Colchicine, an antimicrotubule agent, has enhanced the androgenic response of cruciferous and graminaceous species. Our objectives were to determine if colchicine treatments enhanced androgenic embryo production in diploid potato, and if colchicine treatments during anther culture affected the ploidy of anther-derived plants. An additional study, analysis of genetic identity among regenerated anther-derived plants based on RAPD markers, was also conducted. Anther culture of a hybrid between <i>Solanum chacoense</i> and <i>S. phureja</i> (clone CP2) was conducted with five colchicine treatments (0, 25, 50, 100, and 200 mg/L) for 24 h. Anthers containing late uninucleate pollen were cultured on modified liquid LS medium. A mean of 0.25 embryos per anther was obtained; however, there was no significant difference among the colchicine treatments, which ranged from a mean of 0.17 to 0.28 embryos per anther for 200 mg/L colchicine and control, respectively. In total 56 plants were regenerated of which 11% were monoploid. The second experiment using the same colchicine treatments was conducted with 54 hybrids derived by crossing six anther-derived doubled monoploids with three heterozygous pollinators. A mean of 1.78 embryos per anther was obtained; again, there was no significant difference among the colchicine treatments, which ranged from 1.27 to 2.75 for 50 mg/L colchicine and control, respectively. In total 679 plants were regenerated. The 312 plantlets for which ploidy was determined, 75% were monoploid. The third experiment was conducted to study various durations (0, 90 sec vacuum infiltration, 24, 48, and 72 h) of high colchicine treatment (200 mg/L) applied to <i>S. phureja</i> family DM 13-14 202 x ID 5. There was nearly significant difference among treatments (α = 0.05). Mean embryos per anther ranged from 0.96 to 1.90 for 48 h and 90 sec vacuum infiltration, respectively. In an additional study, 26 regenerated anther-derived monoploid plants, from the DM 13-14 202 x ID 5 family, from the second experiment, were genetically characterized based on RAPD markers. Forty-three loci were scored from 13 primers for groups of monoploids derived from the same flasks of anther culture. From one flask, two pairs of monoploids were found to be genetically identical. From a second flask 6 of 7 were genetically identical, and from a third 3 of 7 were genetically identical. The presence of genetically identical individuals within three monoploid populations indicates the prevalence of secondary embryogenesis during anther culture such that a single embryogenic microspore can generate many anther-derived plants. / Master of Science

Orientador: Robinson Antonio Pitelli / Banca: Janete Apparecida Desidério Sena / Banca: Ricardo Victoria Filho / Resumo: Estudos laboratoriais foram realizados com o objetivo de estudar a caracterização genética de plantas de Euphorbia heterophylla, provenientes de áreas em que a resistência aos herbicidas inibidores da ALS (Acetolactase) estava caracterizada (Santo Ângelo, RS), em processo de desenvolvimento (Sonora, MS) e onde nunca havia sido aplicado herbicida com este mecanismo de ação (Jaboticabal, SP). Testes preliminares comprovaram elevada resistência para as plantas provenientes de Santo Ângelo, moderada resistência para plantas de Sonora e elevada susceptibilidade para plantas provenientes de Jaboticabal. A análise dos resultados através das isoenzimas revelou que existem pequenas diferenças entre as três populações estudadas. A análise do DNA dos indivíduos das diferentes populações através de marcadores moleculares RAPD, permitiu a construção do Dendrograma de Cluster, que mostra uma similaridade mínima de 88% e máxima de 99% entre os diferentes indivíduos, quer pertencentes à mesma população, quer pertencentes às diferentes populações. Tal análise permitiu inferir que, apesar dos indivíduos analisados mostrarem grande similaridade entre si, apresentam variabilidade genética entre os indivíduos