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Evidence Guide: AHCSDT407 - Perform an endophytic seed test

Student: __________________________________________________

Signature: _________________________________________________

Tips for gathering evidence to demonstrate your skills

The important thing to remember when gathering evidence is that the more evidence the better - that is, the more evidence you gather to demonstrate your skills, the more confident an assessor can be that you have learned the skills not just at one point in time, but are continuing to apply and develop those skills (as opposed to just learning for the test!). Furthermore, one piece of evidence that you collect will not usualy demonstrate all the required criteria for a unit of competency, whereas multiple overlapping pieces of evidence will usually do the trick!

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AHCSDT407 - Perform an endophytic seed test

What evidence can you provide to prove your understanding of each of the following citeria?

Prepare for endophyte testing

  1. Review test request to identify samples to be tested, test methods and equipment
  2. Clean and prepare equipment and working space
  3. Prepare endophyte staining solution and alkaline or acid solution depending on method to be used
  4. Prepare working sample from Pure Seed
Review test request to identify samples to be tested, test methods and equipment

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Clean and prepare equipment and working space

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare endophyte staining solution and alkaline or acid solution depending on method to be used

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare working sample from Pure Seed

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Test seed for endophyte presence

  1. Soften seed for recommended time in the prepared solution at the prescribed temperature
  2. Rinse seed thoroughly in running tap water
  3. 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
  4. Place cover glass on seed, applying gentle pressure and blot up excess stain
  5. Examine seed with compound microscope and score as positive if endophytic hyphae are present, or negative if absent
Soften seed for recommended time in the prepared solution at the prescribed temperature

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Rinse seed thoroughly in running tap water

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

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

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Place cover glass on seed, applying gentle pressure and blot up excess stain

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Examine seed with compound microscope and score as positive if endophytic hyphae are present, or negative if absent

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Test seedlings for endophyte presence

  1. Take 200-250 seeds from working sample at random
  2. Sow seeds in good quality commercial potting mix at low density to allow for good growth
  3. Examine well-developed seedlings from the germinated sample after growing for a minimum of 42 days
  4. Cut seedling away from remnant seed and remove outermost sheath from the base of the seedling
  5. Discard sheaths which are discoloured or have developed chlorophyll
  6. Isolate a 3-5 mm wide longitudinal section of the sheath and place section on a microscope slide with the epidermis side down
  7. Stain section immediately with the endophyte staining solution for prescribed time period
  8. Place cover glass over the stained sheath section, press gently and blot of any excess staining solution
  9. Examine the section with compound microscope and score as positive or negative if endophytic hyphae are present or absent
Take 200-250 seeds from working sample at random

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Sow seeds in good quality commercial potting mix at low density to allow for good growth

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Examine well-developed seedlings from the germinated sample after growing for a minimum of 42 days

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Cut seedling away from remnant seed and remove outermost sheath from the base of the seedling

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Discard sheaths which are discoloured or have developed chlorophyll

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Isolate a 3-5 mm wide longitudinal section of the sheath and place section on a microscope slide with the epidermis side down

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Stain section immediately with the endophyte staining solution for prescribed time period

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Place cover glass over the stained sheath section, press gently and blot of any excess staining solution

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Examine the section with compound microscope and score as positive or negative if endophytic hyphae are present or absent

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Record and report test results

  1. Enter results under ‘other determinations’
  2. Record sample size used for testing and whether seeds or seedlings were tested
  3. Report results in terms of the percentage of seeds or seedlings in which endophyte was detected
Enter results under ‘other determinations’

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Record sample size used for testing and whether seeds or seedlings were tested

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Report results in terms of the percentage of seeds or seedlings in which endophyte was detected

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Assessed

Teacher: ___________________________________ Date: _________

Signature: ________________________________________________

Comments:

 

 

 

 

 

 

 

 

Instructions to Assessors

Evidence Guide

Element

Performance criteria

Elements describe the essential outcomes.

Performance criteria describe the performance needed to demonstrate achievement of the element.

