Inês Chamel José

Effect of temperature on polyunsaturated fatty acid accumulation in soybean seeds

Soybean oil contains around 60 % of polyunsaturated fatty acids, which are responsible for the low oxidative stability of soy-derived products. Soybean lines with low linolenic acid content can be obtained by genetic manipulation; however, a high proportion of the variation in fatty acids content is due to environmental factors. This work aimed to determine the effect of temperature on oil composition and on the activity of the enzymes CDP-choline:1,2-diacylglycerolcholine phosphotransferase (CPT) and acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT), responsible for maintenance of polyunsaturated fatty acids in the cytoplasmic acyl-CoA pool, that is used for oil synthesis in the seeds. CAC-1, a soybean variety with linolenic acid content of about 8 % and CC4, a BC3F4 CAC-1 derived line, with about 4 % linolenic acid, were used. The lines were cultivated under two temperature: 34/28oC or 22/13oC - day/night. The seeds were collected along seven development stages, according to their fresh weight. Fatty acid analysis was carried out by gas chromatography and CPT and LPCAT activities were determined by measuring the radioactivity incorporated in their products, phosphatidyl-[14C]choline and phosphatidylcholine-[14C]oleoyl, respectively. Linolenic acid contents were 3.89 and 6.92 % for line CC4 and 7.39 and 12.49 % for variety CAC-1, when submitted to high and low temperature conditions, respectively. Both enzymes were more active, in the development stages analyzed, in seeds produced under low temperature. Kinetics characterization of CPT and LPCAT were conducted previously.

Caracterização bioquímica de linhagens de soja com alto teor de proteína

O objetivo deste trabalho foi caracterizar bioquimicamente duas isolinhas de soja com alto teor de proteina. O aumento do teor de proteina nas isolinhas foi acompanhado por reducao no teor de oleo e de carboidratos totais. Em relacao a composicao aminoacidica, o aumento do teor de proteina promoveu acrescimo em todos os aminoacidos, exceto glicina, alanina, metionina, cisteina e tirosina, mantendo a relacao enxofre/nitrogenio. A quantificacao dos polipeptidios mostrou que o aumento do teor de proteina manteve inalterado o teor das proteinas 7S, promoveu aumento no teor das proteinas 11S e, consequentemente, da relacao 11S/7S. Pode haver melhoria na qualidade do farelo de soja das isolinhas, uma vez que as proteinas 11S tem melhor qualidade nutricional do que as proteinas 7S.The objective of this work was to characterize high protein soybean near isogenic lines. The increasing of protein was followed by reducing of oil and carbohydrate. In respect to aminoacid composition, increasing of protein promoted a rising in all aminoacids, except for glycine, alanine, methionine, cysteine and tyrosine, although the ratio S/N has been kept. The measure of polypeptides showed that the increasing of protein did not alter the quantity of 7S proteins, provided increasing of 11S proteins and 11S/7S ratio. An improvement of meal quality in these lines can occur once the 11S proteins have a better nutritional quality than 7S proteins.

Inheritance of isoflavone contents in soybean seeds

SummaryWe studied the genetic basis of isoflavone content inheritance in soybean seeds. The progenitors BARC-8 (low isoflavone content), IAC-100 (high isoflavone content), the F1 and F2 populations derived from reciprocal crosses, and backcross populations were analyzed for isoflavone content and composition. Six isoflavones were detected: daidzin (DZ), genistin (GT), glycitin (GC), malonyldaidzin (MDZ), malonylgenistin (MGT) and malonylglycitin (MGC). DZ, GT, MDZ and MGT contents were influenced by the cytoplasm and the nuclear genes of the maternal parent. For this reason, a genetic model was considered that included the cytoplasmic effect and epistasis between nuclear and cytoplasmic genes. Except for GT, the additive effect was the most important one. For GT content the cytoplasmic effect was the most important. Except for MDZ, the epistatic effects were significant for all the isoflavone forms. Our data indicate that genetic improvement for these traits should explore the additive genetic variances in superior lines or the cytoplasmic effect and the epistatic interactions between cytoplasmic and nuclear genes to obtain the largest selection gains.

