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Stable transformation of maize: the impact of feeder cells on protoplast growth and transformation efficiency

✍ Scribed by Leszek A. Lyznik; Kathryn K. Kamo; Howard D. Grimes; Randy Ryan; Ka-Lai Chang; Thomas K. Hodges

Publisher: Springer
Year: 1989
Tongue: English
Weight: 612 KB
Volume: 8
Category: Article
ISSN: 0721-7714
DOI: 10.1007/bf00274133

No coin nor oath required. For personal study only.

✦ Synopsis

The importance of cell culture conditions, including the use of feeder cells, on protoplast growth and transformation in maize (Zea mays L.) was investigated. Total GUS activity, measured two days after transformation, was five-fold higher in protoplasts cultured on feeder cells compared to those grown in the absence of feeder cells. Since the specific activity of GUS was only slightly higher in the transformed protoplasts plated over feeder cells, the stimulation in transient gene expression resulted mainly from the improved environment provided by the feeder system. For stable transformation, either PEG treatment or electroporation of protoplasts was used to introduce the neo gene. When PEG was used, over 85% of the putative transformants (resistant to kanamycin) contained the neo gene. The combination of PEG transformation and the optimized cell culture protocol using feeder cells enabled the selection of about 100 stably transformed lines per gFW of cells. Electroporation was less efficient.

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