RESUMO
Fluorides released during the manufacture of aluminum, fertilizers, glass and ceramics are considered to be highly toxic to plants, causing damages in relatively small concentrations. This study aimed to evaluate fluoride (F) accumulation and its effects on Vitex polygama Cham. (Verbenaceae) exposed to simulated rain containing potassium fluoride in concentrations of 5, 10, 15 and 20 mg L-1. Vitex polygama presented no visual symptoms of foliar injury and accumulated a relatively small amount of F-, significant only for the treatments with higher concentrations of the pollutant. F- promoted an increase in nonphotochemical quenching (qN) and in the coefficient due to non-photochemical extinction (NPQ), which indicates higher dissipation of radiant energy in the heat form. Damages in the chloroplast structure, in cellular membranes and in epicuticular waxes, besides cytoplasm granulation and irregularities in the epidermal cell walls were also detected in plants exposed to the pollutant. The results show a considerable resistance of Vitex polygama to F and reinforce the prognostic value of physiological, anatomical and ultrastructural analysis in the detection of damages caused by the pollutant in the leaf structure of this species.(AU)
Fluoretos, liberados durante a manufatura de alumínio, fertilizantes, vidro e cerâmica, são considerados altamente tóxicos para as plantas, causando danos em concentrações relativamente pequenas. Este estudo teve como finalidade avaliar o acúmulo de flúor (F) e seus efeitos sobre Vitex polygama Cham. (Verbenaceae) em plantas expostas a chuva simulada contendo fluoreto de potássio em concentrações de 5, 10, 15 e 20 mg L-1. Vitex polygama não apresentou sintomas visíveis de injúria foliar e acumulou uma quantidade relativamente pequena de F-, significativa apenas para os tratamentos com maiores concentrações do poluente. O F- promoveu aumentos no coeficiente devido à extinção não fotoquímica (qN) e no coeficiente devido à extinção não fotoquímica (NPQ), que indicam maior dissipação de energia radiante na forma de calor. Danos na estrutura dos cloroplastos, em membranas celulares e nas ceras epicuticulares, além de granulação do citoplasma e irregularidades na parede celular de células epidérmicas, foram também detectados em plantas expostas ao poluente. Os resultados demonstram uma considerável resistência de Vitex polygama ao F e reforçam o valor prognóstico de análises fisiológicas, anatômicas e ultraestruturais na detecção de danos causados pelo poluente na estrutura foliar dessa espécie.(AU)
Assuntos
Vitex/anatomia & histologia , Vitex/crescimento & desenvolvimento , Vitex/fisiologia , Flúor/análiseRESUMO
Vitex trifolia is a shrub species with popular use as a medicinal plant, for which leaves, roots and flowers have been reported to heal different distresses. The increasing exploitation of these plants has endangered its conservation, and has importantly justified the use of biotechnological tools for their propagation. Our aim was to present an efficient protocol for plant regeneration through organogenesis; and simultaneously, to analyze the genetic homogeneity of the established clonal lines by Randomly Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers. Plantlet regeneration was achieved in callus cultures derived from stem, leaf and petiole explants of V. trifolia on a differently supplemented Murashige & Skoog medium, and incubated at 25 +/-2 degrees C under a light intensity of 61 micromol/m2s from cool white fluorescent lamps and a 16 h photoperiod. The rate of shoot bud regeneration was positively correlated with the concentration of hormones in the nutrient media. Shoot buds regenerated more rapidly from stem and petiole explants as compared to leaf explants on medium containing 11.10 microM BAP in combination with 0.54 microMNAA. Addition of 135.74-271.50 microM adenine sulphate (Ads) and 0.72-1.44 microM gibberellic acid (GA3) to the culture medium increased the growth of shoot buds. The highest rate of shoot bud regeneration responses was obtained in stem explants using 11.10 microM BAP in combination with 0.54 microM NAA, 271.50 microM Ads and 1.44 microM GA3. In vitro rooting of the differentiated shoots was achieved in media containing 1.23 microM indole butyric acid (IBA) with 2% (w/v) sucrose. Regenerated plantlets were successfully established in soil with 86% survival under field condition. Randomly Amplified Polymorphic DNA and Inter Simple Sequence Repeat markers analyses have confirmed the genetic uniformity of the regenerated plantlets derived from the second up to fifth subcultures. This protocol may help in mass propagation and conservation of this important medicinal plant of great therapeutic potential.
