ANATOMICAL STUDIES OF IN VITRO ORGANOGENESIS INDUCED IN LEAF-DERIVED EXPLANTS OF PASSIONFRUIT

With the aim of studying the organogenesis in vitro in Passiflora edulis Sims f. flavicarpa Deg., the passionfruit, leaf-derived explants were cultured on media containing NAA or BAP and incubated either in continuous darkness or in light. The histological events leading to de novo organ formation were evaluated. Darkness induces rhizogenesis in the presence of NAA, whereas direct shoot regeneration is stimulated by light and BAP. This latter condition is recommended for passionfruit micropropagation as several adventitious shoot buds were formed from meristemoids of parenchymal origin.

Plant regeneration from protoplast of Brazilian citrus cultivars

A procedure is described to regenerate plants from protoplasts of Brazilian citrus cultivars, after isolation, fusion and culture. Protoplasts were isolated from embryogenic cell suspension cultures and from leaf mesophyll of seedlings germinated in vitro. The enzyme solution for protoplast isolation was composed of mannitol (0.7 M), CaCl2 (24.5 mM), NaH2PO4 (0.92 mM), MES (6.15 mM), cellulase (Onozuka RS - Yakult, 1%), macerase (Onozuka R10 - Yakult, 1%) and pectolyase Y-23 (Seishin, 0.2%). Protoplast culture in liquid medium after chemical fusion lead to the formation of callus colonies further adapted to solid medium. Somatic embryo formation occurred spontaneously after two subcultures, on modified MT medium supplemented with 500 mg/L of malt extract. Well defined embryos were germinated in modified MT medium with addition of GA3 (2.0 muM) and malt extract (500 mg/L). Plant regeneration was also achieved by adventitious shoots obtained through direct organogenesis of not well defined embryos in modified MT medium with addition of malt extract (500 mg/L), BAP (1.32 muM), NAA (1.07 muM) and coconut water (10 mL/L). Plantlets were transferred to root medium. Rooted plants were transferred to a greenhouse for further adaptation and development.

Resposta in vitro e suscetibilidade ao Agrobacterium de duas cultivares de Stylosanthes guianensis

As cultivares Bandeirantes e Mineirão de Stylosanthes guianensis (Leguminosae) foram avaliadas quanto à capacidade de formação de brotos adventícios in vitro e quanto à suscetibilidade ao Agrobacterium. Para obter regeneração, foram utilizados vários tipos de explantes, e o meio basal MS foi suplementado com diferentes concentrações de ácido naftalenoacético (NAA) e 6-benzilanimopurina (BAP). Observou-se regeneração de brotos a partir de explantes cotiledonares e hipocotiledonares, nas duas cultivares. O Stylosanthes mostrou-se suscetível ao Agrobacterium selvagem, pois a formação de tumores foi induzida. Expressão transiente do gene uidA foi observada em tecidos infectados de Stylosanthes. Experimentos sobre a ação dos antibióticos cefotaxima e tetraciclina, usados em ensaios de transformação para eliminação do Agrobacterium, mostraram que a cefotaxima (250 µg mL-1) tende a reduzir a regeneração de brotos em explantes da cv. Bandeirantes.

Somatic embryogenesis and in vitro plant regeneration from pejibaye adult plant leaf primordia

The aim of this work was to evaluate a protocol for plant regeneration by means of somatic embryos obtained from isolated adult pejibaye leaf primordia, and to describe histological origin of embryos and morphogenetic response. Explants were cultivated in modified MS medium. Mesophyll parenchymatous cells originated meristemoids (preembryonic complex formation) induced with 7.1 µM BAP in the first two subculture periods. After polarized structures with 12.9 µM NAA and 3.55 µM BAP were formed in the third subculture, somatic embryos developed and regenerated normal plants. The mesophyll parenchymatous cells display high capacity of direct response to the auxin and cytokinin.

