CRYOPRESERVATION OF MENTHA PIPERITA L. GERMPLASM AND CONFIRMATION OF GENETIC STABILITY AFTER CRYO-STORAGE
S. Galatali1, M. Ceylan1, E. Ogur2, N. Adanacioglu2 and E. Kaya*1
1Molecular Biology and Genetics Department, Faculty of Science, Mugla Sitki Kocman University, 48000, Kotekli, Mugla, Turkey
2Biodiversity and Genetic Resources Department, Aegean Agricultural Research Institute, Menemen-Izmir, Turkey
*Corresponding author’s e-mail address: ergunkaya@mu.edu.tr (Ergun Kaya)
ABSTRACT
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 M. piperita 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 M. piperita 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 M. piperita 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.
Key words: DMSO, encapsulation, ISSR-PCR, PVS2, vitrification
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