ABSTRACT The use of silicates in plant tissue culture has resulted in positive effects regarding the morphological, anatomical and physiological characteristics of in vitro plants. However, biological responses to this mineral are dose and genotype dependent. The objective of the present study was to evaluate the influence of silicon supplementation on the somatic embryogenesis of oil palm (Elaeis guineensis Jacq.). The study was carried out in three stages of the cloning process, namely: Regeneration (i) and germination (ii) of somatic embryos and during ex vitro acclimatization (iii) of regenerated plantlets. The culture media were supplemented with four doses (0.0, 0.5, 1.0 and 1.5 g L-1) of calcium silicate (CaSiO3). The resulting plantlets were transplanted and acclimatized in a greenhouse. The regression analysis showed that the dose of 0.63g L-1 of CaSiO3 induced a higher rate of regeneration, which would allow a 58% increase in the number of somatic embryos formed. However, the germination of the plantlets, as well as their morphometric characters, were not affected by the different doses of CaSiO3, although the presence of this compound promoted an increase in the thickness of the epidermis and a reduction in the amount of epicuticular wax. Overall, silicon does contribute to the cloning of E. guineensis. morphological plants However dependent Elaeis Jacq.. Jacq Jacq. . Jacq.) process namely i (i ii (ii iii (iii 0.0, 00 0 (0.0 05 5 0.5 10 1 1. 15 L1 L CaSiO3. CaSiO (CaSiO3) greenhouse 063g 63g L- regeneration 58 formed characters wax Overall E 0.0 (0. 0. (CaSiO3 (0 (CaSiO (

RESUMO O uso de silicatos em cultura de tecidos tem resultado em efeitos positivos no que tange os caracteres morfológicos, anatômicos e fisiológicos das plantas in vitro. Contudo as respostas biológicas a este mineral são dose e genótipo-dependente. Nesse sentido, objetivou-se avaliar a influência da suplementação de silício no processo de embriogênese somática da Elaeis guineensis Jacq. O estudo foi realizado em três etapas do processo de clonagem, sendo elas: Regeneração (i) e germinação (ii) dos embriões somáticos e durante a aclimatação ex vitro (iii) das plântulas. Os meios de cultivo foram suplementados com quatro doses (0.0, 0.5, 1.0 e 1.5 g L-1) de silicato de cálcio (CaSiO3). As plântulas resultantes foram transplantadas e aclimatizadas em casa de vegetação. A análise de regressão evidenciou que a dose de 0.63g L-1 de CaSiO3 induziu maior taxa de regeneração, o que possibilitaria um aumento de 58% no número de embriões somáticos formados. Contudo, a germinação das plântulas, bem como, seus caracteres morfométricos não foram afetados pelas diferentes doses de CaSiO3, embora a presença deste composto tenha promovido aumento da espessura da epiderme e redução na quantidade de cera epicuticular. No geral, o silício contribui para a clonagem de E. guineensis. morfológicos genótipodependente. genótipodependente genótipo dependente. dependente genótipo-dependente sentido objetivouse objetivou se Jacq elas i (i ii (ii iii (iii 0.0, 00 0 (0.0 05 5 0.5 10 1 1. 15 L1 L CaSiO3. CaSiO . (CaSiO3) vegetação 063g 63g L- regeneração 58 formados como epicuticular geral E 0.0 (0. 0. (CaSiO3 (0 (CaSiO (

Resumo A propagação seminífera de Dietes bicolor é dificultada pela provável dormência física e/ou morfofisiológica. Objetivou-se analisar a eficácia de diferentes métodos de superação de dormência em sementes de D. bicolor a fim de determinar possível tratamento pré-germinativo para a espécie. Foram realizados dois experimentos: (I) Avaliação da quebra de dormência física, em que os tratamentos de escarificação mecânica utilizando-se lixa nº 100; escarificação química com ácido sulfúrico (H2SO4) por 5 e 10 minutos e imersão em água quente (70 ºC), também, em 5 e 10 minutos foram avaliados. A semente intacta foi considerada testemunha. (II) Avaliação da dormência morfofisiológica, no qual as sementes foram submetidas a tratamentos combinados de estratificação de temperaturas quentes (20-35 °C) e/ou temperatura fria (9 ºC), distribuídas nos períodos de 0, 1 e 2 semanas, totalizando 9 tratamentos. Em ambos os experimentos se determinou a porcentagem de germinação (G), índice de velocidade de germinação (IVG) e primeira contagem (PC), utilizando-se o delineamento em blocos casualizados e avaliados pelo teste de Scott-Knott a 1% e Dunnett à 5%. O uso H2SO4 por 10 minutos induziu 42% de germinação, 0,52 de índice de velocidade de germinação e 18% de primeira contagem, todavia somente PC mostrou-se significativamente distinto em relação à testemunha. No segundo ensaio, o tratamento que passou somente duas semanas em temperatura (20-35 ºC) apresentou a maior germinação (30%), em relação à testemunha (G = 22%). Esses resultados constituem informações relevantes para a compreensão da fisiologia da germinação de D. bicolor, além de contribuir com a otimização de práticas pré-germinativas para essa importante espécie ornamental.

