Taxonomy and cladistic analysis of the subgenus Xenomorellia Malloch (Diptera: Muscidae: Morellia Robineau-Desvoidy) with description of two new species

Xenomorellia Malloch, a subgenus of Morellia Robineau-Desvoidy, is revised to include two new species, Morellia (Xenomorellia) inca Nihei and Carvalho sp. nov. from South America, and M. (X.) maia Carvalho and Nihei sp. nov. from Costa Rica and Mexico. Diagnoses for M. (X.) holti (Malloch) and M. (X.) montanhesa (Albuquerque) are provided, as well as an identification key to the four species of the subgenus. A cladistic analysis was performed to test the monophyly of Xenomorellia and to recover the phylogenetic relationships among its species. Tree searches resulted in one single most-parsimonious cladogram, wherein the monophyly of Xenomorellia is supported, as well as a sister-group relationship with the Neotropical subgenus Trichomorellia Stein. Xenomorellia was divided into two clades: one with Caribbean-Andean species (maia + inca), and another with species from southeastern South America (holti + montanhesa).

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare is the causative agent of columnaris disease in freshwater fish, implicated in skin and gill disease, often causing high mortality. The aim of this study was the isolation and characterization of Flavobacterium columnare in tropical fish in Brazil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) and cascudo (Hypostomus plecostomus) were examined for external lesions showing signs of colunmaris disease such as greyish white spots, especially on the head, dorsal part and caudal fin of the fish. The sampling comprised 50 samples representing four different fish species selected for study. Samples for culture were obtained by skin and kidney scrapes with a sterile cotton swabs of columnaris disease fish and streaked onto Carlson and Pacha (1968) artificial culture medium (broth and solid) which were used for isolation. The strains in the liquid medium were Gram negative, long, filamentous, exhibited flexing movements (gliding motility), contained a large number of long slender bacteria and gathered into ‘columns'. Strains on the agar produced yellow-pale colonies, rather small, flat that had rhizoid edges. A total of four Flavobacterium columnare were isolated: 01 Brycon orbignyanus strain, 01 Piaractus mesopotamicus strain, 01 Colossoma macropomum strain, and 01 Hypostomus plecostomus strain. Biochemical characterization, with its absorption of Congo red dye, production of flexirubin-type pigments, H2S production and reduction of nitrates proved that the isolate could be classified as Flavobacterium columnare.

Morfoanatomia da flor de cinco espécies de Galipea Aubl. e seu significado na evolução de flores tubulosas entre as Rutaceae neotropicais

A maioria dos gêneros da subtribo neotropical Galipeinae (tribo Galipeeae, Rutoideae) tem flores tubulosas, com várias formas e graus de conação e adnação. Galipea e outros gêneros na subtribo apresentam apenas duas anteras férteis mais cinco ou mais estaminódios, o que é intrigante porque na tribo predominam flores pentâmeras isostêmones. Visando elucidar a condição anatômica dessas características e estabelecer estados acurados para caracteres em análises filogenéticas, um estudo morfoanatômico de flores de cinco espécies de Galipea foi realizado, buscando os padrões de vascularização, posição, e união dos segmentos da flor. Destacam-se os resultados: 1) um tubo floral genuíno se forma no terço basal da flor por conação dos filetes e adnação desse tubo estaminal às pétalas; 2) as pétalas são distalmente coerentes umas às outras e aderentes aos filetes por meio de entrelaçamento de tricomas densos - um caso de pseudossimpetalia; 3) dentre as cinco (às vezes seis) estruturas tratadas como estaminódios, apenas as três externas são de fato homólogas a estames esterilizados, as demais surgem como ramificações adaxiais das pétalas; 4) os carpelos são peltados, congenitalmente conatos axial e lateralmente da base do ovário até o nível das placentas, e no estilete e estigma; na zona mediana e superior do ovário eles são unidos apenas posgenitalmente, com a epiderme diferenciada de carpelos contíguos e suturas evidentes na região ventral de cada carpelo; 5) a vascularização do disco sugere origem receptacular. As implicações desses dados para o entendimento da evolução das flores tubulosas em Galipea e grupos relacionados são discutidas.

