RESUMEN
In Arabidopsis thaliana, EF-1 alpha proteins are encoded by a multigene family of four members. Three of them are clustered at the same locus, which was positioned 24 cM from the top of chromosome 1. A region of DNA spanning 63 kb around these locus was sequenced and analyzed. One main characteristic of the locus is the mosaic organization of both genes and intergenic regions. Fourteen genes were identified, among which only four were already described, and other unidentified are most likely present. Functionally diverse genes are found at close intervals. Exon and intron distribution is highly variable at this locus, one gene being split into at least 20 introns. Several duplications were found within the sequenced segment both in coding and noncoding regions, including two gene families. Moreover, a sequence corresponding to the 5' noncoding region of the EF-1 alpha genes and harboring a 5' intervening sequence is duplicated and found upstream of several genes, suggesting that noncoding regions can be shuffled during evolution.
Asunto(s)
Arabidopsis/genética , Genes de Plantas/genética , Familia de Multigenes/genética , Oxidorreductasas , Factores de Elongación de Péptidos/genética , Proteínas de Plantas/genética , Secuencia de Bases , Mapeo Cromosómico , Cromosomas Artificiales de Levadura , Glutarredoxinas , Datos de Secuencia Molecular , Factor 1 de Elongación Peptídica , Plantas Tóxicas , Proteínas/genética , Ricinus/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido NucleicoRESUMEN
The comparative analysis of plant hormones was undertaken on a 1-naphthaleneacetic acid tolerant mutant and normal tobacco (Nicotiana tabacum cv Xanthi) plantlets. The mutant plantlet was scrubby and impaired in its root morphogenesis. Degeneration of the root meristem was studied on tissue sections; it appeared very fast (as early as the 3rd or 4th day after sowing), after which the root was further transformed into a callus. Indoleacetic acid (IAA), abscisic acid (ABA), and the isopentenyladenine (iP)- and trans-zeatin(Z)-type cytokinin levels were measured in terminal buds and root tips 13 days after sowing, by enzyme linked immunosorbent assay of high performance liquid chromatography fractions. Some differences appeared between the apical buds of the two genotypes, but the mutant tobacco differed from the wild type mainly by the presence of higher levels of IAA, ABA, and iP + isopentenyladenosine (iPA) in its small root. Thus, the IAA, ABA, and iP + iPA contents were increased by a factor of 15, 7, and 24 times, respectively, in mutant root compared to wild-type tobacco roots. Previous work has shown that the mutation impairs membrane polarization effects induced by auxin at the cell level. The present results would favor the hypothesis that the mutation has also affected the control of growth regulator accumulation in tissues.
RESUMEN
A solid phase enzyme immunoassay was developed for isopentenyladenine (iP) and isopentenyladenosine (iPA) quantitation in HPLC purified plant extracts. It was performed on antigen-coated microtitration plates on which bound antibodies were indirectly labeled by the means of a biotinylated goat anti-rabbit antibody and an avidin-alkaline phosphatase conjugate. Less than 3 femtomoles of iP or iPA were easily detected and the measuring range extended from 3 femtomole to 1 picomole. The reproducibility has been tested and intra- and interassay variations did not exceed 5.0%. The specificity of iPA antibodies was good, as determined by cross-reactivity measurements with other adenylic compounds. The specificity of the measurements for iP and iPA was demonstrated by analysis of the immunoreactivity of fractions obtained by HPLC separation of a methanolic tobacco leaf extract.