ABSTRACT. The aim of the study was to assess the effect of plant regulators (gibberellic acid and naphthalene acetic acid) and adjuvant LI 700® with and without hand pollination on the fruiting and quality of Annona squamosa L. cultivars. The experiment was conducted in the experimental orchard of the Florida Tropical Research and Education Center (TREC/UF) in Homestead, Florida, USA. The experimental design in random blocks comprised 14 treatments, 10 repetitions and 3 flowers per treatment and per tree for ‘Red’ and ‘Lessard Thai’ cultivars. To assess quality, 9 fruit from each treatment were used, with 3 repetitions and 3 fruit per repetition. High instability was observed in fruit fixation rates between cultivars. The application of growth regulators did not increase fruit fixation in the tested cultivars. The application of naphthalene acetic acid had a deleterious effect on the flowers of the sugar apple cultivars. The most uneven (asymmetrical) fruit was obtained in the treatments with the application of growth regulators. The application of the adjuvant showed toxicity, causing damage to the skin of the fruit. The fruit with the greatest symmetry had the lowest levels of soluble solids and the highest toxicity. The application of growth regulators associated with hand pollination was not efficient, presenting a low percentage of fruit fixation for Annonaceae cultivars. None of the studied treatments produced parthenocarpic fruit (without seeds). The utilization of an adjuvant in the applied concentration caused damage to the fruit peels (darkening) due to toxicity in both cultivars.

RESUMO - O aperfeiçoamento de técnicas de propagação in vitro, com auxílio da escolha adequada da fonte de luz artificial e das formulações do meio de cultivo, são importantes, pois viabilizam a produção em massa de indivíduos com grande variabilidade genética. Assim, a presente pesquisa teve por objetivos estudar fontes de luz a partir de lâmpadas fluorescentes e diodos emissores de luz (LEDs) e formulações de meio de cultivo na germinação e no desenvolvimento inicial da espécie de orquídea Brassavola perrinii. O experimento foi instalado em delineamento inteiramente casualizado em esquema fatorial 5 × 4, sendo cinco fontes de luz: lâmpada fluorescente (100%); LED branco (100%); LED azul (100%); LED vermelho (100%) e LED azul (50%) + vermelho (50%), e quatro formulações de meio de cultivo, sendo MS, ½ MS, VW e K, com quatro repetições e média de 125 sementes por parcela. Aos 90 dias após a semeadura foram avaliadas a porcentagem de germinação, protocormos clorofilados e o desenvolvimento dos protocormos através das classes: P1, P2, P3 e P4 para cálculo do índice de desenvolvimento protocormos. Os dados foram submetidos à análise de variância e as médias foram comparadas pelo teste de Tukey p < 0,05. Para Brassavola perrinii, concluiu-se que o percentual de germinação não foi influenciado pelos espectros de luz, mas foi maior nas formulações ½ MS e VW. O maior índice de desenvolvimento dos protocormos de B. perrinii foi observado quando submetidos ao espectro de luz da fonte de iluminação com lâmpadas fluorescentes em meio de cultivo VW.

ABSTRACT - The improvement of in vitro propagation techniques using appropriate artificial light sources and culture medium formulations is important to allow the mass production of individuals with high genetic variability. Thus, this research aimed to study the effect of light sources from fluorescent lamps and light-emitting diodes (LEDs) and culture medium formulations on the germination and initial development of the orchid species Brassavola perrinii. The experiment was set up in a completely randomized design in a 5 × 4 factorial scheme, consisting of five light sources – fluorescent lamp (100%), white LED (100%), blue LED (100%), red LED (100%), and blue LED (50%) + red LED (50%) – and four culture medium formulations – MS, ½ MS, VW, and K –, with four replications and a mean of 125 seeds per plot. The percentage of germination, chlorophyll protocorms, and protocorm development through the classes P1, P2, P3, and P4 to calculate the protocorm development index were evaluated at 90 days after sowing. The data were subjected to analysis of variance and the means were compared by the Tukey test (p < 0.05). The percentage of germination for B. perrinii was not influenced by light spectra but was higher in the ½ MS and VW formulations. The highest protocorm development index of B. perrinii was observed when the species was submitted to the light spectrum of the lighting source with fluorescent lamps in the VW culture medium.