e as populações estudadas, de acordo com a conclusão obtida quando da utilização das isoenzimas. Crescendo em condições similares, o biótipo da Santo Ângelo apresentou maior absorção de fósforo em comparação com os demais, e maior absorção de potássio em relação ao biótipo de Jaboticabal. Comparando a densidade estomática, houve diferença estatística entre os três biótipos, sendo maior para o biótipo mais tolerante ao herbicida e menor para o susceptível. Não foi possível estabelecer qualquer relação confiável entre as características morfológicas e de crescimento das plantas e a resistência ao imazethapyr. / Abstract: Laboratory studies were carried out aim to characterize genetically Euphorbia heterophylla plants were collect in three regions. At Santo Ângelo (RS) region this plant is highly resistant to ALS inhibitors herbicides, but moderately at Sonora (MS) and susceptible at Jaboticabal region. Greenhouse tests confirmed the plants reaction in face of imazethapyr spraying. The isoenzymes studies showed small differences between the three populations. The DNA analysis using molecular markers make feasible the Cluster dendrogram showing 88-99% of similarity comparing plants, regardless the plant origin. Besides the high similarity index between the plants, it was possible to determine lower genetic variation in Jaboticabal and Santo Ângelo populations using isoenzymes technique. The nitrogen and potassium contents in the plants shoot was higher in the Santo Ângelo byotipe, although there was no difference between the K contents when the Jaboticabal and Sonora byotipes were compared. The stomata and trichomes densities decreased in the same order of the plant tolerance to herbicide: Santo Ângelo > Sonora > Jaboticabal. None correlation between biotype resistance to imazethapyr and the plant morphology features for the three biotypes studied. The differences in the plant feature may be attributable to the adaptative mechanism of the plant to the regional characteristics they were collected. / Mestre

Experiments were conducted to determine the outcrossing rate, the inheritance of markers and establish a basic linkage map in grasspea, <i> Lathyrus sativus </i>L. The outcrossing rate in a white-flowered line of grasspea ranged from 1.7 to 2.7% among eight combinations of gene frequency and location. The outcrossing rate in this study (2.2 ± 0.7%) suggests that individual lines of grasspea should be maintained in isolation to maintain their genetic integrity. Inheritance and linkage were determined for one morphological, 11 isozyme and 72 RAPD markers in five F₂ populations (all RAPD markers were in one F₂ population). The inheritance of flower colour was monogenic with colour dominant over white. The isozymes, ACO-1, ACO-2, AAT-1, AAT-2, EST-6, FDH, LAP-1, PGD-2, SKDH and TPI-1, were codominantly expressed with monogenic inheritance. The isozymes LAP-1 and PGD-2 segregated in a non-Mendelian ratios in the crosses PI 426891.1.3 x PI 283564c.3.2 and PI 426891.1 x PI 172930.4, respectively. The isozymeEST-3 was monogenically inherited and dominantly expressed. Most RAPD markers segregated in a 3:1 ratio. Marker UBC368_425/655 segregated in a co-dominant fashion. The RAPD markers UBC304₈₃₁, UBC304₉₆₄, UBC308₉₉₀, UBC322₁₄₃₂, UBC328₈₃₁, UBC332₁₁₁₈, UBC3321₁₅₈₁, UBC333₆₁₇, UBC349<sub>752<?sub>, UBC365₁₀₁₃ and UBC388₄₅₉ showed distorted segregation. In two F₂ populations, PI 283564c.3 x PI 426885.2 and PI 358601.5 x PI 173714.5, a linkage between AAT-2 and SKDH was reconfirmed. In the cross PI 426891.1.3 x PI 283564c.3.2, one morphological, three isozyme and 71 RAPD markers were mapped resulting in the delineation of 14 linkage groups including 69 markers (1 morphological, 3 isozyme and 65 RAPD markers). The total genome length covered by these 75 markers (69 linked and six unlinked) was about 864 cM. Considering cost, simplicity and abundance, RAPD analysis was more efficient than isozyme analysis in developing linkage map.

