Hamburg, Germany
University of Hamburg
Identification of genes induced during early kernel development in Zea mays (L.) --Lorbiecke, R, Kukula, J, Paul, C, Wienand, U The identification of seed specific genes from maize endosperm is currently of growing interest to provide more efficient approaches for plant improvement. To get more insight into the regulation of endosperm and kernel development we performed a PCR-based subtractive hybridization based on the method described by Buchanan-Wollaston and Ainsworth (Plant Mol. Biol. 33:821-834, 1997). cDNAs for driver and target populations were synthesized using mRNAs from kernels 0 and 8 days after pollination. The enriched cDNA fragments were cloned and further screened for differentially regulated genes by dot blot hybridization using driver and target cDNAs as probes. Based on this screening we estimated that about 30-40% of the subtraction-enriched cDNAs represent differentially regulated genes. So far, all cDNA fragments cloned are different in sequence.

Northern analysis confirmed an induced, transient expression pattern of the genes analyzed, showing the efficiency of the subtraction and screening procedure. In addition, most of the isolated genes showed strongest expression in developing kernels and weak or no expression in other tissues analyzed, i.e. tassels, silks, leaves or young plants. Database searching led to the identification of new genes involved in lipid metabolism and pathogen response as well as genes already known to be endosperm specific, i.e. BETL2 Hueros et. al (1999).

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