Clonal micropropagation of peppermint (Mentha piperita L.) varieties of Ukrainian breeding

Abstract

Purpose. Developing technology for clonal micropropagation of peppermint (Mentha piperita L.) plants of Ukrainian breeding based on the complex of methods of isolated tissue and organ culture in vitro. Methods. During the experiment, such methods as isolated tissue and organ culture in vitro, clonal micropropagation, detached scion grafting, chemotherapy with adding of virucide Ribavirin to the nutrient medium, biometric and statistical ones were used. Results. The stepped procedure of sterilization that we have developed allows to receive 88–100% of sterile explants. For M. piperita L. introduction into culture and clonal micropropagation, Murashige and Skoog (MS) nut­rient medium appeared to be optimal supplemented with 6-benzylaminopurine (0.75 mg/l), adenine (0.05 mg/l), indole-3-acetic acid (IAA) (0.05 mg/l) and gibberellic acid (0.5 mg/l) on which the reproduction ratio on the 28th day ranged between 1:7 and 1:15. For recovery of plants from viral infection, virucide Ribavirin at concentration of 10 mg/l was added to the nutrient medium. The proposed nutrient medium for rhizogenesis, that contained IAA (0.5 mg/l) and indole butyric acid (IBA) (0.5 mg/l), allows to obtain the frequency of rhizogenesis up to 84–100%. Regenerated plants were adapted to the conditions in vivo on substrate peat : universal soil : perlite : sand in the ratio 2:1:1:1. The survival rate for peppermint varieties amounted to 96–100%. Conclusions. Biotechnological scheme was developed that permits to get healthy, purebred planting material and intensively propagate plants for supplying breeding programs of the Experimental Station for Medicinal Plants of the Institute of Agroecology and Environmental Management of the National Academy of Agrarian Sciences of Ukraine, among which such varieties as ‘Lebedyna pisnia’ and ‘Ukrainska pertseva’ were selected as the most promising for clonal micropropagation.