-The importance in folk medicine, combined to threats in their environment, becomes necessary to carry out studies involving large-scale propagation of Lippia genus. Although the tissue culture propagation is widely disseminated for medicinal plants, for L. rotundifolia any article was published yet. The present study aimed to establish an efficient protocol for micropropagation of L. rotundifolia. Nodal segments, taken from plants collected in the Espinhaço Range, were disinfected, and cultures were initiated on MS medium with PVPP (1 g L -1), sucrose (3%) and agar (0.7%). The culture were maintained in a growth room at controlled conditions. Disinfestation procedures and the supply of PVPP on culture media resulted in both reduced contamination and phenol oxidation rates, with more than 90% of viable cultures. In the multiplication phase were tried different BAP and NAA combinations supplied to the MS medium. The treatment that resulted in highest multiplication rates was 0.33 M BAP. The effects of NAA were evaluated for in vitro rooting. At 0.44 M, rooting was 70% higher than that observed in the control. The acclimatization was held in trays with substrate, coated with translucent plastic and kept under shade. The plantlets were transferred to the greenhouse after 15 days and transplanted to plant beds after 30 days. The acclimatized plantlets bloomed one year after the transference to field conditions, showing that the in vitro culture did not affect the vegetative and reproductive development, which confirms the potential of micropropagation to reduce the extinction risk of L. rotundifolia.Key words: Micropropagation. In vitro multiplication. In vitro rooting. Ex vitro acclimatization. Biodiversity conservation.RESUMO -A importância na medicina popular, assim como as ameaças em seu ambiente, tornam necessária a realização de estudos envolvendo a propagação em larga escala de plantas do gênero Lippia. Embora a propagação in vitro seja amplamente disseminada para plantas medicinais, nenhum artigo foi publicado ainda com L. rotundifolia. O presente estudo teve como objetivo estabelecer um protocolo eficiente para a micropropagação dessa espécie. Segmentos nodais, retirados de plantas coletadas na Cadeia do Espinhaço, foram desinfetados antes da inoculação em meio MS suplementado com PVPP (1 g L -1), sacarose (3%) e ágar (0,7%). Os tubos de ensaio foram mantidos em sala de crescimento sob condições controladas. O tratamento de desinfestação e a adição de PVPP aos meios de cultura resultaram na redução da infecção microbiológica assim como nas taxas de oxidação fenólica, proporcionando mais de 90% de culturas assépticas e viáveis . Na fase de multiplicação foram testadas diferentes combinações de BAP e ANA adicionadas ao meio MS. O tratamento que resultou em maiores taxas de multiplicação foi de 0,33 M de BAP. Os efeitos do ANA foram avaliados no enraizamento in vitro. A 0,44 M de ANA, o enraizamento foi 70% superior ao observado no controle. A aclimatização foi realizada em bandejas com substrato, rev...

