A publication by the National Academy of Sciences, USA (1979) outlined some of the research need for a great variety of economically important woody species whose remaining genetic resources need urgently to be collected and conserved. A viable regeneration system was established via tissue and cell suspension culture for Albizia falcataria and A. lebbeck, two important wood yielding leguminous tree species. The culture medium was standardized after several trials to obtain callus from the leaflet explants of these two tree species. The optimum use of casein hydrolysate (w/v) and coconut milk (v/v) in addition to 6-Benzylaminopurine and Indole-3-butyric acid could induce morphogenesis and somatic embryogenesis in the cultured tissue. This reports the first observation on somatic embryogenesis ofA. lebbeck using leaflets as the explants. Scanning Electron Microscopy and histological studies were done on the different stages plant development following standard techniques. Embryogenesis in suspension culture followed regeneration of plantlets in A. lebbeck. In A.falcaaria the regenerative process followed via organogenesis from the shoot buds developed on the leaf explants. After hardening the regenerated plants were transferred to the greenhouse. Some of the trees grew more than 25 feet tall within a few months outside the greenhouse. Karyotype of the three leguminous trees Albizia lebbeck, A. falcataria, and Dalbergia sissoo was analyzed. In D. sissoo, various chromosomal anomalies were observed in the cultured tissue. The abnormality indices and ploidy level varied with the age and the frequency of the subculture. In the aged culture the regenerative potential declined but was reinstated to some extent with the addition of two complex growth factors, coconut milk and casein hydrolysate. Seed anatomy of 26 species of 4 leguminous genera was studied with SEM. The main distinguishing anatomical features observed in the seed sections were uniseriate or multiseriate epidermis, epidermal projections, and number of rows and nature of columns of hypodermal layer, especially the nature of endosperm. Three species of Dalbergia, Acacia and Cassia and two species of Albizia are difficult to distinguish externally even with seed coat study under SEM, but this study with cross sections provided enough characteristic features to distinguish one from the other.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Mate (Ilex paraguariensis) is an important species in Southern Brazil for its economical and cultural values. The present work had the objectives of analyzing the propagation of mate by cuttings, disinfection and in vitro establishment of nodal segments from adult plants and micropropagation of plants originated from immature embryos. Mate cutting were submitted to three (0, 4000 and 8000 mg L-1) IBA doses and two lengths (3 and 10 cm) of cutting. The cuttings were evaluated at 135 days for survival, rooting and formation of roots, leaves and shoots. In the disinfection experiments, nodal segments of one-year-old shoots were immersed in alcohol solution of 70% for two and four min., in 2% of NaOCl for 15, 25 and 35 min. and inoculated in ¼ of MS medium. The percentage of contaminated, oxidized and healthy explants were evaluated at 15 days. In micropropagation, mate seedlings from immature embryos were submitted to different culture mediums (MS, ¼ MS and WPM) for 30 days and evaluated for number of leaves and length of seedlings. For the multiplication, different doses (0; 0,01; 0,1; 1,0 and 2,0 mg L-1) of BAP were tested in ¼ MS culture medium. At 50 days, they were evaluated for the number of shoots, leaves and internodes, shoots length and height. Different doses (0; 0,5; 1,0 and 1,5 mg L-1) of IBA were evaluated for rooting and kept explants either for 15 or 30 days in the medium. In vitro plants of mate were acclimatized in different (vermiculite, sand or coconut shells) substrates, with controlled temperature and light environmental conditions. The highest percentage of rooted cuttings, in both evaluated length, were with 4000 mg L-1 of IBA. Cuttings with 3 cm length produced the highest percentage of rooted cuttings. Nodal segments of one-year-old shoots have the highest survival, with the disinfection with alcohol 70% for four minutes and NaOCl 