title: Arabidopsis emb175 and other ppr knockout mutants reveal essential roles for pentatricopeptide repeat (PPR) proteins in plant embryogenesis.
abstract: Pentatricopeptide repeat proteins (PPRPs) constitute one of the largest superfamilies in plants, with more than 440 identified in the Arabidopsis thaliana (L.) Heynh genome. While some PPRPs are known to take part in organelle gene expression, little is known about the broader biological contexts of PPRP gene function. Here, using developmental- and reverse-genetic approaches, we demonstrate that a number of PPRPs are essential early in plant development. We have characterized the Arabidopsis embryo-defective175 mutant and identified the EMB175 gene. Emb175 consistently displays aberrant cell organization and undergoes morphological arrest before the globular-heart transition. The emb175 mutation disrupts an intronless open reading frame encoding a predicted chloroplast-localized PPR protein- the first to be rigorously associated with an early embryo-lethal phenotype. To determine if other PPRP genes act in embryogenesis, we searched Arabidopsis insertion mutant collections for pprp knockout alleles, and identified 29 mutants representing 11 loci potentially associated with embryo-defective phenotypes. We assessed gene structures, T-DNA insertion position, and allelism for these loci and were able to firmly establish essential functions for six PPRP genes in addition to EMB175. Interestingly, Nomarski DIC microscopy revealed diverse embryonic defects in these lines, ranging from early lethality to dramatic late-stage morphological defects such as enlarged shoot apices and stunted cotyledons. Together, emb175 and these pprp knockout mutants establish essential roles for PPRPs in embryogenesis, thus broadening the known organismal context for PPRP gene function. The diversity of emb-pprp knockout phenotypes indicates that mutation of different PPRPs can, directly or indirectly, have distinct impacts on embryo morphogenesis.
authors.affiliation: Department of Biology, and Program in Biochemistry, Biophysics and Molecular Biology, Whitman College, Walla Walla, WA 99362, USA.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Cushing
authors.forename: Nancy R
authors.initials: NR
authors.lastname: Forsthoefel
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Gestaut
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Vernon
mesh_terms: D000483:Alleles; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000196:embryology / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics*; D001483:Base Sequence; D002874:Chromosome Mapping; D004269:DNA, Bacterial / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D010641:Phenotype; D020449:Repetitive Sequences, Amino Acid / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000235:genetics / Q000254:growth & development; D017386:Sequence Homology, Amino Acid
chemical_list: D029681:Arabidopsis Proteins; D004269:DNA, Bacterial; C088087:GNOM protein, Arabidopsis; D020662:Guanine Nucleotide Exchange Factors; C036483:T-DNA