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Multiple outcomes for PI3K/Akt/mTOR targeting in non-Hodgkin lymphomaMüller, Anja 25 August 2015 (has links)
Wachstumsfaktor bedingte Aktivierung des PI3K/Akt/mTOR Signalweg wirkt positiv auf Vermehrung und Überleben. Konstitutive Aktivierung des Signalweges in NHL ist jedoch an Tumorprogression und Therapieresistenz beteiligt. Am Zelllinienmodell wurden zwei mögliche Therapiestrategien der PI3K/Akt/mTOR Inhibition erprobt, PI3K Inhibition mit BKM120 und horizontale Kombination von Zytostatika mit PI3K/Akt/mTOR Inhibitoren Erstens, BKM120 hat Antitumoraktivität in NHL und induziert Zelltod. Auf molekularer Ebene führt BKM120 vermittelte Dephosphorylierung von CDK1 an Y15 zur Aktivierung des M-Phase Komplex CDK1/Zyklin B und Eintritt in die Mitose. Parallel erlaubt die Degradation von Zyklin A und Hochregulation von Zyklin B Progression bis zur Metaphase, hemmt jedoch die Transition in die Anaphase. Anhaltender Metaphasearrest bewirkt programmierten Zelltod über den intrinsischen Signalweg der Apoptose durch Hochregulation der BH3-onlys Puma und Hrk, Aktivierung von Bax/Bak und proteolytische Spaltung von Caspase 9. Verlust von Bax/Bak oder Caspase Inhibition schützt vor BKM120 vermitteltem Zelltod. Bax/Bak defiziente Zellen, welche zusätzlich p53 Mutationen aufweisen, werden polyploid. Die Polyploidie ist ATM-MEK1/2 abhängig und kann mit Caffeine oder U0126blockiert werden. Zur Vermeidung von Polyploidie bedingter Tumorprogression, sollte BKM120 nur in Verbindung mit MAPK/ATM Inhibitoren verwendet werden. Zweitens. Horizontale Kombination PI3K/Akt/mTOR Inhibitoren mit cytotoxischen Substanzen schützt vor Apoptose. Der Schutzeffekt tritt auschließlich bei niedrigen Konzentration auf und ist unabhängig von der Art des Inhibitors bzw. Ebene der Inhibition. Das Onkogen und NFkB Target Pim-2 ist möglicherweise am Schutzmechanismus beteiligt. Durch die PI3K/Akt/mTOR vermittelte Pim-2 Regulation ergibt sich eine neue Rückkopplungsschleife. Im Fazit erschwert die Komplexizität des PI3K/Akt/mTOR Signalweges die Etablierung von Therapien. / Growth factor mediated activation of the PI3K/Akt/mTOR pathway positively regulates proliferation and survival. Constitutive activation in NHL, however, is correlated with tumor progression and therapeutic resistance. Therefore, two possible strategies were tested in a cell line model system, Inhibition of PI3K with BKM120 and PI3K/Akt/mTOR Inhibition in addition to cytostatic drug administration. First, it is demonstrated that the pan PI3K inhibitor BKM120 has antitumor activity in NHL and induces cell death. On molecular level, BKM120 mediated dephosphorylation of CDK1 on Y15 causes activation of the M-phase complex CDK1/Cyclin B and entry into mitosis. In parallel, degradation of Cyclin A and Upregulation of Cyclin B enables progression into metaphase but inhibits transition into anaphase. Prolonged metaphase arrest induces programmed cell death via the intrinsic apoptosis pathway by upregulation of the BH3-onlys Puma and Hrk, activation of Bax/Bak and proteolytic cleavage of caspase-9. Loss of Bax/Bak or caspase inhibition protects from BKM120 induced apoptosis. Bax/Bak deficient cells with additional p53 mutation become polyploid. This polyploidy is ATM-MEK1/2 dependent and can be blocked with Caffeine or U0126. To prevent polyploidy related tumor progression, BKM120 should administered only in combination with ATM or MEK inhibitors. Second, combination of PI3K/Akt/mTOR inhibitors with cytotoxic agents protects from apoptosis. The protective effect is only detectable with low PI3K/Akt/mTOR inhibitor concentrations and independent of inhibitor type or cascade level. The oncogene and NFkB target is possibly involved in apoptosis protection and inhibition of NFkB neutralizes the protective effect. PI3K/Akt/mTOR mediated Pim-2 regulation reveals a new feedback loop within the pathway. In conclusion, the complexity of the PI3K/Akt/mTOR pathway impedes therapeutic targeting.
