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Plant Physiology 135:1162-1169 (2004)
© 2004 American Society of Plant Biologists

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BREAKTHROUGH TECHNOLOGIES

Ice-Cap. A High-Throughput Method for Capturing Plant Tissue Samples for Genotype Analysis1

Patrick Krysan*

Department of Horticulture and Genome Center of Wisconsin, University of Wisconsin, Madison, Wisconsin 53706

High-throughput genotype screening is rapidly becoming a standard research tool in the post-genomic era. A major bottleneck currently exists, however, that limits the utility of this approach in the plant sciences. The rate-limiting step in current high-throughput pipelines is that tissue samples from living plants must be collected manually, one plant at a time. In this article I describe a novel method for harvesting tissue samples from living seedlings that eliminates this bottleneck. The method has been named Ice-Cap to reflect the fact that ice is used to capture the tissue samples. The planting of seeds, growth of seedlings, and harvesting of tissue are all performed in a 96-well format. I demonstrate the utility of this system by using tissue harvested by Ice-Cap to genotype a population of Arabidopsis seedlings that is segregating a previously characterized mutation. Because the harvesting of tissue is performed in a nondestructive manner, plants with the desired genotype can be transferred to soil and grown to maturity. I also show that Ice-Cap can be used to analyze genomic DNA from rice (Oryza sativa) seedlings. It is expected that this method will be applicable to high-throughput screening with many different plant species, making it a useful technology for performing marker assisted selection.

1 This work was supported by the U.S. Department of Agriculture (NRI, CSREES; USDA grant no. 2003–02593).

www.plantphysiol.org/cgi/doi/10.1104/pp.104.040774.

* E-mail fpat{at}biotech.wisc.edu; fax 608–262–4743.

Received February 10, 2004; returned for revision March 31, 2004; accepted April 1, 2004.


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Plant Physiol. 2004 135: 1147-1148. [Full Text]  





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