NDLI: A simple and sensitive high-throughput GFP screening in woody and herbaceous plants

Content Provider	Springer Nature Link
Author	Hily, Jean Michel Liu, Zongrang
Copyright Year	2008
Abstract	Green fluorescent protein (GFP) has been used widely as a powerful bioluminescent reporter, but its visualization by existing methods in tissues or whole plants and its utilization for high-throughput screening remains challenging in many species. Here, we report a fluorescence image analyzer-based method for GFP detection and its utility for high-throughput screening of transformed plants. Of three detection methods tested, the Typhoon fluorescence scanner was able to detect GFP fluorescence in all Arabidopsis thaliana tissues and apple leaves, while regular fluorescence microscopy detected it only in Arabidopsis flowers and siliques but barely in the leaves of either Arabidopsis or apple. The hand-held UV illumination method failed in all tissues of both species. Additionally, the Typhoon imager was able to detect GFP fluorescence in both green and non-green tissues of Arabidopsis seedlings as well as in imbibed seeds, qualifying it as a high-throughput screening tool, which was further demonstrated by screening the seedlings of primary transformed T0 seeds. Of the 30,000 germinating Arabidopsis seedlings screened, at least 69 GFP-positive lines were identified, accounting for an approximately 0.23% transformation efficiency. About 14,000 seedlings grown in 16 Petri plates could be screened within an hour, making the screening process significantly more efficient and robust than any other existing high-throughput screening method for transgenic plants.
Starting Page	493
Ending Page	501
Page Count	9
File Format	PDF
ISSN	07217714
Journal	Plant Cell Reports
Volume Number	28
Issue Number	3
e-ISSN	1432203X
Language	English
Publisher	Springer-Verlag
Publisher Date	2008-12-18
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	GFP Fluorescence image analyzer High-throughput screening Arabidopsis Apple Plant Biochemistry Biotechnology Plant Sciences Cell Biology
Content Type	Text
Resource Type	Article
Subject	Plant Science Medicine Agronomy and Crop Science

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