This study aimed to evaluate the variation in the levels of proline, oxidative metabolism and photosynthetic pigments in plants of Pitcairnia encholirioides grown in vitro under different conditions and after acclimatization. The analyses were performed after 150 days of in vitro cultivation in MS media supplemented with 10 µM GA 3 or 0.2 µM NAA, sucrose at 15 or 30 g L -1 , in test tubes which allowed gas exchange or in a hermetically sealed system, and 180 days after acclimatization. The in vitro maintenance in hermetically sealed flasks, with GA 3 and 15 g L -1 sucrose had adverse metabolic effects, which was demonstrated by the lower proline and photosynthetic pigments accumulation and by the increase in antioxidant enzymes activities. After acclimatization, differences for proline and photosynthetic pigments were no longer found and the enzymatic activities ranged unevenly. The results suggest that the in vitro cultivation in media with 0.2 µM NAA and 30 g L -1 sucrose, in test tubes capped with closures which allowed gas exchange, is more suitable for micropropagation of P. encholirioides, providing a prolonged maintenance of in vitro cultures and plantlets with superior quality for ex vitro development.Keywords: enzymatic activity, oxidative stress, micropropagation, proline. Niveis de prolina, do metabolismo oxidativo e dos pigmentos fotossintéticos durante o crescimento in vitro e aclimatização de Pitcairnia encholirioides L.B. Sm. (Bromeliaceae) ResumoEste trabalho objetivou avaliar a contribuição da prolina, do metabolismo oxidativo e dos pigmentos fotossintéticos na propagação in vitro e aclimatização de Pitcairnia encholirioides, uma bromélia criticamente ameaçada de extinção. As análises foram realizadas após 150 dias de cultivo in vitro em meio MS suplementado com 10 µM de GA 3 ou 0,2 µM de ANA, 15 ou 30 g L -1 de sacarose, em tubos de ensaio que permitiam trocas gasosas ou em sistema hermeticamente vedado, e também 180 dias após aclimatização. A manutenção in vitro em frascos hermeticamente fechados, com GA 3 e 15 g L -1 de sacarose apresentou efeito metabólico adverso, demonstrado pelo menor acúmulo de prolina e pigmentos fotossintéticos e também pelo aumento das atividades de enzimas antioxidantes. Após aclimatização, as diferenças para prolina e pigmentos fotossintéticos não foram mais encontradas e as atividades enzimáticas variaram de maneira desuniforme. Os resultados sugerem que o cultivo in vitro em meio com 0,2 µM de ANA e 30 g L -1 de sacarose, em tubos fechados com tampas que permitem trocas gasosas, é mais adequado para a micropropagação de P. encholirioides, proporcionando uma manutenção prolongada das culturas in vitro e plântulas com qualidade superior para o desenvolvimento ex vitro.Palavras-chave: atividade enzimática, estresse oxidativo, micropropagação, prolina.

In bryophytes, establishment can occur by a sexual or asexual process, but the production of spores enables colonization of a wider range of habitats and substrates than can asexual propagules. Successful germination is critical for establishment in a new environment. Th is paper addresses germination and sporeling development in Frullania ericoides, a leafy liverwort species. Fresh spores were inoculated in vitro in diff erent culture strengths of Knop's nutrient solution (one-fourth strength, half strength, full strength, one and a half strength and double strength), in order to evaluate the eff ects of this solution on spore germination and on the development of external protonema. On the fi rst assessment, spore germination was observed at all the concentrations. Germination was endosporic, with cell division and proliferation, resulting in a globular protonema, within the spore wall. Beginning at the fourth week, the development of tightly concave primordial leaves was observed in all but the double-strength medium. Th roughout the period of study, the treatments with lower concentrations exhibited external protonema with greater lengths. Th e double-strength treatment was statistically diff erent from other treatments in at least two parameters. Th e results of this study demonstrate the potential of in vitro culture techniques for bryophyte spore studies and germplasm preservation.

This work describes an efficient micropropagation protocol for Verbena litoralis and the study of the antinociceptive and antioxidant activities in extracts of this species. For the establishment in vitro, surface-sterilization procedures and PVPP showed high efficiency in fungal-bacterial contamination and phenol oxidation controls. Nodal segments cultivation in MS medium supplemented with 6-benzyladenine (7.5 µM)/α-naphthaleneacetic acid (NAA; 0.005 µM) induced multiple shoots. Elongated shoots were rooted with IAA (0.2 µM). Acclimatization rates were elevated and the plants showed the typical features of this species. The hexanic fraction (HF) of powdered leaves presented a radical scavenging activity with IC(50) = 169.3 µg mL(-1). HF showed a non-dose dependent analgesic activity in the writhing test; its antinociceptive activity in the hot plate test was restricted to 500 mg kg(-1), which is the highest dose. The results of this study showed the potential of tissue culture on conservation and large scale multiplication and confirmed the traditional folk medicine use of V. litoralis.

