PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: 95pg1

GENETIC AND MOLECULAR ANALYSIS OF EMP2 AND DSC1: TWO NOVEL MAIZE GENES FUNCTIONING IN KERNEL DEVELOPMENT.

*Michael J. Scanlon, #Philip S. Stinard, #Donald S. Robertson and *Alan M. Myers, *Department of Biochemistry and Biophysics, Department of Zoology and Genetics, Iowa State University, Ames, IA 5001 1.


A transposon tagging experiment using the Robertson's Mutator (Mu) maize transposable element system has generated over 100 recessive mutations causing various defects in kernel development. Among these were two novel defective kernel (dek) mutants, empty pericarp 2 (emp2) and discolored 1 (dsc1). The emp2 mutation causes an empty pericarp, germless phenotype that is homozygous lethal. Classical genetic techniques were used to place emp2 to chromosome 2L, approximately 18.5 cM proximal to w3. emp2 mutant kernels are first distinguishable from wild type sibling kernels 14 days after pollination (DAP). Embryos homozygous for emp2 are developmentally arrested, and cannot be rescued on enriched tissue culture medium. A genomic DNA fragment corresponding to the mutation emp2 was obtained through phage lambda cloning of a Mu transposon-tagged restriction fragment. Maize genomic DNA flanking the Mu insertion in the emp2 clone was used as a probe in Northern hybridization analyses to identify an RNA that is expressed in developing kernels. Efforts are now underway to isolate an Emp2 cDNA clone from a 14 DAP cDNA library. A similar approach was used to clone a Mu- tagged restriction fragment corresponding to the novel kernel mutation dsc1. This lethal mutation causes a discolored, germless seed and was mapped to approximately 4 cM from su1. dsc1 mutants are first distinguishable at 8 DAP, and mutant embryos are developmentally retarded. Embryos dissected from 20 DAP dsc1 kernels germinate on enriched tissue culture medium, producing a seedling that dies at the single leaf stage. A genomic DNA fragment flanking the Mu1 insertion in the dsc1 clone was used as a probe in Northern analyses to identify a kernel specific transcript that is expressed 6 through 9 days after pollination. A 7 DAP kernel library is being screened to isolate a Dsc1 DNA clone.


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