In Vitro Propagation and Genetic Stability Assessment Using the ISSR Markers of Stachys byzantina K. Koch, a Promising Ornamental Species

In this study, a reliable and efficient micropropagation protocol was developed for Stachys byzantina, a valuable and promising ornamental species. For the initial in vitro cultures on the Murashige and Skoog (MS) medium, shoot tips were used as explants. The addition of 5 μM of kinetin (KIN) resulted in the production of multiple (6.0 shoots/explant) and elongated (3.6 cm) shoots. The MS medium supplemented with 10 μM of a-Naphthaleneacetic acid (NAA) proved efficient for the in vitro rooting (73.3%) of the microshoots. For the ex vitro rooting of the microshoots, the treatment with 0.5 g L−1 of Indole-3-butyric acid potassium salt (K-IBA), before planting in 1:1 (v/v) peat and perlite substrate and placed in a fog system, led to 86.7% rooting. The acclimatization stage was successful, and 96.7% survival was recorded for the ex vitro-rooted plantlets. Inter Simple Sequence Repeat (ISSR) markers were employed to examine the genetic uniformity of the in vitro-derived plantlets with the mother S. byzantina plants. The monomorphic banding pattern in the micropropagated plants and the mother plant confirmed the genetic uniformity of the in vitro-derived plantlets and revealed the reliability of the proposed in vitro protocol for S. byzantina. As far as we know, this is the first study on a combined micropropagation and genetic uniformity assessment of the species, the findings of which could be further used to apply new in vitro cultivation techniques or to produce elite genotypes of S. byzantina.