Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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In Vitro Production of Indian Citrs Ringspot Virus-Free Plants of Kinnow Mandarin (Citrus nobilis Lour X C. deliciosa Tenora) by Ovule Culture

  • Singh B. (Department of Botanical and Environmental Sciences, Guru Nanak Dev University) ;
  • Sharma S. (Department of Botanical and Environmental Sciences, Guru Nanak Dev University) ;
  • Rani G. (Department of Botanical and Environmental Sciences, Guru Nanak Dev University) ;
  • Zaidi A.A. (Molecular Plant Virology Laboratory, Floriculture Division, Institute of Himalayan Bioresource Technology) ;
  • Hallan V. (Molecular Plant Virology Laboratory, Floriculture Division, Institute of Himalayan Bioresource Technology) ;
  • Nagpal A. (Department of Botanical and Environmental Sciences, Guru Nanak Dev University) ;
  • Virk G.S. (Department of Botanical and Environmental Sciences, Guru Nanak Dev University)
  • Published : 2005.12.01
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Abstract

Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.

Keywords

References

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