RESUMO
Fluorescent in situ hybridization (FISH) with multiple probes was used to analyze mitotic and meiotic chromosome spreads of Avena sativa cv 'Sun II' monosomic lines, and of A. byzantina cv 'Kanota' monosomic lines from spontaneous haploids. The probes used were A. strigosa pAs120a (a repetitive sequence abundant in A-genome chromatin), A. murphyi pAm1 (a repetitive sequence abundant in C-genome chromatin), A. strigosa pITS (internal transcribed spacer of rDNA) and the wheat rDNA probes pTa71 (nucleolus organizer region or NOR) and pTa794 (5S). Simultaneous and sequential FISH employing pairs of these probes allowed the identification and genome assignation of all chromosomes. FISH mapping using mitotic and meiotic metaphases facilitated the genomic and chromosomal identification of the monosome in each line. Of the 17 'Sun II' lines analyzed, 13 distinct monosomic lines were found, corresponding to four monosomes of the A-genome, five of the C-genome and four of the D-genome. In addition, 12 distinct monosomic lines were detected among the 20 'Kanota' lines examined, corresponding to six monosomes of the A-genome, three of the C-genome and three of the D-genome. The results show that 19 chromosomes out of 21 of the complement are represented by monosomes between the two genetic backgrounds. The identity of the remaining chromosomes can be deduced either from one intergenomic translocation detected on both 'Sun II' and 'Kanota' lines, or from the single reciprocal, intergenomic translocation detected among the 'Sun II' lines. These results permit a new system to be proposed for numbering the 21 chromosome pairs of the hexaploid oat complement. Accordingly, the A-genome contains chromosomes 8A, 11A, 13A, 15A, 16A, 17A and 19A; the C-genome contains chromosomes 1C, 2C, 3C, 4C, 5C, 6C and 7C; and the D-genome consists of chromosomes 9D, 10D, 12D, 14D, 18D, 20D and 21D. Moreover, the FISH patterns of 16 chromosomes in 'Sun II' and 15 in 'Kanota' suggest that these chromosomes could be involved in intergenomic translocations. By comparing the identities of individually translocated chromosomes in the two hexaploid species with those of other hexaploids, we detected different types of intergenomic translocations.
Assuntos
Avena/genética , Cromossomos de Plantas/classificação , Cromossomos de Plantas/genética , Hibridização in Situ Fluorescente , Monossomia/genética , Terminologia como Assunto , Avena/citologia , Mapeamento Cromossômico , Haploidia , Cariotipagem , Metáfase , Poliploidia , Sequências Repetitivas de Ácido Nucleico/genéticaRESUMO
Sequences analogous to plant resistance genes of the NBS-LRR class were cloned from the genomic DNA of 11 Avena species with different genomes and levels of ploidy. Three pairs of degenerate primers were used, based on conserved DNA sequence motifs belonging to the NBS domain, and 33 sequences were identified. These were subdivided into 7 classes depending on nucleotide sequence identity. Despite the high level of degeneracy, the primers behaved in a highly selective way; the majority of sequences from the different species obtained with every primer combination showed strong identity and were considered homologous. For most species, only one sequence of each class was identified in each genome, suggesting that duplicated sequences are fairly divergent. The strong identity among specific NBS sequences precludes any conclusions being made on the evolution of these species. The genomic organization of the RGA sequences was explored using those of A. strigosa as probes in Southern blots involving digested DNA from 15 Avena species. The hybridization patterns showed wide diversity both among sequences within a species and among species for each sequence. However, the dendrogram generated using the RFLPs showed relationships among species to be in good agreement with those previously established using other molecular markers.
Assuntos
Genes de Plantas/genética , Doenças das Plantas/genética , Poaceae/classificação , Poaceae/genética , Clonagem Molecular , Genoma de Planta , Filogenia , Polimorfismo de Fragmento de RestriçãoRESUMO
Degenerate primers based on conserved regions of the nucleotide binding site (NBS) domain (encoded by the largest group of cloned plant disease resistance genes) were used to isolate a set of 15 resistance gene analogs (RGA) from the diploid species Avena strigosa Schreb. These were grouped into seven classes on the basis of 60% or greater nucleic acid sequence identity. Representative clones were used for genetic mapping in diploid and hexaploid oats. Two RGAs were mapped at two loci of the linkage group AswBF belonging to the A. strigosa x A. wiestii Steud map, and ten RGAs were mapped at 15 loci in eight linkage groups belonging to the A. byzantina C. Koch cv. Kanota x A. sativa L. cv. Ogle map. A similar approach was used for targeting genes encoding receptor-like kinases. Three different sequences were obtained and mapped to two linkage groups of the hexaploid oat map. Associations were explored between already known disease resistance loci mapped in different populations and the RGAs. Molecular markers previously linked to crown rust and barley yellow dwarf resistance genes or quantitative trait loci were found in the Kanota x Ogle map linked to RGAs at a distance ranging from 0 cM to 20 cM. Homoeologous RGAs were found to be linked to loci either conferring resistance to different isolates of the same pathogen or to different pathogens. This suggests that these RGAs identify genome regions containing resistance gene clusters.
