941 resultados para Poa alpina, transplantation, altitudinal gradient, genetic diversity, phenotypic plasticity
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ABSTRACT: The present study intended to characterize the phenotypic and genetic diversity of Brazilian isolates of Chromobacterium violaceum from aquatic environments within the Amazon region. Nineteen isolates showed morphological properties of C. violaceum and the majority grew at 44°C. Low temperatures, in contrast, showed to be inhibitory to their growth, as eleven isolates did not grow at 10ºC and nine did not produce pigmentation, clearly indicating an inhibition of their metabolism. The largest variation among isolates was observed in the citrate test (Simmons), in which 12 isolates were positive, and in the oxidation/fermentation of sucrose, with six positives isolates. Chloramphenicol, gentamicin and sulfonamides efficiently inhibited bacterial growth. Amplified products of the recA gene were digested with HindII or PstI, which produced three or four restriction fragments patterns, respectively. The combined analysis arranged the isolates into six genospecies. The higher diversity observed in Belém (genotypes C, D, E and F) may be a consequence of intense human occupation, pollution of the aquatic environment or due to the higher diversity of the environments sampled in that region. In conclusion, a high level of genetic and phenotypic diversity was observed, and four new genospecies were described.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Population genetics theory predicts loss in genetic variability because of drift and inbreeding in isolated plant populations; however, it has been argued that long-distance pollination and seed dispersal may be able to maintain gene flow, even in highly fragmented landscapes. We tested how historical effective population size, historical migration and contemporary landscape structure, such as forest cover, patch isolation and matrix resistance, affect genetic variability and differentiation of seedlings in a tropical palm (Euterpe edulis) in a human-modified rainforest. We sampled 16 sites within five landscapes in the Brazilian Atlantic forest and assessed genetic variability and differentiation using eight microsatellite loci. Using a model selection approach, none of the covariates explained the variation observed in inbreeding coefficients among populations. The variation in genetic diversity among sites was best explained by historical effective population size. Allelic richness was best explained by historical effective population size and matrix resistance, whereas genetic differentiation was explained by matrix resistance. Coalescence analysis revealed high historical migration between sites within landscapes and constant historical population sizes, showing that the genetic differentiation is most likely due to recent changes caused by habitat loss and fragmentation. Overall, recent landscape changes have a greater influence on among-population genetic variation than historical gene flow process. As immediate restoration actions in landscapes with low forest amount, the development of more permeable matrices to allow the movement of pollinators and seed dispersers may be an effective strategy to maintain microevolutionary processes.
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Este trabalho resume os dados de florística e fitossociologia de 11, das 14 parcelas de 1 ha, alocadas ao longo do gradiente altitudinal da Serra do Mar, São Paulo, Brasil. As parcelas começam na cota 10 m (Floresta de Restinga da Praia da Fazenda, município de Ubatuba) e estão distribuídas até a cota 1100 m (Floresta Ombrófila Densa Montana da Trilha do rio Itamambuca, município de São Luis do Paraitinga) abrangendo os Núcleos Picinguaba e Santa Virgínia do Parque Estadual da Serra do Mar. Na Restinga o solo é Neossolo Quartzarênico francamente arenoso, enquanto que na encosta o solo é um Cambisolo Háplico Distrófico argilo-arenoso, sendo que todas as parcelas apresentaram solo ácido (pH 3 – 4) com alta diluição de nutrientes e alta saturação de alumínio. Na Restinga e no sopé da encosta o clima é Tropical/Subtropical Úmido (Af/Cfa), sem estação seca, com precipitação média anual