Aniba Rosaeodora Ducke Research Articles

The Famous Amazonian Rosewood Essential Oil: Characterization and Adulteration Monitoring by Electrospray Ionization Mass Spectrometry Fingerprinting

Direct infusion of samples via electrospray ionization mass spectrometry (ESI-MS) is shown to characterize unequivocally genuine samples of Amazonian Aniba rosaeodora Ducke (Rosewood) essential oils obtained either from the wood or leafs. The ESI-MS also distinguishes the essential oils from synthetic linalool; hence, adulteration by the synthetic oil is also clearly detected. The analysis requires no pretreatment or preseparation, and the most polar components of the essential oil are extracted with an acidified 1:1 methanol/water solution. This simple extract is then analyzed by direct infusion ESI-MS in the positive ion mode, which provides characteristic fingerprintings of the sample composition. The ESI-MS fingerprinting can be used therefore as a simple and fast (few minutes) method for authenticity and quality control of this famous Amazonian essential oil.

Linalool from Rosewood (Aniba rosaeodora Ducke) Oil Inhibits Adenylate Cyclase in the Retina, Contributing to Understanding its Biological Activity

Rosewood oil (RO) (Aniba rosaeodora Ducke) is rich in linalool, a monoterpene alcohol, which has well studied anxiolytic, sedative and anticonvulsant effects. The inhibition of the increases in cAMP protects against seizures in a diversity of models of epilepsy. In this paper, the principal aim was to investigate the effects of RO, (±)-linalool and (-)-linalool) on adenylate cyclase. They were tested in chick retinas and forskolin was used to stimulate the enzyme target. The phosphodiesterase inhibitor, 4-(3-butoxy-4-methoxybenzyl)-imidazolidin-2-one, and the non-selective adenosine receptor antagonist 3-isobutyl-methyl-xanthine (IBMX), were used to control the participation of phosphodiesterase and adenosine receptors in the resulting effects, respectively. The cAMP accumulation was measured by enzyme immune assay (EIA). Rosewood oil, (-)-linalool and (±)-linalool inhibited exclusively the cAMP accumulation stimulated by forskolin, even when adenosine receptors were blocked with IBMX. The IC(50) values (in μ m concentration range) calculated from their concentration response-curves were not statistically different, however, the compounds presented a different relative efficacy. These results extend the range of subcellular mechanisms underlying the relaxant action of linalool on the central nervous system.

Efeito de diferentes reguladores de crescimento na regeneração in vitro de pau-rosa (Aniba rosaeodora Ducke)

O objetivo deste trabalho foi estabelecer um protocolo para a regeneração in vitro de pau-rosa (Aniba rosaeodora Ducke), utilizando brotações apicais e segmentos nodais inoculados em meio de cultura com distintas concentrações de diferentes reguladores de crescimento. Explantes esterilizados com soluções de benomyl (4,0 g.L-1) por 24 horas e hipoclorito de sódio a 20% + tween 20 por 20 minutos, foram submetidos a um experimento de indução de broto, raiz e calo em meio MS1 acrescido de 30g.L-1 de sacarose e 9g.L-1 de agar, suplementado com BAP (0,0 e 4,0 mg.L-1), ANA, AIA e 2,4-D (0,0; 3,0 e 6,0 mg.L-1), e suas respectivas combinações. O delineamento utilizado foi o inteiramente casualizado em esquema fatorial 7 X 2, com 14 tratamentos e 15 repetições cada, onde foram analisados o número médio de brotos, raízes e calo. Após 90 dias, os resultados mostraram que a presença de auxinas é fundamental para a formação dos parâmetros induzidos nos explantes de pau-rosa. O meio de cultura contendo 4,0 mg.L-1 de BAP + 6,0 mg.L-1 de AIA apresentou a melhor média para a brotação com 2,13 brotos/explante. Para o enraizamento o meio contendo 3,0 mg.L-1 de ANA foi o mais eficiente, apresentando uma média de 2,53 raízes/explante. Em relação à indução de calo, todos os tratamentos apresentaram calogênese, porém o meio suplementado com 4,0 mg.L-1 de BAP + 6,0 mg.L-1 de 2,4-D, apresentou a melhor média, 1,67 calos/explante.

