Embryogenic Callus Differentiation in Short-Term Callus Derived from Leaf Explants of Alfalfa Cultivars

Abstract

Alfalfa is a forage crop that accounts for one of the best sources of protein and is commonly grown all over the world. In vitro callus and embryogenic induction of alfalfa have been investigated previously; however, most of the investigations were almost limited with callus formation. In this study, leaves of 5 Medicago sativa L. cultivars (Alsancak, Sazova, Plato Iside, and Bilensoy) have been used. Influences of culture medium contents and hormones applications on callus and embryogenic callus induction were determined to optimize in vitro culture mediums of alfalfa (0.0125 g kinetin 2,4 D 1 mg mL-1; 1 g kinetin 2,4 D 1 mg mL-1 ; 0.25 g kinetin 2,4 D 2 mg mL-1; 0.5 g kinetin 2,4 D 2 mg mL-1). Callus formation was detected at a rate of 74% in 5 different cultivars used in the experiment. The five alfalfa cultivars were classified into four categories in terms of embryogenic differentiation capacity. The tested alfalfa cultivars varied in their callus formation and embryogenic callus differentiation. Sazova, Alsancak, and Bilensoy were detected for better callus formation; similarly, the same cultivars responded with better embryogenic callus formation in the culture mediums including various hormone concentrations. The present study shows that our methods have beneficial impacts on the somatic embryo induced by alfalfa. However, it depends strongly on genotype, hormone concentrations and the other medium components.