Abstract Cryopreservation of pineapple shoot tips has been established from various protocols, including droplet vitrification. Thus, this work aimed to evaluate the morphoanatomical conditions of the starting material over different times (30, 45 and 60 days) of culture before freezing and its correlation with the survival percentage of the cryopreserved shoot tips. Four accessions, Ananas comosus var. comosus (BGA-009); var. bracteatus (BGA-119); var. parguazensis (BGA-376), var. erectifolius (BGA-750) from the Pineapple Active Germplasm Bank (BGA Pineapple) and two hybrids from the Embrapa Genetic Breeding Program, FIB-ROX1 (var. bracteatus X var. erectifolius) and FIB-ROX2 ( var. erectifolius X var. bracteatus), recently introduced in the field from in vitro storage, were used. Histological sections before freezing and the percentages of survival after freezing were obtained taking into account the different times of cultivation of the donor plants. The results showed a significative interaction between genotypes (accessions and hybrids) and the culture period. The accessions BGA-009 and BGA-119 showed the highest survival rates, with 95% and 90% respectively for the 30-day culture time. Different results were obtained for each genotype, showing the need for improvements in the standardization of starting material, which would allow better repeatability of the protocol.

Abstract The objective of this study was to evaluate the stigma morphoanatomy and receptivity in ten promising papaya accessions, to expand knowledge useful for genetic improvement of the culture. The morphoanatomy was analyzed by light and scanning electron microscopy, and the stigma receptivity was investigated by application of hydrogen peroxide and α-naphthyl-acetate with fast blue B salt, in pre-anthesis, anthesis and post-anthesis. The papaya accessions presented dry stigma surface, presence of one to six erect stigmatic lobes, with structures joined to the upper base of the style and covered with numerous elongated unicellular tubular papillae, distributed on both faces of the epidermis. The morphoanatomy had a similar pattern in all the accessions, differing only in the timing of floral development. The stigma receptivity in some accessions occurred even before floral opening, continuing with greater intensity in anthesis and post-anthesis. Pre-anthesis is the stage least propitious for controlled hybridizations due to the weak or absent stigmatic receptivity. The results obtained provide information on the floral stage that is most propitious for fertilization, as well as supporting future investigations of the botanical morphology of the species.

Abstract Vriesea bahiana, Hohenbergia castellanosii and Encholirium spectabile are endemic Brazilian species that are considered endemic or endangered. Development of strategies to conserve these species is important to prevent irreversible genetic erosion. The objective of this study was to evaluate the post-seminal development and seed cryopreservation of three endemic or in danger of extinction bromeliad species in Brazil, to obtain a protocol that can safeguard the genetic variability of these species. In the seed cryopreservation assay, we evaluated five desiccation periods. The seeds in the cryotubes were taken from the desiccator and immediately plunged into liquid nitrogen. For the analysis of post-seminal development, seeds in different germination stages were collected and evaluated by light and scanning electron microscopy. Vriesea bahiana seeds frozen in liquid nitrogen presented almost 100% germination, indicating dormancy break of this species. Vriesea bahiana can be cryopreserved with 5.9% water content after being dried for 24 hours. Hohenbergia castellanosii and E. spectabile seeds did not need to be desiccated before being cryopreserved. The most relevant morphological traits for differentiation of genera and subfamilies of Bromeliaceae are the shape and type of seed appendages. In this study, all three species presented well-differentiated size and shape of their structures.

