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First published online April 27, 2007; 10.1104/pp.107.096933 Plant Physiology 144:836-845 (2007) © 2007 American Society of Plant Biologists OPEN ACCESS ARTICLE
Temporal and Spatial Expression of the Major Allergens in Developing and Germinating Peanut Seed1,[C],[OA]Department of Biology, University of Utah, Salt Lake City, Utah 84112 (I.-H.K.); Agronomy Department (P.S., M.G.), Plant Molecular and Cellular Biology Program (M.G.), and the Genetics Institute (M.G.), University of Florida, Gainesville, Florida 32610 (P.S., M.G.); and Department of Horticulture, University of Georgia, Tifton, Georgia 31793 (P.O.-A.)
Peanut (Arachis hypogaea) seed proteins Ara h 1, Ara h 2, and Ara h 3 are considered to be the major peanut allergens. However, little is known about their temporal and spatial expression during seed development and upon germination and seedling growth. In this study, transcript levels of the three major peanut allergen genes, ara h 1, ara h 2, and ara h 3, and their corresponding proteins were found in all cultivars. Expression patterns were heterogeneous depending on the specific peanut allergen gene and the cultivars tested. However, ara h 3 expression patterns among the cultivars were more variable than ara h 1 and ara h 2. Transcripts were tissue specific, observed in seeds, but not in leaves, flowers, or roots, and were undetectable during seed germination. In situ hybridizations and immunotissue prints revealed that both embryonic axes and cotyledons expressed the allergens. However, more ara h 1 and ara h 3 messenger RNA was detected in cotyledons relative to embryonic axes. Allergen polypeptide degradation patterns were different in embryonic axes compared with cotyledons during germination and seedling growth, with levels of Ara h 1 and Ara h 2 dramatically reduced compared to the Ara h 3 polypeptides in embryonic axes. These characterization studies of major peanut allergen genes and their corresponding seed storage proteins can provide the basic information needed for biochemical and molecular approaches to obtain a hypoallergenic peanut.
1 This work was supported by the U.S. Department of Agriculture (Cooperative State Research, Education, and Extension Service-administered special grant no. 00344209178), by the Georgia Peanut Commodity Commission, by the Florida Peanut Producers Association, and by the Institute of Food and Agricultural Sciences at the University of Florida. The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Maria Gallo (mgm{at}ufl.edu). [C] Some figures in this article are displayed in color online but in black and white in the print edition. [OA] Open Access articles can be viewed online without a subscription. www.plantphysiol.org/cgi/doi/10.1104/pp.107.096933 * Corresponding author; e-mail mgm{at}ufl.edu; fax 3522738284. Received January 31, 2007; accepted April 13, 2007; published April 27, 2007. Related articles in Plant Physiol.:
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