Repositório RCAAP

Com o objetivo de estudar a cinética da despolimerização dos microtúbulos e a dinâmica da indução de micronúcleos, usaram-se dois herbicidas com ação antimitótica, amiprofós-metil (APM) e oryzalin (ORY), em células de Helianthus maximiliani. A aplicação de ambos os herbicidas em células com ativo crescimento resultou num elevado número de células com micronúcleos, em virtude das modificações mitóticas causadas pela ausência de microtúbulos. Primeiramente, cromossomos isolados ou grupos de 2 ou mais cromossomos espalharam-se pelo citoplasma. Duas a três horas após a aplicação dos herbicidas, os microtúbulos, estavam despolimerizados e os cromossomos metafásicos condensaram-se, formando diretamente os micronúcleos sem ter havido a divisão do centrômero e a separação das cromátides. A concentração mais eficiente de ORY foi 20 µM, ao passo que para APM foi 60 µM, para um período de incubação de 36 e 48 h respectivamente. O prolongamento do período de tratamento elevou a freqüência de micronúcleos deformados, bem como aumentou a morte celular. A adição de citocalasina-B (CB) (20 µM), após 24 h da aplicação dos herbicidas, aumentou significativamente a micronucleação e estabilização dos micronúcleos nas células. O controle dos microtúbulos de células em suspensão torna-se uma ferramenta eficiente para a indução de micronúcleos, bem como representa um método elegante para a transferência genômica parcial.

Com este trabalho objetivou-se investigar a aquisição da capacidade germinativa e o estabelecimento da dormência em sementes em desenvolvimento de Bixa orellana L. (urucum). Foram caracterizados 7 estádios de desenvolvimento das sementes, baseados nas características morfológicas, massa fresca, massa seca e teor de umidade dos frutos e das sementes. Sementes imaturas de B. orellana não germinaram durante os estádios iniciais de seu desenvolvimento. As sementes adquiriram a capacidade germinativa máxima quando os níveis de umidade declinaram para valores em torno de 60%. Nessa fase, as sementes apresentaram acúmulo máximo de massa seca. A dormência foi estabelecida quando as sementes exibiam um conteúdo de água em torno de 10%.

Leaf lipoxygenases (LOX) are involved with important physiological processes such as plant growth and development, senescence, biosynthesis of regulatory molecules, and response to pathogens and insects. We did a biochemical evaluation of the LOX pathway of soybean leaves submitted to wounding in a normal genotype (IAC-100) and its counterpart lacking seed LOX (IAC-100 TN). Our results indicate that LOX activities in the different pHs and temperatures tended to be higher in the wounded plants compared to their respective controls. The K M app values at 168 h after wounding reached a minimum in both genotypes indicating that the plants respond by changing the leaf LOX pool. There was an increase on protease inhibitor levels in all time points after wounding, for both cultivars. The levels of hexanal and total aldehydes are similar for the wounded plants at different times after wounding and their respective controls for both genotypes. Our results strongly suggest that the LOX pathway is activated during the wound response leading to jasmonate by the initial action of hydroperoxide cyclase. In addition, the results show that the genetic removal of seed LOX does not interfere with the plant’s ability to respond to wound via the LOX pathway.

Citric and malic acid excretion in the medium and malic acid accumulation in seedling roots and embryo-derived calli as possible mechanisms of aluminum (Al) resistance and the effects of a 17-h Al stress period on root growth in Oryza sativa have been studied. Four-day-old seedlings and embryo-derived calli of Al-resistant (IRAT 112 and IR6023) and Al-sensitive (Aiwu and IKP) cultivars were treated with 250 and 500 µM {Al2(S04)3.18H20 }of total aluminum or without Al for 36 hours. After 3 to 36 hours of stress, seedlings and calli were removed from the flasks and concentration of citric and malic acids was estimated in the Al and control solutions. Malic acid was also assayed in roots tips and in callus tissues. After 17-h of Al stress, inhibition of root growth was a typical effect of Al in rice and the extent of the inhibition depended on both cultivar and Al concentration. At 500 µM of Al, strong reduction of root elongation occurred in all cultivars while at 250 µM of Al, only IRAT was unaffected, when compared to their control. In the absence of Al, all varieties excreted comparable amounts of citric and malic acid. Al treatments, were without effect upon citrate excretion in both Al-resistant and Al-sensitive cultivars. Al treatment, for periods from 3 to 24h, slightly stimulated the excretion of malic acid from seedlings, in all cultivars. Malic acid concentrations in root apices, in the presence or absence of aluminum, were not correlated with aluminum resistance. No differences in malic excretion and internal concentrations were detected between Al-treated and untreated rice calli of the same four cultivars. It is therefore concluded that, in our experimental conditions, differences in Al resistance in our rice cultivars cannot be attributed to citric and malic acids. Further research needs to be carried out to examine other possible mechanisms of Al-resistance in rice and to determine whether organic acids such as succinic and oxalic acid are implicated.

