ABSTRACT. Baccharis dracunculifolia DC. is a pioneer plant native from Brazil that has great importance due to its secondary metabolites (essential oil and Brazilian green propolis) and its potential in the recovery of degraded areas. The main goal of this study was to evaluate the propagation techniques of B. dracunculifolia by sexual reproduction and asexual propagation of male and female stem cuttings. For reproduction, female individuals from a natural population were periodically monitored for their reproductive development and their diaspores were collected to evaluate the maximum dry mass accumulation to determine physiological maturity. The germination test was performed by testing four temperatures (20, 25, 30, and 20-30°C), in addition to the germination speed index (GSI) and first count. For vegetative propagation, stem cuttings (8 cm) of male and female individuals were treated with an indole butyric acid (IBA) solution at 0, 1,500, 3,000, 4,500, and 6,000 mg L−1. The cuttings were evaluated for mortality, survival, rooting, sprouting, leaf retention, callogenesis, number of roots, and average length of roots (cm) after 120 days. The physiological maturity of diaspores occurred at 40 days after anthesis, which was the best time for collection in the field. The temperatures of 25 or 20-30°C should be used in the germination tests of the species. The first count was identified four days after sowing and the last count after 11 days. The rooting of B. dracunculifolia cuttings is very low. Sex did not influence the evaluated parameters, but increasing IBA doses positively influenced rooting, number of roots, and average length of roots and negatively influenced calluses formation.

Resumo O objetivo deste trabalho foi avaliar os compostos bioativos e a anatomia foliar de morfotipos de erva-mate (Ilex paraguariensis). A capacidade antioxidante, os compostos fenólicos totais, as metilxantinas e os ácidos cafeoilquínicos foram determinados a partir do extrato aquoso das folhas dos seguintes morfotipos de erva-mate: sassafrás, verde-escuro, verde fosco, cinza e peludinha. A microscopia de luz e a microscopia eletrônica de varredura foram utilizadas para observar a diferenciação anatômica. O morfotipo verde-escuro apresenta os maiores teores de cafeína (20,4 mg g-1) e difere significativamente apenas do morfotipo peludinha (8,8 mg g-1). Todos os morfotipos apresentaram atividade antioxidante semelhante e alta (886,0 e 588,1 μmol de atividade antioxidante equivalente a Trolox por grama de amostra, para ABTS e DPPH, respectivamente) e elevado conteúdo de compostos fenólicos totais (73,9 mg de equivalente a ácido gálico por grama de amostra). Embora a anatomia do mesofilo da folha seja semelhante, a deposição de cera e a configuração do estriamento da cutícula na superfície adaxial diferem entre os cinco morfotipos.

Abstract The objective of this work was to evaluate the bioactive compounds and foliar anatomy of yerba mate (Ilex paraguariensis) morphotypes. The antioxidant capacity, total phenolic compounds, methylxanthines, and caffeoylquinic acids were determined from the aqueous extract of leaves of the following yerba mate morphotypes: “sassafras”, dark green, dull green, gray, and “peludinha”. Light microscopy and scanning electron microscopy were used to observe the anatomical differentiations. The dark-green morphotype showed the highest contents of caffeine (20.4 mg g-1), differing significantly only from the “peludinha” morphotype (8.8 mg g-1). All morphotypes showed similar and high antioxidant activity (886.0 and 588.1 μmol Trolox equivalent antioxidant activity per gram of sample for ABTS and DPPH, respectively), as well as high total phenolic compounds content (73.9 mg gallic acid equivalent per gram of sample). Although the anatomy of the leaf mesophyll is similar, the wax deposition and cuticle striation configuration on the adaxial surface differ between all five morphotypes.

HIGHLIGHTS Essential oils from populations of B. dracunculifolia were investigated. β-pinene and (E)-nerolidol were the main compounds in B. dracunculifolia populations. The difference in the chemical profile of the essential oils is quantitative only. There is a negative correlation between the antioxidant activity and spathulenol.

Abstract Ilex paraguariensis (yerba mate) is a native species from South America and is a rich source of bioactive compounds. There is a lack of research efforts on the phytochemical investigation of callus culture from this species. In the present study, an effort was made to optimize callus culture conditions and to identify secondary compounds. Calli were induced from 10 genotypes using leaf explants and the best genotype was selected to evaluate the effects of cytokinin types and concentrations on callus induction and biomass accumulation. The best genotype and cytokinin treatment were used to conduct one last experiment with sucrose concentrations in culture media and its effects on calli biomass, antioxidant activity and secondary compounds accumulation. Callus initiation was genotype dependent, and the 6-156-6 line had the best response. Zeatin supplemented medium showed higher callus induction rate (82%) and higher biomass accumulation after 120 days (328.2 mg). Higher biomass and secondary compounds accumulation were observed for calli on 3% sucrose medium. Antioxidant activity was not affected by sucrose concentrations. Yerba mate callus culture allowed the accumulation of chlorogenic acid, cryptochlorogenic acid, neochlorogenic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, theobromine and caffeine.

