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Cadmium (Cd) is one of the most threatening soil and water contaminants in agricultural settings. In previous studies, we observed that Cd affects the metabolism and physiology of tomato (Solanum lycopersicum) plants even after short-term exposure. The objective of this research was to use cross-genotype grafting to distinguish between root- and shoot-mediated responses of tomato genotypes with contrasting Cd tolerance at the early stages of Cd exposure. This study provides the first report of organ-specific contributions in two tomato genotypes with contrasting Cd tolerance: Solanum lycopersicum cv. Calabash Rouge and Solanum lycopersicum cv. Pusa Ruby (which have been classified and further characterized as sensitive (S) and tolerant (T) to Cd, respectively). Scion S was grafted onto rootstock S (S/S) and rootstock T (S/T), and scion T was grafted onto rootstock T (T/T) and rootstock S (T/S). A 35 μM cadmium chloride (CdCl

This work aimed to investigate the partial K-replacement by Na supply to alleviate drought-induced stress in Eucalyptus species. Plant growth, leaf gas exchange parameters, water relations, oxidative stress (H2O2 and MDA content), chlorophyll concentration, carbon (C) and nitrogen (N) isotopic leaf composition (δ13C and δ15N) were analyzed. Drought tolerant E. urophylla and E. camaldulensis showed positive responses to the partial K substitution by Na, with similar dry mass yields, stomatal density and total stomatal pore area relative to the well K-supplied plants under both water conditions, suggesting that 50% of the K requirements is pressing for physiological functions that is poorly substituted by Na. Furthermore, E. urophylla and E. camaldulensis up-regulated leaf gas exchanges, leading to enhanced long-term water use efficiency (WUEL). Moreover, the partial K substitution by Na had no effects on plants H2O2, MDA, δ13C and δ15N, confirming that Na, to a certain extent, can effectively replace K in plants metabolism. Otherwise, the drought-sensitive E. saligna species was negatively affected by partial K replacement by Na, decreasing plants dry mass, even with up-regulated leaf gas exchange parameters. The exclusive Na-supplied plants showed K-deficient symptoms and lower growth, WUEL, and δ13C, besides higher Na accumulation, δ15N, H2O2 and MDA content.

The Eucalyptus grandis L. plantations for industrial purposes (e.g., wood charcoal and pulp) are largely adopted worldwide, mainly in acidic and poorly fertile soils. The application of lime to soil is recommended for eucalyptus plantation, but lime reduces the availability of metallic micronutrients to plants. Therefore, the aim of this study was to investigate the performance of field-grown Eucalyptus grandis in Ferralsol amended with lime at different rates (0.0 and 1.6 and 3.7 Mg ha-1) and Cu plus Zn [0.0 and 0.0 kg ha-1 (-CuZn) and 0.5 and 1.5 kg ha-1 (+CuZn)]. The soil chemical attributes, shoot biomass, and nutrient accumulation were evaluated, while nutrient use efficiency, chlorophyll (Chl) and flavonoid (Flav) contents, and enzymatic antioxidant performance [superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), ascorbate peroxidase (APX), guaiacol peroxidase (GPOX), and glutathione peroxidase (GPX)] were assayed to assess the nutritional status of the plants. Despite the lime application rate increasing Ca and Mg soil availabilities, effective cation exchange capacity (eCEC), base saturation (BS), and pH values, the available Cu, Fe, and Zn concentrations decreased at 6 and 30 months after planting. Conversely, Mn soil availability increased with lime application rates. Liming augmented Cu-, Fe-, Mn-, and Zn-use efficiency, while +CuZn supply increased enzymatic antioxidant performance in 30-month-old plant. Overall, the combined application of lime and +CuZn enhanced shoot biomass relative to that of non-treated plants. However, there was no observed difference in shoot biomass in 30-month-old limed and non-limed eucalyptus trees that were supplied with +CuZn. This result can be partially explained by the improvement in soil chemical attributes, such as soil pH, eCEC, and BS. In conclusion, the use of integrative approaches and temporal evaluations provided new insights into wood yield, biochemical traits, antioxidant metabolism, and the nutritional status of field-grown eucalyptus under lime and +CuZn application rates.

