Br 2.1 Phoma lingam 2017-12-11T10:07:13+00:00

National Seed Health System

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Br 2.1 Phoma lingam

VERSION: 1.1
DATE: 2001
PATHOGEN: Phoma lingam (sexual: Leptosphaeria maculans)
HOST: Brassica sp.
COMMON NAME: black leg; canker; dry rot
METHOD: Br 2.1 Freezing Blotter Method (STA)
METHOD CLASS: TEMPORARY STANDARD (B)
SAMPLE: 1000 – 10,000 seeds

REVISION HISTORY: Version 1.1: Procedure #4. Botran solution correction

PROCEDURE:

1. Prepare a solution of 0.05% Botran.

2. Place 1 sterilized 10×15 inch brown blotter paper on each 11×16 inch styrofoam tray.

3. On top of the brown paper, place two sterilized 11×9 inch white blotter papers side by side, slightly overlapping.

4. Add enough of the Botran (0.05%) solution to each tray to thoroughly wet the blotters.

5. Distribute the seed evenly over each of the trays by hand or with a vacuum planter.

6. Punch holes in each plastic ziploc bag.

7. Place each tray into a 14×20 inch plastic ziploc bag.

8. Incubate the trays at room temperature for a minimum of 8 hours.

9. Freeze the sample for 8-24 hours.

10. Remove the trays from the freezer and incubate under NUV/Fluorescent light with a 12 hour photoperiod at 20-28 C° for 10 days.

REFERENCES:

STA Laboratories