Community Research and Development Information Service - CORDIS

A diagnostic assay for cold hardiness in pine seedlings

Based on the expression patterns of two dehydrin genes a test system was developed for assessment of hardiness of pine seedlings. The test is based on the principle of RT-PCR (cDNA synthesis followed by amplification of specific gene fragments) combined with immunodetection of the amplified gene fragments. For immunodetection the Nucleic Acid Lateral Flow Immuno Assay (NALFIA) was used. A mixture containing the amplified fragments is applied to the lateral flow system. At specific locations on the membrane antibodies are immobilized that capture specific labels attached to the amplified fragments. A second carbonated compound that binds to each amplified fragment is used to visualize the fragments on the membrane. Use of this assay system is simple, cheap and results are clear. The change in ratio of the signal intensity of both dehydrin genes indicates when the seedlings have become frost tolerant. a constitutively expressed gene serves as a technical control. Preliminary data, obtained from pine trees of Dutch origin, suggests that needle tissue can be used equally well as bud tissue. The use of gene expression as monitoring tool for seedling vitality is highly innovative.

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Agrotechnology & Food Innovations BV
P.O. Box 17
6700 AA Wageningen
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