Heat stress can impact crop development and yield and amino acids play diverse essential roles in plants. This work aimed to study the long-term effects of foliar spray with L-arginine in antioxidant machinery, physiology, nutrition, productivity and fruit quality of tomato plants subjected to transient heat stresses. Six concentrations of L-arginine were sprayed on the plants: 0 (control), 0.10, 0.25, 0.50, 1.0 and 2.0 g·L–1. The content of hydrogen peroxide (H2O2), a reactive oxygen species, decreased concurrently to the increasing arginine concentration. The ascorbate peroxide (APX) activity had an inverse behavior to that observed for H2O2 content (r = –0.79), not only indicating that arginine is able to modulate APX, but also suggesting that this enzyme plays an important role on the mitigation of H2O2 generation under heat stress. Ascorbate peroxide and catalase (CAT) activities had a positive correlation (r = 0.82), showing that these enzymes may work in tandem. The influence of arginine on photosynthesis activity and gas exchange was generally weak and depended mainly on the plant developmental stage. Yield was increased by 19.8 and 23.1% in plants that received 1.0 and 0.5 g·L–1 of arginine, respectively, when compared to control plants. In conclusion, the use of exogenous L-arginine can protect tomato plants against oxidative imbalance under transient heat within protected environments.

Cadmium (Cd) is a heavy metal considered as one of the main toxic pollutants in the environment and affects several metabolic processes in plants and animals, justifying scientific efforts to study Cd bioremediation and biochemical, physiological, and molecular mechanisms of plant response and tolerance to cope with Cd toxicity. Phytochelatins (PCs) are nonribosomally synthesized small metal-binding peptides, structurally related to the tripeptide glutathione and efficiently induced by Cd in several plant species. Several studies have demonstrated that PCs contribute to the modulation of Cd response and tolerance in plants, in addition to potential applications of PC properties and functions to improve and optimize phytoremediation. The present chapter embodies information on the existing knowledge about direct and indirect contribution of PCs and their synthesis to modulation of Cd tolerance and accumulation in plants, by highlighting studies focused on gene expression, PC isoforms, peptide quantification, and genetic transformation of plants under Cd exposure, besides proteomic studies showing some differentially expressed glutathione metabolism-related proteins.

The objective of the present study was to assess the response of tomato cultivars with different fruit colors to exposure to increasing Cd levels in the substrate by measuring the impacts of Cd on the oxidative stress indicators and physicochemical features of fruits, as well as plant development and yield components. A completely randomized experiment in a 3 × 3 factorial design [tomato cultivar (which produces purple, red, or white fruits) vs Cd level in the substrate (0, 3.6, or 12 mg kg−1)] was performed. The cultivation of plants in substrate containing 3.6 mg kg−1 Cd did not affect yield, but fruits exhibited nonpermissive Cd concentrations in both peel and mesocarp across all cultivars. By contrast, yield was decreased in plants with red and white fruits after their cultivation in substrate containing 12 mg kg−1 Cd, while the productivity of plants with purple fruits was maintained under such conditions. The hydrogen peroxide content in the fruit mesocarp depended only on cultivar. However, an increased lipid peroxidation level was detected in the mesocarp of purple fruits at the highest Cd concentration. No parameters of fruit quality [i.e., diameter, length, °Brix, pH, titratable acidity, color (L*, a*, and b*), and concentrations of lycopene and β-carotene in mesocarp] were affected by long-term exposure to Cd at 12 mg kg−1. In conclusion, the results of this study suggested that the potential Cd side effects on diverse tomato quality features can be buffered at the fruit level because these features were maintained at the usual values despite high Cd concentrations in tomato peel and pulp. Moreover, these buffering mechanisms are independent of lycopene and β-carotene concentrations in fruit peel, since the three tomato cultivars that were evaluated in the present study (white fruits, possessing no or negligible concentrations of these carotenoids, and red and purple tomato, possessing high lycopene and β-carotene concentrations) were able to sustain several fruit quality parameters after long-term exposure to high Cd concentrations in the substrate.

