ABSTRACT Ultraviolet B radiation has been described as a potential elicitor agent of the synthesis of secondary metabolites in plants. Thus, the present study aimed to investigate the production of betalains and total flavonoids, as well as the antioxidant activity, of Alternanthera sessilis, A. brasiliana, A. tenella and A. philoxeroides exposed to different periods of UV-B radiation (280-315 nm). Plants of these four species were exposed to UV-B radiation for 0, 2, 4, 6, and 8 hours, which amounts to 0, 10, 20, 30, and 40 J cm−2 of radiation, respectively. Significant increases in betacyanin and betaxanthin levels were observed in A. sessilis and A. brasiliana during the period of UV-B exposure, while no differences were observed for the others species. The highest estimated flavonoid levels were for A. sessilis exposed to UV-B radiation for 8 h, followed by a 24-h recovery period. In conclusion, the action of UV-B radiation as an inducer of defence responses in plants is influenced by increasing exposure periods followed by a recovery period. Both increase the levels of these composts, yet this increase is different among the four Alternanthera species, having a greater influence on the species A. sessilis and A. brasiliana.

Salinity is a limiting factor fot growth and development, since it affects several physiological processes in plants. This trial aimed to assess the levels of photosynthetic pigments, betacyanins, lipid peroxidation and activity of antioxidant enzymes in Alternanthera philoxeroides under salt stress. Plants originated from an in vitro culture were acclimatized and irrigated with sodium chloride (0, 200 and 400 mM) for 30 d. The experimental design was completely randomized with a 3×2 factorial arrangement of treatments: three concentrations of NaCl and two plant tissues (stems and leaves). The experimental data was subjected to an analysis of variance and means where compared using Tukey test (p≤0.05). The levels of chlorophylls a, chlorophylls b and total carotenoids presented similar responses, decreasing their values according to salt concentrations, while the chlorophyll a/b ratio of the plants that were submitted to the highest salt concentration presented a significant increase when compared to the control. Higher levels of betacyanin were observed on stems, when compared to the leaves, in the highest salt concentrations. On the leaves, there was a significant increase of lipid peroxidation and superoxide dismutase activity, whereas roots showed an increase of the enzymes catalase and ascorbate peroxidase. Salt stress caused greater degradation in the photosynthetic pigments, increment of betacyanin synthesis in stems, and damage to the cell membranes of the leaves. However, the increase of antioxidant enzymes activity stimulated the protective system against oxidative stress. It is concluded that Alternanthera philoxeroides (Mart.) Griseb. plants present reduction in levels of photosynthetic pigments, increased synthesis of betacyanins and increased activity of antioxidant enzymes in leaves and roots when exposed to salt stress.

La salinidad es un factor limitante para el crecimiento y el desarrollo, ya que afecta varios procesos fisiológicos en las plantas. Este estudio tuvo el objetivo de evaluar los niveles de pigmentos fotosintéticos, betacianinas, peroxidación lipídica y actividad de enzimas antioxidantes en Alternanthera philoxeroides bajo estrés salino. Las plantas se originaron de un cultivo in vitro y se aclimataron e irrigaron con cloruro de sodio (0, 200 y 400 mM) durante 30 d. El diseño experimental fue completamente aleatorio con un arreglo factorial 3×2 de los tratamientos: tres concentraciones de NaCl y dos tejidos vegetales (tallos y hojas). Los datos experimentales se sometieron a un análisis de varianza y las medias se compararon usando la prueba de Tukey (p≤0.05). Los niveles de clorofilas a, clorofilas b y carotenoides totales presentaron respuestas similares, disminuyeron sus valores con las concentraciones de sal, mientras que la relación de clorofila a/b de las plantas que se expusieron a la concentración mayor de sal presentó un incremento significativo respecto al control. Niveles mayores de betacianinas se observaron en los tallos, en comparación con las hojas, para las concentraciones de sal más altas. En las hojas hubo un aumento significativo de peroxidación lipídica y de actividad de superóxido dismutasa, mientras que las raíces mostraron un aumento en las enzimas catalasa y ascorbato peroxidasa. El estrés salino causó degradación mayor de los pigmentos fotosintéticos, un aumento en la síntesis de betacianinas en los tallos, y daño a las membranas celulares de las hojas. Sin embargo, el incremento de actividad de las enzimas antioxidantes estimuló el sistema protector contra el estrés oxidativo. Se concluye que las plantas de Alternanthera philoxeroides (Mart.) Griseb. presentan una reducción en los niveles de pigmentos fotosintéticos, mayor síntesis de betacianinas y mayor actividad de enzimas antioxidantes en las hojas y raíces cuando están expuestas a estrés salino.

