Development of the in vitro micropropagation protocol for the conservation of the species Pulsatilla grandis Wend

Abstract

Pulsatilla grandis Wend, known as the greater pasque flower, is a perennial species from the Ranunculaceae family, originating in Europe and Western Asia. In the Republic of Moldova, it is a species with endangered plant status. The distribution of P. grandis Wend in our country is limited, with some representatives found in certain protected areas and nature reserves. The plant holds pharmacological value, being utilized in folk medicine as a sedative, analgesic, diuretic, expectorant, etc. Due to its vulnerable status, conserving the natural habitats of P. grandis Wend is crucial. One of the conservation methods is in vitro micropropagation, which allows for the fast multiplication of plants and their reintroduction into nature or botanical gardens. In a recent study, P. grandis Wend seeds were used for micropropagation, utilizing the ½ Murashige-Skoog medium. It was found that the addition of benzyladenine (BA) to the culture medium improved the micropropagation coefficient, although a second stage was necessary for inducing rhizogenesis. Supplementation of the culture medium with NAA (1-Naphthaleneacetic acid) and IBA Indole-3-butyric acid (each at 0.1 mg/L) facilitates rooting and improves shoot robustness.