Bioquímica e Biologia Molecular
URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11837
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Item Distyly and variation in floral traits in natural populations of Psychotria ipecacuanha (Brot.) Stokes (Rubiaceae)(Brazilian Journal of Botany, 2005-04) Oliveira, Luiz Orlando de; Vieira, Milene F.; Rossi, Ana Aparecida B.Psychotria ipecacuanha is a perennial, medicinal herb that grows in clusters in the understory of humid, shady areas of the Atlantic Rain Forest of southeastern Brazil. The present study characterized the variation in floral traits among 35 clusters from three natural populations of this plant species. Field observations showed that the clusters are isomorphic, that is, a given cluster will either set long-styled or short-styled flowers. Stigmas and anthers are reciprocally placed in each morph, a dimorphism characteristic of distyly. The populations are isoplethic, that is, a given population exhibits an equilibrium 1:1 ratio of floral morphs. Morphometric analyses revealed that anther length, stigma length, corolla diameter, and pollen grain diameter were consistently greater in short-styled flowers, regardless of the population investigated. Significant differences for floral traits in the short-styled morph were found among populations. Floral traits in the long-styled morph also showed some significant differences among populations, but not for stigma height and corolla length. Controlled pollinations carried out in natural populations showed that fruit production was higher after inter-morph pollination. Nevertheless, observations of pollen tube growth in style, and also fruit production after spontaneous self-pollination and intra-morph pollination, indicated partial intramorph compatibility in this plant species.Item Specific boundaries between the causal agents of the soybean stem canker(Tropical Plant Pathology, 2014-07) Oliveira, Luiz Orlando de; Guillin, Eduardo A.; Grijalba, Pablo E.; Gottlieb, Alexandra M.Pathogens within the Diaporthe complex cause seed decay, stem blight and stem canker on soybean, representing a serious threat for this crop species. We herein utilize worldwide sequence data retrieved from Genbank in order to assess the species boundaries between the soybean stem canker causal agents, and define whether or not they should be regarded as members of the same biological species. These studies were complemented with compatibility tests, in order to validate our findings from a biological standpoint. Species delimitation assays supported the occurrence of a speciation event between D. caulivora and D. phaseolourm var. meridionalis. A speciation hypothesis between D. aspalathi and D. phaseolourm var. meridionalis was also supported, based on three reciprocally monophyletic substitutions at locus EF1-α. Compatibility tests further validated species delimitation assays indicating that D. caulivora has developed barriers to gene exchange with D. phaseolorum var. meridionalis. Clarification of the specific boundaries of the SSC pathogens and related entities will be an important asset to future research in soybean pathology, epidemiology and breeding.Item Specific boundaries between the causal agents of the soybean stem canker(Tropical Plant Pathology, 2014-07) Oliveira, Luiz Orlando de; Guillin, Eduardo A.; Grijalba, Pablo E.; Gottlieb, Alexandra M.Pathogens within the Diaporthe complex cause seed decay, stem blight and stem canker on soybean, representing a serious threat for this crop species. We herein utilize worldwide sequence data retrieved from Genbank in order to assess the species boundaries between the soybean stem canker causal agents, and define whether or not they should be regarded as members of the same biological species. These studies were complemented with compatibility tests, in order to validate our findings from a biological standpoint. Species delimitation assays supported the occurrence of a speciation event between D. caulivora and D. phaseolourm var. meridionalis. A speciation hypothesis between D. aspalathi and D. phaseolourm var. meridionalis was also supported, based on three reciprocally monophyletic substitutions at locus EF1-α. Compatibility tests further validated species delimitation assays indicating that D. caulivora has developed barriers to gene exchange with D. phaseolorum var. meridionalis. Clarification of the specific boundaries of the SSC pathogens and related entities will be an important asset to future research in soybean pathology, epidemiology and breeding.Item Reproductive studies in ipecac (Psychotria ipecacuanha (Brot.) stockes; Rubiaceae): pollen development and morphology(Brazilian Archives of Biology and Technology, 2008-09) Oliveira, Luiz Orlando de; Souza, Margarete Magalhães; Martins, Ernane Ronie; Pereira, Telma Nair SantanaThe aim of this work was to carry out the reproductive studies on Brazilian accessions of ipecac, Psychotria ipecacuanha. It presented heterostyly, with brevistylous and longistylous flowers. The pollen development was observed from the sections of the anthers embedded in resin. Anther development was normal as usually observed in dicotyledones, displaying four layers: outer epidermis, endothecium, middle layer and inner tapetum. The pollen was bicellular and filled with starch at the microspore stage. Pollen