Biometrical analyses of linolenic acid content of soybean seeds

The genetic reduction of linolenic acid levels increases the quality and stability of soybean oil. The objective of this study was to determine the inheritance and evaluate the nature and magnitude of gene effects on soybean seed linolenic acid level. Means and variances of F1, F2, and F3 generations were made from the cross between accession BARC-12 (low linolenic acid content) and the commercial Brazilian cultivar CAC-1 (normal linolenic acid content). The results demonstrated that linolenic acid content in soybean is under the genetic control of a small number of genes. The additive model explained the means for the three generations and for the parents. Non-allelic gene interactions had little effect on the determination of genotypic values for the individuals. The generation means and population variation analyses demonstrated that the dominance deviations contribute little to the trait. These results showed that backcross breeding programs can be used to introduce the low linolenic acid content trait into soybean seeds, since it is possible to identify with very high accuracy the desired genotypes in segregating populations.

Caracterização da via das lipoxigenases em plantas de soja resistentes e susceptíveis a Diaphorte phaseolorum f.sp. meridionalis, agente causal do cancro-da-haste

Lipoxygenases (linoleate: oxygen oxido-reductases, EC 1.13.11.12) are a class of enzymes that catalyze the addition of molecular oxygen to 1,4 - cis,cis - pentadiene motifs of unsaturated fatty acids to form hydroperoxide products. Plant lipoxygenases use linoleic (18:2) or linolenic (18:3) acids as substrates and are involved in important physiological process such as biosynthesis of regulatory molecules, growth and development, senescence, responses to wounding, vegetative storage protein and may directly mediate host resistance to insect, fungal and bacterial pathogens. When plant tissues are damaged, by pathogens or mecanically injured, a series of lipid degradation occurs, via lipoxygenase pathway, giving rise to hydroperoxydes. Among several products formed it can be found traumatin, jasmonic acid, volatile aldehydes and oxyacids. In this work we evaluated the capacity of the soybean plant to respond to the attack of stem canker fungi (Diaporthe phaseolorum (Cke. e Ell.) f. sp. meridionalis), through the lipoxygenase pathway. Leaves from resistant (FT-Cristalina RCH) and susceptible (FT-Cristalina) cultivars were used for kinetic evaluation of lipoxygenase and biochemical quantitation of products from the lipoxygenase pathway. Plants from both cultivars, infected with the pathogen or mecanically injured, showed values of lipoxygenase specific activity higher than the controls in differents pH and temperature values. The lipoxygenase pool of both cultivars showed kinetic of Michaelis-Menten within the range of substrate concentration assayed. There were an increase in protease inhibitors and of total aldehydes contents, which are products of the lipoxygenase pathway. The biochemical and kinetic parameters suggest a response of the soybean plant through the lipoxygenase pathway.

Simulation of the transgenic soybean tolerant to glyphosate through explant cultivation

The objective of this experiment consisted in simulation in vitro of the transgenic soybean tolerant to glyphosate through explant cultivation in culture medium containing aromatic amino acids. The effects of glyphosate on soybean (Glycine max (L.) Merrill) were evaluated in seeds harvested from explants of cv UFV-16. The soybean explants were cultivated in liquid medium culture with pH about 5.0. Each explant consisted of one fruit completely expanded, containing two seeds of 100 mg approximately, and connected to a stem segment of 45 mm length. The explants were submitted to the following treatments: A = glutamine (Gln); B = Gln + phenylalanine (Phe) + tyrosine (Try) + tryptophan (Trp); C = Gln + glyphosate; D = Gln + Phe + Tyr + Trp + glyphosate. The experiment was conducted under irradiance of 80 mmol-2s-1 at 25o for 204 hours. The treatments with aromatic amino acids and glutamine avoided the herbicide effect on seed fresh and dried weights and had no effect on their biochemical constituents (proteins, oil, fatty acids, carbohydrates and chlorophylls). Thus, an exogenous supplementation of aromatic amino acids eliminates phytotoxic effects of glyphosate on soybean explants. Therefore, it was developed an in vitro system that allows to study the mode of action and the metabolism of glyphosate in soybean seeds, since the explants behaved in a similar manner as the transgenic soybean unsusceptible to the herbicide.