superior a 2.200 mm e temperatura média anual de 22 °C. Subindo a encosta mantêm-se a média de precipitação, mas há um gradativo resfriamento, de forma que a 1.100 m o clima é Subtropical Úmido (Cfa/Cfb), sem estação seca, com temperatura média anual de 17 °C. Destaca-se ainda que, quase diariamente, a parte superior da encosta, geralmente acima de 400 m, é coberta por uma densa neblina. Nas 14 parcelas foram marcados, medidos e amostrados 21.733 indivíduos com DAP ≥ 4,8 cm, incluindo árvores, palmeiras e fetos arborescentes. O número médio de indivíduos amostrados nas 14 parcelas foi de 1.264 ind.ha–1 (± 218 EP de 95%). Dentro dos parâmetros considerados predominaram as árvores (71% FOD Montana a 90% na Restinga), seguidas de palmeiras (10% na Restinga a 25% na FOD Montana) e fetos arborescentes (0% na Restinga a 4% na FOD Montana). Neste aspecto destaca-se a FOD Terras Baixas Exploradas com apenas 1,8% de palmeiras e surpreendentes 10% de fetos arborescentes. O dossel é irregular, com altura variando de 7 a 9 m, raramente as árvores emergentes chegam a 18 m, e a irregularidade do dossel permite a entrada de luz suficiente para o desenvolvimento de centenas de espécies epífitas. Com exceção da FOD Montana, onde o número de mortos foi superior a 5% dos indivíduos amostrados, nas demais fitofisionomias este valor ficou abaixo de 2,5%. Nas 11 parcelas onde foi realizado o estudo florístico foram encontradas 562 espécies distribuídas em 195 gêneros e 68 famílias. Apenas sete espécies – Euterpe edulis Mart. (Arecaceae), Calyptranthes lucida Mart. ex DC. e Marlierea tomentosa Cambess (ambas Myrtaceae), Guapira opposita (Vell.) Reitz (Nyctaginaceae), Cupania oblongifolia Mart. (Sapindaceae) e as Urticaceae Cecropia glaziovii Snethl. e Coussapoa microcarpa (Schott) Rizzini – ocorreram da Floresta de Restinga à FOD Montana, enquanto outras 12 espécies só não ocorreram na Floresta de Restinga. As famílias com o maior número de espécies são Myrtaceae (133 spp), Fabaceae (47 spp), 125 Fitossociologia em parcelas permanentes de Mata Atlântica http://www.biotaneotropica.org.br/v12n1/pt/abstract?article+bn01812012012 http://www.biotaneotropica.org.br Biota Neotrop., vol. 12, no. 1 Introdução A Mata Atlântica sensu lato (Joly et al. 1999) é a segunda maior floresta tropical do continente americano (Tabarelli et al. 2005). A maior parte dos Sistemas de Classificação da vegetação brasileira reconhece que no Domínio Atlântico (sensu Ab’Saber 1977) esse bioma pode ser dividido em dois grandes grupos: a Floresta Ombrófila Densa, típica da região costeira e das escarpas serranas com alta pluviosidade (Mata Atlântica – MA – sensu stricto), e a Floresta Estacional Semidecidual, que ocorre no interior, onde a pluviosidade, além de menor, é sazonal. Na região costeira podem ocorrer também Manguezais (Schaeffer-Novelli 2000), ao longo da foz de rios de médio e grande porte, e as Restingas (Scarano 2009), crescendo sobre a planície costeira do quaternário. No topo das montanhas, geralmente acima de 1500 m, estão os Campos de Altitude (Ribeiro & Freitas 2010). Em 2002, a Fundação SOS Mata Atlântica em parceria com o INPE (Instituto..., 2002) realizaram um levantamento que indica que há apenas 7,6% da cobertura original da Mata Atlântica (s.l.). Mais recentemente Ribeiro et al. (2009) refinaram a estimativa incluindo fragmentos menores, que não haviam sido contabilizados, e concluíram que resta algo entre 11,4 e 16% da área original. Mesmo com esta fragmentação, o mosaico da Floresta Atlântica brasileira possui um dos maiores níveis de endemismos do mundo (Myers et al. 2000) e cerca da metade desses remanescentes de grande extensão estão protegidos na forma de Unidades de Conservação (Galindo & Câmara 2005). Entre os dois centros de endemismo reconhecidos para a MA (Fiaschi & Pirani 2009), o bloco das regiões sudeste/sul é o que conserva elementos da porção sul de Gondwana (Sanmartin & Ronquist 2004), tido como a formação florestal mais antiga do Brasil (Colombo & Joly 2010). Segundo Hirota (2003), parte dos remanescentes de MA está no estado de São Paulo, onde cerca de 80% de sua área era coberta por florestas (Victor 1977) genericamente enquadradas como Mata Atlântica “sensu lato” (Joly et al. 1999). Dados de