Pinus nigra subsp. pallasiana is one of the most widespread and economically important forest tree species in Turkey. Primary objective of the present study was to to reveal the effects of forestry practices by determining genetic diversity of natural and managed seed sources by means of RAPD markers. Secondly, two varieties were also investigated to reveal their pattern of genetic variation. Seed stands, seed orchards and plantations were screened against 11 RAPD primers and generated 152 polymorphic DNA loci. Two varieties were compared with a reference seed source and 4 natural seed sources. Seven primers generated 66 polymorphic DNA loci. An overall average for effective number of alleles was 1.68&plusmn / 0.030 / observed heterozygosity was 0.49&plusmn / 0.024 / expected heterozygosity was 0.38&plusmn / 0.014 and proportion of polymorphic loci was 93% for all seed sources considered. Results revealed that there was no considerable variation between seed source categories but some degree of variation was observed within seed orchards and plantations. Mean FST value estimated for the natural populations revealed that 94% of the total genetic variation was within populations. Nei&rsquo / s genetic distance values were also estimated for seed source categories (0.03-0.14). Nevertheless, varieties&rsquo / genetic distance values were considerably higher than other natural seed sources (0.07-0.19). Their dendrogram also claimed that two varieties are genetically different from natural populations. The extent of genetic diversity explored by RAPD markers revealed that forestry practices caused no major changes in the managed populations with respect to natural populations. Moreover, further study is needed to illustrate genetic divergence of varieties.

Made available in DSpace on 2014-06-11T19:26:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-04-18Bitstream added on 2014-06-13T19:13:01Z : No. of bitstreams: 1 amaral_al_me_jabo.pdf: 755163 bytes, checksum: 38fef006637923d725907da11533302e (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Estudos laboratoriais foram realizados com o objetivo de estudar a caracterização genética de plantas de Euphorbia heterophylla, provenientes de áreas em que a resistência aos herbicidas inibidores da ALS (Acetolactase) estava caracterizada (Santo Ângelo, RS), em processo de desenvolvimento (Sonora, MS) e onde nunca havia sido aplicado herbicida com este mecanismo de ação (Jaboticabal, SP). Testes preliminares comprovaram elevada resistência para as plantas provenientes de Santo Ângelo, moderada resistência para plantas de Sonora e elevada susceptibilidade para plantas provenientes de Jaboticabal. A análise dos resultados através das isoenzimas revelou que existem pequenas diferenças entre as três populações estudadas. A análise do DNA dos indivíduos das diferentes populações através de marcadores moleculares RAPD, permitiu a construção do Dendrograma de Cluster, que mostra uma similaridade mínima de 88% e máxima de 99% entre os diferentes indivíduos, quer pertencentes à mesma população, quer pertencentes às diferentes populações. Tal análise permitiu inferir que, apesar dos indivíduos analisados mostrarem grande similaridade entre si, apresentam variabilidade genética entre os indivíduos e as populações estudadas, de acordo com a conclusão obtida quando da utilização das isoenzimas. Crescendo em condições similares, o biótipo da Santo Ângelo apresentou maior absorção de fósforo em comparação com os demais, e maior absorção de potássio em relação ao biótipo de Jaboticabal. Comparando a densidade estomática, houve diferença estatística entre os três biótipos, sendo maior para o biótipo mais tolerante ao herbicida e menor para o susceptível. Não foi possível estabelecer qualquer relação confiável entre as características morfológicas e de crescimento das plantas e a resistência ao imazethapyr. / Laboratory studies were carried out aim to characterize genetically Euphorbia heterophylla plants were collect in three regions. At Santo Ângelo (RS) region this plant is highly resistant to ALS inhibitors herbicides, but moderately at Sonora (MS) and susceptible at Jaboticabal region. Greenhouse tests confirmed the plants reaction in face of imazethapyr spraying. The isoenzymes studies showed small differences between the three populations. The DNA analysis using molecular markers make feasible the Cluster dendrogram showing 88-99% of similarity comparing plants, regardless the plant origin. Besides the high similarity index between the plants, it was possible to determine lower genetic variation in Jaboticabal and Santo Ângelo populations using isoenzymes technique. The nitrogen and potassium contents in the plants shoot was higher in the Santo Ângelo byotipe, although there was no difference between the K contents when the Jaboticabal and Sonora byotipes were compared. The stomata and trichomes densities decreased in the same order of the plant tolerance to herbicide: Santo Ângelo > Sonora > Jaboticabal. None correlation between biotype resistance to imazethapyr and the plant morphology features for the three biotypes studied. The differences in the plant feature may be attributable to the adaptative mechanism of the plant to the regional characteristics they were collected.