-The objective of this work was to establish an efficient protocol for in vitro multiplication and rooting, as well as ex vitro acclimatization of Aegiphila verticillata, a woody species found in Brazilian rocky fields. Aseptic cultures were established by seeds and two multiplication analyses were performed. In the first, we employed 6-benzylaminopurine (BAP -0, 2.5, 5 and 7.5 μM) + α-naphthalene acetic acid (NAA -0, 0.2, 0.4 and 0.6 μM) and, in the second, were studied adenine sulfate, kinetin and thidiazuron (0, 5, 7.5, 10 and 12.5 μM). After 90 days, we assessed the quantitative and qualitative shoot propagation. There were more than 90% seed germination and low contamination (2%). In multiplication phase, the culture medium that promoted the best quantitative and qualitative culture development was supplemented with 7.5 μM BAP + 0.4 μM NAA. In the rooting assay, were used NAA, indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) (0, 0.1, 0.2, 0.3 or 0.4 μM). After 90 days, the root number and rooting quality were evaluated. In this analysis, differences were not found between the control and the other treatments. Rooted plantlets were acclimatized in styrofoam trays for 30 days, after which they were transferred to pots in the greenhouse. Only 3% of the plants subjected to initial acclimatization died and 70% of the plants transferred to the field conditions survived and showed normal development. The results founded in this work are the first involving in vitro propagation and ex vitro acclimatization of Aegiphila verticillata and provide a continuous supply of this medicinal native species, endangered due anthropogenic activities.Keywords: In vitro propagation; Ex vitro culture; Rocky fields. 2,5; 5; e 7,5 μM) + ácido α-naftaleno acético 0,2; 0,4; e 0,6 μM) e, no segundo, sulfato de adenina, cinetina ou thidiazuron (0; 5; 7,5; 10; e 12,5 μM MICROPROPAGAÇÃO E ACLIMATIZAÇÃO DE

Devido à intensa utilização industrial, o cromo é considerado um importante poluente ambiental. O presente trabalho objetivou determinar os teores de pigmentos hidro e lipossolúveis em plantas de salvínia expostas a concentrações crescentes de Cr, visando estabelecer parâmetros bioquímicos para utilização dessa macrófita em programas de biomonitoramento e/ou fitorremediação da poluição causada por esse poluente metálico em ambientes aquáticos. As plantas foram submetidas a concentrações crescentes de Cr e avaliadas após quatro, seis e dez dias de tratamento. Os resultados dos ensaios permitiram concluir que plantas de salvínia sob condições de estresse por Cr apresentam reduções nas concentrações das clorofilas a, b e total e, em contraste, aumentos nas concentrações de antocianinas totais. Embora a concentração de carotenoides totais não tenha sido alterada em resposta ao Cr, as variações nas concentrações dos demais pigmentos lipossolúveis e dos pigmentos hidrossolúveis observadas nas folhas das plantas de salvínia podem ser utilizadas como parâmetros bioquímicos de biomonitoramento da poluição causada por esse elemento metálico em ambientes aquáticos.

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This study aimed to establish and propagate in vitro plants of Bouchea fluminensis, a medicinal species known in Brazil as gervão-falso ("false verbena"), evaluating the influences of different growth regulators on in vitro multiplication and rooting stages, as well as examining ex vitro acclimatization of rooted plants. Explants were established on Murashige and Skoog medium at half strength of salts and vitamins without growth regulators. For multiplication, the explants were subjected to combinations of 6-benzyladenine (BA; 0, 2.5, 5.0 and 7.5 μM) and α-naphthalene-acetic acid (NAA; 0, 0.2, 0.4 and 0.6 μM). The medium found to induce the greatest number of shoot was that containing 5 μM of BA (NAA-free). For rooting, we evaluated three auxins (NAA, indole-3-acetic acid and indole-3-butyric acid; 0.1, 0.2, 0.3 and 0.4 μM), as well as a control. No differences were observed between the control and the other treatments. The auxin-free medium was deemed the most suitable, because it ensures the lowest cost in the micropropagation procedures. We obtained 100% survival of the acclimatized seedlings, and the plants showed normal vegetative and reproductive development, suggesting that the micropropagation did not alter the biological cycle of this species. The results show the importance and potential of micropropagation for biodiversity conservation of Bouchea fluminensis.