Downregulation of a Mitochondrial NAD+ Transporter (NDT2) Alters Seed Production and Germination in Arabidopsis.

Despite the fundamental importance of nicotinamide adenine dinucleotide (NAD+) for metabolism, the physiological roles of NAD+ carriers in plants remain unclear. We previously characterized the Arabidopsis thaliana gene (At1g25380), named AtNDT2, encoding a protein located in the mitochondrial inner membrane, which imports NAD+ from the cytosol using ADP and AMP as counter-exchange substrates for NAD+. Here, we further investigated the physiological roles of NDT2, by isolating a T-DNA insertion line, generating an antisense line and characterizing these genotypes in detail. Reduced NDT2 expression affected reproductive phase by reducing total seed yield. In addition, reduced seed germination and retardation in seedling establishment were observed in the mutant lines. Moreover, remarkable changes in primary metabolism were observed in dry and germinated seeds and an increase in fatty acid levels was verified during seedling establishment. Furthermore, flowers and seedlings of NDT2 mutants displayed upregulation of de novo and salvage pathway genes encoding NAD+ biosynthesis enzymes, demonstrating the transcriptional control mediated by NDT2 activity over these genes. Taken together, our results suggest that NDT2 expression is fundamental for maintaining NAD+ balance amongst organelles that modulate metabolism, physiology and developmental processes of heterotrophic tissues.

The mitochondrial NAD+ transporter (NDT1) plays important roles in cellular NAD+ homeostasis in Arabidopsis thaliana.

Nicotinamide adenine dinucleotide (NAD+ ) is an essential coenzyme required for all living organisms. In eukaryotic cells, the final step of NAD+ biosynthesis is exclusively cytosolic. Hence, NAD+ must be imported into organelles to support their metabolic functions. Three NAD+ transporters belonging to the mitochondrial carrier family (MCF) have been biochemically characterized in plants. AtNDT1 (At2g47490), focus of the current study, AtNDT2 (At1g25380), targeted to the inner mitochondrial membrane, and AtPXN (At2g39970), located in the peroxisomal membrane. Although AtNDT1 was presumed to reside in the chloroplast membrane, subcellular localization experiments with green fluorescent protein (GFP) fusions revealed that AtNDT1 locates exclusively in the mitochondrial membrane in stably transformed Arabidopsis plants. To understand the biological function of AtNDT1 in Arabidopsis, three transgenic lines containing an antisense construct of AtNDT1 under the control of the 35S promoter alongside a T-DNA insertional line were evaluated. Plants with reduced AtNDT1 expression displayed lower pollen viability, silique length, and higher rate of seed abortion. Furthermore, these plants also exhibited an increased leaf number and leaf area concomitant with higher photosynthetic rates and higher levels of sucrose and starch. Therefore, lower expression of AtNDT1 was associated with enhanced vegetative growth but severe impairment of the reproductive stage. These results are discussed in the context of the mitochondrial localization of AtNDT1 and its important role in the cellular NAD+ homeostasis for both metabolic and developmental processes in plants.

Ecofisiologia da germinação de sementes de Campomanesia pubescens

The gabiroba bush [Campomanesia pubescens (DC.) O. Berg. (Myrtaceae)], a native species of the Cerrado is considered a plant with melliferous, ornamental and medicinal potential. Aiming to evaluate the influence of abiotic factors on seed germination, two experiments were conducted. The first was conducted in a completely randomized design in a factorial design(4x3x2), composed of four temperatures (15-25, 20-30°C alternate, 25 and 30°C constant), 3 substrates (paper, sand and vermiculite) and two light conditions (light and dark). The second experiment aimed to evaluate the tolerance to desiccation of fresh seeds dried superficially, with 35% of water, by drying it in oven with forced ventilation of air at 30°C for 30, 60, 90, 120, 150, 180, 260, 360, 750 and 810 minutes. To evaluate the physiological quality of seeds gabiroba, shrub planting on paper, in the dark and 30°C, enables a better expression of the force. The seeds of bush gabiroba are intolerant to desiccation and can be classified as recalcitrant.