Composição química de óleos essenciais de espécies de Aniba e Licaria e suas atividades antioxidante e antiagregante plaquetária

Leaves and stems from Aniba panurensis (Meisn.) Mez, Aniba rosaeodora Ducke and Licaria martiniana (Mez) Kosterm. were collected in the Reserva Florestal Adolpho Ducke-AM and their essential oils were obtained by hydrodistillation procedures. The oils were analyzed by GC-FID and GC-MS resulting on fifty and six compounds being identified. The major components were linalool in A. rosaeodora, and β-caryophyllene in A. panurensis and L. martiniana. At qualitative assays the oils showed antioxidant and antiplatelet activities, but only weak activities were found at quantitative spectrometric assays.

Chemical Variability of Rosewood (Aniba rosaeodora Ducke) Essential Oil in French Guiana

Eighty-two samples of Aniba rosaeodora Ducke were collected from ten localities in French Guiana. Essential oils were extracted from different parts of the tree (trunk wood, branch wood, roots, leaves) and analyzed. Yield measurements were performed by hydrodistillation; total linalool content and percentages of (-)-linalool or (+)-linalool were obtained by gas chromatography. Variations in essential oil yield were correlated with various parameters such as part of the tree, age of the tree, season, phenological status and geographic origin. Oil yields from wood samples (trunk, branch, shoot) ranged from 0.4–3.2%, while yields from roots and leaves varied very little with an average of 0.15% and 0.43%, respectively. In young trees, wood yield was 1.6%, higher than in older trees (1.3%). Collection time had no obvious influence on wood and leaf yields. Nevertheless, fruit-bearing trees gave a higher yield than fruitless ones. For a given tree, a decreasing yield gradient was observed from trunk wood to branches, smaller branches, and leaves. Linalool percentage in oil ranged from 73–99%. All trunk wood oils contained a percentage of (-)-linalool close to 100%, except for two trees from Paracou station where a low proportion of (+)-linalool was present. Oils from small branches contained from 5–28% of (+)-linalool. All leaf oils showed a high percentage (78–89%) of (+)-linalool. The linalool form in oil from shoot, bark and root samples was purely (-)-linalool. Measures performed on different parts of the same tree showed that the linalool from trunk wood and thick branches was purely (-)-linalool, whereas the proportion of (+)-linalool increased with the thinness of the branches and reached 85% in leaves.

Eficiência de Inseticidas no Controle de Pragas em Sementes e Mudas de Pau-rosa (Aniba rosaeodora Ducke), em Viveiros, Manaus, Amazonas

O pau-rosa (Aniba rosaeodora) é conhecido pela presença do álcool linalol, que fornece um odor forte e perfumado em todas as partes da planta. A sua exploração na Amazônia ao longo dos últimos anos, para utilização do linalol como fixador de perfumes, coloca o pau-rosa em risco de extinção, a sua frutificação irregular aumenta a dificuldade em se obter sementes para a produção de mudas e, além disso, parte de sua produção é perdida face ao ataque de pragas nas sementes. Em razão desses fatores o presente estudo teve como objetivo realizar a aplicação de inseticidas nas sementes e mudas em viveiro a fim de evitar ou minimizar o ataque de pragas, aumentando, assim, a produção de mudas. Utilizou-se um total de 300 sementes, distribuídas em 2 tratamentos e 1 testemunha. Cada tratamento (n = 100) foi constituído por 4 repetições (bandejas) com 25 sementes. O inseticida utilizado nos tratamentos foi acetamiprid e fipronil nas concentrações de 0,09 mg/planta e 0,18 mg/planta, respectivamente, para o tratamento 1 (T1) e o tratamento 2 (T2), distribuídos na forma de sachês. O T1 recebeu a aplicação de 1 sachê e o T2 a aplicação de dois sachês, enterrados com areia lavada nas sementeiras. Para avaliar os resultados foi feita uma análise de Variância (ANOVA), usando o programa Systat 9 e as médias comparadas pelo teste de Tukey em nível de 5%. As taxas de germinação foram: testemunha com 62%; T1 com 73% e T2 com 79%. Houve ataque de insetos em sementes apenas na testemunha com 23% do total (n = 100) e os resultados foram estatisticamente significativos (F = 31,263; p < 0,001). Houve diferenças significativas apenas entre a altura das plântulas nos tratamentos T1 e T2 e a testemunha (F = 15,090; p < 0,001). A principal praga identificada atacando as sementes foi Heilipus odoratus (Coleoptera: Curculionidae). Recomenda-se, portanto, a concentração de 0,09 mg de acetamiprid e fipronil distribuídos em sachês nas sementeiras e no transplantio para diminuir o ataque de pragas em sementes e mudas de pau-rosa em viveiros.