RESUMO Flores que apresentam androceu com estames de alturas diferentes (heterodínamos) podem também apresentar características distintas entre os grãos de pólen em cada tamanho de estame. As espécies de Aeschynomene L. apresentam estames didínamos (cinco longos e cinco curtos), mas até o momento pouco se sabe sobre o dimorfismo polínico. Diante da carência de estudos palinológicos do gênero, o objetivo do presente trabalho foi de incrementar o conhecimento sobre as características polínicas do gênero Aeschynomene L., enfatizando as possíveis diferenças morfológicas entre os grãos de pólen dos grupos de estames longos e curtos, contribuindo com os estudos ecológicos e taxonômicos. Os grãos de pólen dos dois grupos de tamanho de estames de 10 espécies foram analisados separadamente utilizando-se a metodologia padrão em morfologia polínica, além de análises de variância e de comparação da média e índice de base. Os resultados inéditos mostraram que as características de âmbito, forma, tipo de endoabertura e ornamentação da sexina não variaram entre os grãos de pólen dos estames longos e curtos num mesmo espécime, mas sim entre as espécies. No entanto, com relação ao tamanho dos grãos de pólen dos dois grupos de estames, quatro espécies apresentaram diferenças significativas quanto ao tamanho dos eixos polar e equatorial. Os dados morfopolínicos adquiridos com relação ao heteromorfismo dos estames podem contribuir para o conhecimento da dinâmica reprodutiva do gênero e para os estudos sistemáticos.

ABSTRACT Flowers with heterodynamous stamens can present differences in the pollen grains of each stamen size group. Species of Aeschynomene L. present didynamous stamens (five long and five short) but little is known about their pollen dimorphism. The objective of this study was to increase the knowledge about the pollen characteristics in Aeschynomene and emphasize the possible morphological differences between the pollen grains of long and short stamens in order to contribute to ecological and taxonomic studies. Pollen grains from the two groups of stamens size of ten species were analyzed separately, according to the standard methodology for studies of pollen morphology. In addition, analyses of variance, comparison of means and base index were performed. The results showed that the amb, shape, endoaperture type and sexine ornamentation did not vary in the pollen grains of the long and short stamens in the same specimen, but they varied among the species. However, in relation to the size of the pollen grains of the two groups of stamens, four species presented significant differences regarding the size of the polar and equatorial axes. The pollen morphological data obtained considering the heteromorphism of the stamens can contribute to the knowledge of the repro ductive dynamics of the genus, and to the systematic studies.

Abstract Stevia rebaudiana belongs to the Asteraceae family with high economic and medicinal potential. This article describes and illustrates morphological and histological aspects of leaves and reproductive organs, and the germination process, to provide detailed information on this species and to contribute to taxonomic, phylogenetic and pharmacobotanical projects. The fruit is a cypsela, small, simple, dry, indehiscent, monospermic, light or dark colored, with aristate pappus, and the seed presents a spatulate axile embryo. Germination is phaneroepigeal with a pivotal root system and many absorbing root hairs. The leaves are simple, elliptical to obovate, with two types of trichomes (glandular and tector), with a short petiole, exhibiting an opposite decussate phyllotaxy. Our results showed 37.5% germination after 12 days, only in the dark cypsela, the light colored being considered unviable. The inflorescence is paniculate and the florets are grouped in capitula with isomorphic ends, monoclinous (bisexual), dichlamydeous, heterochlamydeous, pentamerous calyx and corolla, gamossepalous and gamopetalous. The androecium is gamostemone comprised of five stamens with free filaments, isodynamous and epipetalous stamens, synandrous and rimose anthers. The flower presents an inferior ovary, bicarpelar, unilocular and ovules with a basal placentation. The pollen grains are small, isopolar, radial symmetry, tricolporate, with echinate ornamentation.

ABSTRACT The genus Passiflora is the most diversified of the Passifloraceae, and its palynology presents wide morphological variability. The objective of the study was to evaluate the pollen morphology of 18 Passiflora species in order to identify informative pollinic characteristics to contribute to the taxonomic classification of the genus. The morphology of the pollen grains and the exine structure were investigated using light microscopy and scanning electron microscopy. Differences in the pollen morphology were found in the studied species, mainly in terms of shape, pollen aperture and ornamentation pattern of the exine. Most of the species belonging to the subgenus Passiflora presented ornamented 6-syncolpate pollen grains with an oblate-spheroidal shape and an exine with large lumens. In the subgenus Decaloba the pollen grains were 6-colporate, 12-colpate and 12-colporate, with subprolate to prolate-spheroidal shape, as well as an exine with smaller lumen size and few ornamented. The pollen morphology of the species of the subgenera Passiflora and Decaloba has shown palynological characteristics that have specific diagnostic value, thus allowing a better understanding of the taxonomy of the genus Passiflora.

