Quality Control

Quality control of all CICG cultures involves different procedures at all stages of the process, from cleaning the greenhouse where the cultures remain during growth to the moment they are stored.

Greenhouse

Two greenhouses are available to CICG for growing AMF cultures. One of them is used only for trap cultures or for experiments or trials that contain field soil. This house has an area of ​​24 m², built with concrete blocks, honeycomb polycarbonate sheets and ceramic flooring. It is cleaned monthly with water and sodium hypochlorite.

The second greenhouse is exclusively for pure cultures. It was built in the same way as the previous greenhouse, with an area of ​​12 m² and yellow supplementary lights. Cleaning with water and sodium hypochlorite is done monthly.

Checking Cultures

The spores are extracted from cultures in a place suitable for this task, starting with the cleaning of the sieves, which are exclusively used for the analysis of pure cultures. After centrifugation, spores are stored in Petri dishes in the refrigerator and the cultures examined under a dissecting microscope. This analysis is carried out by the curator of the collection, who takes notes and makes the decision on how to proceed with the culture: a) let it dry and collect, b) reseed and grow it for another cycle, c) discard.

This information is written in spreadsheets (for physical backup) and later entered into a database in the Access program. Soon, this information will be transferred to the database in File Maker Pro.

The spores recovered in this first analysis are collected, cleaned and stored in sodium azide and mounted on semi-permanent slides (in PVLG and PVLG+Melzer). At the time of culture collection, a new extraction is performed to count the number of spores, whose information is placed on the label of each culture.

Drying cultures

If sporulation by AMF species was verified after checking the cultures, the irrigation is suspended and the plants are left to dry in the greenhouse. Care is taken not to let the substrate dry too much, so that it can affect the quality of the spores.

Storing Cultures

Pure cultures and trap cultures are stored separately in 4^o C refrigerators. Pure cultures are kept in 3 refrigerators at 4^o C, in “zip lock” plastic bags. Two labels are attached to each bag: one printed containing the species name, isolate code, number of spores and storage date and the other on a card stappled in the upper left corner containing only the isolate code (for easy and quick viewing). Cultures are arranged alphabetically inside the refrigerators. No other soil samples are allowed to be stored in these refrigerators.

Trap cultures are kept in a commercial refrigerator at 4^o C, in a separate room from the one where pure cultures are found. They are kept in plastic bags containing a label that informs the culture code and the main species sporulating in it.

Refrigerator shelves are periodically cleaned and plastic bags are cleaned with a wet cloth to remove any dust before being stored.