Função hepática e renal de frangos de corte alimentados com dietas com aflatoxinas e clinoptilolita natural

O objetivo deste trabalho foi avaliar os minerais séricos e as funções hepática e renal de frangos de corte, alimentados com dietas com aflatoxinas e dietas com aflatoxinas e argila clinoptilolita natural. Foram utilizados 528 frangos de corte machos, da linhagem Ross, distribuídos em seis tratamentos com quatro repetições cada: T1 - testemunha: ração sem aflatoxina ou clinoptilolita; T2 - ração com 5 ppm de aflatoxinas; T3 - ração com 0,25% de clinoptilolita; T4 - ração com 5 ppm de aflatoxinas e 0,25% de clinoptilolita; T5 - ração com 0,5% de clinoptilolita; e T6 - ração com 5 ppm de aflatoxinas e 0,5% de clinoptilolita. As aves foram submetidas aos tratamentos do 1º ao 42º dia de idade. Foram abatidos 72 animais, e foram analisadas as concentrações séricas de cálcio, fósforo, uréia, creatinina, ácido úrico, colesterol, triglicerídeos e gamaglutamil-transferase (GGT). As aflatoxinas diminuem o nível sérico de colesterol. A clinoptilolita diminui o nível sérico do ácido úrico, quando na concentração de 0,25%. As aflatoxinas com 0,25% de clinoptilolita diminuem os níveis séricos da creatinina, do ácido úrico e do colesterol. As aflatoxinas acrescidas por 0,5% de clinoptilolita diminuem os níveis séricos da creatinina e do colesterol e elevam os de gamaglutamil-transferase.

Tissue culture parameters in sweet orange cultivars

The objective of this work was to establish tissue culture parameters for gene transfer in sweet orange cultivars. Epicotyl explants with different ages were cultured with 6-benzylaminopurine (BAP), kanamycin and hygromycin. Shoots were cultured with alpha-naphthaleneacetic acid (NAA) alone or in combination with indole-3-butyric acid (IBA). The requirement of BAP for shoot development was genotype-specific. Epicotyl explants from 35-day-old seedlings produced significantly more shoots per explant in 'Pêra'. Kanamycin inhibited shoot regeneration for the most cultivars. The percentage of shoots that produced roots in 'Pêra' was significantly higher in medium with NAA and IBA than with NAA alone.

Growth regulators, culture media and antibiotics in the in vitro shoot regeneration from mature tissue of citrus cultivars

The objective of this study was to evaluate the effects of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA) combinations, basal media and beta-lactam antibiotics on in vitro organogenesis from mature stem segments of 'Pêra', 'Valência' and 'Bahia' sweet oranges and 'Cravo' rangpur lime. For induction of shoot regeneration, the segments of the four cultivars were placed on Murashige and Skoog (MS) medium containing the following BAP/NAA concentrations: 0.0/0.0; 0.25/0.0; 0.25/0.25; 0.5/0.0; 0.5/0.5; 1.0/0.0; 2.0/0.0; 2.0/0.25; 2.0/0.5; and 2.0/1.0 mg L-1. In order to test the influence of the culture media on shoot-bud induction, (MS), Murashige and Tucker (MT), and woody plant medium (WPM) formulations were evaluated, associated with the best combination of plant growth regulators obtained in the previous experiment. The influence of four beta-lactam antibiotics (timentin, cefotaxime sodium salt, meropenem trihydrate and augmentin) on shoot regeneration was determined. Better regeneration responses were achieved when internodal segments were cultured onto MS-based medium with 500 mg L-1 cefotaxime with the following BAP/NAA concentrations: 0.5 + 0.25 mg L-1 for 'Cravo', 1.0 + 0.25 mg L-1 for 'Valência' and 'Bahia', and 1.0 + 0.5 mg L-1 for 'Pêra'. Genotype, growth regulators, basal media and beta-lactam antibiotics affect the morphogenetic response in mature tissues of citrus.

In vitro organogenesis from internodal segments of adult sweet orange plants

The objective of this work was to optimize in vitro plant regeneration via organogenesis from tissues of adult 'Hamlin', 'Pêra', and 'Valência' sweet orange plants. Explants were grown in EME culture medium with different concentrations of 6-benzylaminopurine (BAP) and naphthaleneacetic acid (NAA), at 27ºC in the absence of light for 50 days, followed by a 16-hour photoperiod for 20 days. Regeneration was assessed 50 and 70 days after in vitro culture. Organogenesis in cultivars Hamlin and Valência was promoted by EME supplemented with BAP, while NAA showed no apparent effect.