ABSTRACT. In vitro culture systems based on liquid culture media are considered to be more effective than semisolid culture medium systems. Liquid culture media systems provide better nutrient availability for plant tissues, easier culture handling, and the potential for scaling up and automation. However, in vitro liquid culture requires more careful handling due to the potential for contamination and the possibility of negative effects, such as hyperhydricity or vitrification, that hinder the growth and development of the plant material. Temporary immersion bioreactors have emerged as a workable alternative for capturing the benefits of liquid media, though semisolid systems are still traditional. Many studies have shown that silicon (Si) is a beneficial plant nutrient. Silicon might have a positive effect in both semisolid and liquid in vitro systems. The objective of this study was to evaluate the effect of silicon on the micropropagation and acclimatization of banana plants cultivated in vitro by comparing liquid temporary immersion bioreactor technology and semisolid traditional culture systems. Different silicon concentrations (0 and 1 mL L-1) and culture systems (liquid temporary immersion bioreactor and semisolid traditional culture) were evaluated over a 36-day period. The growth characteristics plant size, fresh and dry weight, and number and length of leaves and roots were evaluated. After the 36-day in vitro growth period, plants were transferred to a greenhouse for acclimatization and were evaluated after 30 days for the same growth characteristics used in the in vitro studies. The temporary immersion bioreactor system resulted in greater growth of banana plants compared to the traditional semisolid system. Temporary immersion bioreactors also showed a positive interaction with Si and resulted in higher values for all growth characteristics in the acclimatization phase.

Resumo O objetivo deste estudo foi avaliar a eficiência da solução crioprotetora (PVS2) combinada com floroglucinol para a criopreservação de sementes de duas espécies de orquídeas, Encyclia cordigera e Epidendrum ciliare. Sementes de Encyclia cordigera apresentaram viabilidade inicial de 91,03% e germinação de 91,99%. O tratamento das sementes com PVS2 a 0 °C com 1% de floroglucinol por 60 min promoveu a recuperação de 93,79% da viabilidade e 91,01% da germinação após a recuperação do nitrogênio líquido, resultando consequentemente no desenvolvimento mais rápido dos protocormos. Para Epidendrum ciliare, a viabilidade inicial das sementes foi de 85,65% e germinação de 85,90%. E quando suas sementes foram tratadas com solução PVS2 a 0 °C com 1% floroglucinol por 180 min atingiram viabilidade de 39,23% e germinação de 37,88%. Apesar de observada menor porcentagem de germinação para o referido tratamento, 78,90% dos protocormos atingiram estágio de desenvolvimento P3, quando avaliados 45 dias após a semeadura, não apresentando diferença significativa comparada ao controle 1, e apresentando protocormos com desenvolvimento normal. Esses resultados indicam que a utilização da solução crioprotetora de PVS2 combinado ao floroglucinol 1% foi eficiente na recuperação de sementes criopreservadas de Encyclia cordigera e Epidendrum ciliare. O presente estudo também indica que a resposta à criopreservação após o armazenamento a frio é específica da espécie e requer ajustes no tempo de exposição ao PVS2 a 0 °C antes da imersão em nitrogênio líquido.