ABSTRACT Pollen morphology characterization is important in taxonomy, conservation and plant breeding, and pollen viability studies can support breeding programs. This study investigated pollen morphology and male fertility in 18 species of Bromeliaceae with ornamental potential. For morphological characterization, pollen grains were acetolyzed and characterization of exine was done using scanning and transmission electron microscopy. Pollen viability was investigated by in vitro germination and histochemical tests. Species belonging to Aechmea and Ananas genera presented medium size pollen, except for Ae. fasciata, with large pollen. Al. nahoumii, P. sagenarius and the Vriesea species analyzed showed large pollen, except for V. carinata, with very large pollen. Pollen of Aechmea, Ananas and P. sagenarius presented bilateral symmetry, diporate, exine varying from tectate to semitectate. Al. nahoumii and Vriesea species presented pollen with bilateral symmetry, monocolpate; exine was semitectate, reticulate and heterobrochate. Germination percentage and tube growth were greater in SM and BKM media. Histochemical tests showed pollen viability above 70% for all species, except for Ananas sp. (40%). Pollen morphology is important for the identification of species, especially in this family, which contains a large number of species. High rates of viability favor fertilization and seed production, essential for efficient hybrid production and conservation.

A maioria das doenças dos animais de natureza e cativeiro encontra-se associada à proximidade humana, que resulta da fragmentação e degradação do habitat destes animais, no isolamento das espécies e no contato mais próximo entre estes e animais domésticos e o homem. Foram estudados os sincrânios de 104 mãos-peladas (Procyon cancrivorus) por meio de avaliação direta, preenchimento de ficha odontológica veterinária e documentação fotográfica, que relataram anormalidades encontradas, as quais foram classificadas e contabilizadas para fins estatísticos. Os achados deste trabalho aludem que os animais de cativeiro foram mais acometidos com as lesões relacionadas à doença periodontal, como cálculo, reabsorção óssea alveolar, deiscência, fenestração, exposição de furca, além de maloclusão, apinhamento dentário e os níveis mais graves de desgaste dentário. Os animais de vida livre apresentaram mais altos índices de fraturas, perdas dentárias ante-morte e escurecimento dentário, que caracterizam maior trauma dentário, durante o processo alimentar. Pretendeu-se, assim, estabelecer um parâmetro do estado de saúde oral da espécie estudada, sua frequência e se esta apresenta as mesmas enfermidades orais em vida livre e em cativeiro, relacionando a prevalência de afecções orais com características da ecologia da espécie.

Most diseases of animals in nature and captivity is linked to human proximity, resulting from habitat fragmentation and degradation of these animal habitats, the isolation of the species and the close contact between them and domestic animals and manhood. We studied 104 crab-eating raccoons (Procyon cancrivorus) skulls by direct assessment of sheet filling veterinary dental and photographic documentation reporting these abnormalities, which were classified and recorded for statistical purposes. The findings show that the captive animals were most affected with injuries related to periodontal disease, such as calculus, alveolar bone resorption, dehiscence, fenestration, furcation exposure, as well as malocclusion, crowding and severe levels of tooth wear. The free-living animals showed the highest rates of fractures, ante-mortem tooth loss and tooth blackout, which feature larger dental injuries during the feeding process. The intention was therefore to establish a parameter of oral health status of the species studied, their frequency and whether it has the same oral diseases in the wild life and in captivity, relating the prevalence of oral diseases with ecological characteristics of the specie.