Abstract Seminiferous propagation of Dietes bicolor is hindered by the probable physical and/or morphophysiological dormancy. The objective was to analyze the efficacy of different methods of overcoming dormancy in D. bicolor seeds to determine a possible pre-germination treatment for the species. Two experiments were conducted: (I) Evaluation of the breaking of physical dormancy, in which the treatments of mechanical scarification were carried using sandpaper #100; chemical scarification with sulfuric acid (H2SO4) for 5 and 10 minutes and immersion in hot water (70 ºC), also, for 5 and 10 minutes, were evaluated. The intact seed was considered a control. (II) Evaluation of morphophysiological dormancy, in which the seeds were submitted to combined treatments of stratification of hot temperature (20-35 °C) and/or cold temperature (9 ºC), distributed in periods of 0, 1, and 2 weeks, totalizing 9 treatments. In both experiments, germination percentage (G), germination speed index (GSI), and first count (FC) were determined, using a randomized block design, and evaluated by the Scott-Knott test at 1% and Dunnett’s test at 5%. The use of H2SO4 10 minutes induced 42% germination, 0.52 GSI, and 18% FC, but only FC was significantly different from the control. In the second trial, the treatment that spent only two weeks in hot temperatures (20-35 ºC) showed the highest germination (30%), compared to the control (G = 22%). These results provide relevant information for understanding the physiology of D. bicolor germination, in addition to contributing to the optimization of pre-germination practices for this important ornamental species.

RESUMO Reguladores de crescimento (RCs) e tipos de vedação de frascos que permitem as trocas gasosas podem favorecer o desenvolvimento de sistemas de propagação in vitro de plantas. O objetivo do presente estudo foi avaliar os efeitos da suplementação de citocinina na indução de multibrotações e a influência das trocas gasosas no desenvolvimento in vitro de microestacas de Jacaranda cuspidifolia Mart. Segmentos nodais foram cultivados em meio suplementado com diferentes concentrações (0,25; 0,5; 1,0 e 2,0 mg L-1) de 6-benziladenina (BA). No tratamento controle não houve adição de RCs. Após 30 dias de cultivo, as microestacas foram isoladas e transferidas para um meio de enraizamento suplementado com 1,0 mg L-1 de ácido indol-3-butírico. Nesta etapa, os frascos de cultura foram vedados com tampas rígidas de polipropileno com membrana porosa (1 M; 21 μL L-1 s-1 CO2) ou sem membrana (0 M; 14 μL L-1 s-1 CO2). A suplementação de citocinina induziu a ativação de gemas axilares de J. cuspidifolia. Os maiores números de gemas foram observados nos explantes cultivados na presença de 0,5 – 1,0 mg L-1 BA. As microestacas mantidas em frascos com maior nível de troca gasosa (1M) apresentaram maior porcentagem de enraizamento e maiores comprimentos de raiz e de parte aérea. A análise microscópica mostrou maior diferenciação dos tecidos foliares nas plantas mantidas em frascos com maior nível de troca gasosa (1M). Essas plantas também apresentaram maior espessura do mesofilo e consequentemente maior espessura da lâmina foliar. Os resultados obtidos fornecem novas informações para o estabelecimento de um sistema eficiente de propagação in vitro para J. cuspidifolia.

