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The feasibility of using two different methods of assaying for DNA polymer phisms has been assessed. They were Restriction fragment length polymorphisms (RFLPs) as revealed by a range of characterised Brassica cDNA sequences and Random amplified polymorphic DNA (RAPD). These techniques have been shown to reveal DNA polymorphisms between varieties of Brassica napus L. Further more, the sequence and organisation of a Hind III family of highly repetitive DNA sequences were also studied on Brassica napus L. RFLPs associated with rape extensin, ext A, and Brassica oleraceae self- incompatibility genes were observed when DNA samples from 19 commercial varieties of B. napus were analysed using the cDNA probes pRR566 (coding for root-specific extensin) and pBOS2 (coding for S(_5) self-incompatibility allele in B. oleraceae). Both cDNA clones were able to reveal RFLP patterns with varying degrees of polymorphism depending on the restriction enzymes used in the digestion of genomic DNAs. Although both probes could generate complex RFLP band patterns, those revealed by pB0S2 were generally easier to analyse and more suitable for DNA fingerprinting while those revealed by pRR566 were less distinct as a result of extensive background hybridisations. The probe pRR566, with certain restriction enzymes generated simpler RFLP band patterns that were more suitable for segregation analyses. Segregation analysis of F(_1) individuals revealed additive RFLP band patterns of both parental varieties, while that of F(_2) individuals revealed RFLP band patterns of each parental varieties as well as the additive pattern. When analysed for possible association with varietal glucosinolate content, none of the RFLP band patterns showed such linkage. A cDNA library was constructed from pod material of a high glucosinolate variety in an attempt to obtain clones which could reveal RFLP patterns associated with glucosinolate content. Differential screening using total cDNAs from pod materials of high and low glucosinolate varieties failed to isolate any cDNA clones useful as RFLP markers. Another DNA polymorphism assay studied, RAPD, was able to detect inter- and intraspecific variation in Brassica sp. Analysis of six phylogenetically-related but distinct Brassica sp. revealed extensive variation in the RAPD band patternsof amplification products; with some amphidiploid species sharing conserved band patterns with their ancestral species. RAPD analysis on 17 varieties of rape revealed polymorphic as well as highly conserved RAPD band patterns depending on the primer used. One of the primers was able to amplify a polymorphic band which could be associated with low glucosinolate varieties i.e. present almost exclusively in low glucosinolate varieties. Species-specific as well as variety-specific band patterns were also observed during the RAPD analysis. Finally, sequence and organisation of a Hind III family of repetitive sequences was studied. The monomeric and polymeric forms (trimer and tetramer) of the repetitive sequences were successfully cloned into pUCl8. Sequence analysis of the two clones containing the polymeric forms revealed that the monomers were arranged in tandem array and that all internal Hind III recognition sites were lost due to point mutation(s) which occurred within the six basepair recognition site. A consensus monomeric sequence was deduced from sequence comparison of the 8 copies of the monomeric sequences present in the 3 clones and the deviation from the consensus sequence of each of the eight monomers was less than 3%. No two monomeric sequences were identical. It was estimated that the number of copies of the monomeric sequences in a haploid genome was approximately 0.3 million copies. Estimates of the proportional representation of each of the polymeric sequences based on the number of copies of the monomers in each polymer were also calculated.