2% for 25 min. The ¼ of MS salts medium can be used as basal medium for in vitro culture of mate plants. The basal medium with 2 mg L-1 of BAP increases the number of adventitious shoots and promotes shoot proliferation after some sub cultivations, enabling in vitro plant maintenance as microstump, as shoot supply. In vitro rooting can be achieved in one cycle of 30 days of cultivation, with the addition of 3 mg L-1 of IBA to the medium with ¼ of MS salts. In vitro plants of mate can be acclimatized in coconut shells or sand in controlled temperature and light conditions. / A erva-mate (Ilex paraguariensis) é uma espécie de grande importância no sul do Brasil, pelo valor econômico e cultural que apresenta. O presente trabalho teve por objetivos analisar a propagação de erva-mate por estaquia, o estabelecimento e desinfestação in vitro de segmentos nodais de plantas adultas e a micropropagação de plantas oriundas de embriões imaturos. Para a estaquia, testaram-se três doses de AIB (0, 4000 e 8000 mg L-1) e dois comprimentos de estacas (3 e 10 cm). As estacas foram avaliadas quanto à sobrevivência, o enraizamento, o número de raízes, de folhas e de brotos e o comprimento de raízes e de brotos, aos 135 dias. No experimento de desinfestação, segmentos nodais de brotações de ano de erva-mate foram imersos em etanol 70%, por 2 e 4 min. e, depois, em NaOCl 2%, por 15, 25 e 35 min e, então inoculados em meio com ¼ do MS. Aos 15 dias avaliaram-se a percentagem de explantes contaminados, oxidados e sadios. Na micropropagação, plântulas de erva-mate, obtidas de embriões imaturos, foram submetidas a diferentes meios de cultura (MS, ¼ MS e WPM) por 50 dias, com avaliação do incremento obtido para as variáveis número de folhas e comprimento de plântulas. Para a multiplicação dessas plantas, foram testadas a adição de diferentes doses (0; 0,01; 0,1; 1,0 e 2,0 mg L-1) de BAP ao meio de cultura com ¼ do MS. Foram avaliados o número de brotos, folhas e entrenós, o comprimento dos brotos e a altura das plantas, aos 50 dias de cultivo. No enraizamento de brotos de erva-mate foram utilizadas doses (0; 0,5; 1,0; 1,5; 3,0 e 6,0 mg L-1) de AIB, por 15 e 30 dias. As avaliações constaram da percentagem de enraizamento e de calo obtidas, do número e comprimento de raízes, do número de folhas e altura das plantas, aos 30 dias de cultivo. Plantas de erva-mate, produzidas in vitro, foram aclimatizadas em diferentes substratos (vermiculita média, areia e casca de coco) com temperatura e luminosidade controladas. A maior percentagem de estacas enraizadas, em ambos os comprimentos, foi com a dose de 4000 mg L-1 de AIB, sendo que as estacas de 3 cm obtiveram uma maior percentagem de enraizamento. Segmentos nodais de brotações de ano de erva-mate apresentam a maior sobrevivência com a desinfestação com álcool 70% por 4 min. e em NaOCl 2% por 25 min. O meio com ¼ dos sais do MS pode ser utilizado como o meio de cultura base para o cultivo in vitro de plantas de erva-mate. A adição de 2 mg L-1 de BAP ao meio de cultura base aumenta o número de brotos adventícios em erva-mate, além de formar tufos após alguns subcultivos, possibilitando a manutenção de plantas como microcepas, para o fornecimento de brotos. O enraizamento in vitro de erva-mate pode ser realizado em apenas uma fase de 30 dias, com a adição de até 3 mg L-1 de AIB ao meio com ¼ dos sais do MS. Plantas de erva-mate, produzidas in vitro, podem ser aclimatizadas nos substratos casca de coco ou areia em ambiente com temperatura e luminosidade controladas.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Although the louro-pardo (Cordia trichotoma Vell.) is native forest specie with high timber potential, there are still scarce studies that approach the production of seedlings of this species by vegetative propagation. The objective of this work was to evaluate the vegetative propagation of the louro-pardo by the techniques of cuttings and micropropagation. It was tested the presence or absence of 8000 mg L-1 of indolbutiric acid (IBA) and two types of cuttings (basal and apical). At 40 days, the cuttings were evaluated for survival, rooting, the presence of callus and shoots, the number and length of shoots and the number of leaves. For the establishment of aseptic seedlings, seeds of louro-pardo were treated with 2% or 5% of