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LABORATORY PROCEDURES FOR PLANT CELL LINES

REFERENCE NO.: PC/1998/2/3.2


TITLE: PRELIMINARY QUALITY CONTROL: TEST FOR FUNGAL OR YEAST CONTAMINANTS


To provide unequivocal proof for sterility in addition to macroscopic or microscopic inspection isolation of fungi or yeasts on test agar plates is attempted. Control medium and incubation conditions have been selected to allow growth to a broad range of fungi and yeasts.

PROCEDURE

1. Fungal test agar is prepared according to a standard laboratory procedure (PC/1998 /2/3.2 Appendix 1)

2. Immediately after arrival a control plate is inoculated with a small sample of the plant cell culture. In case of a callus culture a small amount of both cells and agar is suspended (forceps) in 2 ml of physiological saline. The suspension is poured onto the control agar and spread. In case of a suspension culture a small aliquot is transferred (pipette with cut tip) onto the control agar and spread.

3. The inoculated petri-dishes sealed with parafilm are incubated for one week at 30 C.

4. Afterwards the control plates are inspected for growth of yeasts or fungi.

5. If growth of yeast or fungi is be detected the depositor is informed and the culture is discarded (standard letter PC/1998/2/3 Appendix 1).


Guidelines prepared for CABRI by DSMZ, 20 Jan. 1998
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