Neste trabalho, analisamos as respostas bioquímicas inerentes ao sistema antioxidante, assim como propriedades morfológicas e anatômicas de mutantes fotomorfogenéticos e hormonais de tomateiro. Comparados ao não mutante Micro-Tom (MT), observamos que o conteúdo de malondialdeído (MDA) aumentou nos mutantes diageotropica (dgt) e lutescent (l), enquanto os maiores níveis de H2O2 foram encontrados nos mutantes high pigment 1 (hp1) e aurea (au). Análises de enzimas antioxidantes mostraram que todos os mutantes reduziram a atividade de catalase (CAT) quando comparado a MT. A guaiacol peroxidase (GPOX) aumentou em ambos os mutantes sitiens (sit) e notabilis (not), enquanto no mutante not houve aumento da atividade de ascorbato peroxidase (APX). Baseado nas análises de PAGE, as isoformas III, IV, V e VI de glutationa redutase (GR) aumentaram nas folhas do mutante l, enquanto a isoforma III de superóxido dismutase (SOD) reduziu nas folhas dos mutantes sit, epi, Never ripe (Nr) e green flesh (gf). Análises por microscopia revelaram que os mutantes hp1 e au tiveram um aumento nos espaços intercelulares de folhas, enquanto o mutante sit mostrou um decréscimo. Micrografias eletrônicas revelaram que a superfície foliar dos mutantes au e hp1 apresenta reduzida distribuição de tricomas. A caracterização destes mutantes é essencial para que possam ser utilizados em estudos de desenvolvimento e ecofisiologia, tais como estresses abióticos e bióticos no metabolismo oxidativo.

Biochemical responses inherent to antioxidant systems as well morphological and anatomical properties of photomorphogenic, hormonal and developmental tomato mutants were investigated. Compared to the non-mutant Micro-Tom (MT), we observed that the malondialdehyde (MDA) content was enhanced in the diageotropica (dgt) and lutescent (l) mutants, whilst the highest levels of hydrogen peroxide (H2O2) were observed in high pigment 1 (hp1) and aurea (au) mutants. The analyses of antioxidant enzymes revealed that all mutants exhibited reduced catalase (CAT) activity when compared to MT. Guaiacol peroxidase (GPOX) was enhanced in both sitiens (sit) and notabilis (not) mutants, whereas in not mutant there was an increase in ascorbate peroxidase (APX). Based on PAGE analysis, the activities of glutathione reductase (GR) isoforms III, IV, V and VI were increased in l leaves, while the activity of superoxide dismutase (SOD) isoform III was reduced in leaves of sit, epi, Never ripe (Nr) and green flesh (gf) mutants. Microscopic analyses revealed that hp1 and au showed an increase in leaf intercellular spaces, whereas sit exhibited a decrease. The au and hp1 mutants also exhibited a decreased in the number of leaf trichomes. The characterization of these mutants is essential for their future use in plant development and ecophysiology studies, such as abiotic and biotic stresses on the oxidative metabolism.

Linepithema humile Mayr is an ant species originally native to South America that has been spread accidently throughout the globe through international trade. It is a serious urban and crop pest. Despite its economic importance, little is known about the larvae of this species apart from a brief description based on a few specimens. The present investigation is aimed at describing every immature stage of L. humile. Three larval instars were determined through the frequency distribution of the maximum width of head capsules from a sample of 525 larvae. The morphological descriptions were based on 150 eggs, 70 larvae, and 90 pupae examined by light and scanning electron microscopy. Some morphological characteristics reported to be typical of Linepithema Mayr larvae were confirmed - dolichoderoid body shape, presence of dorsal protuberance, sparse simple body hairs, presence of nine pairs of spiracles and dolichoderoid mandibles. We concluded that an earlier published description was based on queen larvae, and that the protuberance is only present in worker larvae. The information provided in this study may aid ant systematics and phylogenetics, as well provide a better understanding of the biology of this species.

