José Raniere Ferreira de Santana

Micropropagation and in vitro conservation of Neoglaziovia variegata (Arr. Cam.) mez, a fiber producing bromeliad from Brazil

Neoglaziovia variegata (Arr. Cam.) Mez is a Bromeliaceae native to the Caatinga, used for fiber extraction in the Northeast Region of Brazil. The antropic activity has place this species among the threatened ones. The objective of the work was to establish an in vitro propagation and conservation of caroa. Seeds were cultivated in MS medium in the presence or absence of light. In vitro germinated seedlings were multiplied in MS medium supplemented with the combinations 0.05 and 0.50 µM NAA and 2.2 and 4.4 µM BAP and KIN. The best percentages of germination were obtained with the seeds incubated in the presence of light. The highest multiplication ratio was obtained for the NAA (0,5 µM) + BAP (4,4 µM) treatment and the number of roots, with NAA (0.5 µM) + KIN (2.2 µM). Plant acclimatization presented differentiated results regarding the substrates tested. The conservation was established.

Effect of cytokinins on in vitro development of autotrophism and acclimatization of Annona glabra L.

The role of cytokinins in the differentiation of the photosynthetic apparatus in micropropagated plants and their effect on the plant’s ability to transition from a heterotrophic to an autotrophic condition during acclimatization was investigated. Annona glabra L. shoots were cultured on woody plant medium supplemented with sucrose and different cytokinins to evaluate leaf tissue for chloroplast development, chloroplast numbers, photosynthetic pigmentation, total photosynthetic potential, and soluble sugar content. Plants were transferred to the rooting medium in the presence or absence of sucrose and then acclimatized. Kinetin and benzyladenine (BAP) stimulated chloroplast differentiation. Inclusion of zeatin in the medium induced the formation of greater numbers of chloroplasts in the leaves, while plants cultivated in the presence of only kinetin and BAP demonstrated greater chlorophyll a and carotenoid content. The use of kinetin and BAP during in vitro culture promoted accumulation of dry matter during the acclimatization phase, especially in plants rooted under autotrophic conditions (without sucrose). Kinetin and BAP promoted development of more leaf area and greater plant survival rates in plant acclimatization on both autotrophic and heterotrophic media. The inhibitory effects of thidiazuron on the differentiation of chloroplasts, accumulation of chlorophyll a, and photosynthetic potential were examined.

In vitro morphogenesis of Syngonanthus mucugensis Giul: subsp. mucugensis

Syngonanthus mucugensis Giul. subsp. mucugensis e uma herbacea com grande potencial de utilizacao no comercio de flores secas ornamentais. A ocorrencia restrita ao municipio de Mucuge-BA, Brasil, associado a exploracao extrativista tem levado essa especie ao risco de extincao. Neste estudo, objetivou-se avaliar o potencial organogenico de tres diferentes tipos de explantes de S. mucugensis subsp. mucugensis visando ao desenvolvimento de um metodo alternativo para a sua propagacao. A capacidade morfogenica de caule, folha e raiz foi testada utilizando o meio de cultura Murashige e Skoog com metade da concentracao salina e diferentes concentracoes dos reguladores de crescimento benzilaminopurina - BAP (0,00; 2,22 e 4,44 µM) e acido naftaleno acetico - ANA (0,00; 1,34 e 2,68 µM). Os eventos morfoanatomicos que levaram a formacao dos brotos foram descritos. O caule demonstrou ser a melhor fonte de explante, apresentando 58,75% de regeneracao de brotos via organogenese direta, em meio livre de regulador de crescimento; e 32,18 e 47,55% de regeneracao de brotos por organogenese indireta na presenca de 2,22 e 4,44 µM de BAP, respectivamente. Para folha, foi obtida a formacao de calos sem regeneracao de brotos, nao sendo observada morfogenese, quando se utilizou raiz como explante. As analises histologicas mostraram que a regeneracao de brotos em S. mucugensis subsp. mucugensis ocorreu tanto por via indireta, pela diferenciacao do tecido calogenico desorganizado, quanto por via direta pela retomada da atividade meristematica em celulas maduras diferenciadas e proliferacao de gemas pre existentes.

