Elements and Performance Criteria
- Prepare for endophyte testing
- Test seed for endophyte presence
- Soften seed for recommended time in the prepared solution at the prescribed temperature
- Rinse seed thoroughly in running tap water
- Remove lemma, palea and endosperm from seed and place the remaining tissue on a microscope slide in a drop of endophyte staining solution crushing if necessary
- Place cover glass on seed, applying gentle pressure and blot up excess stain
- Examine seed with compound microscope and score as positive if endophytic hyphae are present, or negative if absent
- Test seedlings for endophyte presence
- Take 200-250 seeds from working sample at random
- Sow seeds in good quality commercial potting mix at low density to allow for good growth
- Examine well-developed seedlings from the germinated sample after growing for a minimum of 42 days
- Cut seedling away from remnant seed and remove outermost sheath from the base of the seedling
- Discard sheaths which are discoloured or have developed chlorophyll
- Isolate a 3-5 mm wide longitudinal section of the sheath and place section on a microscope slide with the epidermis side down
- Stain section immediately with the endophyte staining solution for prescribed time period
- Place cover glass over the stained sheath section, press gently and blot of any excess staining solution
- Examine the section with compound microscope and score as positive or negative if endophytic hyphae are present or absent
- Record and report test results