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

• Published in: Plant cell, tissue and organ culture tissue and organ culture, 2021, Vol.144 (1), p.67-72
• Main Authors: Sharma, Neelam, Gowthami, R., Devi, S. Vimala, Malhotra, Era Vaidya, Pandey, Ruchira, Agrawal, Anuradha
• Format: Article
• Language: English
• Subjects: Biomedical and Life Sciences; Cold treatment; Cryopreservation; Cryopreservation and In Vitro Banking; Dehydration; Droplets; Endangered plants; Endangered species; Endemic plants; Genetic resources; Genotypes; Gentiana; Germplasm; Herbal medicine; Leaves; Life Sciences; Medicinal plants; Original Article; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plant resources; Plant Sciences; Plant species; Regeneration; Simple sequence repeats; Stability analysis; Tips; Vitrification
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1007/s11240-020-01879-2

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.