Caracterización molecular y fenotipica de mutantes apc5 de Arabidopsis Thaliana.

Abstract

Cell cycle regulation is essential for the growth and development of eukaryotic organisms. The anaphase promoter/cyclosome complex (APC/C) is a key E3 ubiquitin ligase that governs cell cycle transitions by degrading regulatory proteins. This study investigates the effects of disruption of the APC5 gene, which encodes a subunit of the APC/C complex, in Arabidopsis thaliana. The research focused on plants of the SALK_024997 line, characterized by a T-DNA insertion in intron 9 of the APC5 gene. The experimental design consisted of genotypic characterization of 16 plants by polymerase chain reaction (PCR) to identify the presence of the T-DNA insertion, followed by phenotypic analyses including root length measurement and embryo viability assessment. In addition, quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was performed to quantify APC5 gene expression levels, and GUS histochemical assays combined with in silico analysis were used to determine its spatial expression. The results confirmed that fifteen (15) of the plants were heterozygous for the T-DNA insertion, one was wild-type for the gene and no viable homozygous mutants were found, underlining the essential role of APC5. Gene expression analyses revealed a slight reduction in APC5 transcript levels in heterozygous plants. Phenotypic evaluations showed no significant differences in root length; however, embryo viability was markedly reduced to 50 % in mutant plants, compared to 97 % observed in wild-type controls. Spatial expression analysis showed that APC5 is active in roots, leaves, and reproductive tissues, suggesting its involvement in diverse developmental processes. This research highlights the fundamental role of APC5 in Arabidopsis thaliana, particularly in reproductive development, and provides crucial information for future studies on the specialized functions of the APC/C complex in plants.