ABSTRACT Plant growth regulators (PGRs) and types of flasks sealing that allow gas exchange can favor the development of in vitro plant propagation systems. The objective of the present study was to evaluate the eff ects of cytokinin supplementation on the induction of shoot proliferation and the influence of gas exchange on the in vitro development of Jacaranda cuspidifolia Mart microcuttings. Nodal segments were cultured in medium supplemented with diff erent concentrations (0.25; 0.5; 1.0 and 2.0 mg L-1) of 6-benzyladenine (BA). In the control treatment, there was no addition of PGRs. After 30 days of culture, the microcuttings were isolated and transferred to a rooting medium supplemented with 1.0 mg L-1 indole-3-butyric acid. At this stage, the culture fl asks were sealed with rigid polypropylene lids with a porous membrane (1 M; 21 μL L-1 s-1 CO2) or with no membrane (0 M; 14 μL L-1 s-1 CO2). Cytokinin supplementation induced the activation of axillary buds of J. cuspidifolia. The highest numbers of shoots were observed in explants cultivated in the presence of 0.5–1.0 mg L-1 BA. The microcuttings kept in flasks with a higher level of gas exchange (1M) had a higher percentage of rooting and greater root and shoot lengths. Microscopic analysis showed a greater differentiation of leaf tissues in plants kept in flasks with a higher level of gas exchange (1M). These plants also showed greater mesophyll thickness and, consequently, greater leaf blade thickness. The results provide new information for establishing an efficient in vitro propagation system for J. cuspidifolia.

RESUMO: A morfometria de frutos e sementes é uma informação útil e importante para fenotipagem e caracterização genética de espécies vegetais com potencial econômico e, para a conservação de espécies ameaçadas, como é o caso de Butia purpurascens. Nesse estudo, determinamos comparativamente as características morfométricas de frutos e pirênios entre morfotipos amarelo e magenta de B. purpurascens, localizados em quatro localidades diferentes (populações). Beneficiamos 1380 frutos, coletados em 69 indivíduos. Avaliamos o comprimento, largura e o peso dos frutos e pirênios, a massa fresca e seca da polpa, e a porcentagem de umidade da polpa. O morfotipo amarelo apresentou frutos maiores e mais pesados do que o morfotipo magenta. Diferenças morfométricas também foram notórias entre as quatro populações, sugerindo que a fragmentação de ecossistemas pode causar efeitos genéticos deletérios em B. purpurascens ao longo do tempo.

ABSTRACT: Morphometry of fruits and seeds is a useful and important information for phenotyping and genetic characterization of crop plant species and, for conservation of endangered species, as is the case of Butia purpurascens Glassman. In this study, we determined comparatively the morphometric features of fruits and pyrenes between yellow and magenta B. purpurascens morphotypes, located in four different sites (populations). We processed 1380 fruits collected from 69 individuals. We evaluated the length, width and weight of the fruits and pyrenes, the fresh and dry pulp mass, and percentage of pulp moisture. Yellow morph showed larger and heavier fruits than magenta morph. Morphometric differences were also evident among the four populations, suggesting that ecosystems fragmentation can cause deleterious genetic effects in B. purpurascens in long-term.

ABSTRACT Colleters produce a secretion composed of hydrophilic and/or lipophilic substances which lubricates and protects the shoot apical meristem against biotic and abiotic agents. Little attention has been given to these structures in Clusiaceae. In the present study, the structure and development of the leaf colleters of Clusia criuva subsp. criuva were described and variations in the exudate composition of the colleters at different stages of leaf development were identified. The samples were collected and processed according to techniques for light and scanning electron microscopy. Colleters are of the standard type and not vascularized, and during leaf development, changes in color, structure, and secretion abundance were observed. Asynchrony in the development was noticeable in the leaf primordia and young leaves, from colleters in early formation to those in early senescence. In early phases there was an abundance of polysaccharide, lipid, and protein secretions, whereas adult and senescent leaves revealed an accumulation of phenolic compounds and cell degradation. The secretion was released by the rupture of the cuticle. The structural changes and secretion composition during leaf development emphasize the role of colleters in protecting meristems and developing organs.

ABSTRACT The Cerrado biome has several species that can generate income, and the cajá-manga is one of these native species with great potential but with scarce information about its morphological characterization. Thus, this work aimed to characterize the fruit, endocarp, seeds, and seedlings of cajá-manga morphologically. The fruits were evaluated by the characteristics of the length, width, and diameter of the fruit, endocarp, and seeds, the number of loculi, and the weight of fruits and endocarp. The plantlets were evaluated for the number of seedlings per endocarp, number of leaflets, number of leaflets of the 3rd and 4th leaves, stem diameter, length of the shoot and root, root and shoot dry matter and data analyzed by descriptive analysis. The morphometric characteristics of the fruits, endocarp, and seeds were analyzed using the Scott-Knott test. The morphological characteristics of the fruits, the endocarps, the seeds, and the plantlets of cajá-manga varied within the mother plants. These results can be used as a basis for future selection of plants with desirable characteristics on breeding programs.