INTRODUÇÃO: A ampliação da via de saída do ventrículo direito (VSVD), com ou sem condutos, é frequente no tratamento de doenças cardíacas congênitas. Entretanto, a durabilidade desses condutos é restrita e o implante de stent se torna uma boa opção para desobstrução da VSVD. O objetivo deste estudo é relatar a experiência de um centro terciário no implante de stent não-valvado e seus resultados a curto e médio prazos. MÉTODO: Entre setembro de 2000 e janeiro de 2009, 11 pacientes com indicação de reintervenção cirúrgica receberam stents para desobstrução da VSVD. A média de idade no momento do procedimento foi de 12,5 ± 8,3 anos (1-28 anos) e o peso médio foi de 35 ± 20,9 kg (6-62 kg). A média de seguimento do estudo foi de 12,5 ± 6 meses (0-48 meses). RESULTADOS: A média da pressão sistólica no ventrículo direito diminuiu de 99 ± 23 mmHg (pré-procedimento) para 57 ± 12 mmHg após o implante do stent (p < 0,001), assim como, ao ecocardiograma transtorácico, o gradiente ventrículo direito-artéria pulmonar (VD-AP) caiu de 69 ± 19 mmHg para 33 ± 20 mmHg (p < 0,01). Em 40% dos casos houve necessidade de reintervenção, em média, 22 meses após o procedimento. Houve um caso de fratura de stent, um caso de insucesso e um óbito não relacionado ao procedimento. CONCLUSÃO: A angioplastia da VSVD com stent não-valvado é uma medida paliativa, altamente efetiva e segura, capaz de retardar a necessidade de reintervenção cirúrgica. Além disso, permite o crescimento somático dos pacientes jovens e a redução do dano ao ventrículo direito para futuro implante de stents valvados para tratamento da insuficiência pulmonar livre.

BACKGROUND: Right ventricle outflow tract (RVOT) reconstruction is frequently used to repair cardiac malformations. However, the lifetime of these conduits is short and stent implantation is a good option to relieve obstructed RVOT. The objective of this study is to report the experience of a tertiary center with non-valved stent implantation and its short and medium term results. METHODS: Between September 1990 and January 2009, 11 late postoperative patients were submitted to stent implantation due to RVOT obstruction. Mean age at the time of procedure was 12.5 ± 8.3 years (1-28 years), and mean weight was 35 ± 20.9 kg (6-62 kg). Mean follow-up time was 12.5 ± 6 months (0-48 months). RESULTS: Mean systolic right ventricular pressures decreased from 99 ± 23 mmHg before to 57 ± 12 mmHg (p < 0.001) after stent implantation. At echocardiogram, RV to PA gradient decreased from 69 ± 19 mmHg to 33 ± 20 mmHg (p < 0.01). Forty percent of the patients required percutaneous or surgical reintervention during a mean time of 22 months after the procedure. There was one case of stent fracture, relieved with the implantation of another stent, one case of failure, and one death not related to the procedure. CONCLUSION: Non-valved stent implantation in obstructed RVOT is a highly effective and safe palliative approach. Moreover, it allows the somatic growth of young patients and does not preclude future percutaneous implantation of valved stents to treat free pulmonary regurgitation.

Folhas, frutos e sementes de melão (Cucumis melo) Amarelo foram inoculados com Acidovorax avenae subsp. citrulli (Aac1) com o objetivo de estudar a colonização e penetração desse patógeno, utilizando microscopia eletrônica de varredura. Nas folhas coletadas 24, 48, 72, 96 e 120 h após a inoculação foram observadas células bacterianas agregadas e interligadas por fibrilas e muito raramente isoladas. Células bacterianas foram localizadas predominantemente nos flanges cuticulares da epiderme abaxial, na base dos tricomas tectores, próximas aos estômatos diacíticos e no interior dos poros estomáticos. Nos frutos com 37 dias de idade, 24 h após a inoculação, bactérias foram observadas na superfície e concentradas ao redor de estômatos e lenticelas, e nas amostras com 48 h, na polpa do fruto. Nas demais amostras (72 e 96 h) foram encontradas poucas ou até mesmo nenhuma bactéria na superfície dos frutos. Nos frutos com 51 dias de idade foi constatada uma expressiva colonização da superfície, estômatos e lenticelas até 72 h após a inoculação. Na polpa, a bactéria só foi encontrada nas amostras com 96 h. Os resultados sugerem que A. avenae subsp. citrulli penetrou nos frutos através de estômatos e lenticelas. Nas sementes a bactéria colonizou tegumentos interno e externo, embrião e endosperma.