Assuntos
Plantas Medicinais/fisiologia , Regeneração/fisiologia , Vitex/fisiologia , Repetições de Microssatélites , Reguladores de Crescimento de Plantas/farmacologia , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Plantas Medicinais/classificação , Plantas Medicinais/efeitos dos fármacos , Técnica de Amplificação ao Acaso de DNA Polimórfico , Regeneração/efeitos dos fármacos , Vitex/classificação , Vitex/efeitos dos fármacosRESUMO
Vitex trifolia is a shrub species with popular use as a medicinal plant, for which leaves, roots and flowers have been reported to heal different distresses. The increasing exploitation of these plants has endangered its conservation, and has importantly justified the use of biotechnological tools for their propagation. Our aim was to present an efficient protocol for plant regeneration through organogenesis; and simultaneously, to analyze the genetic homogeneity of the established clonal lines by Randomly Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers. Plantlet regeneration was achieved in callus cultures derived from stem, leaf and petiole explants of V. trifolia on a differently supple mented Murashige & Skoog medium, and incubated at 25±2ºC under a light intensity of 61µmol/m2s from cool white fluorescent lamps and a 16h photoperiod. The rate of shoot bud regeneration was positively correlated with the concentration of hormones in the nutrient media. Shoot buds regenerated more rapidly from stem and petiole explants as compared to leaf explants on medium containing 11.10µM BAP in combination with 0.54µMNAA. Addition of 135.74-271.50µM adenine sulphate (Ads) and 0.72-1.44µM gibberellic acid (GA3) to the culture medium increased the growth of shoot buds. The highest rate of shoot bud regeneration responses was obtained in stem explants using 11.10µM BAP in combination with 0.54µM NAA, 271.50µM Ads and 1.44µM GA3. In vitro rooting of the differentiated shoots was achieved in media containing 1.23µM indole butyric acid (IBA) with 2% (w/v) sucrose. Regenerated plantlets were successfully established in soil with 86% survival under field condition. Randomly Amplified Polymorphic DNA and Inter Simple Sequence Repeat markers analyses have confirmed the genetic uniformity of the regenerated plantlets derived from the second up to fifth subcultures. This protocol may help in mass propagation and conservation of this important medicinal plant of great therapeutic potential.
Vitex trifolia es una especie arbustiva de uso popular como planta medicinal, sus hojas, raíces y flores se han reportado para la cura de diferentes aflicciones. El aumento de la explotación de estas plantas ha puesto en peligro su conservación y ha justificado el uso de herramientas biotecnológicas para su propagación. El objetivo de esta investigación fue presentar un protocolo eficiente para la regeneración de estas plantas a través de la organogénesis, y analizar la homogeneidad genética de las líneas clonales establecidas por ADN polimórfico amplificado aleatoriamente (RAPD) mediante la repetición de marcadores de inter secuencia simple (ISSR). La regeneración de plántulas se logró en cultivos de callos derivados de explantes de tallo, hoja y pecíolo de V. trifolia en un medio diferenciado Murashige & Skoog, que se incubaron a 25±2ºC bajo una intensidad de luz de 61μmol/m2s con lámparas fluorescentes blancas y un fotoperíodo de 16h. La tasa de regeneración de brotes se correlacionó positivamente con la concentración de las hormonas en el medio nutritivo. Los brotes se regeneraron más rápidamente a partir de explantes de tallo y pecíolos en comparación con explantes de hoja. La mayor tasa de regeneración de brotes se obtuvo en los explantes de tallo utilizando 11.10μM BAP en combinación con 0.54μM NAA, 271.50μM Ads y 1.44μM GA3. Este protocolo puede ayudar a la propagación masiva y conservación de esta importante planta medicinal de gran potencial terapéutico.