RESUMO Passiflora actinia Hook (nome popular: ‘maracujá-do-mato’) é uma espécie medicinal relevante devido às suas propriedades sedativas e ansiolíticas. Para uma exploração comercial da planta, no entanto, são necessários estudos de propagação, especialmente visando melhorar o enraizamento em estacas caulinares. O presente estudo foi desenvolvido para avaliar o efeito do extrato da alga marinha parda (Ascophyllum nodosum (L.) Le Jol.) aplicado na base de estacas caulinares de P. actinia. Foram avaliadas cinco concentrações do extrato diluído em água destilada: 0% (água destilada pura), 10%, 20%, 30% e 40%. O delineamento experimental foi o inteiramente casualizado com 4 repetições e 12 estacas por unidade amostral. As estacas foram avaliadas 45 dias após o plantio. Os dados foram analisados por meio de análise de regressão polinomial e coeficientes de correlação de Pearson foram calculados para todas as variáveis. Em média, verificou-se 51,27% de enraizamento. A porcentagem de enraizamento aumentou linearmente de acordo com as concentrações de extrato de alga marinha. Quando comparado ao tratamento controle, observou-se aumento de 10% no enraizamento quando as estacas foram tratadas com 40% de extrato de alga marinha. A resposta de retenção foliar às concentrações de A. nodosum também foi ajustada em modelo linear positivo. Um aumento de 15,6% na retenção foliar foi observado na concentração de 40% de extrato de alga quando comparado ao controle. Correlações positivas foram observadas para retenção de folhas e porcentagem de enraizamento e retenção de folhas e comprimento de raízes. A imersão das bases de estacas de P. actinia em extrato de A. nodosum em uma concentração de 40% pode promover efeitos positivos no enraizamento e facilitar a propagação desta espécie.

ABSTRACT Passiflora actinia Hook (common name: ‘maracujá do mato’) is an important medicinal species due to significant sedative and anxiolytic activities. In order to commercially exploit the plant, however, studies on propagation to improve rooting in stem cuttings are needed. The present study was designed to evaluate the effect of the brown seaweed (Ascophyllum nodosum (L.) Le Jol.) extract when applied on P. actinia stem cuttings bases. Five concentrations of the extract in distilled water were evaluated: 0% (pure distilled water), 10%, 20%, 30% and 40%. The experimental design was completely randomized with 4 repetitions and 12 cuttings per experimental unit. Cuttings were evaluated 45 days from planting. Data were analyzed through polynomial regression analysis and Pearson's correlation coefficients were calculated for all the variables. On average, 51.27% rooting was achieved. Rooting percentage increased linearly according to the brown seaweed extract concentrations. When compared to the control treatment, about 10% increase in rooting was observed at the treatment with 40% seaweed extract. Leaf retention response to A. nodosum increasing concentrations was also adjusted in a positive linear model. A 15.6% increase in leaf retention was observed at the 40% seaweed concentration when compared to the control. Positive correlations were observed for leaf retention and rooting percentage and leaf retention and roots length. The immersion of P. actinia stem cuttings bases in A. nodosum extract at a concentration of 40% promote positive effects on rooting and facilitate the species propagation.

RESUMO Pinus tecunumanii é uma das espécies tropicais mais importantes do gênero Pinus no Brasil. Este trabalho teve por objetivo desenvolver um protocolo de micropropagação para Pinus tecunumanii a partir de gemas apicais de plantas jovens mantidas em casa de vegetação. Para a assepsia foi avaliado o efeito de 0,05 ou 0,1% de cloreto de mercúrio em combinação com 0 ou 1 g L-1 de fungicida e NaOCl. O efeito de 2 ou 4 µM BA (6-benzil adenina) foi avaliado na multiplicação in vitro ao longo de três subcultivos. O efeito da adição de 1,5 g L-1 de carvão ativado foi avaliado no meio de alongamento. Para o enraizamento, o efeito dos sais WV5 meia força e de 20 g L-1 de sacarose foi avaliado em meio suplementado com 2.68 µM ácido naftaleno acético (ANA) e 0.44 BA. A assepsia dos explantes com 0,05% de cloreto de mercúrio, hipoclorito de sódio, com ou sem fungicida foi eficiente no estabelecimento in vitro de até 83,3% dos explantes. Em meio suplementado com 2 ou 4 µM BA ocorreu multiplicação de brotações adventícias e após o terceiro subcultivo, 70,83% dos explantes formaram novos brotos em meio suplementado com 2 µM BA, com uma média de 3,7 novos brotos por explante. Em meio suplementado com carvão ativado foi observado alongamento em 59,76% dos explantes, enquanto no meio sem carvão 33,33% dos explantes alongaram. Foi observado enraizamento em 14,8% dos explantes em meio suplementado com sais WV5.