sodium hypochlorite, for 0, 5, 10, 15 or 20 min. The seeds were inoculated in WPM basic culture medium. At 30 days, the percentages of disinfection, fungal contamination and / or bacteria and the average time of germination were evaluated. For multiplication, apical segments were inoculated in basic medium, plus 0; 0,25; 0,50 or 0,75 mg L-1 of 6-benzilaminopurin (BAP). At 45 days were evaluated the length, number of leaves and internodes of the shoots, the percentage of callus and the survival of the explants. In the multiplication, using microstumps kept in vitro, was tested whether or not the addition of 1,5 g L-1 of activated charcoal to the basic medium on the production of shoots. At 30 and 60 days, concerning to the first and second cuts of microstumps, were evaluated the number and length of shoots, number of leaves and internodes of the shoots, and the number of microcutting. Rooting in vitro microcuttings were maintained in basic medium plus 1,5 g L-1 of activated charcoal and IBA (1,5 or 2,0 mg L-1). At 45 days were evaluated the presence of roots and callus, the percentage of survival and sprouting. In cutting, were observed the formation of shoots on the cuttings, but they are not rooted. The cutting type and dose of IBA did not influence the rooting and survival. In micropropagation was found that the sterilization of seeds with 5% of sodium hypochlorite for 5 min. allowed the in vitro establishment of aseptic seedlings. The induction of shoots in the explants was not influenced by different doses of BAP. The presence of activated charcoal in the culture medium favored the formation and growth of shoots in microstumps. The use of 1,5 or 2,0 mg L-1 IBA did not promote the rooting of microshoots of louro-pardo. / Apesar de o louro-pardo (Cordia trichotoma Vell.) ser uma espécie florestal nativa com elevado potencial madeireiro, ainda são escassos os estudos que abordam a produção de mudas dessa espécie pela propagação vegetativa. O objetivo deste trabalho foi avaliar a propagação do louro-pardo pelas técnicas de estaquia e micropropagação. Para a estaquia foi testada a presença ou ausência de 8000 mg L-1de ácido indolbutírico (AIB) e dois tipos de estacas (basais e apicais). Aos 40 dias, as estacas foram avaliadas quanto à sobrevivência, o enraizamento, a presença de calos e de brotos, o número e o comprimento de brotos e o número de folhas. Para o estabelecimento de plântulas assépticas, sementes de louro-pardo foram tratadas com 2% ou 5% de hipoclorito de sódio, por 0, 5, 10, 15 ou 20 min. As sementes foram inoculadas em meio de cultura base WPM. Aos 30 dias foram avaliadas as porcentagens de desinfestação, de contaminação por fungos e/ou bactérias e o tempo médio de germinação. Para a multiplicação, ápices caulinares foram inoculados em meio de cultura base, acrescido de 0; 0,25; 0,50 ou 0,75 mg L-1 de 6-benzilaminopurina (BAP). Aos 45 dias avaliou-se o comprimento, o número de folhas e de entrenós dos brotos, as porcentagens de calo e de sobrevivência dos explantes. Na multiplicação, utilizando microcepas mantidas in vitro, testou-se a adição ou não de 1,5 g L-1 de carvão ativado ao meio de cultura base na produção de brotos. Aos 30 e 60 dias, referentes ao primeiro e ao segundo corte das microcepas, foram avaliados o número e o comprimento dos brotos, o número de folhas e de entrenós dos brotos, e o número de microestacas. No enraizamento in vitro, microestacas foram mantidas em meio de cultura base, acrescido de 1,5 g L-1 de carvão ativado e AIB (1,5 ou 2,0 mg L-1). Aos 45 dias foram avaliadas a presença de raízes e calos, a porcentagem de sobrevivência e de brotação. Na estaquia, foi observada a formação de brotos nas estacas, contudo estas não enraizaram. O tipo de estaca e a dose de AIB utilizada não influenciaram no enraizamento ou na sobrevivência. Na micropropagação foi verificado que a assepsia das sementes com 5% de hipoclorito de sódio, por 5 min., possibilitou o estabelecimento in vitro de plântulas assépticas. A indução de brotos nos explantes não foi influenciada pelas diferentes doses de BAP. A presença de carvão ativado no meio de cultura base favoreceu a formação e o crescimento dos brotos nas microcepas. A utilização de 1,5 ou 2,0 mg L-1 de AIB não promoveu o enraizamento adventício das microestacas de louro-pardo.