morphology was studied using SEM, which showed pollen polymorphism within and between the two floral morphs. Five types of pollen with reticulate or perforate exine were identified. The characteristics showed that the sexual process was as important as the vegetative propagation for the reproduction of this species.Item Teor e composição do óleo essencial de cinco acessos de mentrasto(Química Nova, 2004-01) Oliveira, Luiz Orlando de; Barbosa, Cláudio de Almeida; Ferreira, Francisco Affonso; Silva, Derly José Henriques da; Mosquim, Paulo Roberto; Nascimento, Evandro Afonso; Castro, Henrique Guilhon deThis study aimed to analyse the content and the composition of the essential oil of five accesses of mentrasto (Ageratum conyzoides). Five accesses of mentrasto with three repetitions were used. The accesses were obtained in Mariana-MG (AMA), Piranga-MG (API), Visconde do Rio Branco (ARB) and Viçosa-MG (AVB and AVP). The essential oil was obtained by hydrodistillation and the identification of the oil components by CG and GC/MS. The ARB access presented the higher essential oil content, that is 0.70% (P < 0.05). Eleven chemical compounds were identified. The precocene I was the main constituent in the access API, and the precocene II was the main constituent in the accesses AMA, ARB, AVB and AVP.Item Evolutionary history of Manihot carthagenensis (Euphorbiaceae) and allied species in eastern South America(Molecular Phylogenetics and Evolution, 2019-03) Silveira, Thamyres Cardoso da; Martins, Márcio Lacerda Lopes; Rody, Hugo Vianna Silvapt-BR; Oliveira, Luiz Orlando deIntermittent episodes of climate changes, such as those that occurred during the Pleistocene, likely shaped the diversification of the young genus Manihot Mill. (Euphorbiacheae). One of such recently-derived congeners ─ M. carthagenensis ─ exhibits a widely disjunct distribution across dry environments in Eastern South America. Herein, we used molecular data from four nuclear gene regions (sts, ch_metE, g3pdh, and nia-i3) and seven nuclear microsatellite loci for reconstructing the phylogenetic relationships among M. carthagenensis and allied species and exploring likely phylogeographic scenarios that shaped the diversification and the distribution of gene pools of M. carthagenensis across the Caatinga and Chaco. Our data suggest that M. carthagenensis is not a monophyletic clade, as presently circumscribed. Morphological differences, genealogical relationships, and vegetation associations support three well-differentiated lineages, each of which merits the species rank: M. carthagenensis, M. glaziovii, and M. hahnii. Microsatellite data suggest that the newly circumscribed M. carthagenensis consists of at least three distinct gene pools, which are partly structured according to geography. The three gene pools likely evolved in allopatry, but remained interfertile. Population expansions after climate amelioration contributed to structuring hybrid zones. Moreover, we described two new single-copy gene regions (sts and ch_metE) as sources of molecular variation; they can facilitate the fine-scale probing of other parts of the phylogeny across Manihot.Item Diversidade genética estimada com marcadores entre sequências simples repetidas em cultivos comerciais de Cupuaçuzeiro(Ciência Rural, 2016-01) Oliveira, Luiz Orlando de; Silva, Bruna Mezzalira da; Rossi, Ana Aparecida Bandini; Dardengo, Juliana de Freitas Encinas; Araujo, Vitor Arreguy Amado Correa de; Rossi, Fernanda Saragosa; Clarindo, Wellington RonildoQuinze primers ISSR (entre sequências simples repetidas) foram utilizados para avaliar a diversidade genética entre e dentro de pomares comerciais de Theobroma grandiflorum (Willd. ex Spreng.) K. Schum. Para isso, foram analisados sessenta indivíduos, distribuídos nos três cultivos. Um total de 102 bandas foi amplificado, com uma porcentagem de 52,0% de polimorfismo em nível de espécie e média de 6,8 alelos por primer ISSR. A média do Índice de Conteúdo Polimórfico (PIC) foi de 0,55. Em relação aos índices de diversidade gênica de Nei (H) e de Shannon (I), os cultivos analisados apresentaram os valores: SAR H = 0,114 e I = 0,177; SSL H = 0,108 e I = 0,162 e SEC H = 0,104 e I = 0,156, considerados valores de moderados a baixos. A AMOVA revelou 34,91% da variância total entre os cultivos e 65,09% dentro deles. Os marcadores moleculares ISSR revelaram que há diversidade genética dentro de cada cultivo comercial estudado, portanto é possível selecionar genótipos superiores que poderão ser utilizados para originar cultivos mais uniformes. Esse resultado tem sido considerado de grande relevância, por fornecer ferramentas para a implementação de programas de melhoramento e delineamento de estratégias de conservação ex situ e in situ.Item Genetic entanglement between Cercospora species associating soybean purple seed stain(Mycological Progress, 2017-03-23) Oliveira, Luiz Orlando de; Guillin, Eduardo A.; Grijalba, Pablo E.; Gottlieb, Alexandra M.Soybean purple seed stain (S-PSS) is a destructive, worldwide distributed fungal disease caused by several Cercospora species. This work aims to shed light on the nature of the genealogical and genetic relationships amongst S-PSS causal agents. Fungal isolates were obtained from Argentina and Brazil, which belong to the leading countries in soybean production worldwide. DNA