Genetic diversity in rosewood saplings (Aniba rosaeodora ducke, Lauraceae): an ecological approach

This article takes an ecological approach to the genetic diversity of Rosewood (Aniba rosaeodora Ducke) in a central Amazonian terra firme forest north of Manaus. Planted Rosewood setting, under partial shaded canopy, were assessed in terms of fruiting production, frugivory, and seed dispersal. Using RAPD molecular analysis procedures, the influence of the spatial distribution of adult trees on the genetic diversity (polymorphism) of saplings was assessed with genetic samples from 34 reproductive trees and 60 saplings. The density and distribution patterns the reproductive trees did not modify the sapling"s diversity (1.86%, AMOVA). Two types of adult tree dispersion were identified; i) clumped and ii) more widely dispersed. Polymorphism (77.5%) and gene flow were high between these. Although more sapling genetic variability in areas with a higher density of mature plants was not as high as expected, density did not affect the genetic diversity of samplings, indicating a high incidence of gene flow amongst trees. In planted Rosewood population (surrounded by low disturbed forest), fruiting trees experienced a high level of removal of seeds by toucans (Rhamphastidae), about of 50%. The high gene flow found among native trees suggested that toucans, promoting seed rain at short and long distances from maternal trees, actively contribute to the maintenance of genetic diversity within wild rosewood populations.

Primary metabolite mobilization during germination in rosewood (Aniba rosaeodora Ducke) seeds

This study aimed to characterize protein, oil, starch and soluble sugar mobilization as well as the activity of alpha-amylase during rosewood seed germination. Germination test was carried out at 25°C and the following parameters were analyzed: percentage of germination, initial, average, and final germination time. Seed reserve quantification was monitored in quiescent seeds and during different stages of radicle growth. Starch mobilization was studied in function of a-amylase activity. Germination reached 87.5% at the initial, average, and final time of 16, 21 and 30 days, respectively. Oil mobilization showed a negative linear behavior, decreasing 40% between the first and the last stage analyzed, whereas protein levels increased 34.7% during the initial period of germination. Starch content (46.4%) was the highest among those of the metabolites analyzed and starch mobilization occurred inversely to the observed for soluble sugars; alpha-amylase activity increased until the 15th day, a period before radicle emission and corresponding to the highest starch mobilization. The high percentage of rosewood seed germination may be related to the controlled condition used in the germination chamber as well as to high seed reserve mobilization, in special oil and starch.

Geographic pattern of genetic diversity in natural populations of Rosewood (Aniba rosaeodora), in the Central Amazonia

Rosewood (Aniba rosaeodora Ducke, Lauraceae) is an Amazonian evergreen tree and a source of the purest linalool, the main component of its essential oil, which is very valuable in the international perfumery market. After decades of over-exploitation it is currently considered as threatened. We evaluated the genetic diversity and its distribution in four populations in Central Amazonia. Thirty-five reliable RAPD markers were generated, of which 32 were polymorphic (91.4%). Variation was higher within the populations (76.5%; p < 0.0001) and geographic distribution contributed to population differentiation (23.4%; p < 0.0001). The Amazon River had a small influence on gene flow (3.3%; p < 0.0001), but we identified evidence of gene flow across the river. There were significant differences in marker frequencies (p < 0.05), in agreement with the low gene flow (Nm = 2.02). The correlation between genetic distance and gene flow was - 0.95 (p = 0.06) and between geographic distance and gene flow was -0.78 (p = 0.12). There was a geographic cline of variability across an East-West axis, influenced as well by the Amazon River, suggesting the river could be a barrier to gene flow. Although threatened, these Rosewood populations retain high diversity, with the highest levels in the Manaus population, which has been protected for over 42 years in a Reserve.

RAPD em Pau-rosa (Aniba rosaeodora Ducke): adaptação do método para coleta de amostras in situ, ajuste das condições de PCR e apresentação de um processo para selecionar bandas reprodutíveis