Assuntos
Mapeamento Cromossômico , Imunidade Inata/genética , Doenças das Plantas/genética , Poaceae/genética , Sequência de Aminoácidos , Sequência de Bases , Sítios de Ligação/genética , Cruzamento/métodos , Análise por Conglomerados , Cruzamentos Genéticos , Marcadores Genéticos/genética , Dados de Sequência Molecular , Técnicas de Amplificação de Ácido Nucleico , Doenças das Plantas/microbiologia , Polimorfismo de Fragmento de Restrição , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
Fluorescent in situ hybridization (FISH) employing multiple probes was used with mitotic or meiotic chromosome spreads of Avena sativa L. cv. SunII and its monosomic lines to produce physical chromosome maps. The probes used were Avena strigosa pAs120a (which hybridizes exclusively to A-genome chromosomes), Avena murphyi pAm1 (which hybridizes exclusively to C-genome chromosomes), A. strigosa pAs121 (which hybridizes exclusively to A- and D-genome chromosomes), and the wheat rDNA probes pTa71 and pTa794. Simultaneous and sequential FISH employing two-by-two combinations of these probes allowed the unequivocal identification and genome assignation of all chromosomes. Ten pairs were found carrying intergenomic translocations: (i) between the A and C genomes (chromosome pair 5A); (ii) between the C and D genomes (pairs 1C, 2C, 4C, 10C, and 16C); and (iii) between the D and C genomes (pairs 9D, 11D, 13D, and 14D). The existence of a reciprocal intergenomic translocation (10C-14D) is also proposed. Comparing these results with those of other hexaploids, three intergenomic translocations (10C, 9D, and 14D) were found to be unique to A. sativa cv. SunII, supporting the view that 'SunII' is genetically distinct from other hexaploid Avena species and from cultivars of the A. sativa species. FISH mapping using meiotic and mitotic metaphases facilitated the genomic and chromosomal identification of the aneuploid chromosome in each monosomic line. Of the 18 analyzed, only 11 distinct monosomic lines were actually found, corresponding to 5 lines of the A genome, 2 lines of the C genome, and 4 lines of the D genome. The presence or absence of the 10C-14D interchange was also monitored in these lines.
Assuntos
Avena/genética , DNA de Plantas/genética , Sequências Repetitivas de Ácido Nucleico , Cromossomos de Plantas , Sondas de DNA , Hibridização in Situ FluorescenteAssuntos
Adulto , Humanos , Feminino , Anafilaxia , Anestesia , Fatores Inibidores da Migração de LeucócitosAssuntos
Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Humanos , Masculino , Feminino , Halotano , Histamina , Basófilos , Fatores Inibidores da Migração de LeucócitosAssuntos
9,10-Dimetil-1,2-benzantraceno/efeitos adversos , Neoplasias da Mama/tratamento farmacológico , Metilnitrosoureia/efeitos adversos , Compostos de Nitrosoureia/efeitos adversos , Prolactina/metabolismo , Tamoxifeno/uso terapêutico , Animais , Neoplasias da Mama/induzido quimicamente , Feminino , Ratos , Sulpirida/farmacologiaRESUMO
Se estudia la respuesta inmunologica de catorce pacientes portadores de cancer de mama, mediante tests cutaneos y el analisis de los ganglios regionales (axilares) y alejados (inguinales). Se dividen en cuatro grupos: el primero, de cuatro mastectomizadas de metastasis axiliares, en quienes el predominio linfocitico (PL) estuvo presente en todas, y la histiocitosis sinusal (HS) en la mitad. Un segundo grupo de dos mastectomizadas en metastasis axilares, en donde el ganglio inguinal mostro marcada HS y PL en yna u centros germinales (CG) en la otra. Un tercer grupo de siete mastectomizadas con metastasis axilares y en tratamiento complementario con poliquimioterapia (CMF 48) en quienes los ganglios inguinales presentaron HS en tres PL en tres, una con HS hialinosis y anestimulacion con otra. Un cuarto grupo de dos mastectomizadas e irradiadas, los ganglios inguinales presentaron HS muy pobre y PL en la otra. Se observa un predominio de respuesta mediada por linfocitos T (area paracortical - hiperplasia sinusal), seguida de la de celulas B, concordante con la favorable evolucion
Assuntos
