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Freshly harvested potato tubers, Solanum tuberosum var 'Pukhraj', were inoculated for transformation with Agrobacterium rhizogenes strain Ri1600. Hairy roots were formed after 8 days of co-cultivation and the transformation efficiency was 40 %. The transformants were transferred from Murashige and Skoog medium (MS) to Modified White's medium (MW) and finally on a hormone-free minimal medium (M). The putative transformants were confirmed using rolA and rolB gene specific primers for the polymerase chain reaction (PCR) analysis. The root inducing (Ri) T-DNA transformed potato roots were co-cultured with Glomus intraradices (CMCCROC7) to obtain arbuscular mycorrhizal root organ cultures (AM-ROC dual cultures), which were used for studying the symbiosis with Glomus intraradices and the potential for spore production in vitro. Sporulation was comparable with the existing in vitro carrot-dual culture system. Around 60,250 spores/jar could be harvested with around 38,314 extraradical spores/jar and around 21,936 intraradical spores/jar. The new method using potato is certainly promising for the mass production of mycorrhizal biofertilizers. The viability of the spores when tested on potato roots was nearly 100 % and more than half of the roots were colonized 12 weeks after inoculation.