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Efeito citotóxico causado por emulsão lipídica contendo 7-cetocolesterol (OxLE) em cultura de células tumorais / Cytotoxic effects caused by 7-ketocholesterol-containing emulsion (OXLE) in tumor cells in cultureFavero, Giovani Marino 08 March 2004 (has links)
Oxisteróis são derivados oxigenados do colesterol que podem ser formados por autoxidação ou por atividade de enzimas específicas. Eles apresentam inúmeras atividades biológicas, incluindo inibição da proliferação celular e citotoxicidade. Dentre os oxisteróis, 7-cetocolesterol (7-KC), que difere do colesterol por apresentar um grupamento cetônico no carbono 7, é conhecido por induzir morte celular por diferentes vias. Neste projeto de pesquisa pretendemos avaliar o uso de uma emulsão lipídica contendo oxisterol (OXLE) como veículo para entrega de 7-KC a células tumorais. Os efeitos tóxicos de OXLE foram avaliados nas linhagem celulares RPMI 8226 (mieloma múltiplo) e B16F10 (melanoma). As células foram tratadas com: a) uma emulsão conhecida por atuar funcionalmente como a LDL (LDE); b) OXLE (75 uM até 225 uM de 7-KC); c) 7-KC (100 uM). As distribuições das células nas diferentes fases do ciclo de duplicação celulares foram analisadas por citometria de fluxo com iodeto de propídio. Todos os tratamentos, nestas concentrações, levaram a acúmulo das células de mieloma na fase proliferativa do ciclo celular. Para analisar a resposta apoptótica, nos utilizamos o fluorocromo JC-1, que mede o potencial transmembranar mitocondrial. Ambas a linhagens celulares, quando tratadas com OXLE, apresentaram hiperpolarização do potencial transmembranar associado com progressivo decrescimento do mesmo. Os resultados mostram que os grupos OXLE e 7-KC induziram a uma morte massiva das células de mieloma. OXLE induziu um efeito citostático nas células de melanoma, caracterizadas por alterações de morfologia, com um aumento na polimerização das fibras de actina, hiperpolarização seguida de perda do potencial transmitocondrial (apoptose associada à despolarização) , presença de vacúolos autofágicos como figuras de mielina (morte celular programada do tipo II - Autofagia); e impossibilitando as células de fazer a citocinese, determinado pelo aparecimento de uma população de células poliplóides. Nossos resultados indicam que esta nova emulsão contendo um oxisterol em sua formulação não é apenas um veículo para moléculas hidrofóbicas, mas pode atuar como agente cistotático/citotóxico. A morte mediada por OXLE possui características de apoptose e autofagia. Estes resultados são promissores e o possível use de OXLE in vivo necessita de futuras investigações. / Oxysterols are oxygenated derivatives of cholesterol that may be formed by autoxidation or by action of specific enzymes. They exhibit a number of biologic activities including inhibition of cellular proliferation and cytotoxicity. Among oxysterols, 7-ketocholesterol (7-KC), that differs form cholesterol by a functional ketone group at C7, is known to induce cell death by different ways. Herein we evaluated the use of an oxysterol-containing lipid emulsion(OXLE) as a vehicle to deliver 7-KC to tumor cells. OXLE was evaluated regarding its toxicity on RPMI 8226 (multiple myeloma) and B16F10 (melanoma) cell lines. Cells were treated with: a) an emulsion known to act functionally as LDL (LDE); b) OXLE (75 uM to 225uM of 7KC); c) 7KC (100uM). The distribution of cells in the different phases of cell cycle was analyzed by flow cytometry with propidium iodide. All treatments, according to the concentrations, led the myeloma cells to an arrest in the proliferative phases of the cell cycle. To analyze the apoptotic response, we then used a fluorochrome (JC-1), which