Effect of growth regulators on 'Brookfield' apple gas diffusion and metabolism under controlled atmosphere storage

The objective of this work was to evaluate the effect of growth regulators on gas diffusion and on metabolism of 'Brookfield' apple, and to determine their correlation with quality characteristics of fruit stored in controlled atmosphere. A completely randomized design was used with four replicates. After eight months of storage, the effects of water (control), aminoethoxyvinylglycine (AVG), AVG + ethephon, AVG + naphthaleneacetic acid (NAA), ethephon + NAA, sole NAA, 1-MCP, ethylene absorption by potassium permanganate (ABS), AVG + ABS, and of AVG + 1-MCP - applied at different rates and periods - were evaluated on: gas diffusion rate, ethylene production, respiratory rate, internal ethylene concentration, internal CO2 content, mealiness, and intercellular space. Fruit from the control and sole NAA treatments had the highest mealiness occurrence. Growth regulators significantly changed the gaseous diffusion through the pulp of 'Brookfield' apple, mainly in the treatment AVG + ABS, which kept the highest gas diffusion rate. NAA spraying in the field, with or without another growth regulator, increased ripening metabolism by rising ethylene production and respiration rate, and reduced gas diffusion during shelf life. AVG spraying cannot avoid the ethephon effect during the ripening process, and reduces both the internal space and mealiness incidence, but it is not able to induce ethylene production or to increase respiration rates.

Aplicação de reguladores vegetais no retardamento da abscisão de frutos de laranjeira-'Hamlin'

O trabalho avaliou os efeitos de auxinas e giberelina, combinadas e aplicadas em pré-colheita no desenvolvimento e na porcentagem de queda natural dos frutos de laranjeira-'Hamlin' (Citrus sinensis Osbeck). Foram realizadas 3 aplicações a intervalos de 45 dias dos seguintes tratamentos, via foliar: GA3 + NAA a 12,5mg.L-1 de cada; GA3 + NAA a 25mg.L-1; GA3 + 2,4-D a 12,5mg.L-1; GA3 + 2,4-D a 25mg.L-1; NAA + 2,4-D a 12,5mg.L-1; NAA + 2,4-D a 25mg.L-1; testemunha (água). As variáveis avaliadas foram: porcentagem de queda natural dos frutos (%), massa fresca de frutos (g) e teor de suco no fruto (%). Os resultados obtidos mostram que a utilização de reguladores vegetais atrasa a queda natural de frutos de laranjeira-'Hamlin', podendo-se prolongar o período de colheita, destacando-se o tratamento com GA3 25mg.L-1 +2,4-D 25mg.L-1 e também não influenciaram no aumento da massa fresca e no teor de suco dos frutos.

Somatic embryogenesis from zygotic embryos of Euterpe oleracea Mart.

The aim of this work was to study the morphogenetic responses of zygotic embryos of açai palm (Euterpe oleracea Mart.) submitted to several conditions of in vitro culture. Several research experiments were conducted, in laboratory, using vegetable material collected from açai palm plants at Embrapa Amazon Oriental, Belém-PA, Brazil. It was possible to verify the expression of a direct, repetitive and no-synchronized model of somatic embryogenesis in mature zygotic embryos cultivated in primary MS medium supplemented with 2,4-D (339.36 muM) and transferred to a secondary MS medium in the presence of NAA (0.537 muM) and 2iP (12.30 muM). The conversion of somatic embryos into seedlings was reached after 210 days with the transfer of the cultures to a third medium with sucrose and mineral salts concentrations reduced to a half, without growth regulators.