Abstract The objective of this study was to evaluate the efficiency of cryoprotective solution (PVS2) combined with phloroglucinol for the cryopreservation of seeds of two orchid species, Encyclia cordigera and Epidendrum ciliare. Seeds of Encyclia cordigera had 91.03% initial viability and 91.99% germination. The treatment of the seeds with PVS2 at 0 °C with 1% phloroglucinol for 60 min returned 93.79% viability and 91.01% germination after recovery from LN, consequently resulting in faster development of protocorms. For Epidendrum ciliare, seed viability was 85.65% and germination was 85.90%. Seed exposure to the PVS2 at 0 °C with 1% phloroglucinol for 180 min showed viability of 39.23% and germination of 37.88%. Despite lower germination, 78.90% of the protocorms reached stage P3 of development, when evaluated 45 days after sowing, not significantly different from the control 1, and showed normal development. These results indicate that PVS2 cryoprotective solution is efficient when combined with phloroglucinol for the cryopreservation and successful recovery of seeds of Encyclia cordigera and Epidendrum ciliare. The present study also indicates that response to cryopreservation and success of recovery after cold storage is species-specific and requires adjustments in exposure time to PVS2 at 0 °C prior to immersion in LN.

Abstract: Caçari is a fruit tree that is native to the Amazon region of Brazil. The fruit is considered to have the highest vitamin C content of any edible fruit (13.757 mg 100 g-1 dry weight). The objective of this study was to evaluate the effects of 2,4-D and 6-BAP alone or in combination on in vitro callus induction and pro-embryogenic mass formation in caçari stem segments in different culture media as a precursor for caçari micropropagation. The experiment consisted of two culture media (MS and WPM) and combinations of four concentrations of 2,4-D (0, 1, 2 and 4 mg L-1) and BAP (0, 0.25, 0.5 and 1 mg L-1). The results showed that the highest percentage of callus formation (99%) was obtained in WPM supplemented with 4 mg L-1, 2,4-D and 1 mg L-1 BAP, and this combination resulted in 93% pro-embryogenic mass formation.

Resumo O coqueiro (Cocos nucifera L.) é uma das mais importantes espécies tropicais utilizadas pelo homem, conhecida como “árvore da vida”. Nas últimas décadas, com a expansão das áreas de cultivo, limitações como a ocorrência de pragas, uniformidade de culturas, adaptação a diferentes ecossistemas e outras têm afetado a produção e a longevidade dos plantios. Esta revisão descreverá os principais avanços nas técnicas de propagação do coqueiro, convencionalmente, por sementes e por meio de técnicas de cultura de tecidos de plantas a partir de meados do século XX, bem como aspectos fitossanitários que devem ser considerados para mitigar a disseminação de pragas e doenças.

Abstract Coconut palm (Cocos nucifera L.) is one of the most important tropical species used by man, known as “tree of life”. In recent decades, with the expansion of coconut growing areas, limitations such as the occurrence of pests, uniformity of crops, adaptation to different ecosystems and others have affected the production and longevity of coconut trees. This review describes the major advances in propagation techniques of coconut palm, conventionally, by seeds and through plant tissue culture techniques from the mid-twentieth century onwards, as well as phytosanitary aspects that should be considered to mitigate the spread of pests and diseases.

Abstract Yellow passion fruit (Passiflora edulis Sims) yield and longevity have been drastically reduced by bacterial spot (Xanthomonas axonopodis pv. passiflorae - Xap) and passion fruit woodiness disease (PWD) (Cowpea aphid-borne mosaic virus - CABMV). This study was aimed at evaluating the reaction of 11 genotypes of yellow passion fruit, based on the reaction of their progenies, to both mechanically inoculated Xap and CABMV, under greenhouse conditions. There was a progressive increase in bacterial spot and PWD severity with time. BRS Gigante Amarelo, MAR20#12, and MAR20#34 were selected as the progenies with lowest bacterial spot severity and disease progress over time. MAR20#2005, EC-L-7, UnB2015-1, and EC-3-0 presented the lowest PWD severity and disease progress over time. These progenies, along with individual plants from other progenies in which disease severity was significantly low until the last evaluation, will be cloned and tested again for Xap and CABMV, including other isolates.