measures the mitochondrial transmembrane potential. Both cell lines when treated with OXLE had a hyperpolarization of the transmembrane potential that decrease progressively. The results showed that exposure to either OXLE or 7-KC induced a massive death of myeloma cells. OXLE had an cytostatic effect on melanoma cells characterized by morphologic alterations as increased polymerization of actin fibers; hyperpolarization followed by a dissipation of the mitochondrial transmembrane potential (apoptosis-associated depolarization); presence of IX autophagic vacuoles as myelin figures (programmed cell death type II - Autophagy); and impaired cytokinesis, as determined by the emergence of a polyploid population. Our results indicated that this new oxysterol-containing emulsion is not only a carrier for hydrophobic molecules, but it can act itself as a cytostatic/cytotoxic agent. Cell death mediated by OXLE have both apoptosis and autophagy characteristics. These results are promising and the possible use of OxLE in vivo warrants further investigation
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Disentangling the Reticulate History of Polyploids in <i>Silene </i>(<i>Caryophyllaceae</i>)Popp, Magnus January 2004 (has links)
<p>DNA sequences from the <i>rps16</i> intron and the <i>psbE-petL</i> spacer from the chloroplast genome, the ribosomal nuclear ITS region, and introns from the low copy nuclear genes <i>RPA2</i>, <i>RPB2</i>, <i>RPD2a</i> and <i>RPD2b</i>, are in different combinations used to infer phylogenetic relationships in <i>Sileneae</i> (<i>Caryophyllaceae</i>). Used in concert, the biparentally inherited nuclear regions are useful to distinguish between paralogy due to allopolyploidy and single gene duplications, respectively, because the latter are not expected to give rise to repeated phylogenetic patterns in potentially unlinked sequence regions. In addition, the sequences resolve previously poorly known relationships in the tribe <i>Sileneae</i>. Several independent losses and incomplete concerted evolution are inferred between the two <i>RPD2</i> paralogues in a subgroup of <i>Silene</i>.</p><p>An allopolyploid origin is suggested for the tetraploid <i>S. aegaea</i>, with the maternal ancestor from the diploid <i>S. pentelica</i> lineage, and the paternal contributor from the diploid <i>S. sedoides</i> lineage.</p><p><i>Silene involucrata</i> originated as an allotetraploid with the diploid lineage of Arctic <i>S. uralensis</i> as cytoplasmic donor and the diploid Siberian/Northeast Asian <i>S. ajanensis</i> lineage as pollen donor. A subsequent allopolyploidization with the <i>S. ajanensis</i> lineage as pollen donor and the tetraploid <i>S. involucrata</i> lineage as cytoplasmic donor resulted in the hexaploid lineage of <i>S. sorensenis sensu lato</i>.</p><p>A monophyletic origin of the North American polyploids is rejected. One lineage consists of tetraploid <i>S. menziesii</i> and its diploid allies. A separate lineage leads to a clade consisting of both diploid and polyploid Arctic, European and Asian taxa in addition to the majority of the North American polyploids. The tetraploid <i>S. californica</i> and the hexaploid <i>S. hookeri</i> are derived from separate allopolyploidization events between these two lineages.</p>
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Reticulate Evolution in Diphasiastrum (Lycopodiaceae)Aagaard, Sunniva Margrethe Due January 2009 (has links)