Os aspectos da pesca do espada, Trichiurus lepturus (Linnaeus, 1758) (Trichiuridae), foram descritos e analisados para as artes de pesca artesanal de linha-de-mão em Arraial do Cabo (RJ) e cerco flutuante em Porto Belo (SC), e da pesca comercial de arrasto de fundo com parelhas na costa de São Paulo. Verificaram-se tendências crescentes na produção da espécie, principalmente na pesca artesanal. Foram utilizados marcadores moleculares tipo RAPD para analisar a estrutura genético-populacional da espécie com exemplares de Belém (PA), Natal (RN), Arraial do Cabo (RJ), Guarujá (SP), Porto Belo (SC) e Rio Grande (RS), evidenciando-se uma população distinta em Belém, enquanto as demais não apresentaram estruturação definida. A dinâmica de populações foi analisada para exemplares de Arraial do Cabo, São Paulo e Porto Belo, coletados mensalmente de janeiro/2002 a julho/2003. A desova da espécie é parcelada com período reprodutivo ocorrendo do verão ao início do inverno. As fêmeas maduras migram para a plataforma encontrando-se com os machos maduros que ali permanecem, retornando à costa após a desova. O comprimento médio de primeira maturação sexual variou de 647 a 670mm (Lt) para fêmeas, e de 526 a 650mm para machos. As estimativas dos parâmetros de crescimento foram as seguintes: para fêmeas Loo de 1740 a 2010mm; k de 0,12 a 0,15/ano e t0 de -1,642 a -2,517 anos; para machos Loo de 1373 a 1580mm; k de 0,17 a 0.25/ano e t0 de -1,662 a -1,866 anos. A longevidade foi cerca de 14 anos. As taxas de mortalidade natural variaram de 0,20 a 0,29/ano. O modelo de rendimento relativo por recruta indicou que as taxas de mortalidade por pesca do espada estão próximas ou já ultrapassam os limites do rendimento máximo sustentável. / The fishery aspects of the cutlassfish, Trichiurus lepturus (Linnaeus, 1758) (Trichiuridae), were analyzed for the artisanal fishery of line and hook of Arraial do Cabo (RJ) and floating encirclement of Porto Belo (SC), and of the commercial paired bottom trawlers in the coast of São Paulo. Tendencies in increasing yields were detected, mainly in the artisanal fishery. RAPD markers techniques were employed in order to analyze the genetic-population structure of the cutlassfish from Belém (PA), Natal (RN), Arraial do Cabo (RJ), Guarujá (SP), Porto Belo (SC) and Rio Grande (RS), showing a different population in Belém, while the others did not show a clearly defined structure. The population dynamics were analyzed for fishes of Arraial do Cabo, São Paulo and Porto Belo, collected monthly of January/2002 to July/2003. The cutlassfish spawning is parceled with reproductive period from the summer to the beginning of the winter. Ripe females migrate offshore to mate with ripe males that stay there, returning to the coast just after spawning. The length of first sexual maturity varied from 647 to 670mm (Lt) for females, and 526 to 650mm for males. The estimates of growth parameters were: for females Loo from 1740 to 2010mm; k from 0,12 to 0,15/year and t0 from -1,642 to -2,517 years, and for males Loo from 1373 to 1580mm; k from 0,171 0.255/year and t0 from -1,662 to -1,866 years. The longevity was about 14 years. The natural mortality rate varied from 0,20 to 0,29/year. The relative yield per recruit model showed that fishing mortality rates of the cutlassfish are close or just exceeding the limits of the maximum sustainable yield.