Kronka et al. (2005) mostram que no estado restam apenas 12% de área de mata e menos do que 5% são efetivamente florestas nativas pouco antropizadas. Nos 500 anos de fragmentação e degradação das formações naturais, foram poupadas apenas as regiões serranas, principalmente a fachada da Serra do Mar, por serem impróprias para práticas agrícolas. Usando o sistema fisionômico-ecológico de classificação da vegetação brasileira adotado pelo IBGE (Veloso et al. 1991), a Floresta Ombrófila Densa, na área de domínio da Mata Atlântica, foi subdividida em quatro faciações ordenadas segundo a hierarquia topográfica, que refletem fisionomias de acordo com as variações das faixas altimétricas e latitudinais. No estado de São Paulo, na latitude entre 16 e 24 °S temos: 1) Floresta Ombrófila Densa das Terras Baixas - 5 a 50 m de altitude; 2) Floresta Ombrófila Densa Submontana – no sopé da Serra do Mar, com cotas de altitude variando entre 50 e 500 m; 3) Floresta Ombrófila Densa Montana – recobrindo a encosta da Serra do Mar propriamente dita, em altitudes que variam de 500 a 1.200 m; 4) Floresta Ombrófila Densa Altimontana – ocorrendo no topo da Serra do Mar, acima dos limites estabelecidos para a formação montana, onde a vegetação praticamente deixa de ser arbórea, pois predominam os campos de altitude. Nas últimas três décadas muita informação vem sendo acumulada sobre a composição florística e a estrutura do estrato arbóreo dos remanescentes florestais do estado, conforme mostram as revisões de Oliveira-Filho & Fontes (2000) e Scudeller et al. (2001). Em florestas tropicais este tipo de informação, assim como dados sobre a riqueza de espécies, reflete não só fatores evolutivos e biogeográficos, como também o histórico de perturbação, natural ou antrópica, das respectivas áreas (Gentry 1992, Hubbell & Foster 1986). A síntese dessas informações tem permitido a definição de unidades fitogeográficas com diferentes padrões de riqueza de espécies e apontam para uma diferenciação, entre as florestas paulistas, no sentido leste/oeste (Salis et al. 1995, Torres et al. 1997, Santos et al. 1998). Segundo Bakker et al. (1996) um método adequado para acompanhar e avaliar as mudanças na composição das espécies e dinâmica da floresta ao longo do tempo é por meio de parcelas permanentes (em inglês Permanent Sample Plots –PSPs). Essa metodologia tem sido amplamente utilizada em estudos de longa duração em florestas tropicais, pois permite avaliar a composição e a estrutura florestal e monitorar sua mudança no tempo (Dallmeier 1992, Condit 1995, Sheil 1995, Malhi et al. 2002, Lewis et al. 2004). Permite avaliar também as consequências para a floresta de problemas como o aquecimento global e a poluição atmosférica (Bakker et al. 1996). No Brasil os projetos/programas que utilizam a metodologia de Parcelas Permanentes tiveram origem, praticamente, com o Projeto Rubiaceae (49) e Lauraceae (49) ao longo de todo gradiente da FOD e Monimiaceae (21) especificamente nas parcelas da FOD Montana. Em termos de número de indivíduos as famílias mais importantes foram Arecaceae, Rubiaceae, Myrtaceae, Sapotaceae, Lauraceae e na FOD Montana, Monimiaceae. Somente na parcela F, onde ocorreu exploração de madeira entre 1960 e 1985, a abundância de palmeiras foi substituída pelas Cyatheaceae. O gradiente estudado apresenta um pico da diversidade e riqueza nas altitudes intermediárias (300 a 400 m) ao longo da encosta (índice de Shannon-Weiner - H’ - variando de 3,96 a 4,48 nats.indivíduo–1). Diversas explicações para este resultado são apresentadas neste trabalho, incluindo o fato dessas altitudes estarem nos limites das expansões e retrações das diferentes fitofisionomias da FOD Atlântica durante as flutuações climáticas do Pleistoceno. Os dados aqui apresentados demonstram a extraordinária riqueza de espécies arbóreas da Floresta Ombrófila Densa Atlântica dos Núcleos Picinguaba e Santa Virgínia do Parque Estadual da Serra do Mar, reforçando a importância de sua conservação ao longo de todo o gradiente altitudinal. A diversidade desta floresta justifica também o investimento de longo prazo, através de parcelas permanentes, para compreender sua dinâmica e funcionamento, bem como monitorar o impacto das mudanças climáticas nessa vegetação.