RESUMO: A gabirobeira é uma espécie nativa do Cerrado brasileiro com potencial para uso em sistemas de cultivo. O objetivo deste estudo foi avaliar o efeito da 6-benzilaminopurina (BAP) em estacas radiculares de progênies de Gabirobeira (Campomanesia adamantium). As estacas radiculares foram obtidas de progênies de Gabirobeira de um e dois anos de idade. Estes foram segmentados em 5 cm de comprimento e apresentavam entre 1,90 a 3,22 mm de diâmetro, imersos nas concentrações: 0.0; 1.0; 2.0 e 4.0 mg L-1 de BAP por 15 segundos e plantados em bandejas contendo Bioplant®. O delineamento experimental foi o inteiramente casualizado, em esquema fatorial 2x4 (idades das estacas x concentrações de BAP), com quinze repetições por tratamento. Após 140 dias, foram avaliados o número de estacas com brotações, número de brotações, número de folhas e diâmetro da raiz principal. Estacas de raízes de progênies de dois anos de idade apresentaram melhor desenvolvimento. Menores concentrações de citocinina trouxeram melhores resultados de emissão e número de brotações das progênies de ambas as idades.

ABSTRACT: The gabirobeira is a species native to the Brazilian Cerrado with potential for use in cropping systems. This study evaluated the effect of the cytokinin 6-benzylaminopurine (BAP) on root cuttings of gabirobeira (Campomanesia adamantium). The plant material was obtained from gabirobeira progenies of one and two-years-old. The cuttings were segmented in 5 cm length and 1.90 to 3.22 mm diameter, immersed in the following BAP concentrations: 0.0; 1.0; 2.0 and 4.0 mg L-1 for 15 seconds and planted in trays containing the substrate Bioplant®. A complete randomized experimental design was adopted in a factorial scheme 2x4, (cuttings age x BAP concentrations) with fifteen replicates per treatment. After 140 days the number of cuttings with shoots, number of shoots, number of leaves, and diameter of the main root were evaluated. The better development of the cuttings was observed on progenies of two-years-old. The lowest cytokinin concentrations promoted the better emission and number of shoots of the progenies from both ages.

ABSTRACT The availability of reliable and reproducible micropropagation systems is a decisive step to successfully propagate plant species with slow growth and low germination rate. The effect was investigated of gibberellin (GA) and cytokinin (CK) in the in vitro germination and development of Dietes bicolor (Steud.) Sweet ex Klatt. Seeds were inoculated in Water + Agar; in Murashige e Skoog (MS) medium without plant growth regulators (PGRs) and in MS medium supplemented with 1 or 2 mg L-1 gibberellic acid or 6-benzylaminopurine. The germination, growth, and architecture of the plants presented significant alterations according to the treatments. Germination percentage increased at 2 mg L-1 GA. Plants grown on the GA-supplemented medium also showed larger leaves. The presence of CK in the culture medium induced the activation of axillary shoot buds and reduced the root length. CK and GA had antagonistic effects on the thickness of the leaf tissues. The activation of multiple shoots in plants cultured with CK in the medium allowed the in vitro propagation of approximately three plants per germinated seed. The results obtained in the present study bring an unprecedented description of a germination system for a higher yield in D. bicolor micropropagation.

Resumo Condições ambientais de crescimento podem modificar a estrutura foliar e a produção de metabólitos. Plantas de nim produzem uma quantidade elevada de metabólitos medicinais e contém terpenóides biopesticidas com baixa toxicidade. Entretanto, a alta variação genética e a baixa qualidade do material, além das modificações do ambiente alertam para a necessidade de técnicas de biotecnologia para certificar a produção de metabólitos de elevada qualidade. O objetivo foi investigar a estrutura foliar e características histoquímicas de Azadirachta indica cultivadas in vitro, in vivo e em condições de aclimatização. Foram encontradas diferenças anatômicas entre os ambientes, com maior espessura foliar e estômatos mais uniformemente distribuídos associados a condições in vivo. Essas modificações não afetaram a produção de metabólitos e biopesticidas. Terpenos e taninos foram sintetizados em células especializadas chamadas de idioblastos, localizadas no mesofilo e na região da nervura central, respectivamente. Assim, em uma abordagem qualitativa, pode-se afirmar que os diferentes ambientes não modificaram a produção de metabólitos. O aumento da produção destes compostos poderia ser relizada por meio do isolamento e cultivo in vitro de idioblastos como nova fonte de pesquisa em biotecnologia vegetal .