Molecular systematics of Thorea (Rhodophyta, Thoreales) species in Brazil

This study aimed to evaluate species level taxonomy and phylogenetic relationship among Thorea species in Brazil and other regions of the world using two molecular markers - RUBISCO large subunit plastid gene (rbcL) and nuclear small-subunit ribosomal DNA (SSU rDNA). Three samples of Thorea from Brazil (states of Mato Grosso do Sul and São Paulo) and one sample from Dominican Republic (DR) were sequenced. Analyses based on partial sequences of rbcL (1,282 bp) and complete sequences of SSU (1,752 bp) were essentially congruent and revealed that Thoreales formed a distinct monophyletic clade, which had two major branches with high support, representing the genera Thorea and Nemalionopsis. Thorea clade had four main branches with high support for all analyses, each one representing the species: 1) T. gaudichaudii C. Agardh from Asia (Japan and Philippines) - this clade occurred only in the rbcL analyses; 2) T. violacea Bory from Asia (Japan) and North America (U.S.A. and DR); 3) T. hispida (Thore) Desvaux from Europe (England) and Asia (Japan); 4) a distinct group with the three Brazilian samples (sequence identity: rbcL 97.2%, 1,246 bp; SSU 96.0-98.1%, 1,699-1,720 bp). The Brazilian samples clearly formed a monophyletic clade based on both molecular markers and was interpreted as a separate species, for which we resurrected the name T. bachmannii Pujals. Morphological and molecular evidences indicate that the Thoreales is well-resolved at ordinal and generic levels. In contrast, Thorea species recognized by molecular data require additional characters (e.g. reproductive and chromosome numbers) to allow consistent and reliable taxonomic circumscription aiming at a world revision based on molecular and morphological evidences.

Origin of successive cambia on stem in three species of Menispermaceae

The lianas observed in this study, Abuta convexa (Vell.) Diels, Abuta imene (Mart.) Eichler, and Chondrodendron platiphyllum (A. St.-Hil.) Miers, all have successive cambia in their stems. The terminology applied to stem histology in species with successive cambia is as diverse as the interpretations of the origins of this cambial variant. Therefore, this study specifically investigates the origin of successive cambia through a developmental analysis of the above-mentioned species, including an analysis of the terminology used to describe this cambial variation. For the first time, we have identified several developmental stages giving rise to the origins of successive cambia in this family. First, the pericycle originates in 1-3 layers of conjunctive tissue. After the differentiation of the first ring, the conjunctive tissue undergoes new divisions, developing approximately 10 rows of parenchyma cells. In the middle portion, a layer of sclereids is formed, again subdividing the conjunctive tissue into two parts: internal and external. New cambia originate in the internal part, from which new secondary vascular strands will originate, giving rise to the second successive vascular ring of the stem. The external part remains parenchymatous during the installation of the second ring and will undergo new periclinal division, repeating the entire process. New cambia will originate from the neoformed strands, which will form only rays. In the literature, successive cambia are formed by a meristem called "diffuse lateral meristem."However, based on the species of Menispermaceae studied in this report, it is demonstrated that the diffuse lateral meristem is the pericycle itself.

Two-dimensional gel electrophoretic protein profile analysis during seed development of Ocotea catharinensis: a recalcitrant seed species

The aim of the present work was to characterize changes in the protein profile throughout seed development in O. catharinensis, a recalcitrant species, by two-dimensional gel electrophoresis. Protein extraction was undertaken by using a thiourea/urea buffer, followed by a precipitation step with 10% TCA. Comparative analysis during seed development showed that a large number of proteins were exclusively detected in each developmental stage. The cotyledonary stage, which represents the transition phase between embryogenesis and the beginning of metabolism related to maturation, presents the highest number of stage-specific spots. Protein identification, through MS/MS analysis, resulted in the identification of proteins mainly related to oxidative metabolism and storage synthesis. These findings contribute to a better understanding of protein metabolism during seed development in recalcitrant seeds, besides providing information on established markers that could be useful in defining and improving somatic embryogenesis protocols, besides monitoring the development of somatic embryos in this species.

