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

被引：96

作者：

Wang, AS ^{[1
]}

Evans, RA ^{[1
]}

Altendorf, PR ^{[1
]}

Hanten, JA ^{[1
]}

Doyle, MC ^{[1
]}

Rosichan, JL ^{[1
]}

机构：

[1] Novartis Seeds Inc, Appl Biotechnol, Stanton, MN 55018 USA

来源：

PLANT CELL REPORTS | 2000年 / 19卷 / 07期

关键词：

maize; protoplast; transformation; mannose-selection marker;

D O I：

10.1007/s002999900181

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

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-3-chloro-3-indolyl-beta-D-glucuronide (X-GIuc). Stable integration of the transgenes into the maize genome was demonstrated in T-1 and T-2 plants. Results indicate that the mannose selection system can be used for maize PEG-mediated protoplast transformation.

引用

页码：654 / 660

页数：7

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