Abstract Environmental conditions of grow can modify leaf structure and metabolite production. Neem plants produce a high amount of medicinal metabolites and contain biopesticide terpenoids with low toxicity. However, the high genetic variation and the low material quality, besides the environmental modifications warn to the need of biotechnological techniques to ensure the production of high quality metabolites. The aim was to investigate leaf structural and histochemical characteristics of Azadirachta indica grown in vitro, in vivo and acclimatized condition. It was found anatomical differences among the environments, with higher leaf thickness associated to in vivo conditions, as well as were more evenly distributed stomata. Those modifications did not qualitatively affect the production of medicinal metabolites and biopesticides. Terpenes and tannins were observed in specialized cells called idioblasts, located in the mesophyll and in the midrib region, respectively. Thus, in a qualitative approach, we can affirm that the different environments do not modify metabolites production. Increased production of these bioactive compounds could be achieved by isolation and in vitro culture of idioblasts as a new source of research in plant biotechnology.

Resumo P. grandiflora é uma das plantas ornamentais com abundante floração de coloração variada com formas distintas e pétalas simples, dobradas e/ou múltiplas. O objetivo do trabalho foi induzir a regeneração pela via organogênica e o florescimento in vitro de P. grandiflora. Segmentos nodais de plântulas germinadas in vitro foram utilizados como fonte de explante para a regeneração. Cinetina (KIN) e 6-Benzilaminopurina (BA) foram utilizadas para a indução de organogênese de novo. Os tratamentos suplementados com 1.0, 1.5 mg L−1 de BA induziram a formação de maior número de brotações adventícias com número médio de 7.0 (±1,55) e 5.4 (±0.83), respectivamente. As microestacas obtidas a partir de gemas regeneradas produziram botões florais. Os botões florais localizaram-se nas regiões axilar e terminal das microestacas e desenvolveram-se em aproximadamente 10 dias de cultivo até a antese. O maior número de botões florais foi observado na presença de 0.75 mg L−1 de ácido giberélico. Este estudo abre novas perspectivas para o estabelecimento de ferramentas biotecnológicas a serem aplicadas para essa importante espécie ornamental.

Abstract P. grandiflora is a known ornamental plant with abundant flowering. The flowers exhibit varied coloration with distinct forms and simple folded petals and/or multiple. The objective of this work was to induce regeneration via organogenesis and in vitro flowering of P. grandiflora. Nodal segments of seedlings germinated in vitro were used as explant source for regeneration. Kinetin (KIN) and 6-Benzylaminopurine (BA) were used for the induction of organogenesis. The treatments supplemented with 1.0 and 1.5 mg L−1 BA induced the highest number of adventitious shoots with an average number of 7.0 (±1.55) e 5.4 (±0.83), respectively. The microcuttings obtained from regenerated shoots produced floral buds. The floral buds were located in the axillary and terminal regions of the microcuttings and developed in approximately 10 days of cultivation until the anthesis. The highest number of flower buds was observed in the presence of 0.75 mg L−1 of gibberellic acid. This study opens new perspectives for the establishment of biotechnological tools to be applied for this important ornamental species.