ABSTRACT The preservation of banana genetic material is usually performed through seedlings. However, most banana cultivars do not produce seed and are propagated vegetatively. Therefore, cryopreservation is a feasible technique that allows the preservation of banana genotypes indefinitely. For the success of cryopreservation protocols, the selection of cryoprotectants and pre-freezing techniques are important factor. Therefore, the objective of this study was to verify the effects of different cryoprotectants with and without 1% phloroglucinol and pre-cooling periods on the development of a protocol for cryopreservation of in vitro rhizomes ofMusa accuminata(AAA) cv Grand Naine banana. The addition of 1% phloroglucinol to the cryoprotective solutions, such as PVS2 enhanced recovery of cryopreserved banana rhizomes. In addition, pre-cooling of explants in ice for 3 hours in PVS2 + 1% of phloroglucinol allowed efficient cryopreservation of banana rhizomes, followed by successful recovery and regeneration of in vitro shoots of banana cv Grand Naine.

ABSTRACT Jatropha is a potential oilseed crop, which requires mitigating factors such as the low genetic variability of the species. The solution runs through the research of Brazilian germplasm. Attention should be given to the germplasm of jatropha the north of Minas Gerais, because this is the oldest national collection and because this region may be a regions of jatropha diversity due to selection pressure arising from environmental adversities. Therefore, the objective of this study was to investigate the genetic diversity of 48 accessions of collection from Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG), using SSR and ISSR markers. The results showed low genetic diversity, but some individuals stood out as J. mollissima (48), J. podagrica (47), Mexican accessions (42, 43, 44 and 45) and some national accessions (28, 29, 41 and 46). Therefore, aiming to increase the genetic variability and improve the effectiveness of jatropha breeding programs, it is suggested to explore such as parental accessions to generate commercial hybrids. This fact implies the possibility to support future production of jatropha, since this culture may be an important source of income, especially for small farmers living in semiarid regions of Brazil.

Jatropha curcas L. tem sido identificada para a produção de biocombustível, mas apresenta produção comercial restrita devido à limitada ramificação e à ausência de uniformidade na produção. O objetivo deste estudo foi avaliar os efeitos de uma única aplicação de etefon ou da combinação de benziladenina (BA) com isômeros A4 e A7 do ácido giberélico (GA4+7) na indução de ramifica ções, florescimento e frutificação, em plantas de jatropha com e sem folhas. Plantas com e sem folhas mostraram diferenças para variáveis de crescimento e reprodutivas. Não houve interação significativa entre a presença de folhas e as concentrações de regulador de crescimento, exceto para a variável vingamento de inflorescência. O número total de flores por inflorescência diminuiu à medida que a concentração de etefon aumentou. O peso seco da semente aumentou com o aumento da concen tração de BA + GA4 +7 . Assim, aplicações de etefon e BA + GA4 +7 pareceram ter um efeito mais pronunciado na produção de flores e sementes do que na indução de ramos. A inabilidade de discernir os efeitos significativos dos tratamentos para a maioria das variáveis pode ter ocorrido devido à grande variabilidade existente nas populações de plantas estudadas, resultando em um tamanho da amostra insuficiente. Portanto, os dados coletados neste estudo foram utilizados para estimativas estatísticas de tamanhos de amostras para fornecer referências a estudos futuros.

Jatropha curcas L. has been identified for biofuel production but it presents limited commercial yields due to limited branching and a lack of yield uniformity. The objective of this study was to evaluate the effects of single application of ethephon or a combination of 6-benzyladenine (BA) with gibberellic acid isomers A4 and A7 (GA4+7) on branch induction, flowering and fruit production in jatropha plants with and without leaves. Plants with and without leaves showed differences for growth and reproductive variables. For all variables except inflorescence set, there were no significant statistical interactions between the presence of leaves and plant growth regulators concentration. The total number of flowers per inflorescence was reduced as ethephon concentration was increased. As BA + GA4 +7 concentration increased, seed dry weight increased. Thus, ethephon and BA + GA4 +7 applications appeared to affect flowering and seed production to a greater extent than branching. The inability to discern significant treatment effects for most variables might have been due to the large variability within plant populations studied and thus resulting in an insufficient sample size. Therefore, data collected from this study were used for statistical estimations of sample sizes to provide a reference for future studies.