Reserve carbohydrates and lipids from the seeds of four tropical tree species with different sensitivity to desiccation

Considering the importance of water content for the conservation and storage of seeds, and the involvement of soluble carbohydrates and lipids for embryo development, a comparative study was carried out among the seeds of Inga vera (ingá), Eugenia uniflora (pitanga), both classified as recalcitrant, and Caesalpinia echinata (brazilwood) and Erythrina speciosa (mulungu), considered as orthodox seeds. Low concentrations of cyclitols (0.3-0.5%), raffinose family oligosaccharides (ca. 0.05%) and unsaturated fatty acids (0-19%) were found in the seeds of ingá and pitanga, while larger amounts of cyclitols (2-3%) and raffinose (4.6-13%) were found in brazilwood and mulungu, respectively. These results, in addition to higher proportions of unsaturated fatty acids (53-71%) in orthodox seeds, suggested that sugars and lipids played important role in water movement, protecting the embryo cell membranes against injuries during dehydration.

Morpho-Anatomical Analysis of the Rhizome in Species of Scleria Berg. (Cyperaceae) from Serra do Cipó (MG)

Aspects related to the nature of stem thickening in monocotyledons have been the subject of many studies. Primary thickening has been attributed to the Primary Thickening Meristem (PTM). According to most authors, it gives rise, besides the adventitious roots, to the vascular tissues and part of the cortex. In other words, it has centripetal and centrifugal activity. For some authors, however, it gives rise only to the vascular system, and for others, only to part of the cortex. However, this work demonstrated that PTM corresponds to the pericycle in the meristematic phase or to the pericycle associated with the endodermis, also with meristematic activity. It was observed that the pericycle was responsible for the formation of the vascular system of the rhizome and of the adventitious roots; the endodermis gave rise to cell layers with radial disposition which comprised the inner portion of the stem cortex, and which corresponded to the region known as the derivatives of the meristematic endodermis (DME). A continuity was also demonstrated between the tissues of the stem and root in species of Scleria Berg. (Cyperaceae).

Revisão taxonômica de Croton sect. Lamprocroton (Müll. Arg.) Pax (Euphorbiaceae s.s.)

Croton is the second bigger and more diverse genus in the family Euphorbiaceae, with about 1,200 species distributed in 40 sections, occurring in all tropical areas, most of them in Americas. In South America, Brazil is the country in which a larger number of taxa are found, ca. 356. According to recent classification, the genus belongs to the tribe Crotoneae, and despite the wide and morphological diversity, it would be a monophyletic taxon. However, a phylogenetic analysis using markers of ITS region from nuclear ribosomal DNA, and of trnL-F from plastidial DNA, showed that Croton, like traditionally circumscribed, is not a monophyletic taxon. A taxonomic revision of Croton section Lamprocroton (Müll. Arg.) Pax is presented here. It is a Neotropical group with most of its species occurring from Southeast and South Brazil to southern South America (Uruguay and Argentina). Morphologically, the members of Lamprocroton are characterized as monoecious or dioecious shrubs or subshrubs, with a lepidote indumentum at least in part of foliage, entire leaves with no glands. The staminate flowers have 9 to 16 stamens and the pistillate flowers may have equal or unequal sepals, reduced to absent petals, and styles once or twice bifid. Overall, are recognized 26 species in the group, three of them new to the science. Identification key, morphological descriptions, illustrations, phenological period, as well as data on geographic distribution and general comments of each species are presented. Four taxa were excluded from C. sect. Lamprocroton because they do not show the morphological features that are diagnostics of the section. Four species that are poorly known were not included in the taxonomic treatment.