The capacity of three maize endosperm opaque mutants (o10, o11 and o13) to accumulate soluble lysine in the seed in relation to their wildtype counterpart, W22+, was investigated. The W22o13 and W22o11 mutants exhibited 278% and 186% increases in soluble lysine, respectively, while for W22o10, a 36% decrease was observed, compared with the wildtype. A quantitative and qualitative study of the N constituents of the endosperm has been conducted and data obtained for the total protein, non-protein N, soluble amino acids, albumins / globulins, zeins and glutelins present in the seed of the mutants. Following 2D–PAGE, a total of 38 different forms of zein polypeptides were detected and considerable differences were noted between the three mutant lines. The metabolism of lysine was also studied by analysis of the enzymes aspartate kinase, homoserine dehydrogenase, lysine 2-oxoglutarate reductase and saccharopine dehydrogenase, which exhibited major changes in activity, depending on the genotype, suggesting that the mutant genes may have distinct regulatory activities.

The objective of this study was to investigate and characterize cowpea (Vigna unguiculata) genotypes for total grain protein content, storage protein fractions (globulin, albumin, prolamin, basic and acid glutelins), and phytate and minerals contents. Eighteen cowpea genotypes were selected. Total grain protein content varied from 21.4% to 29.2%, for BRS Marataoã and Paulistinha genotypes, respectively. The variation in the concentration of each protein fraction was significant (P

We evaluated the mechanisms that control Cd accumulation and distribution, and the mechanisms that protect the photosynthetic apparatus of Brachiaria decumbens Stapf. cv. Basilisk and Panicum maximum Jacq. cv. Massai from Cd-induced oxidative stress, as well as the effects of simulated summer or winter conditions on these mechanisms. Both grasses were grown in unpolluted and Cd-polluted Oxisol (0.63 and 3.6 mg Cd kg

This work aimed to quantify the concentration of soluble amino acids, nitrogen (N) and Cd in the developing seeds of maize plants that were grown in Cd-contaminated soil from seed sowing to the reproductive stages. The specific activities and feedback inhibition properties of lysine (Lys) metabolic enzymes were also determined in developing seeds. The potential maize yield was depressed by Cd exposure. Cadmium concentration in plant organs followed the decreasing order: roots > stems = leaves > developing seeds (37.04, 1.95, 1.46 and 0.22 mg kg−1 Cd, respectively). The relatively low Cd concentration in the remained developing seeds was a result from root- and ear-mediated reductions of the Cd translocation in plants. Plants under Cd exposure presented developing seeds with an increased N concentration (up to 7%) when compared to control plants. Furthermore, the level of soluble amino acids (particularly histidine, glycine, tyrosine, methionine, isoleucine and valine) was increased in the developing seeds of Cd-treated plants. In addition, changes in the feedback properties of dihydrodipicolinate synthase (DHDPS), an enzyme from lysine metabolism, were observed in developing seeds. In conclusion, the current study showed that maternal plant exposure to Cd can alter the concentration of soluble amino acids and the behavior of Lys metabolic enzymes in developing seeds. This study provided not only new information about the influence of the long-term Cd exposure on plants, but also data about protective plant strategies against Cd toxicity.