O Pau-rosa (Aniba rosaeodora Ducke) é uma espécie importante economicamente para a Região Amazônica, porque sua madeira é fonte de linalol, insumo utilizado pelas perfumarias. Esta espécie foi explorada durante décadas e, ainda assim, o conhecimento acerca da diversidade genética intra-específica é muito restrito. Foram objetivos deste trabalho: 1) validar um protocolo para coleta de folhas de pau-rosa que permitisse preservar a integridade do DNA até a estocagem em "freezer"; 2) selecionar um protocolo para extração de DNA em quantidade e qualidade adequadas para geração de bandas RAPD e 3) desenvolver um critério para avaliar o grau de reprodutibilidade que pudesse auxiliar a seleção de bandas RAPD úteis para análises de diversidade genética. Imediatamente após a coleta, as folhas foram acondicionadas em tubos de polietileno com sílica gel e aí permaneceram por até 10 dias. Foram testados três protocolos para a extração de ácidos nucléicos destas folhas, condições ideais para as PCR e a reprodutibilidade dos padrões RAPD. Critérios para a eliminação das bandas que mais contribuíram para o afastamento dos resultados do ideal da reprodutibilidade total foram desenvolvidos e a significância estatística das diferenças geradas pela aplicação dos critérios ao conjunto de dados foi testada. DNA com qualidade e em quantidade suficiente para a geração de padrões RAPD, nas condições ideais definidas para as PCRs, foi obtido. A eliminação de bandas com reprodutibilidade menor que 70% não diferiu do controle. A eliminação de bandas com reprodutibilidade menor que 90% diferiu dos demais tratamentos em todos os arranjos testados (P < 5%).

Biomass accumulation, photochemical efficiency of photosystem II, nutrient contents and nitrate reductase activity in young rosewood plants (Aniba rosaeodora Ducke) submitted to different NO3-:NH4+ ratios

The rosewood (Aniba rosaeodora Ducke) is a native tree species of Amazon rainforest growing naturally in acidic forest soils with reduced redox potential. However, this species can also been found growing in forest gaps containing oxide soils. Variations in the forms of mineral nitrogen (NO3- or NH4+) may be predicted in these different edaphic conditions. Considering that possibility, an experiment was carried out to analyze the effects of different NO3-:NH4+ ratios on the growth performance, mineral composition, chloroplastid pigment contents, photochemical efficiency photosystem II (PSII), and nitrate redutase activity (RN, E.C.1.6.6.1) on A. rosaeodora seedlings. Nine-month-old seedlings were grown in pots with a washed sand capacity of 7.5 kg and submitted to different NO3-:NH4+ ratios (T1 = 0:100%, T2 = 25:75%, T3 = 50:50%, T4 = 75:25%, and T5 = 100:0%). The lowest relative growth rate was observed when the NO3-:NH4+ ratio was equal to 0:100%. In general, high concentrations of NO3- rather than NH4+ favored a greater nutrient accumulation in different parts of the plant. For the chloroplastid pigment, the highest Chl a, Chl b, Chl tot, Chl a/b and Chl tot/Cx+c contents were found in the treatment with 75:25% of NO3-:NH4+, and for Chl b and Cx+c it was observed no difference. In addition, there was a higher photochemical efficiency of PSII (Fv/Fm) when high NO3- concentrations were used. A linear and positive response for the nitrate reductase activity was recorded when the nitrate content increased on the culture substrate. Our results suggest that A. rosaeodora seedlings have a better growth performance when the NO3- concentrations in the culture substrate were higher than the NH4+ concentrations.

Avaliação rebrota da copa das árvores de pau-rosa (Aniba rosaeodora Ducke) em sistema de podas sucessivas

O estudo foi conduzido em um plantio de pau-rosa com 36 anos, localizado na Reserva Florestal Adolpho Ducke, Manaus-AM, Brasil, latitude 03º00' 00 "e 03º08'00" S e longitude 59º52'40 "e 59º58'00" W. Foi determinada a biomassa da rebrota da copa das árvores de pau-rosa (Aniba rosaeodora Ducke, Lauraceae), podadas duas vezes (em 2000 e 2002) e das árvores podadas três vezes (em 1987, 2000 e 20002). Os resultados não indicaram diferenças significativas entre o DAP, altura total, número de brotos/fuste, comprimento e diâmetro dos brotos e peso verde da copa das árvores podadas duas ou três vezes. Os coeficientes de regressão e de correlação de Pearson indicaram que a biomassa da copa das árvores podadas pela terceira vez está fortemente correlacionada com a radiação PAR direta, PAR difusa, PAR total e o índice de área foliar. O elevado número de brotos/fuste revelou boa capacidade da rebrota da copa após sucessivas podas, fato que possibilita o manejo da biomassa das árvores através de podas.

