RESUMEN
BACKGROUND: Pyrostegia venusta (Ker Gawl.) Miers occurs in threatened biodiversity hotspots of Cerrado and Atlantic forest biomes in Brazil and is used in traditional medicine to treat various respiratory and skin diseases. METHODS AND RESULTS: This study (i) examined the genetic diversity and structure of six natural populations of P. venusta from different Brazilian regions using sequence-related amplified polymorphism (SRAP) markers; and (ii) compared the intra- and inter-populational levels of the bioactive component verbascoside using high-performance liquid chromatography. The population from Nova Mutum, Mato Grosso, presented the highest genetic variability (Nei index H = 0.2759; Shannon index I = 0.4170; 85.14% polymorphic loci), whereas the population from Araxá, Minas Gerais, presented the lowest genetic variability (H = 0.1811; I = 0.2820; 70.27% polymorphic loci). The intra-populational variability (79%) was significantly higher (p = 0.001) than the inter-populational variability (21%). The populations were clustered into two groups but their genetic differentiation was not associated with geographical origin (Mantel test, r = 0.328; p > 0.05). The verbascoside content significantly differed (p > 0.05) among the six populations and between the individuals from each population. The highest verbascoside levels (> 200 µg/mg extract) were detected in populations from Araxá and Serrana, while the lowest verbacoside levels were detected in populations from Paranaíta and Sinop. CONCLUSIONS: This is the first report on the use of SRAP markers to analyze genetic variability in the family Bignoniaceae. Our findings shall help to better understand the genetic and chemical diversity of P. venusta populations, as well as provide useful information to select the most appropriate individuals to prepare phytomedicines.
Asunto(s)
Bignoniaceae , Bignoniaceae/química , Bignoniaceae/genética , Variación Genética , Glucósidos , Fenoles , PolifenolesRESUMEN
ABSTRACT: Cochlospermum regium roots are used in popular medicine and its extract has diverse phytochemical molecules some with antimicrobial activity, consequently exposing this specie to genetic erosion risks. Thus, the objective of this study was to develop an in vitro multiplication protocol using chemical sterilization of culture medium. Therefore, explants obtained from apical buds of C. regium seedlings were inoculated into with 0.05mg L-1 NAA and 1mg L-1 BAP sterilized by chemical agent sodium hypochlorite (NaOCl) at 0.001%, 0.003% and 0.005% of active chlorine (Cl). Autoclaved culture medium was used as control. Result showed that the contamination by bacterial at 91 days of cultivation was significantly (P<0.05) controlled by autoclaving, 0.001% and 0.005% Cl. Moreover, the callus induction in the culture medium with 0.001% and 0.005% Cl was, respectively, 30% and 20% major than autoclaving sterilization. There was not significant (P<0.05) in the percentage of shoot induction among the sterilization preparations methods, and 65% of the explants survived in the presence of culture medium with 0.005% Cl. Histological analyses indicated that the Cl did not have any deleterious effects on morphogenic events. These results indicated that the chemical sterilization using 0.001% - 0.005% Cl controlled the fungal and bacterial multiplication in the culture medium and no affected the C. regium explants development, becoming it an alternative to autoclaving method.
RESUMO: As raízes de Cochlospermum regium são usadas na medicina popular, pois seu extrato apresenta diversas moléculas fitoquímicas, algumas com atividade antimicrobiana, que, consequentemente, expõe esta espécie ao risco de erosão genética. Assim, o objetivo deste estudo foi elaborar um protocolo de multiplicação in vitro para explantes de C. regium usando esterilização química do meio de cultura. Gemas apicais obtidas de plântulas de C. regium germinadas in vitro foram inoculadas em meio de cultivo MS suplementado com 0,05mg L-1 de ANA e 1mg L-1 de BAP e esterilizado pela adição do agente químico hipoclorito de sódio (NaOCl) nas concentrações de cloro ativo de 0,001%, 0,003% e 0,005%. O meio autoclavado foi utilizado como controle. Os resultados mostraram que a contaminação por bactérias aos 91 dias de cultivo foi significativamente (P<0,05) controlada pela autoclavagem, 0,001% e 0,005% de cloro ativo. Além disso, a indução de calos no meio de cultura com 0,001% e 0,005% de cloro ativo foi, respectivamente, 30% e 20% maior do que na esterilização por autoclavagem. Não houve diferença significativa (P<0,05) da porcentagem de indução de brotos entre os métodos de esterilização e 65% dos explantes sobreviveram na presença de 0,005% de cloro ativo. Análises histológicas indicaram que o cloro ativo não afetou os eventos morfogênicos. Os resultados indicaram que a esterilização química por meio do uso de 0,001% - 0,005% de cloro ativo auxiliou no controle da proliferação de bactérias e fungos no meio de cultura e não afetou o desenvolvimento dos explantes de C. regium, tornando-a uma alternativa em relação a autoclavagem.