In general, the effects of Cadmium (Cd) on crop quality are evaluated solely by Cd accumulation in the edible plant parts; thus, the potential effects on several nutritional features are not often taken into account. This study evaluated Cd effects on the enzymatic activities of lysine (Lys) metabolism, storage protein and amino acid profile in maize. Stress parameters were also assessed. In vegetative organs, Cd was accumulated in the following decreasing order: roots > stems > leaves. Cadmium accumulation in grains decreased at grain maturity (from 0.25 to 0.07 µg g−1 DW). In leaves, neither hydrogen peroxide content nor superoxide dismutase, catalase, ascorbate peroxide and guaiacol peroxidase activities did not change in Cd-treated plants compared to control plants. Lipid peroxidation was not detected in immature grains and leaves of plants under Cd exposure, indicating that Cd accumulation mainly in the roots is a mechanism to avoid oxidative stress in aboveground parts of the plant. However, Lys metabolism in immature grains was modified, showing increases in the specific activities of aspartate kinase (AK) and dihydrodipicolinate synthase (DHDPS) after Cd exposure. AK sensitivity to Lys feedback inhibition increased, but decreased in DHDPS in Cd-treated plants, suggesting differential regulation for these enzymes. In mature grains, the Lys content did not change, while the proline content increased by 54 % in Cd-treated plants. This is the first report on Cd effects on amino acid profile, storage protein contents and enzymes from Lys metabolism in grains of a cereal plant species.

Made available in DSpace on 2022-04-28T19:10:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-01 Abiotic stresses negatively affect crop development and yield. However, the use of seaweed extracts can alleviate production losses through improvement of the antioxidant system and synthesis of compatible osmolytes. The aim of this study was to assess the effects of Ascophyllum nodosum extract on antioxidant response and yield of common bean plants (Phaseolus vulgaris) under drought stress. Therefore, two methods of application (foliar spray and soil irrigation) and two doses (5 and 10 mL L-1) were tested, whereas untreated plants were used as control. There were no significant differences in malondialdehyde and hydrogen peroxide contents among treatments, indicating that application of seaweed extract did not change the oxidative status in treated plants when compared to the control ones. On the other hand, the use of seaweed extract increased proline content even prior to drought stress (46.3 to 145.4% when compared to the control), and this increment remained until the 10th day of stress (from 60.1 to 201.7% over the reference plants), supporting early evidences that A. nodosum extract is a plant elicitor. The yield parameters were not affected by the extract, probably due to the severe drought stress which plants were submitted. The results suggest that A. nodosum extract affects proline metabolism in Phaseolus vulgaris plants, making the synthesis of this osmolyte more responsive to drought stress because seaweed-treated plants presented a higher proline content than the control ones during the exposure to water deficit. São Paulo State University São Paulo State University

In this study, we are presenting recommendations to the best agricultural use as well as for plant breeding of three millet cultivars namely ENA1 and ENA2, which have African origin, and BRS1501 originally from India. These cultivars were evaluated for growth, yield and grain quality traits. The morphological traits evaluated in this study indicated that the African genotypes ENA1 and ENA2 are better than the Indian genotype BRS1501 for no-till farming or to produce forage with 15% of crude protein at flowering and at harvest to produce stover (around 7% of crude protein content) for livestock feeding. The BRS1501 cultivar exhibited the highest values for total crude protein, albumins and prolamins, phytate and mineral contents in grains. ENA1 and ENA2 exhibited the highest values of globulin and glutelin contents. The electrophoretic patterns for storage proteins were similar across the three millets cultivars, except for a higher intensity of two glutelin bands with 21 and 24 kDa in BRS1501. Together, the results allow us to recommend BRS1501 for grain production and ENA1 and ENA2 for biomass production.