Leaves, fruits and seeds of melon (Cucumis melo) type Yellow were inoculated with Acidovorax avenae subsp. citrulli (Aac1) in order to study the penetration and colonization of this pathogen using scanning electron microscopy. Leaves sampled at 24, 48, 72, 96 and 120 h after inoculation showed clusters of bacterial cells interconnected by fibrils, and very rarely isolated. Bacterial cells were located mainly on the cuticular flange of the abaxial epidermis, on the base of tector trichomes, near the diacytic stomata, and inside the stomat pores. Twenty-four hours after inoculation bacteria were observed mainly on the surface of 37 day-old fruits concentrated around stomata and lenticels; after 48 h bacteria were detected in the fruit pulp as well. In the other samples (72 and 96 h) few or no bacteria were found on fruit surfaces. Fruits 51 days old showed expressive colonization of the surface, stomata and lenticels until 72 h after inoculation. The pathogen was only found in the pulp of the 96-hour samples. These results suggest that A. avenae subsp. citrulli penetrated the fruits through the stomata and lenticels. In seeds, the bacteria colonized the internal and external teguments, embryo and endosperm.

Este trabalho, objetivou avaliar o efeito da aplicação de silício em plantas de milho no desenvolvimento da lagarta-do-cartucho Spodoptera frugiperda (J.E. Smith). O tratamento com silício consistiu na aplicação de 3,2 ml de solução de silicato de sódio [25-28% (p/v) de SiO2], mais 96,8 ml de água, via solo aos cinco dias, 10, 15, 20 e 25 dias após a emergência de plantas de milho mantidas em vasos. Outro grupo de plantas foi cultivado sem aplicação de silício. Avaliou-se o desenvolvimento de lagartas alimentadas com folhas de milho retiradas de plantas tratadas com silício, em comparação com lagartas alimentadas com folhas de milho não tratadas, registrando-se a mortalidade ao final do 2º ínstar larval, a duração do período larval e pupal, a mortalidade larval e pupal e o peso de pupa. Não se observou efeito do silício na duração da fase larval e pupal, peso de pupa e na mortalidade de pupas. Entretanto, verificou-se maior mortalidade e aumento de canibalismo em grupos de lagartas ao final do 2º ínstar, e maior mortalidade de lagartas individualizadas no 2º e 6º ínstares, quando foram alimentadas com folhas de plantas de milho tratadas com silício, em comparação com a testemunha. Observou-se que as mandíbulas das lagartas, nos seis ínstares, apresentaram desgaste acentuado na região incisora quando em contato com folhas com maior teor de silício. A aplicação de silício pode dificultar a alimentação de lagartas, causando aumento de mortalidade e canibalismo e, portanto, tornando as plantas de milho mais resistentes à lagarta-do-cartucho.

The effect of the application of silicon to corn plants on the development of the fall armyworm Spodoptera frugiperda (J.E. Smith) was evaluated. Silicon treatment consisted in applying 3.2 ml of a sodium silicate solution [25-28% (w/v) SiO2] plus 96.8 ml of water into the soil at 5, 10, 15, 20 and 25 days after emergence of corn plants kept in pots. Another group of plants were grown free of silicon. Larvae were fed on leaves obtained from silicon treated plants and their mortality at the end of the 2nd larval instar, length of the larval and pupal period, and pupal weight were compared to those of larvae fed on normal leaves. No effect of silicon was observed on the length of larval and pupal phase, pupal weight and mortality of pupae. However, increased mortality and cannibalism were registered in groups of fall armyworm at the end of the 2nd instar and greater mortality of the larvae was seen at the 2nd and 6th instars fed on silicon-treated corn plant leaves in comparison with the control. The jaws mandibles of fall armyworm at the 6th instar showed marked wear on the incisor region when in contact with leaves with higher silicon content. Silicon application can make fall armyworm feeding difficult, causing higher mortality and cannibalism, thus increasing plant resistance to this species.