PAG-II Plant Genome II Conference

Town & Country Conference Center, San Diego, CA, January, 1994.


PG-II: TRANSPOSON TAGGING, ISOLATION AND CHARACTERIZATION OF A NUCLEAR MALE FERTILITY GENE FROM MAIZE

TRANSPOSON TAGGING, ISOLATION AND CHARACTERIZATION OF A NUCLEAR MALE FERTILITY GENE FROM MAIZE.

Tim W. Fox, Mary R. Trimnell and Marc C. Albertsen, Department of Biotechnology, Pioneer Hi-Bred International, Inc. Johnston, IA 50131


Genetic male sterility can result from a mutation in one of many genes involved in microsporogenesis. These genes are collectively referred to as male fertility genes. In maize, over 20 genetic loci have been identified as being essential for male fertility. Cytological studies of sterile mutants for many of these fertility loci indicate that microspore breakdown is diverse in timing as well as its effect on microspore morphology [M. C. Albertsen and R. L. Phillips, Can. J. Genet. Cytol., 21:195 (1981)]. This suggests that microsporogenesis is a complex process, controlled by more than one mechanism. Despite the number of male sterile mutants described in maize, little progress has been made in characterizing genes responsible for male fertility. Most male sterility research has been descriptive. Recently, we used the transposable element Activator (Ac) to tag a gene responsible for male fertility in Zea mays. This Ac induced mutation inhibits wall formation in developing microspores. The genomic DNA flanking the Ac element from the mutant was amplified using inverse-PCR. The amplified fragment was used to screen a cDNA library constructed from maize tassel MRNA. A putative full length cDNA (1.4Kb) was isolated. Northern analysis indicates the transcript is only present in tassels of fertile plants. The cDNA sequence contains a 1342 base pair open reading frame coding for a deduced polypeptide of 414 amino acid residues. To demonstrate gene function, plants were screened for Ac transposition from the mutant allele. A revertant allele was identified that contains a 6 base pair Ac footprint which restores the reading frame of the gene as well as restoring male fertility to the plant. The deduced polypeptide was compared to the Genbank Protein database. Homology (34% identity over 236 amino acids) was found to strictosidine synthase, an enzyme involved in the formation of strictosidine, a key intermediate in indole-type alkaloid production. This male fertility gene has been designated Ms45.


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