Copper (Cu) deficiency and toxicity cause stresses in citrus orchards and limited information is available about which rootstocks and associated mechanisms would enhance plant resistance to such nutritional disorders. Therefore, this study evaluated the nutritional status and antioxidant system responses of citrus grafted onto selected rootstocks differing in horticultural performance [Swingle citrumelo (SW) or Rangpur lime (RL)], grown in nutrient solution with varying concentrations of Cu (0.015, 0.60 or 24.0 μM). The experiment was carried out in a greenhouse using young sweet orange trees. Once taken up, Cu mostly accumulated in roots (75% of total plant Cu content). Trees grafted onto RL were more responsive to enzyme activities related to oxidative stress and to nitrogen metabolism in leaves when grown in the presence of either the lowest or the highest Cu concentrations used. Those grown in 24.0 μM Cu displayed decreased overall nutrient uptake and accumulation, with the exception of iron, which was predominantly found in roots. Cu/Zn superoxide dismutase activity in leaves was dependent upon signalling regulated by rootstocks, being lower in SW than in RL. Therefore, the use of appropriate rootstock varieties contributes to alleviate the effects of Cu stress on the metabolism and nutritional status of citrus plants.

Cadmium (Cd) is a heavy metal highly toxic to plants and human and animal health. The increased concentration of this metal in the environment in recent decades is the result of anthropogenic activities that include the production of mineral fertilizers, pesticides, intense mining, waste disposal, and foundries. Cd accumulation is known to induce oxidative stress in plant cells due to overproduction of reactive oxygen species, which may result in severe damage at the cellular level, ultimately impairing plant growth and productivity. In response to such stress, different molecules having enzymatic or nonenzymatic actions contribute to mitigate plant cellular damage and balance redox homeostasis in plant cells. This chapter aims to summarize current knowledge about the prospecting of important components of the molecular mechanisms associated with antioxidant machinery in plant response and tolerance to Cd toxicity. Insights into antioxidant response regulation, which includes hormones, signaling molecules, transcription factors, and microRNAs, are also presented.

Cadmium (Cd) toxicity is frequently coupled to its accumulation in plants, but not always the highest Cd concentration triggers the worst damages, indicating that additional events influence the magnitude of Cd side-effects. We investigated the early mechanisms behind the differential Cd-induced impacts on plant development of four tomato accessions with contrasting tolerance to Cd toxicity. At organ level, the highest Cd concentration was not associated with the largest biomass losses. In leaves, changes in superoxide dismutase and catalase activities were not related to differences in Cd concentration, which was unable to provoke H2O2 overproduction on the sixth day of plant exposure to this metal. Further investigation in the mineral profile revealed that magnitude of Cd toxicity depends probably on synergic effects from increased B status, in addition to the own Cd accumulation. Furthermore, disbalances in Mn status (i.e., excess in leaves and deficiency in roots) may enhance Cd toxicity degree. According to data, however, the low magnesium (Mg) status can be linked to tomato tolerance against Cd toxicity. In conclusion, the tomato tolerance degree under short-Cd exposure depends on actively, finely regulation of mineral homeostasis that results in different development of plant organs. The better understanding on the mode of action of Cd toxicity in plants can help in the establishment of strategies to mitigate its impacts on crop yield.

The grain proteins of barley are deficient in lysine and threonine due to their low concentrations in the major storage protein class, the hordeins, especially in the C-hordein subgroup. Previously produced antisense C-hordein transgenic barley lines have an improved amino acid composition, with increased lysine, methionine and threonine contents. The objective of the study was to investigate the possible changes in the regulation of key enzymes of the aspartate metabolic pathway and the contents of aspartate-derived amino acids in the nontransgenic line (Hordeum vulgare L. cv. Golden Promise) and five antisense C-hordein transgenic barley lines. Considering the amounts of soluble and protein-bound aspartate-derived amino acids together with the analysis of key enzymes of aspartate metabolic pathway, we suggest that the C-hordein suppression did not only alter the metabolism of at least one aspartate-derived amino acid (threonine), but major changes were also detected in the metabolism of lysine and methionine. Modifications in the activities and regulation of aspartate kinase, dihydrodipicolinate synthase and homoserine dehydrogenase were observed in most transgenic lines. Furthermore the activities of lysine α-ketoglutarate reductase and saccharopine dehydrogenase were also altered, although the extent varied among the transgenic lines.

