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Genetic fidelity assessment in micropropagated plants using cytogenetical analysis and heterochromatin distribution: a case study with Nepenthes khasiana Hook f.

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Abstract

Rapid clonal propagation of selected genotypes has been one of the most extensively exploited approaches of biotechnology. However, inclusion of somaclonal variations in tissue-culture-derived plants results in the production of undesirable plant off-types which limits its applications in tissue culture industry. Therefore, the most critical concern has been the maintenance of genetic uniformity of micropropagated plants. Assessment of genetic fidelity in tissue-culture-raised plants of three consecutive regenerations of Nepenthes khasiana has been successfully carried out using chromosome counts and heterochromatin distribution pattern wherein changes in the number of chromosomes and the distribution of AT and GC base pairs were recorded. The cells studied in the plantlets of the first regeneration (23.33 %) showed deviant number of chromosome which was increased to 33.33 % and 40 % in the plantlets of the second and the third regenerations, respectively. Also, 4′,6-diamidino-2-phenylindole (DAPI)+ and chromomycin A3 (CMA)+ binding sites, on an average of 5.74 ± 0.47 and 5.00 ± 0.30, were observed in the plantlets of the first regeneration. Subsequently, DAPI+ binding sites were increased to 6.61 ± 0.39 and 6.74 ± 0.57 in the plantlets of the second and the third regenerations, respectively, with a corresponding decrease in the CMA+ binding sites (4.63 ± 0.45 and 4.16 ± 0.47 CMA+ sites in the plantlets of the second and the third regenerations, respectively). The study reveals an increase in cytological variations in the morphologically similar micropropagated plants of N. khasiana with the subsequent regenerations which further necessitate the determination of genetic integrity of micropropagated plants.

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Acknowledgments

S. P. Devi gratefully acknowledges the financial support provided by Research Grant No. BT/PR-7055/BCE/08/437/2006 and No. BT/04/NE/ 2009 from the Department of Biotechnology, Government of India, New Delhi, India.

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All the authors have no objection in the submission of this manuscript to the journal “Protoplasma”

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Authors and Affiliations

  1. Department of Botany, North-Eastern Hill University, Shillong, 793022, India

    Soibam Purnima Devi, Suman Kumaria & Pramod Tandon

  2. Department of Biotechnology and Bioinformatics, North-Eastern Hill University, Shillong, 793022, India

    Satyawada Rama Rao

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  1. Soibam Purnima Devi
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  2. Suman Kumaria
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  3. Satyawada Rama Rao
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  4. Pramod Tandon
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Correspondence to Suman Kumaria.

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Devi, S.P., Kumaria, S., Rao, S.R. et al. Genetic fidelity assessment in micropropagated plants using cytogenetical analysis and heterochromatin distribution: a case study with Nepenthes khasiana Hook f.. Protoplasma 252, 1305–1312 (2015). https://doi.org/10.1007/s00709-015-0763-z

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