The actual (A) and potential (Apot) photosynthetic rates of C3 and C4 tropical crop species grown under greenhouse conditions was compared. The following species were investigated: Oryza sativa, Phaseolus vulgaris, Glycine max, Helianthus annuus, Gossypium hirsutum, Manihot esculenta, Theobroma cacao, Coffea arabica, Hevea brasiliensis, and Eucalyptus urophylla × E. grandis, all from the C3 group, and Amaranthus sp., Panicum maximum, Pennisetum purpureum, Zea mays and Saccharum officinarum, from the C4 group. A, determined under non-limiting light at ambient temperature and CO2, was measured with an infrared gas analyser, whilst Apot, determined under saturating light and CO2 at an optimal temperature (35 ºC for all species), was gauged with a gas-phase oxygen electrode. On an area basis, A varied from 5.0 up to 26.3 mumol CO2 m-2 s-1, whilst Apot was very similar in 14 of the 15 species, with an average rate of 35.0 ± 2.4 mumol O2 m-2 s-1. The value of Apot in T. cacao was approximately half the mean of the remaining species. On a mass basis, variations in A were much larger, and differences in Apot, although not large, emerged. The overall mean Apot per unit mass in the four tree species was 28.0 ± 2.2 mumol O2 g-1 min-1 against 44.6 ± 5.8 mumol O2 g-1 min-1 in the remaining species. As a whole, the results evidenced a conservative behaviour of the photosynthetic apparatus to fix CO2 amongst the species investigated, despite the large differences in A among them.

It has been reported that Coffea arabica seeds contain as the main reserve protein, a legumin-like protein, constituted of two subunits, alpha and beta, of approximately 35 and 20 kDa. In this work the seed proteins of several coffee species and varieties were investigated by SDS-PAGE and gel filtration. No differences were observed in the electrophoretic profiles among varieties of C. arabica, however, marked differences were observed among species, or even among individuals of some species. In general, the molecular weight of the subunits alpha and beta accounted for a monomer of 48 to 62 kDa. However, native molecular weight obtained by gel filtration showed that for most of the species there is association of 6 of such proteins, in a hexamer. The most marked difference was observed for C. canephora and C. racemosa. The former clearly showing isoforms of the subunits, and the later showing absence of the beta subunit. The influence of proteases in this observations is discussed.

Two-dimensional gel electrophoresis analysis was performed on adventitious and storage root in cassava (Manihot esculenta Crantz). Adventitious root lacking swelling formation and swelled storage root were obtained from the accession WU104 grown in the green house of the Department of Biology in Washington University in St. Louis (MO-USA). Saline buffer-soluble proteins were extracted, separated in a high-resolution 2-D electrophoresis system, visualized with silver staining gel procedure, and digital image generate for further analysis. Quantitative and qualitative protein spots analysis was performed with a computer assisted image software system. Results revealed large variation in the complexity of the gel protein profile between the two root systems. About 90% of the protein spots appeared in the pI range value of 4.0 to 6.5 and between 14 to 80 Kda of molecular mass. Detailed computer assisted analysis of this gel allowed us to establish 5 distinct classes of protein based on spot quantification that could be associated with swelling and non-swelling roots. Variation in the complexity of protein pattern was related with different type of root. Whereas the adventitious root showed a more simple profile related to primary growth, the storage root showed to be a more complex profile related to secondary growth and starch accumulation.

