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“Electrotransformation” Transformation System for Root Endophytic Fungus Piriformospora indica

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Piriformospora indica

Part of the book series: Soil Biology ((SOILBIOL,volume 33))

Abstract

Electroporation-mediated transformation (EMT) is widely used for gene delivery in many fungi. However, no such method has been developed for newly discovered fungus Piriformospora indica. Here, we describe an efficient protocol EMT using fungal mycelia and chlamydospores. Fresh grown mycelial fragments and germinated spores were transformed by electroporation. Transgenic colonies obtained were subjected to two rounds of selection on medium containing higher levels of the selective agent to eliminate false-positive colonies. Use of mycelia offers flexible experiment planning and a convenient source of experimental material for fungus that does not produce enough spores. Additionally, we also report the use of RNAi construct as a reporter in P. indica for functional expression. Using this protocol, transgenic fungus can be obtained in 3–6 weeks. This method offers new prospects for the genetic manipulations of this axenically cultivable, plant probiotic fungus.

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Acknowledgments

We are very thankful to Dr. Hitoshi Nakayashiki, Kobe University, Japan, for providing pSD-1G vector. This project was financed by Council of Scientific and Industrial Research, Government of India. MK is thankful to ICMR and JNU for providing research fellowship. AKJ and MD also acknowledge DST-Purse Grant and UGC Resource Networking grant.

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Authors and Affiliations

  1. School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Manoj Kumar, Ruby Sharma & Atul Kumar Johri

  2. School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Meenakshi Dua

  3. Plant Molecular Biology Group, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Narendra Tuteja

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  1. Manoj Kumar
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  2. Ruby Sharma
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  3. Meenakshi Dua
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  5. Atul Kumar Johri
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Correspondence to Atul Kumar Johri .

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Editors and Affiliations

  1. Amity Institute of Microbial Technology, Amity University Uttar Pradesh, E-3 Block, Sector 125, Noida, 201303, Uttar Pradesh, India

    Ajit Varma

  2. , FB Biology, Systematic Botany & Mycology, Philipps University of Marburg, Karl von Frisch Str. 8, Marburg, 35032, Germany

    Gerhard Kost

  3. Inst. Allgemeine Botanik, LS Pflanzenphysiologie, Univ. Jena, Dornburger Str. 159, Jena, 07743, Thüringen, Germany

    Ralf Oelmüller

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Kumar, M., Sharma, R., Dua, M., Tuteja, N., Johri, A.K. (2013). “Electrotransformation” Transformation System for Root Endophytic Fungus Piriformospora indica . In: Varma, A., Kost, G., Oelmüller, R. (eds) Piriformospora indica. Soil Biology, vol 33. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-33802-1_19

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