Plant sources of amazon rosewood oil

The aim of this study is to reevaluate the plant sources of the Amazon rosewood oil which have been named Aniba rosaeodora Ducke and Aniba duckei Kosterm. There is some disagreement on the exact botanical status of these species. Some Lauraceae specialists analyzing available material from both species concluded that there is no basis for regarding them as different. Based on our results we are confirming that the chemical composition of both species is quite different from that previously reported. So we are suggesting to bring back the previous botanical rosewood status as proposed by Adolph Ducke.

Evaluation of Leaf-Derived Extracts as an Environmentally Sustainable Source of Essential Oils by Using Gas Chromatography−Mass Spectrometry and Enantioselective Gas Chromatography−Olfactometry

In consideration of the world's present environmental situation and the threat of species extinction, investigations concerning alternative sustainable sources of natural substances represent an extremely important issue. In this respect, the present research is focused on the analytical evaluation of Brazilian rosewood (Aniba rosaeodora Ducke) leaves, as an alternative source (with respect to wood) of rosewood essential oil and, as such, of natural linalool, which is extensively used in perfumery. Enantioselective-gas chromatography-olfactometry (Es-GC-O) was used as a tool for the simultaneous stereodifferentiation and olfactive evaluation of the volatile optically active components present in the analyzed samples. In addition to Es-GC-O analyses, direct olfactive analyses were also performed, enabling the evaluation of the global aroma exerted by each sample and the influence of each linalool antipode, as also other minor compounds. The samples were also submitted to gas chromatography-mass spectrometric analysis, thus establishing their chemical profiles. The assessment of enantiopure chiral compounds through Es-GC-O, along with direct olfactive analyses, confirmed that the leaves are a potential substituent for wood in the extraction of Brazilian rosewood essential oil, representing a sustainable nonwood source of natural linalool.

Nuclear DNA content and karyotype of Rosewood (Aniba rosaeodora)

Rosewood (Aniba rosaeodora Ducke, Lauraceae) is ecologically and economically important to the Amazon region. As a consequence of its economic importance, rosewood populations have been decimated in the Amazon forest. Species of nine genera of the Lauraceae family have characterized karyotypes with n = x = 12 chromosomes in the gametophytic phase but the genus Aniba is one of the least studied Lauraceae genera with a previously undescribed genome. We used cytogenetic techniques to determine that the A. rosaeodora karyotype contained 12 pairs (2n = 24) of relatively small submetacentric chromosomes with lengths ranging from 1.34 to 2.25 mum and a nucleolar organizer region (NOR) in the short arm of chromosome 7. Flow cytometry gave 2C = 2.32 pg of DNA, equivalent to approximately 2.24 x 10(9) base pairs.

Growth, photosynthesis and stress indicators in young rosewood plants (Aniba rosaeodora Ducke) under different light intensities

Aniba rosaeodora is an Amazonian tree species that belongs to the family Lauraceae. Due to intense exploitation for extraction of essential oils (mainly linalol), A. rosaeodora is now considered an endangered species. On the other hand, there is little information about its ecophysiology which would be useful to support future forest planting programs. Hence, the effect of different light intensities on the growth and photosynthetic characteristics of young plants of A. rosaeodora was studied. Nine-month-old plants were subjected to four light treatments (T1= 10 a 250 µmol.m-2.s-1 / control; T2=500 to 800, T3=700 to 1000 and T4=1300 to 1800 µmol.m-2.s-1 / full sunlight). Allometric variables, gas exchange, contents of pigments and chlorophyll a fluorescence were analysed. As to the relative growth rates, it was found that plants of A. rosaeodora showed higher biomass accumulation when grown under intermediary irradiance conditions (T2). The best photosynthetic performance was achieved under conditions of T3. When growth was correlated with photosynthesis, it was found that plants under treatments T2 and T3 presented better responses in comparison with the lowest (T1) and highest (T4) light extremes. The highest pigment contents were obtained for plants in the shade (T1) and the lowest for those exposed to full sunlight (T4). The photochemical efficiency of photosystem II (Fv/Fm) was found that only plants in the shade treatment (T1) presented no stress from high irradiance. These findings suggest that both treatments (T1 and T4) altered the function of the A. rosaeodora plants, inhibiting photosynthesis and growth. Plants of A. rosaeodora developed photo-protection mechanisms under full sunlight. However, the species presented better photosynthetic response and biomass gain under intermediary irradiance conditions, displaying relative physiological plasticity, during the seedling phase.