In order to evaluate the influence of photoperiod on the accumulation of allantoin in comfrey (Symphytum officinale L.), a well known medicinal plant, an experiment was conducted during July and August, 1998. Cuttings obtained from 90 day old plants were submitted to four photoperiods (8, 12, 16 and 20 hours). After 60 days, allantoin content in roots and rhizomes was evaluated. The results showed that increases in photoperiod promoted an increment in the average content of allantoin in roots (0.06%, 0.303%, 1.213% and 4.78%). On the other hand, in rhizomes, allantoin accumulation decreased (9.65%, 7.14%, 0.55%) when the photoperiod was increased from 8 to 12 and 16 hours, respectively, stabilizing on a 20-hour photoperiod (0.53%). Plants cultivated under field conditions presented 2.55% and 2.63% allantoin content in rhizomes and roots, respectively. Based on the fact that in comfrey the roots are considered to be sites of allantoin synthesis, the results demonstrated that photoperiod could influence both the synthesis of allantoin in these organs as well as its accumulation in the rhizomes.

A leucena (Leucaena leucocephala (Lam.) de Wit), quando usada como cobertura no solo, apresenta a propriedades de controlar plantas daninhas, sendo esse efeito resultante da presença de aleloquímicos, principalmente mimosina, encontrados na parte aérea da planta. A maioria dos estudos em alelopatia refere-se apenas ao efeito do aleloquímico sobre a germinação e o crescimento da planta-teste, sem considerar os eventos celulares relacionados às mudanças fisiológicas. Com este trabalho objetivou-se avaliar o efeito do extrato aquoso da leucena sobre o desenvolvimento, índice mitótico radicular e atividade da peroxidase e suas isoformas na parte aérea e raízes de plântulas de milho. Os bioensaios foram conduzidos em casa-de-vegetação utilizando-se sementes de milho híbrido desenvolvidas nas concentrações 0; 0,4; 0,8; 1,6; 3,2 e 6,4 % do extrato aquoso de leucena. Verificaram-se inibição do crescimento de raízes e redução do índice mitótico proporcional ao incremento na dose do extrato, não sendo observada divisão celular a partir da concentração de 1,6 %. A atividade da peroxidase, tanto em raízes como em folhas, cresceu com o aumento da concentração do extrato, sem, contudo, apresentar diferença nos zimogramas de suas isoformas. A maior atividade dessa enzima, nas raízes, foi correlacionada com a presença das isoformas aniônicas, pI 4,99 e 4,86, o que possivelmente esteja contribuindo para maior espessamento das raízes, o que foi verificado nas doses elevadas do extrato. As análises por HPLC revelaram teor de mimosina crescente com aumentos na concentração do extrato, sugerindo o envolvimento desse aleloquímico no desenvolvimento das plântulas.

Active vegetative and reproductive growth in field-grown trees of Coffea arabica L. in Viçosa (20º45’S, 650 m altitude), south-eastern Brazil, occur concurrently. The overall patterns of branch growth and leaf area gain were to a certain extent altered by fruit removal, with growth rates being remarkably greater in de-fruited trees. The content of N-NO3 was not affected by fruiting, whilst that of amino-N was greater in de-fruited than fruiting trees most of the time, but the differences were not large enough to have significantly contributed to the increased growth rates in de-fruited trees. Leaf nitrate reductase activity was greater in plants bearing fruit than in non-bearing ones most of the time; activity was roughly inversely associated with growth. Although the roots contained much more nitrate than the leaves, the root nitrate reductase activity was much lower and not affected by fruiting. Much of the restrictive effects of fruiting on vegetative growth appeared to be associated to starch exhaustion, in addition to the outstanding effect of supra-optimum temperatures per se.

Young sexually and assexually propagated Barbados cherry plants were submitted to water deficit (20 days without irrigation). During this period the accumulation of proline, water potential of branches, osmotic potential, the relative water content of leaves, the leaf diffusive resistance, the transpiration rate and leaf temperature in the cuvette were determined. In addition, photosynthetically active radiation (PAR) and vapor pressure deficit (VPD) were measured in the porometer cuvette. The concentration of proline for both types of plants began to increase on the fifth day without watering, and reached 38.1 times the concentration in the control plants grown from seeds and 26.4 times the concentration in grafted plants on the tenth day without watering. The lowest levels of leaf water potential in the plants suffering from severe water stress varied from -4.5 to -5.7 MPa, the lowest values being observed in the sexually propagated plants. These plants also showed the highest values for transpiration (0.9 mmol.m-2.s-1) and proline concentration (20.42 mg.g-1 DM), the lowest for relative water content of the leaves (38.4%) and diffusive resistance (940 s.m-1) at the end of the experiment. The Barbados cherry plants developed strategies for surviving drought, with differences between various characteristics, resulting from prolonged stress, which significantly influenced the parameters evaluated, with the exception of leaf temperature.

