of toxic metabolites produced
by the fungus is detrimental if infected forage is used for feed. This study was conducted to compare the precision and efficiency of two methods of endophyte detection, an immunoblot assay and a microscopic method. Three grass species, tall fescue (Festuca arundinacea L.), red fescue
(Festuca rubra subsp.rubra) and perennial ryegrass (Lolium perenne L.) were used. One pre-determined seed lot from each species infected with a high (85-100%), medium (40-70%), or low (5-10%) level of endophyte was selected to compare the two methods. Both methods provided
comparable results of endophyte detection. The immunoblot results were slightly more consistent within replications than the microscopic method in perennial ryegrass (PRG) and tall fescue (TF) but not in red fescue (RF). The time to complete the test (sample size of 100 seeds) by the microscopic
method was four to five hours compared to one and a half to two hours by the immunoblot assay (excluding the incubation time). The cost of the immunoblot assay is slightly higher than the cost of the microscopic method. However, the microscopic method requires a high level of proficiency and
experience to detect the endophyte and the hyphae of the fungus can be missed if the technician lacks sufficient expertise. The immunoblot assay requires less training and large number of samples can be examined per day, although there is some subjectivity in interpretation of color development.
Document Type: Research Article
Publication date: 01 April 2007
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