Propagação in vitro de Erythrina velutina

Erythrina velutina (mulungu) is a native plant of the semi-arid region that can be propagated via tissue culture, which may contribute to the rapid multiplication of this species, reducing the possibility environmental impacts. The objective of this study was to establish a protocol for micropropagation of mulungu using aseptic explants seedlings grown in vitro. For the induction of shoots, , nodal segments and cotyledons were used as explants hypocontyl and were inoculated in a WPM culture medium supplemented with different concentrations of BAP and NAA. The rooting of shoots was achieved in a WPM medium supplemented with 0.0; 2.46; 4.92; 9.88µM IBA. The best answer to the multiplication was obtained in medium supplemented with 17.76µM BAP using nodal segment or cotyledonary node explants. The shoots rooted in all conditions tested, including the culture medium without auxin.

Controle da abscisão foliar e morfogênese in vitro em culturas de Anadenanthera colubrina (Vell.) Brenan var. cebil (Griseb) Altschul

ABSTRACT – This work aimed to control leaf abscission of seedlings of angico using AgNO 3 and CoCl 2 andevaluate the effect of the paclobutrazol on the in vitro behavior of seedlings. The seeds were disinfected andinoculated in Petri plates containing germtest paper, previously sterilized and imbibed in sterile water. Theplates were kept in the dark for two days, until seed germination. Seeds were transferred to test tubes containingWPM medium. In the first experiment, the culture medium was supplemented with different concentrationsof (0.0; 5.0; 10.0; 20.0; 40.0 µM) and types (AgNO 3 and CoCl 2 ) of ethylene inhibitors. In the second experiment,the medium was supplemented with different concentrations (1.7; 3.4; 6.8; 13.6 µM) of paclobutrazol. There Sociedade de Investigacoes Florestais CONTROLE DA ABSCISAO FOLIAR E MORFOGENESE IN VITRO EMCULTURAS DE Anadenanthera colubrina (Vell.) Brenan var. cebil (Griseb)Altschul 1 Cristina Ferreira Nepomuceno 2 , Ana Paula de Souza Rios 3 , Sandra Regina de Oliveira Domingos Queiroz

Variabilidade genética de populações naturais de caroá por meio de marcadores RAPD

The objective of this work was to quantify the genetic variability within and among populations of caroa (Neoglaziovia variegata) using random amplified polymorphic DNA(RAPD) markers. One hundred eighty caroa genotypes from Guanambi, Juazeiro and Valente counties in the state of Bahia, Brazil, were analyzed. A high polymorphism was observed among the caroa populations. The genetic dissimilarities among all genotypes ranged from 0.08 to 0.95 with an average of 0.44. The molecular variance showed that 56% of the total variation was explained by the differences among individuals with in locations.The differences among counties explained 17% of the total variation, while the differences among places within counties explained 26% of the variation.

Anatomia foliar de seis espécies de anonáceas cultivadas in vitro e em casa de vegetação

Micropropagation of Annonaceae can produce homogeneous plants and bring new species into commercial production. Plants cultivated in vitro, however, frequently demonstrate anatomical alterations, and the quantification of these changes should aid in determining more efficient culture protocols. The present work undertook a comparative study of the leaf anatomy of six species of Annonaceae cultivated in vitro and in greenhouses. Annona coriacea was the only species that did not show variations in the density or the dimensions of their stomata when cultivated in vitro, while Annona bahiensis, Annona glabra, Annona squamosa, and Rolinia silvatica demonstrated increases in leaf stomatal density and a reduction of the leaf epidermis under these conditions.

