Rapid plant regeneration through organogenesis and somatic embryogenesis from cultured protoplasts ofBrassica juncea

✍ Scribed by P. B. Kirti; V. L. Chopra

• Publisher: Springer
• Year: 1990
• Tongue: English
• Weight: 211 KB
• Volume: 20
• Category: Article
• ISSN: 0167-6857
• DOI: 10.1007/bf00034759

✦ Synopsis

Protoplasts derived from hypocotyls of seedlings grown on half-strength MS medium containing 1% sucrose were cultured at a density of 5 x 104 ml ~ in Kao's medium supplemented with 1.0 mg 1 ~ 2,4-D, 0.1 mg 1l NAA and 0.5mgl ~ zeatin riboside. After three days of culture in darkness at 25 + I°C, cultures were transferred to light (70 #E m -2 s J) in a 16/8 h liga ht/dark cycle. Cultures were diluted on the 7th, 10th and 13th day with Kao's medium containing 3.4% sucrose, 0.1 mgl -~ 2,4-dichlorophenoxyacetic acid and 1.0mgl -I benzyladenine. On the fifteenth day, microcalli were plated on K 3 medium gelled with 0.5% agarose (Type 1, low EEO, Sigma). After a further period of two weeks, transfers were made to specific media to achieve either organogenesis or somatic embryogenesis. Time taken from plating protoplasts to obtaining plantlets is 8-10 weeks. Using this procedure, several hundred regenerated plants have been hardened in a growth chamber and transferred to soil.