MOLECULAR CHARACTERIZATION OF PgiC IN A TETRAPLOID PLANT AND ITS DIPLOID RELATIVES.

Abstract

Allotetraploid plant species combine the genomes of related diploid species, but little is known about whether homologous genes from the diploid genomes are expressed, how they interact, or whether they evolve differently when in a common tetraploid nucleus. Polyploidy may lead to gene silencing, but few molecular characterizations of silenced genes encoding enzymes in polyploids and related diploids have been reported. Here we describe the PgiC genes in the tetraploid Clarkia gracilis and related diploid species, which are native from California to southern Washington. PgiC encodes the cytosolic isozyme of phosphoglucose isomerase (PGIC; EC 5.3.1.9). The gene was duplicated in the basal stock of Clarkia and now both genes, PgiC1 and PgiC2, are active in about half of the diploid species, whereas only PgiC1 is active in the others. Clarkia gracilis was found to have three PgiC genes: two PgiC1s and a PgiC2. Reverse-transcriptase-polymerase chain reaction (RT-PCR) experiments, starting with mRNAs prepared from seedling leaves of C. gracilis, showed that the three genes are expressed. Analysis of their sequences showed they are evolving at similar rates to their homologues and that they have the same intron-exon structure. The presence of an expressed PgiC2 in C. gracilis was unexpected because all related diploid species, including one identified as a parent, have only active PgiC1s. The donor of the PgiC2 is now presumed extinct, but parsimony analysis identified its phylogenetic position. None of the PgiC genes that were active when C. gracilis arose were silenced. A possible example of gene conversion involving a 300-nuclectide region of one PgiC1 and PgiC2 was identified, but it probably occurred in the diploid parental species rather than in C. gracilis. PgiC2 is the first known example of an active locus in a tetraploid plant species that is no longer expressed in its diploid relatives.