Morphology and anatomy of leaf miners in two species of Commelinaceae (Commelina diffusa Burm. f. and Floscopa glabrata (Kunth) Hassk)

In specialized literature, reports on anatomy of miners in host plants are few in number. These agents trigger excavations, or paths, by consumption of plant inner tissues by larvae of several insects. The aim of this work was to investigate leaf miner occurrence in Commelina diffusa (a cosmopolitan plant) and Floscopa glabrata (an amphibious plant) using anatomical techniques. The place where the plants were collected is subjected to seasonal floods, consequently both the species were exposed to the same weather conditions and seasonal floods. This study showed that members of Agromyzidae and Chironomidae families, which are Diptera endophytophagous larvae types, were responsible for the tunnels. Moreover, in Commelina diffusa Agromyzidae larvae were found, while in Floscopa glabrata three Chironomidae cephalic exuviae were found. The miners, as can be seen from anatomical studies, used only mesophyll parenchyma tissues for feeding, causing the formation of linear mines. In addition, in both the species, the epidermis and the medium-sized vascular units were kept intact, showing no structural modification, such as neoformation of tissues.

Absorption of metals and characterization of chemical elements present in three species of Gracilaria (Gracilariaceae) Greville: a genus of economical importance

Gracilaria Greville is a genus of seaweed that is economically explored by the cosmetic, pharmaceutical and food industries. One of the biggest problems associated with growing Gracilaria is the discharge of heavy metals into the marine environment. The absorption of heavy metals was investigated with the macroalga Gracilaria tenuistipitata Zhang et Xia, cultivated in a medium containing copper (Cu) and cadmium (Cd). In biological samples, EC50 concentrations of 1 ppm for cadmium and 0.95 ppm for copper were used. These concentrations were based on seaweed growth curves obtained over a period of six days in previous studies. ICP-AES was used to determine the amount of metal that seaweeds absorbed during this period. G. tenuistipitata was able to bioaccumulate both metals, about 17% of copper and 9% of cadmium. Basal natural levels of Cu were found in control seaweeds and in G. tenuistipitata exposed to Cd. In addition, the repertoire of other important chemical elements, as well as their concentrations, was determined for G. tenuistipitata and two other important seaweeds, G. birdiae Plastino & Oliveira and G. domingensis (Kützing) Sonder ex Dickie, collected in natural environments on the Brazilian shore.

Diversidade de abelhas (Hymenoptera, Apidae) ao longo de um gradiente latitudinal na Mata Atlântica

A Mata Atlântica é um dos ambientes mais ricos e ameaçados do mundo, o que deveria ter estimulado em muito o estudo e a conservação do Bioma, mas a fauna de Hymenoptera permanece ainda relativamente pouco conhecida. Em especial, a fauna de abelhas da floresta ombrófila densa é pouco estudada em comparação à fauna das áreas abertas brasileiras. O projeto temático "Biodiversidade de Hymenoptera e Isoptera: riqueza e diversidade ao longo de um gradiente latitudinal na Mata Atlântica - a floresta úmida do leste do Brasil", integrante do Programa Biota-Fapesp, foi idealizado com o objetivo de catalogar térmitas, formigas e famílias selecionadas de vespas ao longo da Mata Atlântica, disponibilizando dados que permitam melhor embasar a conservação deste bioma. O protocolo de amostragem aplicada para a coleta de himenópteros (excluindo as formigas) empregou armadilhas Malaise, pratos-armadilha de cor amarela e esforço similar na varredura de vegetação ao longo de 17 localidades selecionadas, representando um gradiente de quase 20° de latitude na Mata Atlântica, dos Estados da Paraíba até Santa Catarina. Este protocolo foi definido para otimizar a coleta de vespas, sendo as abelhas um produto secundário da amostragem (levantamentos de abelhas em geral utilizam captura em flores ao longo das estações do ano). No entanto, devido à escala regional do projeto e ao grande esforço de amostragem, uma expressiva quantidade de abelhas foi coletada durante o projeto, incluindo novos registros de abelhas para a Mata Atlântica. Foi amostrado um total de 797 espécimes distribuídos em 105 espécies de abelhas; o grupo de abelhas mais rico e abundante foi Meliponina. Uma análise de correspondência "destendenciada" ('detrended correspondence analysis') aplicada à uma matriz de presença ou ausência de Meliponina revela a relativamente fraca influência do gradiente latitudinal na composição das assembléias de abelhas de Mata Atlântica. Uma listagem das espécies amostradas por localidade, com suas freqüências relativas, é apresentada e discutida.

