Differential DNA methylation and gene expression during development of reproductive and vegetative organs in Ilex species

• Autores: Cascales, Jimena; Acevedo, Raúl Maximiliano; Paiva, Daniela Ivana; Gottlieb, Alexandra Marina
• Año de publicación: 2021
• Idioma: inglés
• Tipo de recurso: artículo
• Fuente: Journal of Plant Research ( Published: 23 March 2021)
• Materia: Ilex; Ilex paraguariensis; Expresión Génica; Marcadores Genéticos; Análisis Multivariante; Metilación; Gene Expression; Genetic Markers; Multivariate Analysis; Methylation; Marcadores Moleculares; Yerba Mate
• Institución: Instituto Nacional de Tecnología Agropecuaria

Differential epigenetic (DNA cytosine methylation) and gene expression patterns were investigated in reproductive and vegetative organs from Ilex paraguariensis and I. dumosa, at distinct developmental stages. We aimed at contributing towards elucidating major molecular changes underlying the sexual differentiation processes which, in these dioecious species, are completely unknown. Simultaneously, as a first step towards the development of an early sexing system, we searched for promising molecular markers. This was assessed through Methylation Sensitive Amplified Polymorphism (MSAP) and Amplified Fragment Length Polymorphism on cDNA (cDNA-AFLP) techniques, applying discriminant multivariate analyses, and bioinformatic characterization of differential fragments. A significant positive correlation was found between epigenetic and indirect ‘genetic’ information for both species, indicating influence of the genetic background on the epigenetic variation. Higher epigenetic than genetic diversities were estimated. Our outcomes showed up to 1.86 times more representation of mCG subepiloci than mCCG in all organs sampled. Along the maturing stages of floral buds, the frequency of mCG evidenced an incremental trend, whereas mCCG and unmethylated conditions showed opposite tendencies. Reproductive and vegetative samples tended to cluster apart based on epigenetic patterns; at gene expression level, organs exhibited clear-cut distinctive patterns, nonetheless profiles of young leaves and floral primordia resemble. Epigenetic and expression data allowed discrimination of I. dumosa´s samples according to the gender of the donor; more elusive patterns were observed for I. paraguariensis. In total, 102 differentially methylated and expressed fragments were characterized bioinformatically. Forty-three were annotated in various functional categories; four candidate markers were validated through qPCR, finding statistical differences among organs but not among sexes. The methylation condition of epilocus C13m33 appears as indicative of gender in both species. Thirty-three organ-specific and 34 gender-specific methylated markers were discriminated and deserve further research, particularly those expressed in leaves. Our study contributes concrete candidate markers with potential for practical application.
EEA Montecarlo
Fil: Cascales, Jimena. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina
Fil: Cascales, Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Acevedo, Raúl M. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste. Laboratorio de Biotecnología Aplicada y Genómica Funcional; Argentina
Fil: Acevedo, Raúl M. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina
Fil: Paiva, Daniela Ivana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Montecarlo; Argentina
Fil: Paiva, Daniela Ivana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gottlieb, Alexandra Marina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina
Fil: Gottlieb, Alexandra Marina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina