Callus induction and bioactive phenolic compounds production from Byrsonima verbascifolia (L.) DC. (Malpighiaceae)

ABSTRACT -This study developed a methodology for callus induction in leaf segments of B. verbascifolia and evaluated the bioactive phenolic compounds production. Leaf explants were cultured in MS medium with 30 g L -1 sucrose, solidified with 7 g L -1 agar supplemented with 2,4-D (0; 4.52; 9.05; 18.10 M) and BAP (0; 4.44; 8.88; 17.75 M ) in the presence and absence of light. Forty-five days after inoculation we assessed the percentage of callus induction, color, consistency, fresh and dry matter, total phenols, flavonoids, tannins contents, and chromatographic profile by HPLC-DAD method. Callus induction occurred only in medium with growth regulators. Maximal induction (100%) was found in medium containing 2,4-D combined with BAP in the presence and absence of light. We obtained friable and compact callus in yellow, green, and red. Culture media containing 4.52 M 2,4-D + 4.44 M BAP induced 100% of friable callus with higher fresh and dry weight in the absence of light. The callus produced higher amounts of total phenols and flavonoids than the initial explant

AVALIAÇÃO SAZONAL DA ATIVIDADE DA FENILALANINA AMÔNIA-LIASE E DOS TEORES DE FENÓIS E TANINOS TOTAIS EM Stryphnodendron adstringens (Mart.) Coville

We performed analyses of the leaves and stem bark of Stryphnodendron adstringens, a Brazilian Cerrado plant, to examine the seasonal patterns of phenylalanine ammonia-lyase activity and total phenol and tannin contents in 16 adult individuals under field conditions. The measurements of gas exchange parameters, including net photosynthesis, transpiration and stomatal conductance, as well as the photosynthetic photon flux density, leaf temperature, vapor pressure deficit and leaf water potential were also performed. The results were subjected to a principal component analysis. The values obtained for stomatal conductance, net photosynthesis, transpiration, water use efficiency, minimum water potential and photosynthetic photon flux density were higher in the rainy season, while the maximum water potential and vapor pressure deficit in the atmosphere were similar between seasons. The seasonal variations in gas exchange characteristics were influenced by changing weather conditions and were positively correlated with higher air temperatures, rainfall amounts and relative air humidity and negatively correlated with increased insolation. There was no correlation between the maximum leaf water potential and any of the weather variables. Phenylalanine ammonia-lyase activity was similar in the dry and rainy seasons, and higher levels of total phenols and tannins were observed in the dry and rainy seasons, respectively. Phenylalanine ammonia-lyase activity and total phenol contents were negatively influenced by temperature and rainfall in the dry season, and the total tannin contents were mainly correlated with higher temperatures and a lower influence of rainfall in the rainy season.Folhas e cascas do caule de Stryphnodendron adstringens, uma espécie do Cerrado brasileiro, foram utilizadas para examinar os padrões sazonais da atividade da fenilalanina amônia-liase e os teores de fenóis e taninos totais em 16 indivíduos adultos, em condições de campo. Avaliações das trocas gasosas, incluindo fotossíntese líquida, transpiração, condutância estomática, densidade de fluxo de fótons fotossinteticamente ativos, temperatura da folha, deficit de pressão de vapor e potencial hídrico foliar foram realizadas. Os resultados foram submetidos à análise de componentes principais. A condutância estomática, fotossíntese líquida, transpiração, eficiência do uso da água, potencial hídrico mínimo e densidade de fluxo de fótons fotossinteticamente ativos foram maiores na estação chuvosa, enquanto que o potencial hídrico máximo e o deficit de pressão de vapor na atmosfera foram semelhantes entre as estações. As variações sazonais nas características de trocas gasosas foram influenciadas por mudanças nas condições climáticas e se correlacionaram positivamente com as maiores temperaturas, precipitação e umidade relativa do ar e negativamente com o aumento da insolação. Não houve correlação entre o potencial hídrico foliar máximo e as demais variáveis climáticas. A atividade da fenilalanina amônia-liase foi semelhante nas estações seca e chuvosa e os maiores teores de fenóis e taninos totais foram observados nas estações seca e chuvosa, respectivamente. A atividade da fenilalanina amônia-liase e os teores de fenóis totais foram influenciados negativamente pela temperatura e precipitação na estação seca, e os teoresde taninos totais foram correlacionados principalmente com temperaturas mais elevadas, com menor influência da precipitação na estação chuvosa

Leaf anatomy of Cordiera sessilis (Vell.) Kuntze (Rubiaceae)

A study on the leaf anatomy of Cordiera sessilis (Rubiaceae), a native medicinal shrub from Brazilian Cerrado was carried out to contribute for species identification and provide support for future biochemical studies and medicinal application. Leaves were collected, fixed and processed by usual techniques, and studied by light and electron microscopy. Quantitative analyzes of stomata and trichomes were performed. The leaves showed typical characteristics of Rubiaceae family, like hypostomatic leaves, paracytic stomata, dorsiventral mesophyll and collateral bundles. Two types of vascular patterns were identified in the petiole: in distal part, the vascular system is arranged cylindrically surrounded by sclerenchyma sheath and in proximal part, the vascular system is arranged in U-shape coupled to sclerified cells. The micromorphological organization of leaf surface, epicuticular wax types, the petiole pattern and histochemical characteristics as the presence of druses, sand crystals and alkaloids and absence of raphids in the mesophyll, midrib and petiole are considerate representative characteristics of C. sessilis and may be useful in the species identification.A study on the leaf anatomy of Cordiera sessilis (Rubiaceae), a native medicinal shrub from Brazilian Cerrado was carried out to identify features that may be useful in species recognition. Leaves were collected, fixed and processed by usual techniques, and studied by light and electron microscopy. Quantitative analyzes of stomata and trichomes were performed. In addition to the typical anatomical characteristics of Rubiaceae leaves, two types of vascular patterns were identified in the petiole: in distal part, the vascular system is arranged cylindrically surrounded by sclerenchyma sheath and in proximal part the vascular system is arranged in U-shape coupled to sclerified cells. The micromorphological organization of leaf surface, epicuticular wax types, the petiole pattern and histochemical characteristics as the presence of druses, crystal sand and alkaloids and absence of raphides in the mesophyll, midrib and petiole are considerate representative characteristics of C. sessilis and may be useful in the species recognition.