In this thesis relationships and the occurrence of reticulate evolutionary events in the club moss genus Diphasiastrum are investigated. Diphasiastrum is initially established as a monophyletic group within Lycopodiaceae using non recombinant chloroplast sequence data. Support is obtained for eight distinct parental lineages in Diphasiastrum, and relationships among the putative parent taxa in the hypothesized hybrid complexes; D. alpinum, D. complanatum, D. digitatum, D. multispicatum, D. sitchense, D. tristachyum and D. veitchii are presented. Feulgen DNA image densitometry data and sequence data obtained from three nuclear regions, RPB2, LEAFY and LAMB4, were used to infer the origins of three different taxa confirmed to be allopolyploid; D. zanclophyllum from South Africa, D. wightianum from Malaysia and an undescribed taxon from China. The two Asian polyploids have originated from two different hybrid combinations, D. multispicatum x D. veitchii and D. tristachyum x D. veitchii. Diphasiastrum zanclophyllum originates from a cross between D. digitatum and an unidentified diploid taxon. The occurrence of three homoploid hybrid combinations commonly recognized in Europe, D. alpinum x D. complanatum, D. alpinum x D. tristachyum and D. complanatum x D. tristachyum, are verified using the same three nuclear regions. Two of the three hybrid combinations are also shown to have originated from reciprocal crosses. Admixture analyses performed on an extended, dataset similarly identified predominately F1 hybrids and backcrosses. The observations and common recognition of hybrid species in the included populations are hence most likely due to frequent observations of neohybrids in hybrid zones. Reticulate patterns are, however, prominent in the presented dataset. Hence future studies addressing evolutionary and ecological questions in Diphasiastrum should emphasize the impact of gene flow between parent lineages rather than speciation as the result of hybridization.
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Disentangling the Reticulate History of Polyploids in Silene (Caryophyllaceae)Popp, Magnus January 2004 (has links)
DNA sequences from the rps16 intron and the psbE-petL spacer from the chloroplast genome, the ribosomal nuclear ITS region, and introns from the low copy nuclear genes RPA2, RPB2, RPD2a and RPD2b, are in different combinations used to infer phylogenetic relationships in Sileneae (Caryophyllaceae). Used in concert, the biparentally inherited nuclear regions are useful to distinguish between paralogy due to allopolyploidy and single gene duplications, respectively, because the latter are not expected to give rise to repeated phylogenetic patterns in potentially unlinked sequence regions. In addition, the sequences resolve previously poorly known relationships in the tribe Sileneae. Several independent losses and incomplete concerted evolution are inferred between the two RPD2 paralogues in a subgroup of Silene. An allopolyploid origin is suggested for the tetraploid S. aegaea, with the maternal ancestor from the diploid S. pentelica lineage, and the paternal contributor from the diploid S. sedoides lineage. Silene involucrata originated as an allotetraploid with the diploid lineage of Arctic S. uralensis as cytoplasmic donor and the diploid Siberian/Northeast Asian S. ajanensis lineage as pollen donor. A subsequent allopolyploidization with the S. ajanensis lineage as pollen donor and the tetraploid S. involucrata lineage as cytoplasmic donor resulted in the hexaploid lineage of S. sorensenis sensu lato. A monophyletic origin of the North American polyploids is rejected. One lineage consists of tetraploid S. menziesii and its diploid allies. A separate lineage leads to a clade consisting of both diploid and polyploid Arctic, European and Asian taxa in addition to the majority of the North American polyploids. The tetraploid S. californica and the hexaploid S. hookeri are derived from separate allopolyploidization events between these two lineages.
