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Analysis of genetic stability of in vitro propagated potato microtubers using DNA markers

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Abstract

The genetic stability of in vitro propagated potato microtubers was assessed using random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR), simple sequence repeat (SSR) and amplified fragment length polymorphism (AFLP) markers. Microtubers were developed through in vitro from potato microplants using standardized protocols. The microtubers were conserved for 1 year under three different culture media and consequently microplants were regenerated for the DNA analyses. During the study, a total of 38 (10 RAPD, 11 ISSR, 12 SSR and 5 AFLP) primers produced a total of 407 (58 RAPD, 56 ISSR, 96 SSR and 197 AFLP) clear, distinct and reproducible amplicons. Cluster analysis revealed 100 % genetic similarity among the mother plant and its derivatives within the clusters by SSR, ISSR and RAPD analyses, whereas AFLP analysis revealed from 85 to 100 % genetic similarity. Dendrogram analysis based on the Jaccard’s coefficient classified the genotypes into five clusters (I–V), each cluster consisting of mother plant and its derivatives. Principal component analysis (PCA) also plotted mother plant and its genotypes of each cluster together. Based on our results, it is concluded that AFLP is the best method followed by SSR, ISSR and RAPD to detect genetic stability of in vitro conserved potato microtubers. The in vitro conservation medium (T2) is a safe method for conservation of potato microtubers to produce true-to-type plans.

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Acknowledgments

The authors are grateful to the Director, Central Potato Research Institute (Indian Council of Agricultural Research), Shimla and the Head, Division of Crop Improvement, CPRI for providing the necessary facilities for the preparation of the manuscript.

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Author notes

  1. Jai Gopal

    Present address: Directorate of Onion and Garlic Research, Rajgurunagar, Pune, 410 505, Maharashtra, India

Authors and Affiliations

  1. Central Potato Research Institute, Shimla, 171 001, Himachal Pradesh, India

    Jagesh K. Tiwari, Poonam Chandel, Shruti Gupta, Jai Gopal, B. P. Singh & Vinay Bhardwaj

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  1. Jagesh K. Tiwari
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  2. Poonam Chandel
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  3. Shruti Gupta
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Correspondence to Jagesh K. Tiwari.

Electronic supplementary material

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Supplementary Table 1

RAPD, ISSR and SSR primers sequence used in genetic stability analysis of the in vitro propagated potato genotypes (DOCX 19 kb)

Supplementary Table 2

AFLP adapters and primers sequence used for genetic stability analysis of in vitro propagated potato genotypes (DOCX 16 kb)

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Tiwari, J.K., Chandel, P., Gupta, S. et al. Analysis of genetic stability of in vitro propagated potato microtubers using DNA markers. Physiol Mol Biol Plants 19, 587–595 (2013). https://doi.org/10.1007/s12298-013-0190-6

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