Adulto , Pessoa de Meia-Idade , Idoso , Humanos , Feminino , Linfonodos , Imunidade Celular , Neoplasias da MamaRESUMO
Se estudia la respuesta inmunologica de catorce pacientes portadores de cancer de mama, mediante tests cutaneos y el analisis de los ganglios regionales (axilares) y alejados (inguinales). Se dividen en cuatro grupos: el primero, de cuatro mastectomizadas de metastasis axiliares, en quienes el predominio linfocitico (PL) estuvo presente en todas, y la histiocitosis sinusal (HS) en la mitad. Un segundo grupo de dos mastectomizadas en metastasis axilares, en donde el ganglio inguinal mostro marcada HS y PL en yna u centros germinales (CG) en la otra. Un tercer grupo de siete mastectomizadas con metastasis axilares y en tratamiento complementario con poliquimioterapia (CMF 48) en quienes los ganglios inguinales presentaron HS en tres PL en tres, una con HS hialinosis y anestimulacion con otra. Un cuarto grupo de dos mastectomizadas e irradiadas, los ganglios inguinales presentaron HS muy pobre y PL en la otra. Se observa un predominio de respuesta mediada por linfocitos T (area paracortical - hiperplasia sinusal), seguida de la de celulas B, concordante con la favorable evolucion
Assuntos
Adulto , Pessoa de Meia-Idade , Humanos , Feminino , Neoplasias da Mama , Imunidade Celular , LinfonodosAssuntos
Antineoplásicos/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica , Neoplasias da Mama/tratamento farmacológico , Neoplasias Ovarianas/tratamento farmacológico , Antineoplásicos/efeitos adversos , Neoplasias da Mama/patologia , Compartimento Celular/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Ciclofosfamida/administração & dosagem , Ciclofosfamida/uso terapêutico , Doxorrubicina/administração & dosagem , Doxorrubicina/uso terapêutico , Esquema de Medicação , Quimioterapia Combinada , Feminino , Fluoruracila/administração & dosagem , Fluoruracila/uso terapêutico , Humanos , Metotrexato/administração & dosagem , Metotrexato/uso terapêutico , Neoplasias Ovarianas/patologiaRESUMO
Se hace un analisis de la modalidad operativa de la quimioterapia en cancer de mama y ovario a la luz de los actuales conocimientos sobre historia natural y biologia de los tumores. Se pasa revista a la evolucion que han seguido los tratamientos quimioterapicos de estas dos patologias y sus perspectivas con las actuales drogas, considerando su efectividad y reduciendo los efectos colaterales.El material comprende 65 carcinomas de mama y 17 canceres de ovario de los cuales fueron tratados como adyuvantes a la cirugia de mama en 30 pacientes, y en 7 casos adyuvantes a la cirugia del ovario. En 35 casos de canceres avanzados de mama y en 10 cancerosos avanzados de ovario. En mama utilizamos CMF o FAC 36 cada 21 dias y en ovario alquilante o asociados a adriamicina. Las respuestas fueron parciales en 40% y totales en 26% de los canceres de mama avanzados, de 70% las reducciones parciales del cancer de ovario avanzado. Se concluye que la quimio es una de las bases del tratamiento de los tumores genitomamarios
Assuntos
Antineoplásicos , Neoplasias da Mama , Neoplasias Ovarianas , Ciclofosfamida , Fluoruracila , MetotrexatoRESUMO
Se hace un analisis de la modalidad operativa de la quimioterapia en cancer de mama y ovario a la luz de los actuales conocimientos sobre historia natural y biologia de los tumores. Se pasa revista a la evolucion que han seguido los tratamientos quimioterapicos de estas dos patologias y sus perspectivas con las actuales drogas, considerando su efectividad y reduciendo los efectos colaterales.El material comprende 65 carcinomas de mama y 17 canceres de ovario de los cuales fueron tratados como adyuvantes a la cirugia de mama en 30 pacientes, y en 7 casos adyuvantes a la cirugia del ovario. En 35 casos de canceres avanzados de mama y en 10 cancerosos avanzados de ovario. En mama utilizamos CMF o FAC 36 cada 21 dias y en ovario alquilante o asociados a adriamicina. Las respuestas fueron parciales en 40% y totales en 26% de los canceres de mama avanzados, de 70% las reducciones parciales del cancer de ovario avanzado. Se concluye que la quimio es una de las bases del tratamiento de los tumores genitomamarios