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Hancornia speciosa Gomes is a fruit tree native from Brazil that belongs to Apocinaceae family, and is popularly known as Mangabeira. Its fruits are widely consumed raw or processed as fruit jam, juices and ice creams, which have made it a target of intense exploitation. The extractive activities and intense human activity on the environment of natural occurrence of H. speciosa has caused genetic erosion in the species and little is known about the ecology or genetic structure of natural populations. The objective of this research was the evaluation of the genetic diversity and genetic structure of H. speciosa var. speciosa. The genetic variability was assessed using 11 allozyme loci with a sample of 164 individuals distributed in six natural populations located in the States of Pernambuco and Alagoas, Northeastern Brazil. The results showed a high level of genetic diversity within the species (e= 0.36) seeing that the most of the genetic variability of H. speciosa var. speciosa is within its natural populations with low difference among populations (
or = 0.081). The inbreeding values within (
= -0.555) and among populations (
=-0.428) were low showing lacking of endogamy and a surplus of heterozygotes. The estimated gene flow (
m ) was high, ranging from 2.20 to 13.18, indicating to be enough to prevent the effects of genetic drift and genetic differentiation among populations. The multivariate analyses indicated that there is a relationship between genetic and geographical distances, which was confirmed by a spatial pattern analysis using Mantel test (r = 0.3598; p = 0.0920) with 1000 random permutations. The high genetic diversity index in these populations indicates potential for in situ genetic conservation.
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The Atlantic Forest is an excellent case study for the elevational diversity of birds, and some inventories along elevational gradients have been carried out in Brazil. Since none of these studies explain the patterns of species richness with elevation, we herein review all Brazilian studies on bird elevational diversity, and test a geometric constraint null model that predicts a unimodal species-altitude curve, the Mid-domain Effect (MDE). We searched for bird inventories in the literature and also analysed our own survey data using limited-radius point counts along an 800 m elevational gradient in the state of São Paulo, Brazil. We found 10 investigations of elevational diversity of Atlantic Forest birds and identified five different elevational patterns: monotonic decreasing diversity, constant at low elevations, constant at low elevations but increasing towards the middle, and two undescribed patterns for Atlantic Forest birds, trough-shaped and increasing diversity. The average MDE fit was low (r² = 0.31) and none of the MDE predictions were robust across all gradients. Those studies with good MDE model fits had obvious sampling bias. Although it has been proposed that the MDE may be positively associated with the elevational diversity of birds, it does not fit the Brazilian Atlantic Forest bird elevational diversity.
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Primula apennina Widmer is endemic to the North Apennines (Italy). ISSR were used to detect the genetic diversity within and among six populations representative of the species distribution range. High levels of genetic diversity were revealed both at population (PPB = 75.92%, HS = 0.204, Hpop = 0.319) and at species level (PPB = 96.95%, HT = 0.242, Hsp = 0.381). Nei gene diversity statistics (15.7%), Shannon diversity index (16.3%) and AMOVA (14%) detected a moderate level of interpopulation diversity. Principal coordinate and bayesian analyses clustered the populations in three major groups along a geographic gradient. The correlation between genetic and geographic distances was positive (Mantel test, r = 0.232). All together, these analyses revealed a weak but significant spatial genetic structure in P. apennina, with gene flow acting as a homogenizing force that prevents a stronger differentiation of populations. Conservation measures are suggested based on the observed pattern of genetic variability. P. apennina belongs to Primula subsect. Euauricula which includes 15 species distributed on the whole Alps and Apennines. A phylogenetic analysis was carried out using AFLP markers in order both to clarify the relationships among the species of subsection Euauricula that remained unresolved in previous works and to make some hypoteses on their evolutive dynamics. NJ, PCO and BAPS analyses strongly confirmed the monophyly of P. subsect. Euauricula and all the species form strongly supported clades. NJ tree topology suggested a simultaneous fragmentations of ancestral species in a large number of isolated populations that survived in refugia along the unglaciated margins of the Alps in response to the Pleistocene climatic oscillations.