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Context dependency of plant – animal interactionsKönig, Malin A. E. January 2014 (has links)
The strength and direction of interactions between organisms vary spatially across the landscape. Traditionally, the focus has been on how trait variation affects the interactions between species. However, differences in abiotic and biotic environmental factors may also alter the distribution, phenology and behavior of the interacting species. To be able to understand why an interaction varies across the landscape, the effects of trait variation has to be separated from the effects of the environmental context. In this thesis, I try to separate the effects of context and trait differences on plant resistance against herbivory, through experimental and observational studies conducted with two cytotypes of the perennial herb Cardamine pratensis and its main herbivore, Anthocharis cardamines. The results show that differences in plant resistance against oviposition under controlled conditions were mainly mediated by flower size; larger flowers were more attractive to the female butterfly. However, among-populations differences in oviposition under natural conditions were not related to the resistance observed under controlled conditions, or to ploidy type, flowering phenology or plant size. Within populations under natural conditions the oviposition patterns by A. cardamines was affected by the plant traits plant size and flowering phenology. The result of this thesis shows that among-population differences in intensity of plant-herbivore interactions were caused by differences in environmental context rather than by herbivore preferences for any phenotypic plant traits, while host plant selection within population was based on plant traits. This suggests that biotic and biotic context can have important effects on the intensity of plant-herbivore interactions. Although genetic traits influenced the outcome of the interaction within populations, it was the environmental context of the populations that determined largely if the interaction took place or not. / <p>At the time of the doctoral defence the following papers were unpublished and had a status as follows: Paper 2: Manuscript in review in Plos One; Paper 3: Manuscript in review in <em></em>Ecological Entomology; Paper 4: Manuscript</p>
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Establishing genetic diversity of Rwanda highland banana using random amplified polymorphic DNA markers.Nsabimana, Antoine. January 2006 (has links)
The characterization of the banana germplasm collection from Rubona - Rwanda was
investigated using morphological and cytological characteristics of the genomic groups.
Genetic diversity was assessed using Random Amplified Polymorphic DNA analysis.
The survey was conducted to evaluate the distribution of banana cultivars in the four
major growing regions of Rwanda.
A total of 90 accessions from the National Banana Germplasm Collection at Rubona
Rwanda were characterized and six characters of the fingers (length, width, weight,
green life, post green life and length/width ratio) were subjected to principal component
analysis (PCA). The cooking and beer clones were separated. The cooking clones were
further grouped into three clone sets: Musakala, Nakabululu, and one that constitutes
Nakitembe and Nfuuka clone sets. The AAB genomic group was separated from AAA,
AB and ABB genomic groups.
The results from the survey showed that East African Highland bananas are the most
important genotype group in the four major banana growing regions of Rwanda ranging
between 60 - 90% of banana mats counted. Several new Highland banana cultivars
were recorded, such as 'Intokatoke', 'Igihuna', 'Ingenge', 'Ingaju', 'Icyerwa', 'Mitoki',
'Madamu', 'Inkokobora', 'Intokekazi', 'Bugoyi', 'Ishoki'. Amongst these cultivars, some
were classified as cooking and others as brewing bananas. However, in the National
Banana Germplasm Collection at Rubona - Rwanda, the uses of these cultivars are
recorded differently therefore increasing the need for agro-morphological
characterization.
The assessment of ploidy level of accessions from the National Banana Germplasm
Collection at Rubona - Rwanda, by flow cytometry showed misclassification of some
accessions such as 'Pomme', 'Kamaramasenge', 'Gisubi kayinja', 'Gisubi kagongo', and
'Dibis' which were classified as diploid, diploid, triploid, and tetraploid respectively. They
IV
were found to be triploid, triploid, triploid, diploid and triploid. All these bananas were
recently introduced into Rwanda, while the endemic Highland bananas were triploid.
The genomic group and genetic similarities of 49 accessions were investigated using
Random Amplified Polymorphic DNA markers. The genomic group of bananas
assessed were established using OPA-18 (PILLAY et al., 2000) and OPG-17 primers.
These primers showed bands 441 and 443 base pairs (bp) respectively for the
accessions having only the B genome. Whilst they were absent for the accessions
"
having an A genome. The genetic similarity was estimated via a Simple Matching
coefficient which showed the lowest value 0.46 measured between 'Ingumba' and
'Ishika 'and the highest value of 0.85 between 'Kirayenda' and 'Inyabukuwe'. The data
of matrix of coefficient of similarity was subjected to cluster analysis with unweighted
pair group method with arithmetic average (UPGMA). Each accession was clearly
separated demonstrating the usefulness of RAPDs in analysis of genetic diversity. The
results of this study are very important to the Curator of the banana germplasm
collection in Eastern Central Africa and for the future breeding of this crop. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.