Both the scientific community and society have shown interest in improving the content of amino acids, carbohydrates and mineral nutrients in maize because it represents an important staple food in many developing countries. Earlier studies demonstrated that the treatment of seeds using ascorbic acid (AsA-seed priming) enhanced soluble carbohydrates, proteins and soluble amino acids for other species. AsA seed priming in maize showed the potential for reducing abiotic stresses. The effects on grain quality have not been previously demonstrated. This study investigated the impacts of AsA seed priming on maize kernel quality of seeds produced by the plants generated from the primed seeds, based on the amino acid profile and carbohydrate and mineral nutrient contents. AsA seed priming improved the maize kernel quality with respect to the ascorbate content, boron allocation, total carbohydrate content and increased soluble amino acid levels, including serine, tyrosine, alanine, valine, glutamate, arginine, proline, aspartate, lysine and isoleucine, whereas soluble methionine was decreased. Therefore, AsA seed priming can represent a potential technique for improving maize grain quality.

Storage of recalcitrant seeds leads to the initiation of subcellular damage or to the initiation of germination process, and both may result in viability loss. This study aimed to elucidate the biochemical basis of embryos survival of Araucaria angustifolia recalcitrant seeds during storage. After harvesting, seeds were stored at ambient conditions (without temperature and humidity control) and in a cold chamber (temperature of 10 ± 3 °C, and relative humidity of 45 ± 5 %). Moisture content, viability, H2O2 content, lipid peroxidation, protein content, and activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX), at 0, 15, 45 and 90 days of storage, were evaluated. Seed viability reduced about 40 % during the storage period accompanied by a reduction in soluble protein (about 64 % of reduction) in both storage conditions, and increased lipid peroxidation (about 115 % and 66 % for ambient and cold chamber conditions, respectively). H2O2 content used as a marker of oxidative stress was reduced during the period, possibly controlled by the action of CAT and APX, for which increased activities were observed. The results allowed the identification of seven SOD isoenzymes (one Mn-SOD, five Fe-SOD and one Cu/Zn-SOD), whose activities also increased in response to storage. Some biochemical damage resulting from storage was observed, but viability reduction was not due to failure of enzymatic protection mechanisms.

Sugarcane yield and quality are affected by a number of biotic and abiotic stresses. In response to such stresses, plants may increase the activities of some enzymes such as glutathione transferase (GST), which are involved in the detoxification of xenobiotics. Thus, a sugarcane GST was modelled and molecular docked using the program LIGIN to investigate the contributions of the active site residues towards the binding of reduced glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). As a result, W13 and I119 were identified as key residues for the specificity of sugarcane GSTF1 (SoGSTF1) towards CDNB. To obtain a better understanding of the catalytic specificity of sugarcane GST (SoGSTF1), two mutants were designed, W13L and I119F. Tertiary structure models and the same docking procedure were performed to explain the interactions between sugarcane GSTs with GSH and CDNB. An electron-sharing network for GSH interaction was also proposed. The SoGSTF1 and the mutated gene constructions were cloned and expressed in Escherichia coli and the expressed protein purified. Kinetic analyses revealed different Km values not only for CDNB, but also for GSH. The Km values were 0.2, 1.3 and 0.3 mM for GSH, and 0.9, 1.2 and 0.5 mM for CDNB, for the wild type, W13L mutant and I119F mutant, respectively. The Vmax values were 297.6, 224.5 and 171.8 μmol min −1 mg −1 protein for GSH, and 372.3, 170.6 and 160.4 μmol min −1 mg −1 protein for CDNB.