sequences were obtained from eight loci across the collection of isolates. Relationships were evaluated through Bayesian phylogenetic inferences, and distance and character-based network analyses and discriminant analyses. The occurrence of reticulate evolutionary events was tested with recombination tests. The high haplotype diversity (H = 1.0) was arranged in four validated haplogroups. Reticulate network topologies were evident, and 11 recombination events were validated through several tests. Five of these events occurred across species boundaries. Comparison with sequences from 70 Cercospora species indicated that at least five monophyletic groups of S-PSS-causing agents are currently present in South America. The provided evidence supports the hypothesis that interspecific genetic exchange plays a significant role in the evolutionary dynamics of Cercospora species in this region. The occurrence of interspecific recombination has implications for understanding epidemiological threats to soybean production that appear to be more serious than previously anticipated.Item More Cercospora species infect soybeans across the Americas than meets the eye(Plos One, 2015-08-07) Soares, Ana Paula Gomes; Guillin, Eduardo A.; Borges, Leandro Luiz; Silva, Amanda C. T. da; Almeida, Álvaro M. R. de; Grijalba, Pablo E.; Gottlieb, Alexandra M.; Bluhm, Burton H.; Oliveira, Luiz Orlando deDiseases of soybean caused by Cercospora spp. are endemic throughout the world’s soybean production regions. Species diversity in the genus Cercospora has been underestimated due to overdependence on morphological characteristics, symptoms, and host associations. Currently, only two species (Cercospora kikuchii and C. sojina) are recognized to infect soybean; C. kikuchii causes Cercospora leaf blight (CLB) and purple seed stain (PSS), whereas C. sojina causes frogeye leaf spot. To assess cryptic speciation among pathogens causing CLB and PSS, phylogenetic and phylogeographic analyses were performed with isolates from the top three soybean producing countries (USA, Brazil, and Argentina; collectively accounting for ~80% of global production). Eight nuclear genes and one mitochondrial gene were partially sequenced and analyzed. Additionally, amino acid substitutions conferring fungicide resistance were surveyed, and the production of cercosporin (a polyketide toxin produced by many Cercospora spp.) was assessed. From these analyses, the long-held assumption of C. kikuchii as the single causal agent of CLB and PSS was rejected experimentally. Four cercosporin-producing lineages were uncovered with origins (about 1 Mya) predicted to predate agriculture. Some of the Cercospora spp. newly associated with CLB and PSS appear to represent undescribed species; others were not previously reported to infect soybeans. Lineage 1, which contained the ex-type strain of C. kikuchii, was monophyletic and occurred in Argentina and Brazil. In contrast, lineages 2 and 3 were polyphyletic and contained wide-host range species complexes. Lineage 4 was monophyletic, thrived in Argentina and the USA, and included the generalist Cercospora cf. flagellaris. Interlineage recombination was detected, along with a high frequency of mutations linked to fungicide resistance in lineages 2 and 3. These findings point to cryptic Cercospora species as underappreciated global considerations for soybean production and phytosanitary vigilance, and urge a reassessment of host-specificity as a diagnostic tool for Cercospora.Item Molecular evidence of cryptic speciation, historical range expansion, and recent intraspecific hybridization in the Neotropical seasonal forest tree Cedrela fissilis (Meliaceae)(Elsevier Molecular Phylogenetics and Evolution, 2011-09-10) Garcia, Magali Gonçalves; Silva, Roberta Santos; Carniello, Maria Antonia; Veldman, Joseph William; Rossi, Ana Aparecida Bandini; Oliveira, Luiz Orlando deMolecular phylogeography can lead to a better understanding of the interaction between past climate events, large-scale vegetation shifts, and the evolutionary history of Neotropical seasonal forests. The endangered timber tree species Cedrela fissilis is associated with seasonal forests and occurs throughout South America. We sampled C. fissilis from 56 sites across the species’ range in Brazil and Bolivia and obtained sequence data for nuclear and chloroplast DNA. Most specimens (149 out of 169) exhibited intraindividual polymorphism for the nuclear internal transcribed spacer (ITS). Cloning and an array of complementary sequence analyses indicated that the multiple copies of ITS were functional paralogs – concerted evolution in C. fissilis appeared to be incomplete. Independent Bayesian analyses using either ITS or cpDNA data revealed two separate phylogenetic lineages within C. fissilis that corresponded to populations located in separate geographic regions. The divergence occurred in the Early Pliocene and Late Miocene. We argue that climate-mediated events triggered dispersal events and split ancestral populations into at least two large refugial areas of seasonal forest that were located to the east and west of the present day Cerrado. Upon recent climate amelioration, formerly isolated lineages reconnected and intraspecific hybridization gave rise to intraindividual polymorphism and incomplete concerted evolution in C. fissilis.