Efficient regeneration ofBrassica oleraceahypocotyl protoplasts and high frequency genetic transformation by direct DNA uptake
✍ Scribed by Arundhati Mukhopadhyay; Reinhard Töpfer; Akshay K. Pradhan; Yaspal S. Sodhi; Hans-Henning Steinbiß; Jeff Schell; Deepak Pental
- Publisher
- Springer
- Year
- 1991
- Tongue
- English
- Weight
- 850 KB
- Volume
- 10
- Category
- Article
- ISSN
- 0721-7714
No coin nor oath required. For personal study only.
✦ Synopsis
Efficient regeneration (80%) and high frequency genetic transformation (10-33%) were achieved by culturing protoplasts isolated from hypocotyl tissues of six day old Brassica oleracea seedlings and by subjecting these protoplasts to PEG mediated direct plasmid uptake. Three different plasmid vectors carrying marker genes for resistance to methotrexate (dhfr), hygromycin (hpt) and phosphinotricin (bar) were constructed and used for transformation. Large number of normal, fertile transformants were obtained with vectors carrying hpt and bar genes. No transformants could be regenerated for resistance to methotrexate as it severely suppressed shoot differentiation.