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Cooperative Oncogenesis and Polyploidization in Human Cancers: A DissertationHeilman, Susan Ann 09 May 2007 (has links)
A common phenotype observed in most cancers is chromosomal instability. This includes both structural and numerical chromosomal aberrations, which can promote carcinogenesis. The fusion gene CBFB/MYH11 is created by the structural chromosomal inversion(16)(p13.1q22), resulting in the fusion protein CBFβ-SMMHC, which blocks differentiation in hematopoietic progenitor cells. This mutation alone, however, is not sufficient for transformation, and at least one additional cooperating mutation is necessary.
The role of wildtype Cbfb in modulating the oncogenic function of the fusion protein Cbfβ-SMMHC in mice was examined. Transgenic mice expressing the fusion protein, but lacking a wild-type copy of Cbfb, were created to model the effects of these combined mutations. It was found that wild-type Cbfb is necessary for maintaining normal hematopoietic differentiation. Consequently, complete loss of wild-type Cbfb accelerates leukemogenesis in Cbfb/MYH11 mice compared to mice expressing both the fusion and wild-type proteins. While there is no evidence in human patient samples that loss of wild-type Cbfb expression cooperates with the fusion protein to cause transformation, it is apparent from these experiments that wild-type Cbfβ does play a role in maintaining genomic integrity in the presence of Cbfβ-SMMHC. Experiments have also shown that loss of Cbfb leads to accumulation of hematopoietic progenitor cells, which may acquire additional cooperating mutations.
Not unlike CBFB/MYH11, the human papillomavirus (HPV) E6 and E7 proteins are not sufficient for cellular transformation. Instead, high risk HPV E7 causes numerical chromosomal aberrations, which can lead to accumulation of additional cooperating mutations. Expression of HPV-16 E7 and subsequent downregulation of the retinoblastoma protein (Rb) has been shown to induce polyploidy in human keratinocytes. Polyploidy predisposes cells to aneuploidy and can eventually lead to transformation in HPV positive cells.
There are several possible mechanisms through which E7 may lead to polyploidization, including abrogation of the spindle assembly checkpoint, cleavage failure, abrogation of the postmitotic checkpoint, and re-replication. Rb-defective mouse and human cells were found to undergo normal mitosis and complete cytokinesis. Furthermore, DNA re-replication was not found to be a major mechanism to polyploidization in HPV-E7 cells upon microtubule disruption. Interestingly, upon prolonged mitotic arrest, cells were found to adapt to the spindle assembly checkpoint and halt in a G1-like state with 4C DNA content. This post-mitotic checkpoint is abrogated by E7-induced Rb-downregulation leading to S-phase induction and polyploidy.
This dissertation explores two examples of the multi-step pathway in human cancers. While certain genes or genetic mutations are often characteristic of specific cancers, those mutations are often not sufficient for transformation. The genetic or chromosomal abnormalities that they produce often stimulate the additional mutations necessary for oncogenesis. The studies with Cbfb/MYH11 and HPV E7 further exemplify the significance of numerical and structural chromosomal aberrations in multi-step carcinogenesis.
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Taxonomic study of the Eurasian taxa of Tortula muralis (Pottiaceae, Musci) complex / Taxonomic study of the Eurasian taxa of Tortula muralis (Pottiaceae, Musci) complexKOŠNAR, Jiří January 2014 (has links)
The thesis aims at clarifying the taxonomic difficulties in the Tortula muralis complex. For the first time, the group was studied by means of morphometric analysis, ploidy level assessment by flow cytometry, and molecular analysis based on sequencing of ITS region of nuclear ribosomal DNA. Morphometric and cytometric studies found only two well distinguished groups within the complex. These groups corresponded to the markedly variable species T. muralis and to the rather uniform species T. lingulata. Variability in ITS sequences suggested extensive gene flow among some of traditional morphologically defined taxa of the complex. Multiple polytopic autopolyploid origin of polyploids was revealed in some taxa. Changes in taxonomic conception of T. muralis complex were proposed in order to reflect structure of morphological, karyological, and DNA variability in the group. Natural hybridization is probably an important evolutionary mechanism that generated morphological diversity and taxonomic complexity in the mosses of T. muralis complex.