A gabiroba de arbusto [Campomanesia pubescens (DC.) O. Berg. (Myrtaceae)], espécie nativa do Cerrado é considerada uma planta com potencial melífero, ornamental e medicinal. Com o objetivo de avaliar a influência de diferentes temperaturas, substratos e condições de luminosidade e da secagem, na qualidade fisiológica de suas sementes foram conduzidos dois experimentos. No primeiro utilizou-se sementes com 13% de teor de água, o qual foi realizado em delineamento experimental inteiramente casualizado em esquema fatorial (4x3x2), composto por quatro temperaturas (15-25, 20-30°C alternadas, 25 e 30°C constantes), três substratos (sobre papel, entre areia e entre vermiculita) e duas condições de luminosidade (claro e escuro) e, o segundo com sementes recém extraídas dos frutos e secas superficialmente, contendo 35% de teor de água e após a secagem em estufa com ventilação forçada de ar a temperatura de 30°C por 30, 60, 90, 120, 150, 180, 260, 360, 750 e 810 minutos. Para a avaliação da qualidade fisiológica de sementes de gabiroba de arbusto, a semeadura sobre papel, no escuro e a 30°C possibilita uma melhor expressão do vigor. As sementes de gabiroba de arbusto são intolerantes a secagem, podendo ser classificadas como recalcitrantes.

Ecofisiologia da germinação de sementes de Campomanesia pubescens

The gabiroba bush [Campomanesia pubescens (DC.) O. Berg. (Myrtaceae)], a native species of the Cerrado is considered a plant with melliferous, ornamental and medicinal potential. Aiming to evaluate the influence of abiotic factors on seed germination, two experiments were conducted. The first was conducted in a completely randomized design in a factorial design(4x3x2), composed of four temperatures (15-25, 20-30°C alternate, 25 and 30°C constant), 3 substrates (paper, sand and vermiculite) and two light conditions (light and dark). The second experiment aimed to evaluate the tolerance to desiccation of fresh seeds dried superficially, with 35% of water, by drying it in oven with forced ventilation of air at 30°C for 30, 60, 90, 120, 150, 180, 260, 360, 750 and 810 minutes. To evaluate the physiological quality of seeds gabiroba, shrub planting on paper, in the dark and 30°C, enables a better expression of the force. The seeds of bush gabiroba are intolerant to desiccation and can be classified as recalcitrant.

A gabiroba de arbusto [Campomanesia pubescens (DC.) O. Berg. (Myrtaceae)], espécie nativa do Cerrado é considerada uma planta com potencial melífero, ornamental e medicinal. Com o objetivo de avaliar a influência de diferentes temperaturas, substratos e condições de luminosidade e da secagem, na qualidade fisiológica de suas sementes foram conduzidos dois experimentos. No primeiro utilizou-se sementes com 13% de teor de água, o qual foi realizado em delineamento experimental inteiramente casualizado em esquema fatorial (4x3x2), composto por quatro temperaturas (15-25, 20-30°C alternadas, 25 e 30°C constantes), três substratos (sobre papel, entre areia e entre vermiculita) e duas condições de luminosidade (claro e escuro) e, o segundo com sementes recém extraídas dos frutos e secas superficialmente, contendo 35% de teor de água e após a secagem em estufa com ventilação forçada de ar a temperatura de 30°C por 30, 60, 90, 120, 150, 180, 260, 360, 750 e 810 minutos. Para a avaliação da qualidade fisiológica de sementes de gabiroba de arbusto, a semeadura sobre papel, no escuro e a 30°C possibilita uma melhor expressão do vigor. As sementes de gabiroba de arbusto são intolerantes a secagem, podendo ser classificadas como recalcitrantes.