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OBJECTIVES: The aim of this study was to determine the phenotypic and genotypic resistance profiles of methicillin-resistant Staphylococcus pseudintermedius (MRSP) and to examine the clonal distribution in Europe and North America. METHODS: A total of 103 MRSP isolates from dogs isolated from several countries in Europe, the USA and Canada were characterized. Isolates were identified by PCR-restriction fragment length polymorphism (RFLP), antimicrobial susceptibility was determined by broth dilution or gradient diffusion, and antimicrobial resistance genes were detected using a microarray. Genetic diversity was assessed by multilocus sequence typing (MLST), PFGE and spa typing. Staphylococcal cassette chromosome mec (SCCmec) elements were characterized by multiplex PCR. RESULTS: Thirteen different sequence types (STs), 18 PFGE types and 8 spa types were detected. The hybrid SCCmec element II-III described in a MRSP isolate was present in 75 (72.8%) isolates. The remaining isolates either had SCCmec type III (n=2), IV (n=6), V (n=14) or VII-241 (n=4) or were non-typeable (n=2). The most common genotypes were ST71(MLST)-J(PFGE)-t02(spa)-II-III(SCCmec) (56.3%) and ST68-C-t06-V (12.6%). In addition to mecA-mediated beta-lactam resistance, isolates showed resistance to trimethoprim [dfr(G)] (90.3%), gentamicin/kanamycin [aac(6')-Ie-aph(2')-Ia] (88.3%), kanamycin [aph(3')-III] (90.3%), streptomycin [ant(6')-Ia] (90.3%), streptothricin (sat4) (90.3%), macrolides and/or lincosamides [erm(B), lnu(A)] (89.3%), fluoroquinolones (87.4%), tetracycline [tet(M) and/or tet(K)] (69.9%), chloramphenicol (cat(pC221)) (57.3%) and rifampicin (1.9%). CONCLUSIONS: Two major clonal MRSP lineages have disseminated in Europe (ST71-J-t02-II-III) and North America (ST68-C-t06-V). Regardless of their geographical or clonal origin, the isolates displayed resistance to the major classes of antibiotics used in veterinary medicine and thus infections caused by MRSP isolates represent a serious therapeutic challenge.
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Background The European trout (Salmo trutta species complex) occurs across a very wide altitudinal range from lowland rivers to alpine streams. Historically, the major European river systems contained different, evolutionarily distinct trout lineages, and some of this genetic diversity has persisted in spite of extensive human-mediated translocations. We used AFLP-based genome scans to investigate the extent of potentially adaptive divergence among major drainages and along altitudinal gradients replicated in several rivers. Results The proportion of loci showing evidence of divergent selection was larger between drainages than along altitudinal transects within drainages. This suggests divergent selection is stronger between drainages, or adaptive divergence is constrained by gene flow among populations within drainages, although the latter could not be confirmed at a more local scale. Still, altitudinal divergence occurred and, at approximately 2% of the markers, parallel changes of the AFLP band frequencies with altitude were observed suggesting that altitude may well be an important source of divergent selection within rivers. Conclusions Our results indicate that adaptive genetic divergence is common both between major European river systems and along altitudinal gradients within drainages. Alpine trout appear to be a promising model system to investigate the relative roles of divergent selection and gene flow in promoting or preventing adaptation to climate gradients.
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Alpine grasslands are ecosystems with a great diversity of plant species. However, little is known about other levels of biodiversity, such as landscape diversity, diversity of biological interactions of plants with herbivores or fungal pathogens, and genetic diversity. We therefore explored natural and anthropogenic determinants of grassland biodiversity at several levels of biological integration, from the genetic to the landscape level in the Swiss Alps. Differences between cultural traditions (Romanic, Germanic, and Walser) turned out to still affect land use diversity and thus landscape diversity. Increasing land use diversity, in turn, increased plant species diversity per village. However, recent land use changes have reduced this diversity. Within grassland parcels, plant species diversity was higher on unfertilized mown grasslands than on fertilized or grazed ones. Most individual plants were affected by herbivores and fungal leaf pathogens, reflecting that parcels harbored a great diversity of herbivores and pathogens. However, as plant damage by herbivores and pathogens was not severe, conserving these biological interactions among plants is hardly compromising agricultural goals. A common-garden experiment revealed genetic differentiation of the important fodder grass Poa alpina between mown and grazed sites, suggesting adaptation. Per-village genetic diversity of Poa alpina was greater in villages with higher land use diversity, analogous to the higher plant species diversity there. Overall, landscape diversity and biodiversity within grassland parcels are currently declining. As this contradicts the intention of Swiss law and international agreements, financial incentives need to be re-allocated and should focus on promoting high biodiversity at the local and the landscape level. At the same time, this will benefit landscape attractiveness for tourists and help preserve a precious cultural heritage in the Swiss Alps.
