Abstract
Protoplast cultures were prepared from 6-day-old hypocotyls of six spring, seven winter cultivars of Brassica napus L. and one line of Brassica campestris L. The molarity of enzyme solution was raised to 0,714 M mannitol resulting in well manipulable, cytoplasm dense protoplasts. In the protoplast purification procedure density gradient centrifugation was used to minimize physical damage of protoplasts. Three different protoplast culture systems —(1) liquid, (2) 2nd day embedded, (3) directly embedded in low melting agarose were compared. The two different protoplast embedding techniques resulted in the same efficiency of cell division as the liquid culture method and over this fact the colony browning was avoided. Using protoplast agarose-embedding and culture techniques, healthy calli were obtained for plant regeneration experiments. Incorporation of silver nitrate into the regeneration medium improved the efficiency of plant regeneration in responsive genotypes and the regeneration was induced in three nonresponsive (without silver nitrate) genotypes, too. The supplement of silver nitrate in regeneration medium was especially advantageous in plant regeneration of B. campestris. Out of fourteen commercial cultivars of Brassica napus and B. campestris, there is only one recalcitrant genotype in obtaining plantlets from protoplast-derived calli.
Highlights
Since several of the most widespread methods of introducing foreign genes into plant cells rely on protoplasts, the establishment of a reproducible protoplast-plant system is of a crucial importance in any plant species of biotechnological interest
Cytoplasm-rich (Fig. lb) protoplasts were well manipulable during isolation procedure and proved to be excellent for different culture procedures
The culture methods applied resulted in difference in initiation of cell division at 24hour age (Table 1)
Summary
Since several of the most widespread methods of introducing foreign genes into plant cells rely on protoplasts, the establishment of a reproducible protoplast-plant system is of a crucial importance in any plant species of biotechnological interest.The first successful! protoplast isolation and plant regeneration experiments have been performed by Wenzel (1973). Protoplast isolation and plant regeneration experiments have been performed by Wenzel (1973). The Brassica species vary considerably in their ability to regenerate plant from protoplast. While the Brassica napus and the Brassica oleracea (Sidney et al 1983, Robertson et al 1984) generally give good results in protoplast regeneration experiments (Kartha et al 1974, Li et al 1982, Glimelius 1984), in case of turnip rape (B. campestris, B. rapa) the plant regeneration from protoplast derived calli remained a sporadic event (Glimelius 1984, Ulrich et al 1980). The aim of the present study was to develop a largely genotype independent, efficient plant regeneration procedure for B. napus L. and B. campestris L. with agronomical importance
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
