A mannose selection system for production of fertile transgenic maize plants from protoplasts |
| |
Authors: | A S Wang R A Evans P R Altendorf J A Hanten M C Doyle J L Rosichan |
| |
Institution: | (1) Novartis Seeds Inc., Applied Biotechnology, 317 330th St., Stanton, MN e-mail: Andy.wang@seeds.novartis.com Fax: +507-6457519, XX |
| |
Abstract: | Maize (Zea mays L.) callus cultures cannot use mannose as a sole carbohydrate source, but can utilize fructose for that purpose. Phosphomannose
isomerase (PMI) can convert mannose to fructose. Transgenic maize plants were obtained by selecting polyethylene glycol (PEG)-mediated
transformed protoplasts on mannose (20 g/l) containing medium. Transgenic calluses and plants carrying the PMI structural
gene, manA, were able to convert mannose to fructose. The PEG-mediated protoplast transformation frequency was 0.06%. Stable transformation
was confirmed by PCR, PMI activity, germination tests, and by histochemical staining with 5-bromo-4-chloro-3-indolyl-β-D-glucuronide
(X-Gluc). Stable integration of the transgenes into the maize genome was demonstrated in T1 and T2 plants. Results indicate that the mannose selection system can be used for maize PEG-mediated protoplast transformation.
Received: 12 July 1999 / Revision received: 11 October 1999 / Accepted: 11 October 1999 |
| |
Keywords: | Maize Protoplast Transformation Mannose-selection marker |
本文献已被 SpringerLink 等数据库收录! |
|