As orquídeas tornaram-se parcela importante do mercado da floricultura nacional. Arundina graminifolia é uma orquídea terrestre, produz flores atraentes e uma espécie que poderia ser uma candidata potencial para o mercado da floricultura, entretanto, sua altura considerável torna difícil seu transporte e comercialização. Alguns reguladores de crescimento de plantas têm sido utilizados para controlar a altura em plantas ornamentais e em outras espécies. O objetivo desse estudo foi avaliar a eficiência dos reguladores de crescimento, paclobutrazol e cloreto de chlormequat no desenvolvimento vegetativo da orquídea A. graminifolia em vasos, visando o controle de altura. Paclobutrazol (Cultar) foi aplicado a 0, 5, 10 e 20 mg L-1, e CCC (Cycocel) foi aplicado a 0; 2000; 4000 e 6000 mg L-1. As plantas foram avaliadas mensalmente para altura de planta e número de brotos por vaso. CCC não teve efeito sobre a altura final das plantas nas concentrações aplicadas. Paclobutrazol foi eficaz no controle de altura da planta na concentração de 5 mg L-1, mas concentrações mais elevadas (10 e 20 mg L-1) provou ser tóxico para as plantas, causando a morte em brotos novos. Paclobutrazol em concentrações mais baixas oferece um meio viável para o controle de altura em A. graminifolia.

Orchids have become an important portion of the international floriculture market. Arundina graminifolia is a terrestrial orchid that produces attractive flowers, and, although the species could be a potential candidate for the floriculture market, its considerable height makes it difficult to transport and commercialize. A number of plant growth regulators have been utilized to control plant height in ornamentals and other species. Thus, the aim of this study was to evaluate the efficiency of growth regulators, paclobutrazol and chlormequat chloride on the vegetative development of containerized A. graminifolia orchid aiming at height control. Paclobutrazol (Cultar) was applied at 0, 5, 10, and 20 mg L-1, and CCC (Cycocel) was applied at 0, 2000, 4000, and 6000 mg L-1. The plants were assessed monthly for the plant height and number of shoots per container. CCC had no effect on the final height of plants at the concentrations applied. In contrast, paclobutrazol was effective in controlling plant height at a concentration of 5 mg L-1, but higher concentrations (10 and 20 mg L-1) proved to be toxic to the plants, causing death to the new shoots. Paclobutrazol at lower concentrations offers a viable means for height control in A. graminifolia.

As orquídeas são plantas exuberantes e apresentam alto valor agregado devido à diversidade e à beleza de suas flores, o que incrementa a sua comercialização. A família Orchidaceae compreende cerca de 35.000 espécies, distribuídas em mais de 1000 gêneros distintos e 100.000 híbridos, totalizando cerca de 8% a 10% de todas as plantas com flores. Com o avanço da agricultura e a constante degradação do seu habitat natural, as espécies de orquídeas são coletadas de maneira indiscriminada por colecionadores e comerciantes. Este extrativismo ameaça muitas espécies de extinção reduzindo drasticamente a sua variabilidade genética na natureza. Portanto, torna-se fundamental a busca de alternativas que viabilizem a preservação destas espécies, com a utilização de técnicas que apresentem baixo custo de manutenção, que proporcionem maior tempo de armazenamento, e que possibilite ao material vegetal boas condições fitossanitárias para utilização comercial. A criopreservação compreende a conservação de material biológico a temperaturas ultrabaixas, geralmente em nitrogênio líquido a -196 ºC, ou em sua fase de vapor a -150 ºC. Esta é a única técnica disponível atualmente para a preservação a longo prazo do germoplasma de espécies de plantas que são propagadas vegetativamente ou que possuem sementes inviáveis, recalcitrantes ou intermediárias. Esta revisão bibliográfica objetivou reportar a importância, métodos e aplicação do método de criopreservação para orquídeas. De acordo com os trabalhos revisados, pode-se verificar que essa é uma área relativamente nova e importante e encontra-se em pleno desenvolvimento, o que gera bastante discussão e requer ainda muita pesquisa em relação ao tipo de tratamento a utilizar para criopreservar e à metodologia a ser aplicada de acordo com a espécie utilizada. Devido à observação, neste trabalho, dos tipos de métodos que são utilizados para criopreservação e das grandes variações nas respostas das orquídeas aos métodos de criopreservação, o desenvolvimento de protocolos mais adequados para preservação das orquídeas torna-se necessário.

