title: Characterization of a potato activation-tagged mutant, nikku, and its partial revertant.
abstract: A potato mutant with a strong stress-response phenotype, and a partial mutant revertant, were characterized. Gene expression patterns and DNA cytosine methylation varied between these and wild-type, indicating a role for DNA cytosine methylation changes in the gene expression and visible phenotypes. Morphological and molecular studies were conducted to compare potato cv. Bintje, a Bintje activation-tagged mutant (nikku), and nikku revertant phenotype plants. Morphological studies revealed that nikku plants exhibited an extremely dwarf phenotype, had small hyponastic leaves, were rootless, and infrequently produced small tubers compared to wild-type Bintje. The overall phenotype was suggestive of a constitutive stress response, which was further supported by the greater expression level of several stress-responsive genes in nikku. Unlike the nikku mutant, the revertant exhibited near normal shoot elongation, larger leaves and consistent rooting. The reversion appeared partial, and was not the result of a loss of 35S enhancer copies from the original nikku mutant. Southern blot analyses indicated the presence of a single T-DNA insertion on chromosome 12 in the mutant. Gene expression studies comparing Bintje, nikku and revertant phenotype plants indicated transcriptional activation/repression of several genes flanking both sides of the insertion in the mutant, suggesting that activation tagging had pleiotropic effects in nikku. In contrast, gene expression levels for many, but not all, of the same genes in the revertant were similar to Bintje, indicating some reversion at the gene expression level as well. DNA methylation studies indicated differences in cytosine methylation status of the 35S enhancers between the nikku mutant and its revertant. In addition, global DNA cytosine methylation varied between Bintje, the nikku mutant and the revertant, suggesting involvement in gene expression changes, as well as mutant phenotype.
authors.affiliation: Department of Horticulture, Virginia Tech, Blacksburg, VA, 24060, USA.
authors.forename: Sukhwinder S
authors.initials: SS
authors.lastname: Aulakh
authors.forename: Richard E
authors.initials: RE
authors.lastname: Veilleux
authors.forename: Guozhu
authors.initials: G
authors.lastname: Tang
authors.forename: Barry S
authors.initials: BS
authors.lastname: Flinn
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism; D001681:Biological Assay; D015139:Blotting, Southern; D003562:Cytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D019175:DNA Methylation / Q000187:drug effects / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics; D018628:Gene Dosage; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D016296:Mutagenesis / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D010641:Phenotype; D035281:Plant Tubers / Q000187:drug effects / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011198:Solanum tuberosum / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D013312:Stress, Physiological / Q000187:drug effects
chemical_list: D004269:DNA, Bacterial; D012333:RNA, Messenger; C036483:T-DNA; D003562:Cytidine; D044503:5-Methylcytosine; C009131:pyrimidin-2-one beta-ribofuranoside