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Root-Organ Cultures

Ri-plasmid transformed root cultures were pioneered by Mugnier and Mosse (1987). The full potential of this culture method has been developed by Yve Piche and colleagues in Canada (Becard and Piche 1989, 1990; Chabot et al. 1992), Wu in Taiwan, and Declerck and colleagues in Belgium. Most of these researchers are part of the organization GINCO and their take on the benefits and disadvantages of this method are documented on their website. Alternatively, you can download a PDF of the website pages. A book on root organ culture can be purchased on Amazon if you’ve got the cash (its not cheap).

The two most widely used species in this system are Rhizophagus intraradices and Gigaspora margarita ( Gi. rosea in the axenic cultures we have examined), but the species list is expanding somewhat. We have had superlative success with Rhizophagus clarus (see below), mostly through the efforts of Sonia Purin when she was a student in our lab conducting anastomosis experiments.

Rhizophagus clarus  Rhizophagus clarus  Rhizophagus clarus

HE METHOD (as done at INVAM)

Sonia Purin, a former PhD student in our lab, was very adept at establishing and growing root organ cultures using immortalized chicory roots purchased from GINCO. Download a PDF of the procedure used to maintain chicory roots axenically for transfer to plates containing excised colonies of mycorrhizal roots.

Below is a video of Sonia carrying out three operations in sequence: (1) transferring mycorrhizal roots from an established culture of R. clarus to new plates (2) adding cooled (but still liquid) MSR medium (with phytagel as the solidifying agent) to the transferred roots (3) starting a new culture by transferring growing chicory roots (3-4 weeks old) to the plate with the transferred mycorrhizal roots now in solidified MSR. medium.


References

Mugnier, J. and B. Mosse. 1987. Vesicular-arbuscular mycorrhizal infection in transformed root-inducing T-DNA roots grown axenically. Phytopathology 77:1045-1050.

Becard, G. and Y. Piche. 1989. New aspects of the acquisition of biotrophic status by a vesicular-arbuscular mycorrhizal fungus, Gigaspora margarita. New Phytologist 112: 77-83.

Becard, G. and Y. Piche. 1990. Physiological factors determining vesicular-arbuscular mycorrhizal formation in host and non-host Ri T-DNA transformed roots. Canadian Journal of Botany 68: 1260-1264.

Chabot, S., G. Becard, and Y. Piche. 1992. Life cycle of Glomus intraradix in root organ culture. Mycologia 84: 315-321.