Resumo A compreensão do comportamento in vitro de explantes de flamboyant cultivados em meio de cultura suplementado com diferentes tipos e concentrações de citocininas pode auxiliar na otimização de sistemas de regeneração estabelecidos para essa espécie vegetal. Objetivou-se com o presente estudo avaliar o comportamento de explantes cotiledonares de flamboyant quando cultivados na presença de elevadas concentrações de 6-benziladenina (BA) ou cinetina (KIN). Segmentos cotiledonares obtidos de plântulas germinadas in vitro foram inoculados em meio de cultura Murashige e Skoog (MS), suplementado com diferentes concentrações (1, 2, 4, 8 mg L-1) de 6-benziladenina (BA) ou cinetina (KIN). No tratamento controle não foi adicionado reguladores de crescimento. Após 40 dias de cultivo, independente da concentração utilizada, os tratamentos suplementados com BA apresentaram maiores porcentagens de formação de calos e massa fresca, em comparação aos tratamentos suplementados com KIN. A formação de gemas adventícias foi observada principalmente nos tratamentos suplementados com BA. Análises histológicas evidenciaram que a formação de gemas adventícias ocorreu na periferia de calos formados a partir de células do mesofilo, nas regiões de seccionamento do explante. Os resultados obtidos fornecem novas informações para o estabelecimento de um sistema de micropropagação de flamboyant, uma importante espécie arbórea ornamental.

Abstract Understanding the in vitro performance of royal poinciana explants cultured in media supplemented with different types and concentrations of cytokinins may aid in the optimization of regeneration systems established for this woody species. In the present study we evaluate the in vitro performance of royal poinciana cotyledonary explants cultured in the presence of high concentrations of 6-benzyladenine (BA) and kinetin (KIN). Cotyledonary segments obtained from in vitro germinated seedlings were inoculated in Murashige and Skoog (MS) medium, supplemented with different concentrations (1, 2, 4, 8 mg L-1) of BA or KIN. In the control treatment, no plant growth regulators (PGRs) were added. After 40 days of culture, regardless of the concentration used, the treatments supplemented with BA presented higher calli percentage and fresh mass compared to treatments supplemented with KIN. Adventitious shoots were mainly observed in BA-treatments. Histological analysis showed that adventitious shoots formed at the periphery of callus formed from mesophyll cells in the regions of the explant sectioning. The results obtained provide new information for the establishment of a micropropagation system for royal poinciana, an important ornamental tree species.

Resumo O setor de flores ornamentais vem crescendo ao longo dos últimos anos em todo o mundo com potencial expansao. Entre as plantas ornamentais, espécies de Passiflora vem ganhando espaço no mercado, principalmente nos países da Europa e América do Norte. Contudo, o uso destas espécies na ornamentação ainda é pouco explorado. A inclusão da flor do maracujazeiro, na lista de plantas ornamentais está relacionada às características peculiares da flor como sua estrutura complexa, capacidade de floração durante o ano todo e também pela abundância e exuberância das folhas, que em muitas espécies agrega valor ornamental. Entre as ferramentas biotecnológicas para a produção de plantas ornamentais, cultura de tecidos tem sido proeminente na clonagem de genótipos elite, com alta qualidade fitossanitária e produção em larga escala. Além disso, oferece possibilidades de produzir novas variedades com características peculiares ao mercado de plantas ornamentais. A diversidade de espécies silvestres abre perspectivas para o mercado e produção de cultivares de Passiflora ornamental.

Abstract The ornamental flower sector has growing over the past years worldwide with potential for further expansion. Among the ornamental plants, Passiflora species have been gaining ground in the market, mainly in European and North American countries. However, the market aiming the use of these species in ornamentation is still poorly explored. The inclusion of passion flower in the list of ornamental plants is related to the peculiar characteristics of the flower as it is complex structure, capacity of flowering all year long and also by the abundance and exuberance of the leaves, which in many species adds an ornamental value. Among the biotechnological tools for the production of ornamental plants, tissue culture has been outstanding in the cloning of elite genotypes, with high phytosanitary quality and large scale production. In addition, it offers possibilities of producing new varieties with characteristics peculiar to the market of ornamental plants. The diversity of wild Passiflora opens perspectives to the conservation, market and production of ornamental Passiflora cultivars.

ABSTRACT The present study reports a shoot organogenesis-based system for in vitro regeneration of Passiflora miniata, an Amazonia passion fruit species. Root segments were cultured in Murashige and Skoog (MS) medium supplemented with different concentrations (range 2-9 µM) of 6-benzyladenine (BA); thidiazuron (TDZ) or kinetin (KIN). Plant growth regulators were not added to the control treatment. Root explants have showed a high regenerative potential. After 30 days of in vitro culture, the root explants showed several shoots formed direct and indirectly. TDZ provided the best response in the differentiation adventitious shoots, mainly in the presence of 6.8 µM. The cytokinins BA and KIN responded producing a reduced number of shoots. After 120 days, rooted regenerated plants were transferred to a greenhouse for acclimatization. This regeneration system opens new perspectives for micropropagation and conservation of this wild Amazonic passion fruit species.

