Abstract Vascular epiphytes contribute significantly to the biodiversity of tropical forests. We aimed to identify the epiphyte species occurring in the Muriqui Trail at Serra do Brigadeiro State Park (SBSP), a priority area for Atlantic forest conservation in southeastern Brazil. We selected 10 phorophytes, from which we sampled epiphytes and took light intensity measurements at three strata: up to 4 m, 4 to 8 m, and above 8 m. Phorophytes represented eight species, eight genera, and six families. We found 25 epiphytic species from 17 genera and nine families. Ferns showed highest richness, especially Polypodiaceae. We report herein important data on epiphyte diversity in eastern Minas Gerais state (Zona da Mata), including the occurrence of species rarely cited in surveys. Fomenting local conservation along with environmental education may be pivotal for counteracting anthropic pressure on SBSP. forests SBSP, SBSP , (SBSP) Brazil 1 phorophytes strata m families 2 richness Polypodiaceae Zona Mata, Mata Mata) surveys (SBSP

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.

ABSTRACT The objective of this study was to evaluate the effects of toxic levels of aluminum (Al) on the growth and external morphology of root tips in two maize genotypes with differential Al tolerance. The maize genotypes UFVM-100 (Al-sensitive) and UFVM-200 (Al-tolerant) were treated with 0 and 50 mM Al in a 0.5 mM CaCl2 solution at pH 4.5; root elongation, Al content and the external morphology of the root tips were evaluated. Chemical analysis, hematoxylin staining and energy dispersive X-ray spectrometry (EDS) showed greater Al accumulation in the root tips of the UFVM-200 genotype. The inhibition of root elongation in the UFVM-100 genotype, however, was much stronger than in the UFVM-200 genotype. Both maize genotypes exhibited visible and intense alterations in external micromorphology of the root tip, especially the Al-sensitive UFVM-100 genotype. Scanning electron micrographs showed intense cell disorganization and transverse ruptures of the protodermic and outer cortex layers of the cells in both genotypes. The ruptures were deeper and wider and reached the inner cortex layers in the UFVM-100 genotype. The EDS analysis showed that, in addition to Al accumulation, there was a proportional increase in the P concentration in the root tips of the UFVM-200 genotype. This is indicative of possible precipitation and/or immobilization of Al in the root tip apoplast of this genotype, which contributes to symplastic detoxification.

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.

The use of cell and plant tissues culture techniques to produce economically important active metabolites has been growing. Among these substances, azadirachtin (AZA), produced by the neem tree (Azadirachta indica), has received considerable attention due to its bioinsecticide action. The main goal of this work was to analyze the AZA levels in neem cotyledonary calli. The calli were grown in agitated Woody Plant Medium (WPM) liquid medium, supplemented with glucose (Gl), hydrolyzed casein (HC) and methyl jasmonate (MeJ) as elicitor agent. An interaction was observed between these substances, depending on in vitro cultivation time with orbital agitation. The highest concentrations (average of 0.2470 µg g-1) of AZA were produced in the first and second weeks of culture when the cell mass was grown in a medium with 2% Gl v/v, 500 mg L-1 HC and 100 µM of MeJ. This corresponded to approximately 57% of the AZA content stored in the donor plants seeds, used as a source of explants to induce in vitro callus formation. It was concluded that the nutrition, as well as the concentration of MeJ as signal transduction of secondary metabolism in neem cells, might influence the AZA content produced in vitro.

Experimentos foram conduzidos para avaliar o grau de susceptibilidade e determinar as injúrias causadas pela chuva ácida simulada na anatomia e micromorfologia foliar de Genipa americana. Plantas foram expostas à chuva com pH 3,0 durante 10 dias consecutivos. No tratamento controle utilizou-se apenas água destilada (pH 6,0). Amostras foliares foram coletadas e fixadas para microscopia de luz e eletrônica de varredura. Foram observados nas folhas expostas à chuva ácida: necroses pontuais intervenais, colapso das células do mesofilo e da epiderme; hipertrofia do parênquima lacunoso e acúmulo de compostos fenólicos e grãos de amido. A análise micromorfológica evidenciou, nas áreas necrosadas, plasmólise das células-guarda e ruptura da cutícula e da crista estomática. Alterações anatômicas ocorreram antes que sintomas visuais fossem observados nas folhas. Estes resultados comprovam a importância de dados anatômicos na diagnose precoce da injúria e na determinação da sensibilidade de G. americana à chuva ácida.