The possibility of improving nitrogen use efficiency in maize was investigated using S1 endogamic families of the populations Sol da Manhã NF and Catetão. A simple 10 X 10 lattice design was adopted and the trials carried out at the experimental field of MITLA AGRÍCOLA LTDA, in Uberlândia, State of Minas Gerais, during the 1994/95 planting season. Based on grain production figures, the three best and three worst performing S1 endogamic families were selected for this study. These were pooled to form four sub-populations denominated NFB, NFR (the best and worst families, respectively, of the Sol da Manhã NF variety), CATB and CATR (the best and worst families, respectively, of the Catetão variety). Each of these sub-populations was evaluated under greenhouse conditions. The experimental design was factorial with treatments arranged in randomized blocks. Sample replicates consisted of pots with four plants. Feeding with modified Hoagland’s nutrient solution began on the seventh day after sowing. The study involved four nitrogen regimes, where varying proportions of NO3- and NH4+ were formulated, such that the nutrient solution contained the following mixtures: 75% NO3- : 25% NH4+; 25% NO3- : 75% NH4+; 50% NO3- : 50% NH4+ (all high N mixtures) and 5% NO3- : 5% NH4+ (low N mixture). Twenty-five days after planting, the activities of the enzymes nitrate reductase and glutamine synthetase (transferase and synthetase assays) were determined for the leaves using the third topmost expanded leaf of the four plants in each pot. The data show that glutamine synthetase (transferase assay) and nitrate reductase activities were efficient in discriminating the S1 endogamic families and could therefore be useful biochemical parameters in breeding programs seeking nitrogen use efficiency.

The paper proposes the classification of seeds in relation to the forms of phytochrome instead of classical photoblastism. On the basis of published data all seeds have phytochrome and the term photoblastism should be replaced by forms of phytochrome that control germination: 1. Positive photoblastic seeds have phyB (and, to a lesser extent, phyD and phyE) controlling the germination process through Low Fluence Responses (LFR); 2. Negative photoblastic seeds have phyA controlling germination through High Irradiance Responses (HIR) and, when the pre-existing Pfr level is high enough to induce germination in darkness, through LFR by phyB; and 3. light insensitive seeds have phyA controlling germination through Very Low Fluence Responses (VLFR).

The induction and control of in vitro somatic embryogenesis is dependent of a number of factors. The genotype of the mother plant donor of explants and the kind of auxin play essential roles in conferring embryogenic competence. Once obtained somatic embryos may be encapsulated in alginate originating synthetic seeds that can be stored at low temperatures. Considering this the present work aimed at the assessment of five different genotypes, five periods of 2,4-D shock (20 muM for 1, 2, 4, 8 and 16 weeks) (), and different substances supplemented to sodium alginate to obtain synthetic seeds. Zygotic embryos of Feijoa sellowiana were inoculated in basal LP medium (von Arnold and Erikson, 1981) supplemented with Morel’s vitamins, sucrose (3%) and phytagel (0.2%). The highest rates of somatic embryogenesis induction were observed in the genotypes 50-4 and 101. Shock of two weeks in 2,4-D conferred embryogenic competence in the same levels obtained with the shock of 8 weeks. Histological evaluations revealed the direct origin of the somatic embryos from the epidermic surface of cotyledons. Synthetic seeds containing MS salts and sucrose resulted in higher rates of contamination than capsules free of these substances. KNO3 (100 mM) resulted in the opening of 81.2% of the capsules as compared to 0% of opening in the treatment with water.

