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A new system using Solanum tuberosum for the co-cultivation of Glomus intraradices and its potential for mass producing spores of arbuscular mycorrhizal fungi

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Abstract

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.

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Abbreviations

AM:

Arbuscular mycorrhiza

AMF:

Arbuscular mycorrhizal fungus

CMCC:

Centre for Mycorrhizal Culture Collection

CTAB:

Cetyl trimethyl ammonium bromide

INT:

Indo nitro tetrazolium

M:

Minimal medium

MS:

Murashige and Skoog medium

MTT:

3-(4,5-dimethyl–2–thiazolyl)-2,5-diphenyl-2H- tetrazolium bromide

MW:

Modified White’s medium

NBT:

Nitro blue tetrazolium

PCR:

Polymerase chain reaction

Ri:

Root inducing

ROC:

Root organ culture

SPSS:

Statistical package for the social sciences

T-DNA:

Transfer DNA

VAM:

Vesicular arbuscular mycorrhiza

YMA:

Yeast mannitol agar

YMB:

Yeast mannitol broth

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Acknowledgments

We sincerely thank Dr R K Pachauri, Director General, TERI, for providing the necessary infrastructure and a conducive environment for research. Financial contribution of Council of Scientific and Industrial Research, India and partly by Department of Biotechnology, Government of India is acknowledged. The copy editing by Yateendra Joshi and SEM analysis carried by Chandrakant Tripathi, TERI is also acknowledged. The authors thank two anonymous reviewers for very valuable comments and help with editing and improvement of the English.

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  1. Centre for Bioresources and Biotechnology, Teri University, 10, Institutional Area, Vasant Kunj, New Delhi, 110070, India

    Anshul Puri

  2. Centre for Mycorrhizal Research, Biotechnology and Bioresources Division, The Energy and Resources Institute, DS Block, India Habitat Centre, Lodhi Road, New Delhi, 110 003, India

    Alok Adholeya

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  1. Anshul Puri
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Puri, A., Adholeya, A. A new system using Solanum tuberosum for the co-cultivation of Glomus intraradices and its potential for mass producing spores of arbuscular mycorrhizal fungi. Symbiosis 59, 87–97 (2013). https://doi.org/10.1007/s13199-012-0213-z

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