Research objective. The study was aimed to establish development and distribution patterns of cabbage stem weevil (Ceutorhynchus pallidactylus), cabbage seed weevil (C. obstrictus) and Brassica pod midge (Dasineura brassicae) in winter and spring oilseed rape crops, to estimate insect damage, to compare the efficacy of insecticides of different mode of action and to determine the optimal timing of insecticide application. Research tasks. 1. To determine species abundance and diversity of Ceutorhynchus spp. infesting oilseed rape crops. 2. To study cabbage stem weevil’s (C. pallidactylus) biology (migration activity of adult insects, embryonic and post-embryonic development) in winter and spring oilseed rape crops and to estimate insect injury. 3. To establish the patterns of distribution of adults of cabbage seed weevil (C. obstrictus) as well as those of embryonic and post-embryonic development in winter and spring oilseed rape crops and to estimate insect injury. 4. To explore the biology of Brassica pod midge (Dasineura brassicae), to identify distribution and development peculiarities and damage done to winter and spring oilseed rape. 5. To determine biological and economic efficacy and optimal application timing of insecticides of different mode of action against stem (C. pallidactylus) and pod (C. obstrictus and D. brassicae) insect pests in winter and spring oilseed rape. / Tyrimų tikslas. Nustatyti kopūstinių stiebinių paslėptastraublių (Ceutorhynchus pallidactylus), ankštarinių paslėptastraublių (C. obstrictus) ir ankštarinių gumbauodžių (Dasineura brassicae) plitimo ir vystymosi ypatumus žieminiuose ir vasariniuose rapsuose bei žalingumą, palyginti skirtingo poveikio insekticidų efektyvumą ir nustatyti optimalų insekticidų naudojimo laiką nuo šių kenkėjų. Tyrimų uždaviniai. 1. Nustatyti žieminių ir vasarinių rapsų pasėliuose plintančių paslėptastraublių (Ceutorhynchus spp.) rūšių gausumą ir įvairovę. 2. Ištirti kopūstinių stiebinių paslėptastraublių (C. pallidactylus) biologiją (suaugėlių migracijos aktyvumą, embrioninį ir poembrioninį vystymąsi) žieminiuose ir vasariniuose rapsuose, įvertinti šių kenkėjų žalingumą. 3. Nustatyti ankštarinių paslėptastraublių (C. obstrictus) suaugėlių paplitimo, embrioninio ir poembrioninio vystymosi ypatumus žieminiuose ir vasariniuose rapsuose bei jų žalingumą. 4. Ištirti ankštarinių gumbauodžių (Dasineura brassicae) biologiją, nustatyti jų žalingumą bei vystymosi ypatumus žieminiuose ir vasariniuose rapsuose. 5. Nustatyti skirtingo poveikio insekticidų biologinį ir ūkinį efektyvumą ir optimalų jų naudojimo laiką nuo stiebų (C. pallidactylus) ir ankštarų kenkėjų (C. obstrictus ir D. brassicae) žieminiuose ir vasariniuose rapsuose.

Phosphoenolpyruvate (PEP) carboxylase (PEPC) is a tightly controlled enzyme situated at a pivotal branchpoint of plant C-metabolism. Two physically and kinetically distinct oligomeric classes of PEPC exist in the endosperm of developing castor oil seeds (COS). Class-1 PEPC is a typical homotetramer composed of 107-kDa plant-type PEPC (PTPC) subunits, whereas the 910-kDa Class-2 PEPC hetero-octameric complex arises from a tight interaction between Class-1 PEPC and distantly related 118-kDa bacterial-type PEPC (BTPC) subunits. BTPC functions as both a catalytic and regulatory subunit of the allosterically-desensitized Class-2 PEPC, which has been hypothesized to support massive PEP-flux to malate for leucoplast fatty acid synthesis. Previous studies established that BTPC: (i) subunits of COS Class-2 PEPC are subject to inhibitory phosphorylation in vivo, and (ii) at Ser425 and Ser451 within an intrinsically disordered region. This study focuses on characterization of the COS protein kinase (BTPC-K) that phosphorylates BTPC at Ser451. BTPC-K, having a native molecular mass of 63 kDa, was purified ~500-fold from developing COS endosperm. Its activity was absolutely dependent upon the presence of Ca2+ (Ka= 2.7 μM) and millimolar Mg2+. BTPC-K phosphorylated BTPC subunits of Class-2 PEPC strictly at Ser451 (Km= 1.1 μM), as well as histone type III-S (Km= 1.7 μM), but did not phosphorylate a BTPC S451D phosphomimetic mutant, native COS PTPC or sucrose synthase, or α-casein. BTPC-K displayed a broad pH-activity optima of pH 7.3, a Km for Mg2+-ATP of 6.6 μM, and marked inhibition by 3-P-glycerate and PEP. The possible control of BTPC-K by disulfide-dithiol interconversion was suggested by its rapid inactivation and subsequent reactivation when incubated with oxidized glutathione and then dithiothreitol. BTPC-K activity was insensitive to exogenous calmodulin, but potently inhibited by 100 µM trifluoperazine (a calmodulin antagonist). BTPC-K-mediated Ser451 phosphorylation of BTPC subunits of Class-2 PEPC inhibited BTPC activity by ~50% when assayed under suboptimal conditions (pH 7.3, 1 mM PEP with 10 mM L-malate). Overall the results of this study have led to the hypothesis that in vivo phosphorylation of COS BTPC at Ser451 is mediated by a dedicated calcium-dependent protein kinase (CDPK). / Thesis (Master, Biology) -- Queen's University, 2013-08-30 14:23:39.648

