ABSTRACT
UNLABELLED: DNA methylation is an epigenetic regulatory mechanism of gene expression which can be associated with developmental phases and in vitro morphogenetic competence in plants. The present work evaluated the effects of 5-azacytidine (AzaC) and 2,4-dichlorophenoxyacetic acid (2,4-D) on Acca sellowiana somatic embryogenesis (SE) and global DNA methylation levels by high-performance liquid chromatography mass spectrometry (HPLC/MS/MS). 2,4-D-free treatments revealed no somatic embryo formation in both accessions tested. Treatments supplemented with 2,4-D pulse plus AzaC in the culture medium resulted in increased embryo formation. In AzaC-free treatment, HPLC/MS/MS analysis showed a gradual increase in methylation levels in cultures of both accessions tested during SE induction. Treatment with AzaC and 2,4-D-free resulted in a marked decrease in methylation for both accessions, ranging from 37.6 to 20.8 %. In treatment with 2,4-D and AzaC combined, the 85 accession showed increasing global methylation levels. Otherwise, the 101X458 accession, in the same treatment, showed a decrease between 10 and 20 days, followed by an increase after 30 days (39.5, 36.2 and 41.6 %). These results indicate that 2,4-D pulse combined with AzaC improves SE induction. However, the conversion phase showed that although positively influencing SE induction, AzaC had a dysregulatory effect on the stage of autotrophic plant formation, resulting in significantly lower conversion rates. The results suggest that DNA methylation dramatically influences SE in Acca sellowiana, and global DNA methylation dynamics are related to morphogenetic response. KEY MESSAGE: 5-Azacytidine combined with 2,4-D increases the number of Acca sellowiana somatic embryos. Global DNA methylation is directly affected by these compounds.
Subject(s)
2,4-Dichlorophenoxyacetic Acid/pharmacology , Azacitidine/pharmacology , DNA Methylation , Feijoa/drug effects , Plant Somatic Embryogenesis Techniques/methods , Seeds/drug effects , Chromatography, High Pressure Liquid , Culture Media/metabolism , DNA, Plant/genetics , DNA, Plant/metabolism , Epigenesis, Genetic , Feijoa/embryology , Feijoa/genetics , Gene Expression Regulation, Plant , Genes, Plant , Mass Spectrometry , Seeds/embryology , Seeds/geneticsABSTRACT
Araucaria angustifolia (Bert.) O. Kuntze is a dioecious conifer species native of Brazil. The rare occurrence of monoiceous specimens have been attributed to pathogenic infections or other injuries in adult trees. Here, the morphological characteristics of male and female cones and pollen grains of a monoiceous A. angustifolia are described. Male and female cones and pollen grains presented normal morphology, lacking any sort of injuries or infection and suggesting the existence of further grounds for the occurrence of monoicy in this conifer species.
Araucaria angustifolia (Bert.) O. Kuntze é uma conífera primariamente dióica nativa do Brasil. A ocorrência de raros exemplares monóicos tem sido creditada a infecções patogênicas ou outras injúrias em árvores adultas. Neste trabalho, as características morfológicas de cones masculinos e femininos e dos grãos de pólen de uma A. angustifolia monóica são descritas. Os cones masculinos e femininos e os grãos de pólen apresentaram morfologia normal, se a presença de qualquer tipo de injuria ou infecção, sugerindo a existência de outros mecanismos para a ocorrência de monoicia nesta conífera.
Subject(s)
Tracheophyta/anatomy & histology , Ovule/anatomy & histology , Pollen/anatomy & histology , Brazil , Tracheophyta/physiology , Pollination , Reproduction/physiologyABSTRACT
Araucaria angustifolia (Bert.) O. Kuntze is a dioecious conifer species native of Brazil. The rare occurrence of monoiceous specimens have been attributed to pathogenic infections or other injuries in adult trees. Here, the morphological characteristics of male and female cones and pollen grains of a monoiceous A. angustifolia are described. Male and female cones and pollen grains presented normal morphology, lacking any sort of injuries or infection and suggesting the existence of further grounds for the occurrence of monoicy in this conifer species.