In vitro culture of Spondias mombin L. nodal segments

Spondias mombin L. shoot cultures were initiated from nodal explants taken from plants propagated by seeds. Explants coming from 4-6 months old plants, previously disinfected, were cultivated on WPM medium supplemented with a wide range of concentrations of BAP (0.0, 0.22, 0.44, 2.22 and 4.44 muM) and NAA (0.0, 0.27 and 2.70 muM). After four weeks, the responses obtained were axillary shoot and root formation. The first response were preferentially induced with the medium containing only BAP, regardless of the BAP concentration. The addition of NAA on medium reduced significantly axillary shoot formation and induced rhizogenesis. Roots were formed on nodal explant basis, preferentially on medium supplemented with 4.44 muM NAA. The medium supplemented with BAP reduced significantly root formation.

Efeitos de reguladores vegetais no enraizamento de estacas caulinares de Poncirus trifoliata var. monstrosa (T. Ito)

O trabalho objetivou estudar os efeitos de reguladores vegetais no enraizamento de estacas caulinares de Flying Dragon [Poncirus trifoliata var. monstrosa (T. Ito)]. Na metade de cada estação do ano, ramos de citros foram coletados (4 experimentos), a partir dos quais foram retiradas estacas com 10 cm de comprimento com uma folha cortada ao meio. As bases das estacas foram tratadas com os seguintes reguladores vegetais na forma de talco: Testemunha (H2O); IBA 0,5%; NAA a 0,5%; IBA + ácido caféico a 0,5%, e NAA + ácido caféico 0,5%. As estacas foram plantadas em bandejas de isopor contendo fibra de coco e mantidas por três meses em câmara de nebulização. Os resultados mostraram que para a propagação desta espécie por estaquia, a coleta dos ramos deve ser no outono e que as mesmas sejam tratadas com a mistura de NAA + ácido caféico, ambos a 0,5%.

Avaliação da aplicação exógena de poliaminas no crescimento de calos de mangabeira (Hancornia speciosa Gomes)

Este trabalho teve como objetivo estudar o efeito das poliaminas espermidina e espermina no crescimento de calos Hancornia speciosa Gomes. Calos com 0,5 cm de diâmetro foram inoculados em meio Murashige & Skoog (1962) (MS) a 50% + 100 mg L-1 de caseína hidrolisada + 200 mg L-1 de levedura de cerveja, variando os tratamentos:A: 1 mmol de espermina + 2 mg L-1 de 2,4-D (ácido 2,4 diclorofenoxiacético) + 0,5 mg L-1 de NAA (ácido naftalenoacético); B: 1 mmol de espermidina + 2 mg L-1 de 2,4-D + 0,5 mg L-1 de NAA; C: 2 mg L-1 de 2,4-D + 0,5 mg L-1 de NAA. Não houve influência das poliaminas no crescimento dos calos. observou-se, nos calos tratados com espermidina, maior concentração celular de putrescina (582,37 µg g mf-1) aos 60 dias, maior teor de espermidina (502,54 µg g mf-1) e espermina (868,53 µg g mf-1) aos 40 dias de cultivo, quando se aplicou a própria poliamina. Conclui-se que a aplicação exógena de poliaminas em Hancornia speciosa não proporciona aumento no crescimento de calos. A oxidação promovida por longos períodos de cultivo in vitro induz aumento nos níveis de putrescina.

Sour orange bud regeneration and in vitro plant development related to culture medium composition and explant type

In order to evaluate the formation of adventitious buds and in vitro regeneration of sour orange plants (Citrus aurantium L.) two organogenesis-inducing experiments were conducted. In the first experiment, the induction and in vitro regeneration of adventitious buds were tested on epicotyl and internodal segments under the influence of BAP or KIN associated with NAA. The second experiment evaluated the in vitro regeneration of sour orange plants related to different explant types (epicotyls segments, internodal segments of in vitro germinated plantlets and internodal segments of greenhouse cultivated plants). Data collected on both experiments included the percentage of responsive explants (explants that formed buds), and the number of buds per explant. The addition of BAP showed the best organogenic response. In vitro germinated epicotyl segments and internodal segments are recommended as explants for sour orange in vitro organogenesis. Rooting of regenerated shoots was achieved without the need of auxin in the medium.