Orchids are lush and highly valuable plants due to their diversity and the beauty of their flowers, which increases their commercialization. The family Orchidaceae comprises approximately 35,000 species, distributed among more than 1,000 distinct genera and 100,000 hybrids, totaling approximately 8% to 10% of all flowering plants. With the advance of agriculture and the constant destruction of their natural habitat, orchid species are collected in an indiscriminate manner by collectors and vendors, and this extractive activity threatens many species with extinction, drastically reducing their genetic variability in nature. Therefore, it is essential to seek alternatives that make the preservation of such species feasible using techniques with low maintenance costs that provide greater storage time and that enable good phytosanitary conditions for the plant material for commercial use. Cryopreservation involves the conservation of biological materials at ultra-low temperatures, generally in liquid nitrogen at -196 ºC or in its vapor phase at -150 ºC. This is the only technique currently available for the long-term preservation of the germplasm of plant species that are vegetatively propagated or that have unviable, recalcitrant or intermediate seeds. The objective of this bibliographic review is to report on the importance, methods and application of cryopreservation for orchids. According to the studies reviewed, this is an incipient, developing and relevant field that generates a lot of discussion and requires further research relative to the type of treatment to use for cryopreservation and the methodology to be applied according to the species. The types of methods that are used for cryopreservation and the large variation in the responses of orchids to the cryopreservation methods observed in this study emphasize the need for the development of more appropriate protocols for the preservation of orchids.

A diversidade genética da coleção de 64 acessos de pinheira, coletados em diferentes cidades, no norte do Estado de Minas Gerais, foi avaliada por meio do uso de marcadores RAPD. Foram selecionados 20 primers RAPD que geraram 167 fragmentos, dos quais 48 foram polimórficos (28,7%), produzindo uma média de 2,4 fragmentos polimórficos por primer. Baixa percentagem de polimorfismo foi obtida com o conjunto de primers (< 29%), indicando baixa variação genética entre os 64 acessos avaliados. As distâncias genéticas foram estimadas, utilizando-se o coeficiente de similaridade de Jaccard. Acessos de diferentes cidades foram agrupados em um mesmo grupo, indicando que não há correlação entre os agrupamentos moleculares e origem geográfica. O dendrograma revelou cinco grupos. O primeiro grupo reuniu os acessos C19 e G29, coletados nas cidades de Verdelândia e Monte Azul, respectivamente. O segundo grupo reuniu os acessos G16 e B11, coletados nas cidades de Monte Azul e Coração de Jesus, respectivamente. Os acessos remanescentes foram agrupados em três grupos, com oito, 15 e 37 acessos, respectivamente. O marcador RAPD apresentou baixo nível de polimorfismo entre os acessos avaliados.

Genetic diversity in a collection of 64 sugar apple accessions collected from different municipalities in northern Minas Gerais was assessed by RAPD analysis. Using 20 selected RAPD primers 167 fragments were generated, of which 48 were polymorphic (28.7%) producing an average of 2.4 polymorphic fragments per primer. Low percentage of polymorphism (< 29%) was observed by using the set of primers indicating low level of genetic variation among the 64 accessions evaluated. Genetic relationships were estimated using Jaccard's coefficient of similarity. Accessions from different municipalities clustered together indicating no correlation between molecular grouping and geographical origin. The dendrogram revealed five clusters. The first cluster grouped C19 and G29 accessions collected from the municipalities of Verdelândia and Monte Azul, respectively. The second cluster grouped G16 and B11 accessions collected from the municipalities of Monte Azul and Coração de Jesus, respectively. The remaining accessions were grouped in three clusters, with 8, 15 and 37 accessions, respectively. In summary, RAPD showed a low percentage of polymorphism in the germplasm collection.

