Elements and Performance Criteria
- Prepare for endophyte testing
- Test seed for endophyte presence
- Seed is softened in sodium hydroxide for recommended time period.
- Seed is rinsed thoroughly in running tap water.
- Seed is de-glumed, placed on a microscope slide in a drop of endophyte staining solution and slightly crushed.
- Cover glass is placed on seed and gentle pressure is applied.
- Seed is examined with compound microscope and scored as positive if endophytic hyphae are present.
- Test seedlings for endophyte presence
- Seeds from working sample are selected at random and germinated.
- Seedlings from the sample germinated are examined after growing for a minimum of 48 days.
- Outermost sheath is removed from the seedling such that tissue to be examined should have no obvious discoloration from saprophytes and should have as little chlorophyll as possible.
- A longitudinal section of leaf sheath approximately 3–5 mm in width is isolated.
- Section is placed on a microscope slide with the epidermis side down.
- Section is stained immediately with the endophyte staining solution for prescribed time period.
- Excess dye is blotted off with tissue paper and a coverglass is place over section and flooded with water.
- Section is examined with compound microscope and scored as positive if endophytic hyphae are present.
- Record and report test results
- Results are entered under Other Determinations.
- The scientific name of the pathogen which has been tested for and the test method used is recorded in the test results.
- Sample size used for testing is recorded.
- In the case of a negative result where the pathogen was not detected, the results is reported in terms of the tolerance standard.
- In the case of a positive result the report should indicate percentage of infected seed.