In Vitro Preparation and Reproduction of Inoculants of Glomus intraradices Fungus

Document Type : Research Paper

Authors

1 Scientific staff member at Soil Biology Res. Div. of the Soil and Water Res. Institute

2 Professor of herbal medicine at the school of Agr., Tarbiat Modarres University

3 rofessor, school of Agricultural, Tarbiat Modarres University and Director General of the Soil and Water Res. Institute

4 Professor, school of Agr. Tehran University

5 Scientific staff members of Soil Biol. Res. Div. of the Soil and Water Res. Institute, respectively

Abstract

The obligate Arbuscular mycorrhizal fungi that live symbiotically on plant roots can be found on some 80% of plant families. Despite their useful effects, the problems with growing them in synthesized culture media caused by their obligate symbiotic nature poses as the major obstacle in utilizing them for agricultural purposes. So far three methods of industrial scale reproduction of the fungi, namely, the methods of hydroponic, aeroponic, and simultaneous in vitor culturing of roots and fungi have been successfully developed. Of the three methods, the last one is the most effective both in term of inoculant purity as well as economic returns. This investigation was carried out on the basis of the need to develop and expand the method of simultaneous in vitro culturing of the fungi and roots of host plant to be used as pure inoculant as such. For this purpose, the spores of the fungi Glomus intraradices indigenous to the soils of Iran were used in the experiment. Young spores removed from the host plants medium of growth were washed several times with distilled water and Tween 20 solution in order to eliminate any surface contaminations. The spores were then surface sterilized first with a 2 percent solution of chloramin-T and then with a solution contining 200 mg/liter of streptomycin sulfate and 100 mg/liter of gentamycin sulfate. In order to check for any possible contamination, the spores were transferred to a minimum culture medium and maintained for one week at 25oC. Symbiotic relation was obtained by placing decontaminated spores on the end of induced root hairs found on the surface of the minimum culture medium. A simultaneous reproduction of the fungi and the root hairs was achieved using screw cap glass bottles containing 100 ml of the minimum culture medium. By staining the resulting spores with a solution of MTT, it was seen that 95% of the spores were active and capable of growing. Likewise, it was found that 70% of root colonization by the spores was successful. The investigations show that the in vitro method of reproduction used in this experiment is very effective for growing Glomus intraradices and possibly other species of Arbuscular mycorrhizal fungi.

Keywords

References

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Iranian Journal of Soil Research
Volume 20, Issue 2
February 2007
Pages 261-270

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