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In vitro production of Indian citrus ringspot virus (ICRSV) free Kinnow plants employing thermotherapy coupled with shoot tip grafting

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Abstract

Indian citrus ringspot virus (ICRSV) is known to cause serious disease problem in Kinnow (Citrus nobilis Lour × C. deliciosa Tenora) plants. This work reports the elimination of ICRSV by using thermotherapy coupled with shoot tip grafting in vitro. Nodal segments from infected mother plants (indexed by indirect ELISA and RT-PCR) were treated both in water bath and moist hot air at different temperatures viz. 40, 45 and 50°C for 30, 60 and 120 min and cultured on MS medium containing 2-iP (1 mg/l) and malt extract (800 mg/l). Shoot tips were excised from the nodal sprouts and grafted on to rough lemon (C. jambhiri) rootstock under aseptic conditions. Water bath treatment was found to be more effective as compared to moist hot air treatment as maximum number of ICRSV free plants (36.84%) were obtained by grafting the tips (0.7 mm) taken from the nodal segments treated at 50°C in water bath for 2 h. In an alternate treatment regime, 1-year-old infected plants were kept at various temperatures viz.36, 38 and 40°C in a thermotherapy chamber. Maximum of 60% ICRSV free plants were obtained by grafting the tips (0.7 mm) from the plants placed at 40°C followed by the plants placed at 38°C (59.09%) and the least was observed in case of the plants placed at 36°C (40.74%). Only those plants/plantlets were considered virus free, which showed negative reaction both in Indirect ELISA and RT-PCR.

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Acknowledgments

The authors are thankful to the Council of Scientific and Industrial Research (CSIR), New Delhi (India) for providing the financial assistance. Thanks are due to Dr. P.S Ahuja, Director, IHBT, Palampur for providing necessary facilities. Thanks are also due to Dr. S.K Thind and Dr. J.N Sharma, Former Director of Regional Fruit Research Station (RFRS), Abohar and Dr. Gurkanwal Singh, Nodal Officer, Punjab Horticulture Department for their helpful suggestions.

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Authors and Affiliations

  1. Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar, 143 005, Punjab, India

    Sanjeev Sharma, Balwinder Singh, Gita Rani, Avinash Kaur Nagpal & G. S. Virk

  2. Molecular Plant Virology Laboratory, Institute of Himalayan Bioresource and Technology, C.S.I.R, Palampur, 176 061, Himachal Pradesh, India

    A. A. Zaidi & Vipin K. Hallan

Authors
  1. Sanjeev Sharma
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  2. Balwinder Singh
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  3. Gita Rani
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  4. A. A. Zaidi
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  5. Vipin K. Hallan
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  6. Avinash Kaur Nagpal
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  7. G. S. Virk
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Correspondence to Avinash Kaur Nagpal.

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Sharma, S., Singh, B., Rani, G. et al. In vitro production of Indian citrus ringspot virus (ICRSV) free Kinnow plants employing thermotherapy coupled with shoot tip grafting. Plant Cell Tiss Organ Cult 92, 85–92 (2008). https://doi.org/10.1007/s11240-007-9307-3

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  • DOI: https://doi.org/10.1007/s11240-007-9307-3

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