Cryopreservation of shoot tips of Gentiana kurroo Royle – a critically endangered medicinal plant of India

Abstract

Gentiana kurroo Royle, a critically endangered medicinal plant species of India is conserved as in vitro slow growing cultures in the In Vitro Genebank (IVGB) at ICAR-National Bureau of Plant Genetic Resources (NBPGR), New Delhi, India. Shoot tips (about 1 mm in length) excised from 8-weeks-old stock cultures subjected to cold hardening for four weeks at 8 °C (dark) were precultured on Murashige and Skoog (MS) medium supplemented with 5% Dimethylsulfoxide (DMSO) at 8 °C (dark) for two days. Thereafter, shoot tips were dehydrated with PVS2 solution (30 min) at 0 °C and cryopreserved using conventional vitrification (V) and droplet-vitrification (DV) techniques. Average post-thaw regeneration after 4 weeks was ~ 35% by V technique and ~ 60% by DV in all the three accessions tested. Genetic stability assessment on the basis of 40 Inter Simple Sequence Repeats and 30 Expressed Sequence Tagged-Simple Sequence Repeats markers revealed no variation between in vitro-cryopreserved plants and controls, in any of the genotypes. Thus, standardized DV method has the potential for long-term conservation of germplasm of G. kurroo and the three accessions are now safely cryobanked at ICAR-NBPGR.

Key message

Gentiana kurroo is a critically endangered medicinal plant endemic to North-western Himalayas, India. A protocol to cryopreserve in vitro shoot-tips was developed using droplet vitrification technique with high recovery rates and genetically stable regnerants. The protocol is being applied for cryobanking of Gentiana germplasm at ICAR-NBPGR.

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Acknowledgements

We thank Director, ICAR-NBPGR, New Delhi, for facilities and encouragement. Financial support provided by the Department of Biotechnology, Ministry of Science and Technology, is gratefully acknowledged. Thanks are due to Mr Ramesh Chandra and Mr Suresh Mali for technical assistance.

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Correspondence to Neelam Sharma.

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Sharma, N., Gowthami, R., Devi, S.V. et al. Cryopreservation of shoot tips of Gentiana kurroo Royle – a critically endangered medicinal plant of India. Plant Cell Tiss Organ Cult 144, 67–72 (2021). https://doi.org/10.1007/s11240-020-01879-2

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Keywords

  • Conventional vitrification
  • Droplet-vitrification
  • Endangered medicinal plant
  • Gentiana
  • ISSR
  • SSR