Cyrtopodium glutiniferum is an endemic orchid of Brazil with potential medicinal and ornamental applications. As mycorrhizal fungi are essential for the initiation of the orchid life cycle, the aim of this study was to determine the strains of mycorrhizal fungi suitable for seed germination and protocorm development of C. glutiniferum and to characterize the symbiotic development of protocorms. Seeds of C. glutiniferum were inoculated with nine mycorrhizal fungi, Epulorhiza spp., Ceratorhiza spp., Rhizoctonia sp., originally isolated from Brazilian neotropical orchids. Only Epulorhiza isolates promoted seed germination and protocorm development. Three Epulorhiza isolates (M1, M6 = E. epiphytica, M20 = Epulorhiza sp.) promoted protocorm development until leaf production at 63 days. The protocorms are comprised of parenchyma cells delimited by a unistratified epidermis; the parenchyma cells of the upper part of the protocorms are smaller than those located more towards the base. Intact and digested pelotons were observed inside of protocorms implying that the seedlings were capable of mycotrophy. Additionally, the development of a bud primordium only occurred after colonization by fungus. This study suggests that C. glutiniferum has a preference for strains of Epulorhiza and that fungus digestion is essential to protocorm development.

Avaliou-se o potencial prognóstico de parâmetros microscópicos de folhas assintomáticas de . Clusia hilariana Schltdl. expostas à deposição particulada de ferro (2.14 mg cm² dia-1), por 45 dias consecutivos. Amostras de folhas jovens e expandidas sem sintomas foram coletadas e submetidas a técnicas usuais em microscopia de luz e eletrônica de varredura. Foram mensurados a altura das células epidérmicas de ambas as faces da folha e a espessura da hipoderme, do parênquima clorofiliano e da lâmina foliar. Danos micromorfológicos ocorreram na superfície abaxial de folhas jovens e em ambas as faces de folhas expandidas. Erosão da cera epicuticular e ruptura cuticular foram frequentes na face adaxial, enquanto na face abaxial, de ambas as folhas, houve perda da sinuosidade da parede anticlinal das células epidérmicas, deformação e obstrução estomática. Alterações micromorfométricas foram observadas em todos os tecidos foliares, exceto na altura das células epidérmicas em razão, provavelmente, da espessa cutícula e flanges cuticulares proeminentes. A maior diferença na espessura da lâmina foliar foi verificada nas folhas jovens de plantas expostas à SPM Fe, indicando maior sensibilidade ao ferro particulado em comparação com as folhas expandidas. As alterações micromorfológicas e micromorfométricas observadas na lâmina foliar de . Clusia hilariana Schltdl. demonstraram o potencial prognóstico dessas ferramentas na avaliação de impactos causados pela deposição de particulados, em especial na vegetação natural de restinga, onde a exposição é crescente diante da expansão de usinas mineradoras implantadas em sua vizinhança.

This study aims to evaluate the prognostic value of microscopic parameters of asymptomatic leaves of Clusia hilariana Schltdl. subjected to particulate deposition of iron (2.14 mg cm-2 day-1) for 45 consecutive days. Samples of young and expanded leaves without symptoms were collected and subjected to light and scanning electron microscopy techniques. The height of the epidermal cells on both surfaces of the leaf and the thickness of the hypodermis, the chlorophyll parenchyma, and the leaf blade were measured. Micromorphological injury occurred in the abaxial surface of young leaves and on both surfaces of expanded leaves. Erosion of the epicuticular wax and cuticle rupture were frequent on the adaxial surface, while on the abaxial surface of both leaves there was a loss of sinuosity on the anticlinal wall of the epidermal cells, stomatal deformity and obstruction. Micromorphometric alterations were seen in all leaf tissues except in the height of epidermic cells, probably due to the thick cuticle and prominent cuticular flanges. The highest difference in thickness of the leaf blade was seen in young leaves of plants subjected to SPMFe, indicating greater sensibility to particulate iron in comparison to the expanded leaves. The micromorphological and micromorphometric alterations in the leaf blade of Clusia hilariana Schltdl. showed the prognostic potential of these tools on the evaluation of impacts caused by the deposition of particulate matter, especially in the 'Restinga' natural vegetation, where the exposure is increasing due to the presence of iron ore industry in their surroundings.