Estímulo do comportamento fotoautotrófico durante o enraizamento in vitro de Annona glabra L., I. desenvolvimento do sistema radicular e da parte aérea

Conduziu-se este trabalho, com o objetivo de induzir a estimulo do comportamento fotoautotrofico durante o enraizamento in vitro em brotacoes de Annona glabra L. Brotacoes oriundas de cultivo em tubos fechados com tampa e pelicula de PVC (cultivo sem aeracao), com tampa sem a pelicula de PVC e tampao de algodao (cultivo com aeracao) foram inoculadas em meio WPM suplementado com 4,9 µ M de AIB e 164,4mM de carvao ativado na presenca (58,42mM) ou ausencia de sacarose. Durante o enraizamento, foram mantidas as mesmas condicoes de vedacao dos tubos de ensaio em que as brotacoes foram induzidas. Apos a inoculacao, os tubos contendo os explantes foram mantidos em sala de crescimento sob radiacao fotossintetica ativa de 45-56 µ mol.m-2.s-1 a 25±3oC. Os resultados mostraram que o enraizamento das brotacoes de A. glabra nao dependeu do suprimento de sacarose no meio de cultura, em tubos fechados com o tampao de algodao ou tampa plastica sem PVC. A aeracao dos tubos de ensaio trouxe incrementos de significativos (ate 250%) na materia seca radicular. A inducao de raizes secundarias (laterais) em A. glabra so ocorreu em culturas com aeracao, independentemente da presenca ou ausencia de sacarose no meio de cultura. Com os resultado desse experimento, conclui-se que o estimulo do comportamento fotoautotrofico em A.glabra pode ser obtido com sucesso durante a fase de enraizamento in vitro.

Agentes osmóticos e temperatura na conservação in vitro de sempre-viva

The in vitro conservation is an ex situ conservation strategy that ensures the maintenance of genetic and biological integrity of species. The present study evaluated the effects of, osmotic agents and different temperature regimes on the in vitro conservation of Syngonanthus mucugensis Giul. subsp. mucugensis. The shoots were inoculated into half salt strength Murashige and Skoog culture medium (MS ½) containing 7g L-1 of agar. The culture medium was supplemented with 60gL-1 sucrose and with the sucrose concentrations 15, 30 and 45g L-1 combined with 0 and 15g L-1 of sorbytol or mannitol. Two different temperatures were used in these experiments (18 and 25°C). The percentage of plant survival was evaluated monthly and at 180 days were analyzed length of shoot and root, the percentage of green leaves, the percentage of explants with shoots and number of shoots per explants and shoot length. The addition of osmotic agents resulted in decreased growth of the plants and therefore reduced their viability. The averages observed in the experiments undertaken at 18°C were significantly superior to those observed at 25°C for all of the variables analyzed. S. mucugensis subsp. mucugensis can be cultured at 18°C in MS½ culture medium containing 15g L-1 of sucrose, for up to 180 days, without subculturing.

Effect of different culture tube caps and concentrations of activated charcoal and sucrose on in vitro growth and budding induction of Annona glabra L.

Este trabalho foi realizado com o objetivo de avaliar o efeito do tipo de selamento do recipiente de cultura e diferentes concentracoes de sacarose e carvao ativado na inducao e crescimento de brotacoes in vitro de Annona glabra L.; especie frutifera conhecida popularmente como araticum. Segmentos nodais obtidos de plantas de A. glabra mantidas em casa de vegetacao, apos o processo de assepsia, foram inoculados em meio de cultura WPM com 7 g L-1 de agar e sacarose (0,00; 29,21; 58,63 e 116,84 mM) combinado com carvao ativado (0,0 e 2,0 g L-1) e 250 mg L-1 de benomil. Avaliaram-se dois tipos de selamento nos tubos de ensaio: tampa com pelicula de PVC e tampao de algodao. O experimento foi instalado em DIC em arranjo fatorial 4x2x2, com 4 repeticoes/5 tubos por parcela. Avaliou-se o numero brotacoes, a porcentagem de explantes responsivos, o numero de folhas expandidas por brotacao, o comprimento da maior folha, a abscisao foliar, o comprimento e a materia seca das brotacoes. O tipo de selamento influenciou a organogenese em segmentos nodais de A. glabra. Contudo, nao houve inducao de brotacoes na ausencia de sacarose. O maior numero de folhas expandidas foi obtido quando utilizou-se 58,42 mM de sacarose em tubos selados com tampao de algodao e a abscisao foliar foi reduzida a metade na presenca do carvao ativado. Maiores comprimentos e maiores pesos da materia seca das brotacoes foram obtidos em tubos selados com tampao de algodao, tambem na presenca de carvao ativado.