Os aspectos da pesca do espada, Trichiurus lepturus (Linnaeus, 1758) (Trichiuridae), foram descritos e analisados para as artes de pesca artesanal de linha-de-mão em Arraial do Cabo (RJ) e cerco flutuante em Porto Belo (SC), e da pesca comercial de arrasto de fundo com parelhas na costa de São Paulo. Verificaram-se tendências crescentes na produção da espécie, principalmente na pesca artesanal. Foram utilizados marcadores moleculares tipo RAPD para analisar a estrutura genético-populacional da espécie com exemplares de Belém (PA), Natal (RN), Arraial do Cabo (RJ), Guarujá (SP), Porto Belo (SC) e Rio Grande (RS), evidenciando-se uma população distinta em Belém, enquanto as demais não apresentaram estruturação definida. A dinâmica de populações foi analisada para exemplares de Arraial do Cabo, São Paulo e Porto Belo, coletados mensalmente de janeiro/2002 a julho/2003. A desova da espécie é parcelada com período reprodutivo ocorrendo do verão ao início do inverno. As fêmeas maduras migram para a plataforma encontrando-se com os machos maduros que ali permanecem, retornando à costa após a desova. O comprimento médio de primeira maturação sexual variou de 647 a 670mm (Lt) para fêmeas, e de 526 a 650mm para machos. As estimativas dos parâmetros de crescimento foram as seguintes: para fêmeas Loo de 1740 a 2010mm; k de 0,12 a 0,15/ano e t0 de -1,642 a -2,517 anos; para machos Loo de 1373 a 1580mm; k de 0,17 a 0.25/ano e t0 de -1,662 a -1,866 anos. A longevidade foi cerca de 14 anos. As taxas de mortalidade natural variaram de 0,20 a 0,29/ano. O modelo de rendimento relativo por recruta indicou que as taxas de mortalidade por pesca do espada estão próximas ou já ultrapassam os limites do rendimento máximo sustentável. / The fishery aspects of the cutlassfish, Trichiurus lepturus (Linnaeus, 1758) (Trichiuridae), were analyzed for the artisanal fishery of line and hook of Arraial do Cabo (RJ) and floating encirclement of Porto Belo (SC), and of the commercial paired bottom trawlers in the coast of São Paulo. Tendencies in increasing yields were detected, mainly in the artisanal fishery. RAPD markers techniques were employed in order to analyze the genetic-population structure of the cutlassfish from Belém (PA), Natal (RN), Arraial do Cabo (RJ), Guarujá (SP), Porto Belo (SC) and Rio Grande (RS), showing a different population in Belém, while the others did not show a clearly defined structure. The population dynamics were analyzed for fishes of Arraial do Cabo, São Paulo and Porto Belo, collected monthly of January/2002 to July/2003. The cutlassfish spawning is parceled with reproductive period from the summer to the beginning of the winter. Ripe females migrate offshore to mate with ripe males that stay there, returning to the coast just after spawning. The length of first sexual maturity varied from 647 to 670mm (Lt) for females, and 526 to 650mm for males. The estimates of growth parameters were: for females Loo from 1740 to 2010mm; k from 0,12 to 0,15/year and t0 from -1,642 to -2,517 years, and for males Loo from 1373 to 1580mm; k from 0,171 0.255/year and t0 from -1,662 to -1,866 years. The longevity was about 14 years. The natural mortality rate varied from 0,20 to 0,29/year. The relative yield per recruit model showed that fishing mortality rates of the cutlassfish are close or just exceeding the limits of the maximum sustainable yield.