Four papaya genotypes, two from the ‘Solo’ group (Sunrise Solo TJ and Improved Sunrise Solo line 72/12) and two from the ‘Formosa’ group (Tainung 02 and Know You 01) grown in ultisol under field conditions in Macaé, RJ, Brazil, were used in this study. Two different effective depths (ED) were determined in the area using a penetrographer with average soil moisture of 11.2%. The area with ED of 0.35 m with a maximum pressure of 4.12 MPa for penetration was defined as an area with restriction (WR) to root growth, while, the area with minimum ED of 0.60 m and a pressure lower than 2.30 MPa as an area with no restriction (NR). The net CO2 assimilation rate (A), stomatal conductance (g s), leaf temperature (Tl), intercellular partial pressure of CO2 (c i) and intrinsic water use efficiency (IWUE) were evaluated for three consecutive days after irrigation. Mechanical root restriction affected gas exchange of the four papaya genotypes. All genotypes grown in the WR area had lower A, g s and c i and higher Tl than the same genotypes in the NR area. All genotypes grown in the WR area had high IWUE and Sunrise Solo TJ had the highest IWUE.

The effects of temperature on germination and endo-beta-mannanase activity in seeds of Sesamum indicum was investigated. The minimum germination temperature (Tmin) lies between 12.8°C and 13.2°C while the maximum temperature (Tmax) is located between 45.5°C and 46°C. Germinabilities are statistically not different from estimated viability (88%) between 18.8°C and 43.2°C. The Mann-Whitney test indicated the interval 31.9°C to 35.1°C as the optimum temperature (Topt) range for germination rate. When seeds incubated at temperatures at or below the Tmin and close to or above the Tmax were transferred to 30°C, those incubated at lower temperatures achieved high germinability. On the other hand, the higher the pre-incubation temperature above Tmax, the lower the germinability achieved near Topt. Seed endosperm cell wall was found to contain mannose as the main monosaccharide. An increase in endo-beta-mannanase activity in the micropylar endosperm prior to seed germination was observed only at supra-optimum temperature.

This study was carried out in a plantation of a degraded area in Central Amazon, Brazil, in order to assess environmental light effects on leaf pigment concentrations and chlorophyll a fluorescence in mahogany (Swietenia macrophylla King) and tonka bean (Dipteryx odorata Aubl. Willd). Three-year old saplings were grown in an open site (sun) with maximum radiation of approximately 2000 mumol m-2 s-1 and under the shade of a Ochroma pyramidale (Cav. ex Lam. Urb) canopy. A leaf area index of 2.46 was optically determined for the Ochroma foliage with 13% of full sunlight (maximum radiation about 260 mumol m-2 s-1). Carotenoid and chlorophyll concentrations were spectrophotometrically determined. Initial (Fo), maximum (Fm) and variable fluorescence (Fv) were determined with a portable fluorometer. Chlorophyll concentrations (Chl a, Chl b and Chl tot) on a fresh mass basis were greater in shade leaves than in sun acclimated leaves in both species. On the other hand, when comparisons were made on a per area basis, Chl a concentrations were greater in the sun than in shade leaves of tonka bean. In contrast, Chl b concentrations were higher in the shade environment for this species. No differences were observed in Chl concentrations per unit area in mahogany leaves. Carotenoid concentrations were higher for sun-acclimated leaves of mahogany, both on a per unit area and on a fresh mass basis. In tonka bean, higher carotenoid concentrations in shade leaves were only observed on a fresh weight basis. No differences were found between environments when carotenoid concentrations were expressed as a function of leaf area. Fluorescence variables (Fo, Fm, and Fv) were greater in sun leaves than in shade leaves of mahogany. In contrast, Fv and Fm, but not Fo, values were higher in shade acclimated leaves than in full sunlight-adapted leaves of tonka bean. However, there was a higher Fv/Fm ratio value for tonka bean in sunlight environments. We conclude that these species, classified as mid and late successional species, differ markedly regarding the utilization of photosynthetic pigments, increasing either light harvesting efficiency at low irradiance or protection against photosynthetic machinery damage in higher solar radiation environments.