Promotores tecido-específico controlam a transcrição de genes em diferentes tecidos vegetais bem como em diferentes estádios de desenvolvimento da planta, levando à indução de distintos níveis de atividade transiente e/ou estável do gene. Tais promotores podem ser empregados para a expressão seletiva de genes de interesse. O promotor rol A de Agrobacterium rhizogenes, por exemplo, é floema-específico, sugerindo que possa ser empregado em estratégias de defesa de plantas que são infectadas por vírus com replicação restrita ao floema. A expressão do gene marcador da ß-glucuronidase (gus) dirigido pelo promotor rol A (pBRA3) foi observada em plantas transgênicas de batata (cvs. Macaca e Baronesa). Entrenós e secções de folhas foram submetidos ao cocultivo com A. tumefaciens. A atividade do gene gus avaliada em brotações resistentes à canamicina não se restringiu ao floema (alto nível de expressão do gene), mas também se manifestou no xilema dos caules. As expressões transiente e estável são, no entanto, tecido-específicas, localizadas sobretudo no sistema vascular de entrenós e ausente em raízes e folhas. As plantas gus positivas foram micropropagadas, plantadas em casa de vegetação e avaliadas por PCR, utilizando-se 'primers' específicos para o gene npt II. Nenhuma alteração fenotípica foi observada em plantas transgênicas, em relação às não transformadas.

Tissue-especific promoters allow the modulation of gene transcription in different tissue types as well as in different stages of plant development, leading different levels of transient and stable activity of the gene product. These promoters have been employed for selective gene expression. The Agrobacterium rhizogenes rol A gene promoter (BRA3) controls phloem-specific expression indicating that this promoter might have an important role in plant defense strategies against virus which replicated only in the phloem. The expression of ß-glucuronidase (gus) activity drived by BRA3 promoter was observed in transgenic potato plants (cvs Macaca and Baronesa). Entrenodes and leaf explants were infected and cocultivated with A. tumefaciens. The regeneration of putative transformants was in medium with 50 mg.L-1 of kanamycin. High frequency of transformants based upon kanamycin resistance and gus expression was obtained. All gus positive plants showed blue staining in vascular bundle (phloem and xylem). PCR analysis showed the amplification of npt II. No conspicuous phenotipic alteration was observed when comparing transgenic vs non-transgenic plants.

Com o objetivo de estudar a cinética da despolimerização dos microtúbulos e a dinâmica da indução de micronúcleos, usaram-se dois herbicidas com ação antimitótica, amiprofós-metil (APM) e oryzalin (ORY), em células de Helianthus maximiliani. A aplicação de ambos os herbicidas em células com ativo crescimento resultou num elevado número de células com micronúcleos, em virtude das modificações mitóticas causadas pela ausência de microtúbulos. Primeiramente, cromossomos isolados ou grupos de 2 ou mais cromossomos espalharam-se pelo citoplasma. Duas a três horas após a aplicação dos herbicidas, os microtúbulos, estavam despolimerizados e os cromossomos metafásicos condensaram-se, formando diretamente os micronúcleos sem ter havido a divisão do centrômero e a separação das cromátides. A concentração mais eficiente de ORY foi 20 µM, ao passo que para APM foi 60 µM, para um período de incubação de 36 e 48 h respectivamente. O prolongamento do período de tratamento elevou a freqüência de micronúcleos deformados, bem como aumentou a morte celular. A adição de citocalasina-B (CB) (20 µM), após 24 h da aplicação dos herbicidas, aumentou significativamente a micronucleação e estabilização dos micronúcleos nas células. O controle dos microtúbulos de células em suspensão torna-se uma ferramenta eficiente para a indução de micronúcleos, bem como representa um método elegante para a transferência genômica parcial.

In order to verify the kinetics of microtubule depolymerization and dynamics of micronuclei induction, two anti-mitotic herbicides, Amiprophos-methyl (APM) and Oryzalin (ORY), were added to dividing cells of Helianthus maximiliani. The addition of both herbicides to actively dividing cells resulted in a high number of cells with micronuclei through modification of mitosis. Single chromosomes and groups of 2 or more chromosomes were scattered over the cytoplasm. After 2-3 h of herbicide application, microtubules were depolymerized and the metaphase chromosomes changed directly into micronuclei, without centromere division and chromatid separation. The most efficient concentrations were 20 µM ORY and 60 µM APM when incubated for 36 and 48 h, respectively. When the treatment duration was increased, the frequency of cells with deformed micronuclei as well as cell death increased. The presence of citochalasina-B (CB) (20 µM) after 24 h of herbicide application increased the frequency of micronucleation and stabilization of the micronuclei in the cells significantly. The control of microtubules using anti-mitotic toxins provides an efficient tool for micronuclei induction as well as representing an elegant method for partial genome transfer.