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GrainGenes Reference Report: PRT-20-429

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Reference	PRT-20-429
Title	Efficient biolistic transformation of maize (Zea mays L.) and wheat (Triticum aestivum L.) using the phosphomannose isomerase gene, pmi, as the selectable marker
Journal	Plant Cell Reports
Year	2001
Volume	20
Pages	429-436
Author	Wright M [ Show all 10 ]
Abstract	Summary: A selectable marker system for plant transformation that does not require the use of antibiotics or herbicides was developed. The selectable marker consists of the manA gene from Escherichia coli under the control of a plant promoter that encodes for phosphomannose isomerase, pmi. Only transgenic plants were able to metabolize the selection agent, mannose, into a usable source of carbon, fructose. Transgenic plants were produced efficiently after delivery by Biolistics of the pmi gene into maize and wheat tissues, with mean transformation frequencies of 45% for maize and 20% for wheat. Adjustment of the sucrose and mannose levels in the selection medium essentially eliminated escapes. Transgenic events can be identified as early as 2 months for wheat and 4 months for maize. A simple test, a modified chlorophenol red assay, was used for early identification of transgenic events expressing the pmi gene. Transformation frequencies for both crops exceeded those obtained with the bar and pat genes with selection on either Basta or bialaphos
Keyword	[ Hide all but 1 of 43 ] antibiotic assay bar biolistic biolistic transformation carbon coat protein crop delivery escape escherichia coli events frequencies fructose gene herbicide immature embryo maize mannose marker media microprojectile bombardment mosaic virus phosphomannose isomerase plant transformation pmi promoter rapid production resistance selectable marker selection source sucrose system tissue transformation transformation frequency transgenic transgenic plant transgenic wheat triticum triticum aestivum zea mays

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