Orientador: Dirceu Brasil Vieira / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Civil, Arquitetura e Urbanismo / Made available in DSpace on 2018-08-06T07:03:34Z (GMT). No. of bitstreams: 1 Gomes_EvandroMachado_D.pdf: 1096274 bytes, checksum: 9c1ee2a7cb50229fd383352705fa494a (MD5) Previous issue date: 2005 / Resumo: O ensaio foi conduzido nos anos de 2001 a 2003, em duas áreas experimentais: Campo de Pesquisa Hidroagrícola do Pinhal - Unicamp e no Horto Municipal Florestal, ambos no município de Limeira - SP, Brasil, com o objetivo de estudar o efeito da irrigação, sobre alguns parâmetros do girassol (Helianthus annuus L.), cultivar M-742. As semeaduras e colheitas foram realizadas, respectivamente, nos dias 18 de junho e 14 de setembro de 2001, 16 de junho e 8 de outubro de 2002 e 24 de junho e 15 de outubro de 2003. O delineamento experimental foi o de blocos casualizados com três tratamentos e quatro repetições. Os três tratamentos consistiram de: 1)irrigação de acordo com o balanço de umidade no solo (Sempre Irrigado); 2) irrigado nos períodos críticos do desenvolvimento, ou seja, formação de botão floral e enchimento de grãos (Irrigado nas Fases) e 3) sem suplementação hídrica (Sem Irrigação). A irrigação foi realizada por sistema de aspersão convencional, conduzida pelo método gravimétrico. Os principais parâmetros analisados foram: altura de planta, diâmetro do capítulo, peso do capítulo, peso da haste, peso das folhas, peso de mil sementes, teor de óleo e proteína de grãos, produção, distribuição do sistema radicular e demanda hídrica da cultura. Os dados foram submetidos à análise de variância e suas médias comparadas pelo teste de Tukey. Pelos resultados obtidos e para as condições do experimento, as seguintes conclusões se evidenciaram como as mais importantes: a) a planta desenvolvida sob marcante estresse hídrico tende a produzir raÍzes em maior número e se aprofundar mais no solo que aquela sem restrição hídrica; b) há maior concentração de raÍzes no horizonte superficial do solo, até 20 centímetros, independentemente da disponibilidade de água. c) a irrigação nas fases de formação do botão floral e de enchimento de grãos é suficiente para garantir níveis de desenvolvimento da planta e do sistema radicular bem como produção de grãos semelhantes à da planta desenvolvida sem restrição Hídrica; d) ocorre acentuada diminuição na massa seca de haste, capítulo e folha como conseqüência do estresse Hídrico; e) plantas desenvolvidas sob marcante estresse Hídrico produziram cerca de 30% menos em peso de grãos que as Sempre Irrigadas e 20% menos que as Irrigadas nas Fases; t) a redução da área foliar, através da redução na altura da planta, mostrou-se como um dos mecanismos de resposta do girassol à escassez de água; g) a planta sob restrição Hídrica tende a produzir um maior teor de proteína em detrimento da produção de lipídeos; h) o kc encontrado mostrou os menores valores aproximadamente aos 20 dias após a emergência, atingindo os valores mais elevados na fase de formação do botão floral (45-50 dias após a emergência) / Abstract: The study was conducted from 2001 to 2003, in Limeira (Brazil), with the sunflower hybrid M-742. The aim of the present work was to study the effect of the supplementary irrigation on some sunflower (Helianthus annuus L.) parameters. The sowing and harvest were done, respectively in June 18, September 14 in 2001; June 16, October 8 in 2002 and June 24, October 15 in 2003. The experimental design was a randomized blocks with three treatments and four replications. The treatments were: a) irrigation according to soil moisture (Always Irrigated); b) irrigation in the critical periods (Phases Irrigated) and c) without supplementary irrigation (Rainfed). The irrigation was performed by sprinkler and scheduled by gravimetric method. The main analyzed parameters were: head, leaves and stem dry weight and height; 1.000 seed mass; head diameter; oil and protein content; root system distribution; water demand. The data were submitted to analysis of variance and the averages were compared by Tukey test. Based upon the results some conc1usions are feasible: a) sunflower plants under sever water stress produces a higher root number that explore the soil deeply than plants with no water stress; b) the greater root concentration occurred in the superficial horizon, for all treatments. c) water supplementation during flower bud formation and grain filling stages is sufficient to assure good levels of grain yield and root system development; d) the water Stress causes an emphasize decrease in stem, head and leave dry mass; e) plants grown under accentuate water stress yielded 30% less than plants with no stress and 20% less than plants that received water only in the critical periods of bud formation and grain filling; f) the leaf area decrease through the plants height appears to be a mechanism of sunflower's defense to water stress; g) the sunflower under water stress increases protein and reduces oil seed content; h) the kc coefficient was lower at 20 days after emergence and higher at flower bud stage / Doutorado / Recursos Hidricos / Doutor em Engenharia Civil