Os métodos utilizados para determinar a quantidade de clorofila na folha requerem destruição de amostras de tecido e muito trabalho no processo de sua obtenção. O recente desenvolvimento do medidor portátil de clorofila, que permite leituras instantâneas do teor relativo de clorofila na folha sem destruí-la, surgiu como uma nova técnica para estimar esse parâmetro. Com os objetivos de avaliar a relação entre leitura do clorofilômetro com os teores de clorofila extraível e de nitrogênio na folha em três estádios de desenvolvimento da planta de milho e de validar o uso do equipamento como parâmetro para estimar o teor relativo de clorofila na folha, conduziu-se um experimento no município de Eldorado do Sul, na região fisiográfica da Depressão Central do Estado do Rio Grande do Sul, no ano agrícola de 1999/2000. Os tratamentos constaram de dois híbridos de milho (Pioneer 32R21 e Premium) e de oito sistemas de manejo de nitrogênio em cobertura. As leituras com clorofilômetro, a extração de clorofila e a determinação do teor de nitrogênio da folha foram realizadas nos estádios de seis a sete folhas, 10 a 11 folhas e de espigamento do milho. Nos dois híbridos, as quantidades de clorofila total e das clorofilas a e b extraíveis foram significativamente relacionadas às leituras do clorofilômetro, nos três estádios de desenvolvimento avaliados. A leitura realizada com clorofilômetro estimou com boa precisão o teor relativo de clorofila na folha de milho, além de apresentar as vantagens de maior rapidez, menor custo e de não implicar em destruição de folhas. No entanto, o clorofilômetro não é muito preciso para avaliação do nível de N na planta nos estádios iniciais de desenvolvimento de milho (seis a sete folhas).

Tolerância à seca em plantas não é uma característica simples, mas sim um complexo de mecanismos que trabalham em conjunto ou isoladamente para evitar ou tolerar períodos de déficit hídrico. Genótipos que diferem em tolerância ao déficit hídrico devem apresentar diferenças qualitativas e quantitativas na expressão gênica quando submetidos a períodos de seca. Três cultivares de soja (BR-4, BR-16 e MG/BR-46 Conquista), com respostas contrastantes ao déficit hídrico, foram estudadas com o uso da técnica "Differential Display" (DD) para identificar e isolar genes que podem apresentar diferenças de expressão durante períodos de seca entre os genótipos estudados. Um total de 84 fragmentos de DNAc diferencialmente expressos foram detectados. Trinta e cinco fragmentos foram clonados em vetores pGEM-T, onde vinte e oito puderam ser seqüenciados. Comparação das seqüências obtidas com seqüências em bancos de genes mostraram identidade de seqüência com genes conhecidos. Um dos clones, por exemplo, mostrou homologia com um ativador de transcrição encontrado em Arabidopsis thaliana, ao passo que outro clone mostrou-se homólogo a uma subunidade de NADH Desidrogenase de Spinacia oleracea. O ativador de transcrição apresentou-se diferencialmente expresso somente em raízes, e o homólogo à subunidade de NADH Desidrogenase foi expresso somente no genótipo considerado tolerante à seca e somente durante a condição de estresse hídrico. Pelos resultados deste trabalho, foi possível identificar alguns fragmentos potencialmente envolvidos em respostas à condição de seca em soja. Com essas informações, o estudo desses genes pode ser aprofundado, visando à confirmação desses resultados e a auxiliar no desenvolvimento de novos genótipos mais adaptados às condições de seca em soja e possivelmente em outras culturas.

Epiphytic orchids have been considered an interesting model for plant development and may have unusual features related to the hormonal control of shoot and root growth. In the present experiment we have studied two genotypes of Catasetum fimbriatum Lindl. (Orchidaceae) with an inverse pattern of shoot and root dry matter accumulation. Whereas the CFC1 genotype directs 60% of dry matter towards the shoot system, this same amount is directed towards the root system in the CFC4 genotype. Differences in the endogenous content of cytokinins, IAA and ABA, as well as differences in IAA conjugation and oxidation in shoots and roots were observed in the two genotypes. These differences were correlated with their dry matter partitioning patterns. Thus, the genotype that directed 60% of dry matter towards the root system showed a 20-fold higher auxin accumulation in those organs, while the one that accumulated 60% of dry matter in the shoots showed a 10-fold higher cytokinin accumulation in this plant part. Based on the well established information about auxin and cytokinin biosynthesis, conjugation, oxidation, transport, and effect on organ growth, we have presented an integrative working hypothesis to interpret the data obtained in this study. This working hypothesis may also account for the understanding of the complex relationships involving the hormonal control of the relative growth of shoots and roots in other plant models.