Isolation and culture of protoplast from leaves of Lactuca sativa

• Published in: Songklanakarin Journal of Science and Technology (SJST)
• Main Author: Witool Chaipakdee
• Format: Article
• Language: English
• Published: Prince of Songkla University 2007-07-01
• Subjects: protoplasts; lettuce; isolation; microcolony formation
• Online Access: http://www.sjst.psu.ac.th/journal/29-4_online/0125-3395-29-4-0929-0935.pdf

Protoplasts were isolated from leaves of lettuce (Lactuca sativa L.) seedlings after in vitro germination for 25, 30, 40 and 50 days. The leaves were stripped and incubated in various combinations of cellulase and pectinase. Protoplasts were cultured on MS medium containing various kinds and concentrations of plant growth regulators in different culture systems including liquid media, hanging, drop culture and solid media. Results revealed that the highest number of viable protoplasts, 14.1x105 cells per gram of fresh weight, was obtained from 30 day-old leaves of lettuce seedlings and isolated by using 2% cellulase in combination with 1% pectinase. Liquid MS medium supplemented with 0.5 mg/l NAA and 0.5 mg/l BA promoted the highest cell division up to 17.67%. First division of protoplasts was observed at 4 days after culture and microcolony formation occurred at the 4th week after culturing. Unfortunately, neither callus formation nor plantlet regeneration were obtained.