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / A pesquisa foi conduzida em casa de vegetaÃÃo no Campus do Pici da Universidade Federal do CearÃ, no perÃodo de janeiro a marÃo de 2006, com o objetivo de estudar a resposta das culturas da mamona (Ricinus communis L.), gergelim (Sesamum indicum L.) e amendoim (Arachis hypogaea L.) a ciclos de deficiÃncia hÃdrica na fase de crescimento vegetativo. Foi avaliado o comportamento de duas cultivares de cada espÃcie, a saber: amendoim, cultivares PI 165317 (grupo âSpanishâ) e 57422 (grupo VirgÃnia); gergelim, cultivares CNPA-G3 e CNPA-G4 e mamona, cultivares Mirante e BRS 149 Nordestina. As sementes foram semeadas em tubos de PVC com 100 cm de comprimento e 15 cm de diÃmetro, em solo de textura arenosa. As cultivares foram submetidas a dois regimes hÃdricos - irrigaÃÃo diÃria e ciclos de deficiÃncia hÃdrica. Foram determinadas variÃveis relativas Ãs condiÃÃes hÃdricas da planta, trocas gasosas, crescimento da parte aÃrea e crescimento e estratificaÃÃo do sistema radicular. A colheita das plantas ocorreu aos 50, 55, 56 dias apÃs o plantio para as culturas do gergelim, amendoim e mamona, respectivamente. As variÃveis potencial hÃdrico, conteÃdo relativo de Ãgua, transpiraÃÃo foliar, fotossÃntese foliar e condutÃncia estomÃtica foram reduzidas nas trÃs culturas em resposta aos ciclos de deficiÃncia hÃdrica. A relaÃÃo entre a temperatura foliar e a ambiente aumentou em resposta ao estresse aplicado. O uso eficiente de Ãgua nÃo foi alterado nas culturas da mamona e amendoim, apresentando reduÃÃo significativa no gergelim em resposta ao dÃficit hÃdrico. A matÃria seca da parte aÃrea, das raÃzes e total, bem assim o volume radicular foram reduzidos em todas as espÃcies em resposta ao dÃficit hÃdrico. A relaÃÃo raiz/parte aÃrea aumentou em resposta ao estresse hÃdrico nas culturas do amendoim e gergelim, enquanto na mamona essa relaÃÃo permaneceu inalterada. O amendoim apresentou a menor partiÃÃo de matÃria seca para as raÃzes; o gergelim ficou numa posiÃÃo intermediÃria e a mamona destinou a maior proporÃÃo de biomassa para as raÃzes. O amendoim apresentou a maior modificaÃÃo da estratificaÃÃo do sistema radicular em resposta ao dÃficit hÃdrico, com um maior aprofundamento das raÃzes. A mamona apresentou a menor resposta e o gergelim ficou numa posiÃÃo intermediÃria. Independentemente do regime hÃdrico as trÃs culturas apresentaram maior proporÃÃo do sistema radicular nos primeiros 25 cm de solo. O nÃmero de folhas e a Ãrea foliar foram reduzidos nas trÃs espÃcies em resposta ao dÃficit hÃdrico. / A greenhouse research was carried out at the Universidade Federal do CearÃ, Campus do Pici, from January to March 2006, with the objectives of studying the response of castorbean (Ricinus communis L.), sesame (Sesamum indicum L.) and peanut (Arachis hypogaea L.) to cycles of water deficit in the early vegetative growth phase. The following cultivars for each crop were evaluated: peanut cultivars PI 165317 (group Spanish) and 57422 (group Virginia); sesame CNPA G3 and CNPA G4; and castorbean Mirante and BRS 149 Nordestina. The seeds were planted in PVC tubes 100 cm log and 10 cm wide, filled with sand soil. The cultivars received two water regime treatments: daily irrigation and cycles of water stress. Several variables were determined relative to plant water status, gas exchange, top and root growth and root system distribution in the soil profile. Plant harvest was accomplished at 50, 55 and 56 days after planting for sesame, peanut and castorbean, respectively. The variables water potential, relative water content, leaf transpiration, leaf photosynthesis and stomatal conductance were reduced in all three crops in response to water stress. The relation between leaf and ambient temperature increased in response to water stress. The water use efficiency was not affected by the stress imposed in castorbean and peanut. On the other hand, in sesame this variable decreased in response to water stress. Tops, roots and total dry matter as well as the root system volume were reduced in all species in response to water stress. Root shoot ratio increased in response to water stress for sesame and peanut, while for castorbean this relation remained unchanged. Among the crops studied, peanut plants had the smallest dry matter partition to the roots; sesame stayed in an intermediary position and castorbean partitioned the higher proportion of dry matter to roots. Peanut showed a large modification in the root system stratification in response to the water stress, with the increase of proportion of the root system in the deep soil. Castorbean showed the smaller response as far as deepening of the root system and sesame retained an intermediary position. All three crops, no matter the water regime was, had the most proportion of the root system in the first 25 cm layer of soil. The leaf number and leaf area were reduced in the three species in response to water stress.