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• Premise of the study: Because not all plant species will be able to move in response to global warming, adaptive evolution matters largely for plant persistence. As prerequisites for adaptive evolution, genetic variation in and selection on phenotypic traits are needed, but these aspects have not been studied in tropical species. We studied how plants respond to transplantation to different elevations on Mt. Kilimanjaro, Tanzania, and whether there is quantitative genetic (among-seed family) variation in and selection on life-history traits and their phenotypic plasticity to the different environments. • Methods: We reciprocally transplanted seed families of 15 common tropical, herbaceous species of the montane and savanna vegetation zone at Mt. Kilimanjaro to a watered experimental garden in the montane (1450 m) and in the savanna (880 m) zone at the mountain’s slope and measured performance, reproductive, and phenological traits. • Results: Plants generally performed worse in the savanna garden, indicating that the savanna climate was more stressful and thus that plants may suffer from future climate warming. We found significant quantitative genetic variation in all measured performance and reproductive traits in both gardens and for several measures of phenotypic plasticity in response to elevational transplantation. Moreover, we found positive selection on traits at low and intermediate trait values levelling to neutral or negative selection at high values. • Conclusions: We conclude that common plants at Mt. Kilimanjaro express quantitative genetic variation in fitness-relevant traits and in their plasticities, suggesting potential to adapt evolutionarily to future climate warming and increased temperature variability.
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Gene flow is usually thought to reduce genetic divergence and impede local adaptation by homogenising gene pools between populations. However, evidence for local adaptation and phenotypic differentiation in highly mobile species, experiencing high levels of gene flow, is emerging. Assessing population genetic structure at different spatial scales is thus a crucial step towards understanding mechanisms underlying intraspecific differentiation and diversification. Here, we studied the population genetic structure of a highly mobile species – the great tit Parus major – at different spatial scales. We analysed 884 individuals from 30 sites across Europe including 10 close-by sites (< 50 km), using 22 microsatellite markers. Overall we found a low but significant genetic differentiation among sites (FST = 0.008). Genetic differentiation was higher, and genetic diversity lower, in south-western Europe. These regional differences were statistically best explained by winter temperature. Overall, our results suggest that great tits form a single patchy metapopulation across Europe, in which genetic differentiation is independent of geographical distance and gene flow may be regulated by environmental factors via movements related to winter severity. This might have important implications for the evolutionary trajectories of sub-populations, especially in the context of climate change, and calls for future investigations of local differences in costs and benefits of philopatry at large scales.
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La diversidad genética poblacional de maíz en México es muy dinámica y depende de factores biológicos, agroecológicos y socio-económicos, y necesidades familiares. En este trabajo se describió y clasificó la variabilidad morfológica de una colección de 60 muestras poblacionales de maíz, colectadas en 44 municipios de la Mixteca Baja Oaxaqueña (846 msnm a 1842 msnm). Las muestras se sembraron y cultivaron durante el ciclo primavera-verano de 2010, en Santo Domingo Tonala, Oaxaca, bajo un diseño de bloques completos al azar con cuatro repeticiones. Se evaluaron 19 caracteres morfológicos de planta, mazorca, grano y espiga (panoja), y se determinaron diferencias significativas entre poblacionales en estos caracteres. Los caracteres altura de planta y mazorca, días a floración masculina y femenina, y número de granos por hilera en la mazorca fueron determinantes para describir la variabilidad morfológica total. La variación morfológica y fenológica de las poblaciones de maíz se asocia con los patrones altitudinales y geográficos de donde proceden. Se determinaron seis grupos fenotípicos significativamente diferentes con características de mazorca, grano y planta semejantes a las descritas para las razas Celaya, Bolita, Pepitilla, Ancho, y ciertos complejos raciales entre Ancho, Mixteco, Celaya y Bolita.