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Mapeamento genético de marcadores SNPs (Single Nucleotide Polymorphisms) em cana-de-açúcar (Saccharum spp.) / Genetic mapping of SNPs markers (Single Nucleotide Polymorphisms) in sugarcane (Saccharum spp.)Jardim, Priscila Magalhães da Veiga 30 July 2018 (has links)
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Previous issue date: 2018-07-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Sugarcane is an important culture quite relevant to the Brazilian economy. The production is growing as well as a cultivated area is increasing every year. The genome of this culture is still deficient due to complications such as high ploidy and the big genome that presents. Among the different genetic characterization studies, the development of genetic maps is important for providing information about the genome structure of a species. In addition, it can help develop the techniques of interpretation and use of genetic information. They provide more understanding of how genetic information is organized in the genome of sugarcane supplying a lack in the basic element in this culture. The maps constructed for sugarcane, so far, not shown saturated. This work was obtained a map for sugarcane using SNP markers based on genotyping-by-sequencing technology in next-generation sequencing using the target sequencing strategy (RAPiD-Seq). For obtaining the map, 103 clones RB97327 and RB72454 were used. Probes were designed based on sequence similarity using the sorghum genome as a reference. The construction of the binding groups, considering as a binding criterion of a recombination fraction equal 0.20; allowed the identification of 249 binding groups for the biparental population with 1: 1 segregation. A total of 20555 polymorphic were scored in the analysis. The sum of the average sizes of homeologia groups identified, using the sorghum genome as a reference, was 3964.68 cM for the female parent and 3797.05 cM for the male parent. / A cana-de-açúcar é uma cultura de importância bastante relevante para a economia brasileira. A produção de cana-de-açúcar no Brasil é crescente assim como a área cultivada vem aumentando a cada ano. A compreensão do genoma da cana-de-açúcar ainda é deficiente devido a complicações como alta ploidia e o grande genoma que a cultura apresenta. Dentre os diferentes estudos de caracterização genética, o desenvolvimento de mapas genéticos é importante por fornecer informações acerca da estrutura do genoma de uma espécie. Além disso, pode auxiliar no desenvolvimento das técnicas de interpretação e uso da informação genética. Eles possibilitam a compreensão mais abrangente da organização da informação genética no genoma da cana-de-açúcar suprindo uma carência do estudo básico sobre essa cultura. Os mapas construídos para cana-de-açúcar, até agora, não se mostraram completos. Neste trabalho foi obtido um mapa de ligação para cana-de-açúcar utilizando marcadores SNPs baseados na tecnologia de genotipagem por sequenciamento de nova geração utilizando a estratégia de target sequencing (RAPiD-Seq). Para a obtenção do mapa foram utilizados 103 genótipos obtidos do cruzamento entre os clones RB97327 e RB72454. Foram desenhadas sondas baseadas em sequenciamento de semelhança utilizando o genoma de sorgo como referência. A construção dos grupos de ligação, considerando-se como critério de ligação uma fração de recombinação de 0,20; permitiu a identificação de 249 grupos de ligação para a população biparental com segregação 1:1. Foram consideradas 20555 marcas polimórficas nas análises de construção do mapa de ligação. A soma dos tamanhos médios dos grupos de homeologia identificados, utilizando-se o genoma de sorgo como referência, foi de 3964,68 cM para o genitor feminino e de 3797,05 cM para o genitor masculino.
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