In vitro Protoplast Isolation and Regeneration of Solanum tuberosum cv. Binella and Burren

The present study aims to develop simple reliable method for protoplast isolation, culture and regeneration from the leaves of potato cvs. Binella and Burren. Optimal conditions for protoplast culture, callus initiation and regeneration were determined. Moreover, genetic diversity between regenerated and mother plants, was evaluated by Inter Simple Sequence Repeats (ISSR) technique. Protoplasts were isolated from in vitro explants after four weeks multiplication medium containing 50 μM of Silver Thiousulfate (STS). Protoplast division was started after 4 days in the cv. Binella, and after 5 days in the cv. Burren, on medium B containing (Murashig and Skoog, 1962) with 1 or 1.5 mg/L of Zeatin (Z). Maximum rate of cell colonies was recorded when 1.0 mg/L Z was used, with significant differences between the two cvs. Binella and Burren (1.58 ± 0.1%, 1.27 ± 0.09%), respectively. Maximum percentage of shoot regenerated from Potato callus came out, on medium with of 0.1 mg/L naphthalene acetic acid (NAA), with significant differences between cvs. Binella and Burren (51.12 ± 8.77%, 18.96 ± 2.81%), respectively. Maximum shoots regenerated from cv. Binella callus was (50±17.68%), on regeneration medium containing 1.5 mg/L of Z, while in cv. Burren callus did not form shoots. Two months later 92% of plantlets were successfully acclimatized, in green house. Average plant length reached 26.6 cm. Also, tubers were produced with a diameter mean of 13.17 mm and average weight of 2.06g. ISSR technique showed that regenerated plants from cv. Binella callus, were identical to the mother plants by 100%.