1. Prepare for endophyte testing

1.1 Review test request to identify samples to be tested, test methods and equipment

1.2 Clean and prepare equipment and working space

1.3 Prepare endophyte staining solution and alkaline or acid solution depending on method to be used

1.4 Prepare working sample from Pure Seed

2. Test seed for endophyte presence

2.1 Soften seed for recommended time in the prepared solution at the prescribed temperature

2.2 Rinse seed thoroughly in running tap water

2.3 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

2.4 Place cover glass on seed, applying gentle pressure and blot up excess stain

2.5 Examine seed with compound microscope and score as positive if endophytic hyphae are present, or negative if absent

3. Test seedlings for endophyte presence

3.1 Take 200-250 seeds from working sample at random

3.2 Sow seeds in good quality commercial potting mix at low density to allow for good growth

3.3 Examine well-developed seedlings from the germinated sample after growing for a minimum of 42 days

3.4 Cut seedling away from remnant seed and remove outermost sheath from the base of the seedling

3.5 Discard sheaths which are discoloured or have developed chlorophyll

3.6 Isolate a 3-5 mm wide longitudinal section of the sheath and place section on a microscope slide with the epidermis side down

3.7 Stain section immediately with the endophyte staining solution for prescribed time period

3.8 Place cover glass over the stained sheath section, press gently and blot of any excess staining solution

3.9 Examine the section with compound microscope and score as positive or negative if endophytic hyphae are present or absent

4. Record and report test results

4.1 Enter results under ‘other determinations’

4.2 Record sample size used for testing and whether seeds or seedlings were tested

4.3 Report results in terms of the percentage of seeds or seedlings in which endophyte was detected

Required Skills and Knowledge

Element

Performance criteria

Elements describe the essential outcomes.

Performance criteria describe the performance needed to demonstrate achievement of the element.

1. Prepare for endophyte testing

1.1 Review test request to identify samples to be tested, test methods and equipment

1.2 Clean and prepare equipment and working space

1.3 Prepare endophyte staining solution and alkaline or acid solution depending on method to be used

1.4 Prepare working sample from Pure Seed

2. Test seed for endophyte presence

2.1 Soften seed for recommended time in the prepared solution at the prescribed temperature

2.2 Rinse seed thoroughly in running tap water

2.3 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

2.4 Place cover glass on seed, applying gentle pressure and blot up excess stain

2.5 Examine seed with compound microscope and score as positive if endophytic hyphae are present, or negative if absent

3. Test seedlings for endophyte presence

3.1 Take 200-250 seeds from working sample at random

3.2 Sow seeds in good quality commercial potting mix at low density to allow for good growth

3.3 Examine well-developed seedlings from the germinated sample after growing for a minimum of 42 days

3.4 Cut seedling away from remnant seed and remove outermost sheath from the base of the seedling

3.5 Discard sheaths which are discoloured or have developed chlorophyll

3.6 Isolate a 3-5 mm wide longitudinal section of the sheath and place section on a microscope slide with the epidermis side down

3.7 Stain section immediately with the endophyte staining solution for prescribed time period

3.8 Place cover glass over the stained sheath section, press gently and blot of any excess staining solution

3.9 Examine the section with compound microscope and score as positive or negative if endophytic hyphae are present or absent

4. Record and report test results

4.1 Enter results under ‘other determinations’

4.2 Record sample size used for testing and whether seeds or seedlings were tested

4.3 Report results in terms of the percentage of seeds or seedlings in which endophyte was detected

The candidate must be assessed on their ability to integrate and apply the performance requirements of this unit in a workplace setting. Performance must be demonstrated consistently over time and in a suitable range of contexts.

The candidate must provide evidence that they can:

prepare endophyte staining solution

test seed for endophyte presence

test seedlings for endophyte presence

record and report results of tests

The candidate must demonstrate knowledge of:

enterprise standard operating procedures (SOPs)

identification of endophytes

recording and reporting protocols

relevant health, safety and environment requirements

seed biology

Range Statement