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / A mamoneira (Ricinus communis) Ã tolerante a seca e cultivada para produÃÃo de Ãleo para uso industrial. Na regiÃo Nordeste os pequenos produtores cultivam a mamona consorciada algodÃo, feijÃo caupi, gergelim, girassol, milho, sorgo ou amendoim. Dois experimentos de campo foram conduzidos nos anos agrÃcolas 2008, 2009 e 2010, na Fazenda Experimental Lavoura Seca, QuixadÃ, CearÃ, com o objetivo de avaliar o comportamento da mamona e girasssol cultivados em dois padrÃes de consorciaÃÃo. No experimento I avaliou-se o consorcio mamona (Ma) x girassol (Gi) com 1Ma:1Gi; 1Ma:2Gi; 1Ma:3Gi; 2Ma:2Gi; 2Ma:3Gi fileiras, acrescidos da mamona e girassol em monocultivo. No experimento II, avaliou-se o consorcio mamona x girassol com antecipaÃÃo do plantio da mamona em 0, 7, 14 ou 21 dias antes do plantio do girassol. Para cada experimento adotou-se o delineamento de blocos ao acaso com 4 repetiÃÃes. Avaliou-se a produtividade de grÃos, seus componentes e o uso eficiente da terra para a mamona e girassol consorciados e em monocultivo. A produtividade da mamona e girassol no monocultivo foi significativamente superior nos dois sistemas de consÃrcio. A antecipaÃÃo do plantio da mamona 14 e 21 dias em comparaÃÃo ao girassol mostraram vantagens em relaÃÃo as demais datas de plantio. No experimentos I e II verificou-se que a mamona foi dominante sobre ao girassol. PrecipitaÃÃes pluviais inferiores a demanda hÃdrica das culturas da mamona e girassol, proporcionaram reduÃÃo no uso eficiente de terra (UET). / Castor bean (Ricinus communis L.) is an oil seed crop drought tolerant with its oil having several uses in industry. In the Northeast of Brazil it is commonly cultivated by small farmers intercropped with cotton, cowpea, sesamum, sunflower, sorghum or peanuts. Two field experiments were carried during 2008, 2009 and 2010, at â Fazenda Experiental Lavoura Secaâ, Quixada, Ceara, with the objective to evaluate the behavior of castor bean and sunflower under two intercropping arrangements. In experiment I, its was evaluated the intercropping castor bean (CB) x sunflower (SF) with 1CB:1SF; 1CB:2SF; 1CB:3SF; 2SB:2SF; 2CB:3SF rows, added of sole crop of castor bean and sunflower. In the experiment II, was evaluated with the early planting of castor bean in 0, 7, 14 or 21 days in relations to sunflower the intercropping systems. In both experiments it was used a randomized complete block design with 4 replications. In this study it was determined the yield, yield components and land equivalent ratio (LER) for castor bean and sunflower grown in intercropping and sole crop. The grain yield of castor bean and sunflower was significantly higher in sole crop than intercropping in both experiments. Between the intercropping systems the highest yields were observed when early planting of castor bean was of 14 or 21 days before sunflower. In experiments I and II, showed that the castor bean was the dominant crop in relation to sunflower. Rainfall lower the water requirements of castor bean and sunflower crops, showed a reduction in LER.