Experiments were conducted in order to characterize the injuries on leaf structure and micromorphology of G. americana and evaluate the degree of susceptibility of this species to simulated acid rain. Plants were exposed to acid rain (pH 3.0) for ten consecutive days. Control plants were submitted only to distilled water (pH 6.0). Leaf tissue was sampled and fixed for light and scanning electron microscopy. Necrotic interveinal spots on the leaf blade occurred. Epidermis and mesophyll cells collapse, hypertrophy of spongy parenchyma cells, accumulation of phenolic compounds and starch grains were observed in leaves exposed to acid rain. The micromorphological analysis showed, in necrotic areas, plasmolized guard cells and cuticle rupture. Epidermal and mesophyll cells alterations occurred before symptoms were visualized in the leaves. These results showed the importance of anatomical data for precocious diagnosis injury and to determine the sensitivity of G. americana to acid rain.

Os objetivos deste trabalho foram: caracterizar os efeitos do flúor em espécies arbóreas nativas, nos estádios de plântula e muda; identificar injúrias provocadas pelo flúor na estrutura da lâmina foliar da espécie mais sensível; fornecer subsídios para seleção de características a serem utilizadas na bioindicação e contribuir com informações sobre a resistência ou tolerância das plantas, visando ao reflorestamento de áreas impactadas pela chuva com flúor. As espécies analisadas foram Gallesia gorazema Moq. (Phytolaccaceae), Genipa americana L. (Rubiaceae), Joannesia princeps Vell. (Euphorbiaceae), Peltophorum dubium (Spreng.) Taub. (Leguminosae, Caesalpinioideae) e Spondias dulcis Forst. f. (Anacardiaceae). Plântulas e mudas dessas espécies, provenientes do Parque Estadual do Rio Doce (MG), foram submetidas a 20 min diários de chuva com flúor (30 mg.L-1), por 10 dias consecutivos. Necroses apicais e marginais foram observadas em todas as espécies analisadas, logo após a primeira chuva simulada. S. dulcis, no estádio de muda, foi a espécie mais sensível ao flúor, pois apresentou extensas necroses com apenas dois dias de tratamento, enquanto que G. americana foi a espécie mais resistente. Nas mudas, as espécies que acumularam mais flúor foram também as que apresentaram maior sensibilidade a esse poluente; essa relação não foi verificada nas plântulas. A concentração de flúor utilizada promoveu alterações drásticas na lâmina foliar de S. dulcis com extensas áreas necrosadas, danificando toda a sua estrutura anatômica. A sensibilidade ao flúor observada em S. dulcis indica que essa espécie apresenta potencial para ser usada como bioindicadora. Entretanto, estudos detalhados serão necessários para a melhor caracterização das respostas de S. dulcis ao flúor visando a sua utilização em programas de biomonitoramento ambiental.

The objective of this study was to characterize the effects of fluoride on seedlings and young plants of native woody species from Parque Estadual do Rio Doce (MG) and on the leaf blade structure of the most sensitive of the studied tree species. This work was carried out to gain information on diagnosing criteria for bioindication and to throw some light on the mechanisms of plant resistance and tolerance in order to assist in the revegetation programs of highly affected areas. The employed species were Gallesia gorazema Moq. (Phytolaccaceae), Genipa americana L. (Rubiaceae), Joannesia princeps Vell. (Euphorbiaceae), Peltophorum dubium (Spreng.) Taub. (Leguminosae, Caesalpinioideae) and Spondias dulcis Forst. f. (Anarcadiaceae). Seedlings and young plants of those species were submitted to rain with dissolved fluoride (30 mg.L-1), during 20 min daily, for 10 consecutive days. Apical and marginal necrosis occurred in the leaf blades of all species just after the first simulated rain. Two days after treatment, S. dulcis, in the young plant stage, showed to be the most sensitive species by exhibiting large necrotic areas, while G. americana was the most resistant species. At the young plant stage, the species that accumulated more fluoride were also the most sensitive ones; such a relation was not observed in plants at the seedling stage. The pollutant at the level employed caused drastic alterations in the leaf blade of S. dulcis and also damaged all of its anatomical structure. The high susceptibility of S. dulcis to fluoride suggests that this species possess potential features to be employed as a bioindicator. However, for a better characterization of the responses of S. dulcis to fluoride and its usage in biomonitoring programs, more studies are required.