ABSTRACT Pinus tecunumanii is one of the most important tropical species of Pinus in Brazil. This work aimed to develop a micropropagation protocol for Pinus tecunumanii from the apical shoots of young plants grown in a greenhouse. To optimise the aseptic treatment protocol, the effect of 0.05 or 0.1% mercuric chloride in combination with 0 or 1 g L-1 fungicide and NaOCl was evaluated. Next, the effect of 2 or 4 µM BA (6-benzyl adenine) on in vitro multiplication was evaluated over three subcultures. Supplementation with 1.5 g L-1 activated charcoal was tested for elongation. For rooting, the effects of WV5 salts (at half strength) and 20 g L-1 sucrose were evaluated on media supplemented with 2.68 µM naphthalene-acetic acid and 0.44 µM BA. Aseptic treatment of explants with 0.05% mercuric chloride and sodium hypochlorite (with or without fungicide) was effective to establish 83.3% of explants in vitro. On media supplemented with 2 or 4 µM BA, shoot multiplication was promoted. After the third subculture on medium supplemented with 2 µM BA, 70.83% of the explants exhibited new shoots with an average of 3.7 new shoots per explant. On medium containing activated charcoal, elongation was observed in 59.76% of the explants, while on medium without activated charcoal only 33.33% elongated. Rooting was observed in 14.8% of explants on medium supplemented with WV5 salts.

RESUMEN Stevia rebaudiana Bertoni es una especie de alta importancia económica debido a la producción de edulcorantes naturales con bajo valor calórico. La producción de semillas de esta especie es irregular y las técnicas de micropropagación aumentan el costo de la producción de plántulas. Por lo tanto, este trabajo tuvo como objetivo optimizar la propagación vegetativa de un genotipo brasileño de esta especie a través de estacas de tallo. El material vegetal se recogió de plantas maduras con 1 año de edad. Se realizaron dos experimentos para determinar la mejor concentración de ácido indolbutírico (IBA) para mejorar el enraizamiento y evaluar la interacción entre la posición de corte del tallo (apical, mediana y basal) y sustratos (Tropstrato HT®, vermiculita y mezcla de vermiculita y Tropstrato HT®). Las estacas de tallos se vieron afectadas por las concentraciones de IBA. Se recomienda la aplicación de 710 mg L-1 de IBA para mayores porcentajes de enraizamiento. Las estacas de tallos de la región apical de las plantas mostraron mayor porcentaje de enraizamiento que las medianas y basales (medias de 70, 45,8 y 7,5%, respectivamente). Se observaron resultados similares para el número, longitud y masa de raíz fresca y seca. El sustrato Tropstrato HT® promovió mayores porcentajes de enraizamiento de las estacas de tallo y menor mortalidad en comparación con la vermiculita.

ABSTRACT Stevia rebaudiana Bertoni is a species with high economic importance because of the production of natural sweeteners with a low caloric value. The seed production of this species is irregular and micropropagation techniques raise the cost of seedling production. Thus, this study aimed to optimize the vegetative propagation of a Brazilian accession of this species through stem cuttings. The plant material was collected from one-year-old mature plants. Two experiments were carried out to determine the best indolebutyric acid (IBA) concentration to improve rooting and to evaluate the interaction between the stem cutting position (apical, medium and basal) and substrates (Tropstrato HT®, vermiculite, and vermiculite/Tropstrato HT® mixture). The rooting of the stem cuttings was affected by the IBA concentrations. The application of 710 mg L-1 was recommended for higher rooting percentages. The stem cuttings from the apical branch region had higher rooting percentages than the medium and basal areas (means of 70, 45.8 and 7.5%, respectively). Similar results were observed for the root numbers, root lengths and root fresh and dry mass. The substrate Tropstrato HT® promoted higher rooting percentages and lower mortality than vermiculite.