Influence of soil properties on the abundance of plant species in ferruginous rocky soils vegetation, southeastern Brazil

Ferruginous "campos rupestres" are a particular type of vegetation growing on iron-rich primary soils. We investigated the influence of soil properties on plant species abundance at two sites of ferruginous "campos rupestres" and one site of quartzitic "campo rupestre", all of them in "Quadrilátero Ferrífero", in Minas Gerais State, southeastern Brazil. In each site, 30 quadrats were sampled to assess plant species composition and abundance, and soil samples were taken to perform chemical and physical analyses. The analyzed soils are strongly acidic and presented low fertility and high levels of metallic cations; a principal component analysis of soil data showed a clear segregation among sites due mainly to fertility and heavy metals content, especially Cu, Zn, and Pb. The canonical correspondence analysis indicated a strong correlation between plant species abundance and soil properties, also segregating the sites.

Age polyethism in Plebeia emerina (Friese) (Hymenoptera: Apidae) colonies related to propolis handling

Stingless bees collect plant resins and make it into propolis, although they have a wider range of use for this material than do honey bees (Apis spp.). Plebeia spp. workers employ propolis mixed with wax (cerumen) for constructing and sealing nest structures, while they use viscous (sticky) propolis for defense by applying it onto their enemies. Isolated viscous propolis deposits are permanently maintained at the interior of their colonies, as also seen in other Meliponini species. Newly-emerged Plebeia emerina (Friese) workers were observed stuck to and unable to escape these viscous propolis stores. We examined the division of labor involved in propolis manipulation, by observing marked bees of known age in four colonies of P. emerina from southern Brazil. Activities on brood combs, the nest involucrum and food pots were observed from the first day of life of the marked bees. However, work on viscous propolis deposits did not begin until the 13th day of age and continued until the 56th day (maximum lifespan in our sample). Although worker bees begin to manipulate cerumen early, they seem to be unable to handle viscous propolis till they become older.

Seasonal variation of soluble carbohydrates and starch in Echinolaena inflexa, a native grass species from the Brazilian savanna, and in the invasive grass Melinis minutiflora

Echinolaena inflexa (Poir.) Chase is an abundant C3 grass species with high biomass production in the Brazilian savanna (cerrado); Melinis minutiflora Beauv. is an African C4 forage grass widespread in cerrado and probably displacing some native herbaceous species. In the present work, we analysed seasonally the content and composition of soluble carbohydrates, the starch amounts and the above-ground biomass (phytomass) of E. inflexa and M. minutiflora plants harvested in two transects at 5 and 130 m from the border in a restrict area of cerrado at the Biological Reserve and Experimental Station of Mogi-Guaçu (SP, Brazil). Results showed that water soluble carbohydrates and starch amounts from the shoots of both species varied according to the time of the year, whilst in the underground organs, variations were observed mainly in relation to the transects. Marked differences in the pattern of the above-ground biomass production between these two grasses relative to their location in the Reserve were also observed, with two peaks of the invasive species (July and January) at the Reserve border. The differences in carbohydrate accumulation, partitioning and composition of individual sugars concerning time of the year and location in the Reserve were more related to the annual growth cycle of both grasses and possibly to specific physiological responses of M. minutiflora to disturbed environments in the Reserve border.