A new spermophagous species of Heilipus Germar from the Amazonian Region (Coleoptera, Curculionidae, Molytinae)

Heilipus odoratus sp. nov. (type locality: Brazil, Amazonas, Manaus) is described based on adults reared from seeds of Aniba rosaeodora Ducke (Lauraceae), popularly called "pau-rosa". The species is spermophagous and develops from egg to adult inside the seeds. The new taxon is compared with the similar Heilipus draco (Fabricius, 1801). Illustrations of larva, pupa, adult and of important structures for species identification are given, including the male and female genitalia, figured for the first time for a species of Heilipus.

Cultura in vitro de embriões e de gemas de mudas de pau-rosa (Aniba rosaeodora Ducke)

Este trabalho teve como objetivo o estabelecimento in vitro de embriões e de gemas de mudas de pau-rosa (Aniba rosaeodora Ducke) livres de contaminações e de oxidação fenólica. As gemas foram obtidas da rebrota de mudas cultivadas em viveiro e os embriões a partir de sementes em diversos estágios de maturação. Para a assepsia dos explantes foram utilizados dois antibiótico (Ampicilina e Agrimicina), etanol (70%) e hipoclorito de sódio, em concentrações e tempo de exposição variando em função do tratamento. Para o controle da oxidação foram utilizados imersão em ácido ascórbico (250 mg/l) e PVP (Polivinilpirrolidona) no meio Murashige & Skoog (MS). O delineamento estatístico empregado foi o inteiramente ao acaso com tratamentos e repetições em função do tipo de explante. Foi observado 71% de sobrevivência e 53% de germinação de embriões tratados com hipoclorito de sódio (50% e 2% de cloro ativo) por 10 minutos e inoculados em meio MS contendo 20 mg/l de água de côco após 45 dias. As gemas das rebrotas de mudas tratadas com solução de Sulfato de Estreptomicina (Agrimicina) na concentração de 500 mg/l (1h) apresentaram 51% de sobrevivência. Quando submetidas ao pré-tratamento com o emprego de bomba a vácuo (180 mmHg) contendo a Agrimicina (500 mg/l), apresentaram 25% de sobrevivência.

Rosewood Exploitation in the Brazilian Amazon: Options for Sustainable Production

The authors report on ongoing work in the Brazilian Amazon to assess the current and prospective management of rosewood (Aniba rosaeodora Ducke) populations threatened by a half-century of predatory extraction for the valuable essential oil linalool used widely in perfumery. The report synthesizes prior research on rosewood exploitation and markets and recent research to develop new essential oil products derived from rosewood leaves and stems. The study suggests alternative rosewood production systems, to guide investment in management and certification of sustainable rosewood oil supplies.

Influência da cobertura e do sombreamento do canteiro na germinação e desenvolvimento inicial de Plântulas de Pau Rosa (Aniba rosaeodora)

Um experimento foi instalado no viveiro da Reserva Florestal Adolfo Ducke, Manaus, Amazonas, Brasil, com o objetivo de comparar três tipos de cobertura de canteiros empregados em viveiros florestais e testar quatro níveis de sombreamento na germinação, emergência e desenvolvimento de plântulas de pau rosa (Aniba rosaeodora Ducke). Na cobertura dos canteiros utilizou-se palha de arroz, serragem e vermiculita. Os níveis de sombreamento de 30, 50 e 70% foram obtidos por meio de telas de poliolefinas de cor preta, sendo o de 0% obtido por semeadura a pleno sol. O delineamento estatístico foi de parcelas subdivididas em blocos ao acaso. Os tipos de cobertura utilizados e a interação entre os níveis de sombreamento e tipo de cobertura de canteiros não influenciaram na germinação das sementes e no índice de velocidade de emergência. Os níveis de sombreamento não influenciaram na germinação das sementes, mas apresentaram efeito significativo no índice de velocidade de emergência. Os tipos de cobertura dos canteiros não influenciaram no desenvolvimento das plântulas em altura, diâmetro à altura do colo e peso da matéria seca da parte aérea, do sistema radicular e total; a área foliar das plântulas obtidas com a cobertura de vermiculita foi melhor que com serragem. Melhores resultados de crescimento em altura e peso da matéria seca da parte aérea, do sistema radicular e total, foram observados nas plântulas cultivadas sob os níveis de 30 e 50% de sombreamento. As interações de 30% de sombreamento com a cobertura de vermiculita c 50% de sombreamento com a cobertura de palha de arroz proporcionaram maiores valores de crescimento em altura e peso da matéria seca total das plântulas de pau rosa, respectivamente.