The present study investigated the germination and vigor of Caesalpinia echinata (Brazilwood) seeds stored at negative temperatures. Recently harvested seeds were cryopreserved at -18º or -196ºC and periodically evaluated for germination, seed vigor and carbohydrate composition. The temperatures did not influence the germination percentages or vigor. The germination percentage decreased from 88% in recently harvested seeds to 60% after 730 days of storage. The different temperature and storage times tested did not affect the vigor seed germination as indicated by the measures of plant growth and survival. The different temperatures used did not cause changes in the carbohydrate composition. The tegument cell walls were rich in lignin, arabinose and xylose. The cytoplasm of the cotyledons and embryos had high levels of glucose, fructose, and sucrose. The cryopreservation technique here presented was effective in the conservation of Brazilwood seeds for the medium term.

A regulação da calogênese é o primeiro passo para a embriogênese somática indireta in vitro. Nesse sentido, foram investigados os efeitos de diferentes meios de cultura, reguladores de crescimento e fontes nitrogenadas no desenvolvimento de calos em tecidos foliares de Caesalpinia echinata (pau-brasil). Explantes foliares foram cultivados em meios de cultura MS, B5, White e WPM. Testou-se, também, o efeito dos compostos nitrogenados (NH4NO3, KNO3 e glutamina) e a interação entre auxinas (2,4-D, AIA e AIB) e citocininas (BAP e KIN) na desdiferenciação celular indicado pela formação de calos. Os meios MS, B5 e White não foram significantemente diferentes e proporcionaram melhores resultados. Em meio MS contendo as fontes nitrogenadas 2,4 g L-1 NH4NO3 e 1,35 g L-1 glutamina + 4,11 g L-1 de KNO3 na presença de 1,0 mg L-1 2,4-D e 5,0 mg L-1 BAP ocorreu maior proliferação celular indicada pela massa fresca dos calos. As combinações dos reguladores 0,5 mg L-1 2,4-D + 5 mg L-1 BAP e 0,5 mg L-1 AIB + 5,0 mg L-1 BAP suplementado com 2,4 g L-1 NH4NO3 em meio MS estimularam o crescimento dos calos. De maneira geral, os calos apresentaram aspectos friáveis, não embriogênicos, com acúmulo de fenóis e presença de áreas meristemáticas (meristemóides).

The regulation of callus formation is the first step to indirect somatic embryogenesis in vitro. Therefore, we investigated hormonal and nutrient factors on development of callus in leaf tissues of Caesalpinia echinata (Brazilwood). Explants were cultivated in culture media MS, B5, WPM, and White. The effect of nitrogen sources (NH4NO3, KNO3 and glutamine) and the interaction between auxins (2,4-D, IAA and IBA) and cytokinins (BAP and KIN) were also evaluated. Media MS, B5 and White were not significantly different and provided better results. MS medium containing 2.4 g L-1 NH4NO3, 1.35 g L-1 glutamine, and 4.11 g L-1 of KNO3 in the presence of 1 mg.L-1 2,4-D and 5 mg.L-1 BAP increased cellular proliferation as determined by fresh weight of callus. The combination of plant growth regulators at 0.5 mg L-1 2,4-D + 5.0 mg L-1 BAP and 0.5 mg L-1 IBA + 5.0 mg L-1 BAP supplemented with 2.4 g L-1 NH4NO3 in MS medium stimulated growth of callus. In general, callus showed friable, non-embryogenic characteristics, with accumulation of phenolics and the presence of meristematic areas (meristems).