17 S. Galatali M. Ceylan E. Ogur N. Adanacioglu E. Kaya Journal of Animal and Plant Sciences JAPS 20232023/03/24 33 2 345-356 1018-7081 https://doi.org/10.36899/JAPS.2023.2.0625 <p>Peppermint is an important aromatic-medicinal plant species and it has valuable essential oil contents such as menthol, linalool and limonene. The main purpose of the current study was to optimize a protocol for cryopreservation of&nbsp;<em>M. piperita</em>&nbsp;local cultivars (G-74, Candarli and Gomec). The secondary aim was to investigate the genetic stability after cryopreservation using ISSR marker system. Three different single step freezing techniques were compared for long-term preservation of these local cultivars and the optimum regeneration percentages were acquired by using the droplet vitrification technique. The PVS2 treatments of this technique showed succesful long-term preservation of&nbsp;<em>M. piperita</em>&nbsp;cultivars with regeneration percentages of 72% to Candarli cultivar (60 min PVS2), 52% to Gomec cultivar (75 min PVS2) and 62.5% to G-74 cultivar (30 min PVS2) respectively. The ISSR PCR results showed that the genetic stability from cryopreserved&nbsp;<em>M. piperita</em> cultivars were high. While genetic stability percentage was ~99% for the Candarli cultivar, for the Gomec and G-74 cultivars the genetic fidelity was100%. The shoots that come from the cryopreserved shoot tips showed normal and health rooting, and all of them were also easily adapted to greenhouse conditions.</p> DMSO, encapsulation, ISSR-PCR, PVS2, vitrification Pakistan Agricultural Scientists Forum https://thejaps.org.pk/AbstractView.aspx?mid=Biot-21-0068