In Vitro Mass Multiplication and Assessment of Genetic Stability of In Vitro Raised Artemisia absinthium L. Plants Using ISSR and SSAP Molecular Markers

Abstract

The present investigations were made attempting to develop a rapid, reliable, and reproduciblein vitroregeneration protocol forArtemisia absinthiumL., a medicinal plant of Kashmir Himalayas. Out of several auxin-cytokinin combinations tested, Murashige and Skoog’s (MS) medium supplemented with 0.5 mgL−12,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mgL−1kinetin (Kn) was found to be the best for the callus induction. On the other hand, 4.5 mgL−16-benzylaminopurine (BAP) and 0.5 mgL−11-α-naphthaleneacetic acid (NAA) in the medium resulted in maximum shoot induction from the callus. Similarly, BAP and NAA at a concentration of 1.5 mgL−1and 0.5 mgL−1, respectively, proved to be the best for the multiple shoot induction from nodal explants. Numerous shoots were obtained from nodal explants after third subculture.In vitrorooting was maximum on medium containing indole-3-butyric acid (IBA) at 0.5 mgL−1. The genetic stability of thein vitroraised plants ofArtemisia absinthiumwas assessed using the intersimple sequence repeat (ISSR) and sequence-specific amplification polymorphism (SSAP) molecular markers. Both markers were able to detect the somaclonal variations in the callus regenerated plants, while no variation was detected in the plants regenerated from the nodal explants. SSAP has been found to be more useful in detection of variability as compared to ISSR molecular marker. The results of present study concluded that the direct regeneration protocol will be useful for the production of true to type plants of this medicinally important plant. This will go a long way in reducing the pressure on the natural populations for the secondary metabolite production, especially for extraction of essential oils.

EfficientIn VitroPropagation byEx VitroRooting Methods ofArtemisia absinthiumL., an Ethnobotanically Important Plant

“…Table 2). The number of shoots produced in this study is considerably higher in comparison to the previous reports on the same plant species [24][25][26]. The application of NAA in shoot proliferation was reported in Ceropegia bulbosa [27] and Caralluma edulis [28].…”

“…The combined use of BAP and NAA also enhanced proliferation of shoots in Artemisia annua [29]. The addition of NAA with cytokinins plays significant role in multiple shoot production [26]. The higher concentration of BAP and Kn than NAA in the culture medium could induce shoot organogenesis.…”

Efficient in vitro propagation of Artemisia nilagirica var. nilagirica (Indian wormwood) and assessment of genetic fidelity of micropropagated plants

“…a In vitro regenerated plantlets transferred to plastic cups containing potting mixtures (after 4 weeks; Bar = 3.1 cm); b Established plant transferred to pots in green house garden (after 8 weeks; Bar = 6.7 cm); c Plants grown up to 1.25 m (after 30 weeks; Bar = 15.9 cm) Fig. 3 The frequency of ex-vitro survival and growth of acclimatized plant in different potting mixtures paniculata (Dandin and Murthy 2012b), Atremisia absinthium (Kour et al 2014) and Spilanthes oleracea (Dandin et al 2014). …”

Genetic Stability of Banana Plant Regenerated from Floral Axis Organogenesis Assessed by Newly Developed SSR Markers

“…In spite of the advantages of the in vitro propagation in particular organogenesis, a genetic instability occurred as a result of growth regulators. Additionally, repeated subculture activities trigger the emergence of different physiological responses between explant (Butiuc-keul, Farkas, & Cristea, 2016;Kour, Kour, Kaul, & Dhar, 2014). Moreover, high similar pattern on SSR and morphological characters of culture plants and mother plant, suggested a successful micropropagation using floral axis with optimal method to encounter off-totype clones.…”