In vitro germination, callus induction and phenolic compounds contents from Pyrostegia venusta (Ker Gawl.) Miers

A protocol for the in vitro germination and callus induction in Pyrostegia venusta, a medicinal plant species from the Brazilian savannah (Cerrado) is provided. The establishment of cultures of P. venusta was originally obtained from seeds germinated in vitro and induced callus directly from leaf explants of seedlings. Seeds were germinated on MS and WPM media containing 100 and 50% of salt concentration, supplemented with 30 g L-1 sucrose. Callus induction consisted of the inoculation of leaf segments on MS medium plus 2,4-D or BAP in the presence or absence of light. The germination percentage averaged 85% and the aerial parts and roots of seedlings obtained in WPM with 50 and 100% of salt concentration showed elevated contents of total phenolic compounds and flavonoids compared to those obtained on MS medium. Calli induced with lower concentrations of 2,4-D had higher fresh and dry weight values. All treatments resulted in obtaining calli with contents of total phenolic compounds and flavonoids at or above the initial explant, highlighting treatments containing 9.05 mM 2,4-D and 8.88 mM BAP in the presence and absence of light, and 17.75 mM BAP in the absence of light.

Teores de fenóis totais e taninos nas cascas de angico-vermelho (Anadenanthera peregrina)

Este trabalho teve como objetivo quantificar fenóis totais e taninos nas cascas de angico-vermelho (Anadenanthera peregrina) em diferentes classes de diâmetro, além de comparar duas metodologias de quantificação de fenóis. Foram coletadas cascas de 35 árvores, distribuídas em sete classes diamétricas. As cascas foram secas ao ar livre, trituradas e peneiradas, sendo utilizado o material retido na peneira de 60 mesh. Para a extração dos fenóis, utilizou-se metanol:água (50%). O doseamento de fenóis totais foi realizado pelos métodos de Folin-Denis e Folin-Ciocalteau modificados, e para taninos, pelo método difusão radial. Não foi encontrada diferença estatística entre as metodologias de quantificação de fenóis. Verificou-se aumento nos teores de fenóis e taninos com o aumento do diâmetro das árvores

Short-Term Liquid Nitrogen Storage of <i>Pyrostegia venusta</i> Embryos: Effects on Germination, Phenotypic and Biochemical Characteristics, and In Vitro Secondary Metabolite Production

In this study, short-term liquid nitrogen (LN) storage was used as a strategy to conserve Pyrostegia venusta embryos, and its effects on in vitro germination, phenotypic and biochemical characteristics, and in vitro secondary metabolite production were assessed. Embryos stored in LN for 1 and 7 days presented a higher germination rate and germination speed index compared to those of the control (non-cryostored embryos). Short-term LN storage also favored the phenotypic characteristics of seedlings. LN storage significantly affected the proteins (PTN), soluble sugar (SS) and reducing sugar (RS) contents, oxidative metabolism, and phenylalanine ammonia-lyase (PAL) activity, as well as the total phenolic compound, flavonoid, phytosterol, and alkaloid levels in seedlings regenerated from embryos cryostored for 7 days. Benzoic acid derivatives and flavonoids were observed in regenerated non-acclimatized seedlings. LN storage did not affect the survival rate or phenotypic characteristics of seedlings during acclimatization. Acclimatization promoted significant changes in PTN, SS and RS contents, oxidative metabolism, and PAL activity in seedlings from embryos cryostored for 7 days. Roots from acclimatized seedlings exhibited the highest phenolic, phytosterol, and total alkaloid levels. Differences in the chromatographic profiles of the acclimatized seedlings compared with the non-acclimatized seedlings were observed. LN storage can be an effective means of ex situ conservation of P. venusta genetic resources

Callus induction and bioactive phenolic compounds production from Byrsonima verbascifolia (L.) DC. (Malpighiaceae)

ABSTRACT This study developed a methodology for callus induction in leaf segments of B. verbascifolia and evaluated the bioactive phenolic compounds production. Leaf explants were cultured in MS medium with 30 g L-1 sucrose, solidified with 7 g L- 1 agar supplemented with 2,4-D (0; 4.52; 9.05; 18.10 µM) and BAP (0; 4.44; 8.88; 17.75 µM ) in the presence and absence of light. Forty-five days after inoculation we assessed the percentage of callus induction, color, consistency, fresh and dry matter, total phenols, flavonoids, tannins contents, and chromatographic profile by HPLC-DAD method. Callus induction occurred only in medium with growth regulators. Maximal induction (100%) was found in medium containing 2,4-D combined with BAP in the presence and absence of light. We obtained friable and compact callus in yellow, green, and red. Culture media containing 4.52 µM 2,4-D + 4.44 µM BAP induced 100% of friable callus with higher fresh and dry weight in the absence of light. The callus produced higher amounts of total phenols and flavonoids than the initial explant. Total tannins were detected only in callus induced in media containing 17.75 µM BAP and 4.52 µM 2,4-D + 17.75 µM BAP, and were not found in the initial explant. The bioactive phenolic compounds detected are derived from benzoic, p-coumaric, cinnamic, gallic acids, and catechins