Abstract
BACKGROUND: Tricholepis roylei Hook. f. is a bushy annual shrub of the Asteraceae family and point endemic species of the Western Himalaya. T. roylei is a critically endangered species and factors like poor seed germination and habitat destruction are further rendering the species towards extinction. Therefore, the present investigation was designed to document the seed germination potential of the species and to establish a reproducible in vitro propagation and mass multiplication protocol for the ex situ conservation of T. roylei germplasm. RESULTS: Seeds of T. roylei were collected from Sangdha, Himachal Pradesh, India, and were sown in different substrates under open and controlled conditions. Though the overall seed germination potential of the species was reported to be very low and maximum 45% germination was observed in cocopeat substrate after 26 days of incubation. Half-strength Murashige and Skoog (MS) medium enriched with 6-benzylaminopurine (4.4 μM l(-1)) and naphthalene acetic acid (1.0 μM l(-1)) was observed to be the optimum medium for shoot induction in shoot tip explants of T. roylei. Maximum 98.89% shoot formation was observed with 28.42 shoots per culture and 4.4-cm shoot length, respectively. The healthy shoots (4.0 cm) were transferred onto rooting media (1/4, 1/2 and full MS) and roots were developed after 8 weeks of incubation in the half-strength MS medium. Half MS augmented with 4.9 μM l(-1) indole butyric acid was observed to be optimum for the root development and an average of 10.2 roots per shoot with 4.0-cm length was obtained. Rooted plantlets were successfully acclimatized under greenhouse conditions and subsequently established in the field, with a recorded survival rate of 90%. The plants acclimatized to the open environment were also planted in the wild under the habitat rehabilitation and species recovery programme. CONCLUSION: The seed germination study envisages that the seed germination potential of the species is very poor and might be one of the probable factors responsible for the shrinkage of T. roylei population in the wild. The standardized micropropagation protocol can be helpful for the ex situ conservation of germplasm and rehabilitation of species in the wild. Moreover, the study could be helpful in elucidating the phytochemical and molecular analysis of species.