A mannose selection system for production of fertile transgenic maize plants from protoplasts.

Autor: Wang, A. S., Evans, R. A., Altendorf, P. R., Hanten, J. A., Doyle, M. C., Rosichan, J. L.
Zdroj: Plant Cell Reports; Jun2000, Vol. 19 Issue 7, p654-660, 7p
Abstrakt: 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. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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