The total protein content and activity of the enzymes glutathione reductase (GR), superoxide dismutase (SOD) and thioredoxin reductase (TrxR) were evaluated in Acidithiobacillus ferrooxidans LR cells maintained in contact with the metal sulfide chalcopyrite for 1 and 10 days. A significant decrease in total protein content was observed in cells maintained for 10 days in the presence of chalcopyrite, suggesting proteolytic breakdown due to exposure to the metal sulfide. Following 10 days of contact with chalcopyrite, increases in GR, SOD and TrxR activities were detected, suggesting the formation of reactive oxygen species. After ten days, there was a fivefold increase in GR activity, of which, isoenzyme IV represented approximately 82% of the total. An increase in Fe-SOD activity following ten days exposure to chalcopyrite was also determined, as measured on non-denaturing polyacrylamide gels. Also, after 10 days, an approximately 31-fold increase was observed for TrxR activity. The presence of oxidative stress when A. ferrooxidans is in the presence of chalcopyrite could have a negative impact on bioleaching.

Tomato is an important crop worldwide. Cadmium (Cd) concentrations in fruits depend on tomato genotype. This work aimed to study the relation among Cd accumulation, tolerance mechanisms, and fruit features in two tomato cultivars with contrasting tolerance to Cd stress. Tolerant (Yoshimatsu) and sensitive (Tropic Two Orders) plants were grown in control and contaminated soils (0.04 and 3.77mg/kg Cd, respectively) from the seedling stage to fruit production. Both cultivars were able to acclimatize to Cd exposure, probably through mechanisms associated with reductions in the magnesium status. Cadmium concentrations varied according to the following descending order: roots=leaf blades > (peduncle+sepals) > stem=fruits. However, the tolerant cultivar accumulated more Cd than did the sensitive one. Although Cd reached the fruits from the first to the fourth bunches, peduncle and sepals may act as a barrier to Cd entrance in tomato pulp and peel. The Cd-induced changes in the fruit mineral profile varied according to plant cultivar, organ, tomato tissue, and bunch position. Moreover, plant yield was not affected by the Cd stress, which was able to improve fruit size and weight in the tolerant cultivar. In conclusion, new insights about the Cd-induced effects on tomato development and fruit attributes were provided by growing plants in soil, which is the media generally used to cultivate this crop, rather than hydroponics. It was shown that tomato cultivars with contrasting tolerance to Cd toxicity can reach sexual maturity and produce fruits with no yield losses, despite impacts on development from long-term Cd exposure. This study also revealed the role of floral receptacle and its related structures in limiting, even partially, Cd translocation to the fruits. Furthermore, Yoshimatsu's capacity to produce bigger and heavier fruits, in plants under Cd exposure, may probably be associated with enhanced Cd accumulation. Fundacao de Amparo a Pesquisa do Estado de Sao Paulo [2009/54676-0, 2013/15217-5]; Conselho Nacional de Desenvolvimento Cientifico e Tecnologico [303749/2016-4, 476096/2013-8]

O ovo é o produto de eficiente transformação biológica feita pela galinha (Gallus gallus) de postura, com alta qualidade nutricional para o consumo humano. Na gema do ovo encontra-se a proteína denominada fosvitina, que atua como carreadora do ferro se ligando a ele. A deficiência de ferro ocorre normalmente devido ao consumo insuficiente de alimentos fontes de ferro ou à baixa biodisponibilidade. O objetivo da pesquisa foi verificar a concentração de proteínas, em especial a fosvitina/lipovitelina presente em gemas de ovos crus e cozidos nos diferentes ciclos de postura (inicial, intermediário e final) e avaliar a interferência desta proteína na disponibilidade do micronutriente ferro. Os tratamentos foram realizados em diferentes períodos e utilizaram ovos crus e cozidos, sendo gemas cruas de poedeiras em início, meio e fim de postura e gemas cozidas nos três ciclos de postura. Foram realizadas as análises de composição centesimal, concentração de ferro total e dialisável, concentração de proteínas em gemas e eletroforese em gel de poliacrilamida (SDS-PAGE). Houve diferença na concentração de proteínas e diferentes níveis de ferro dialisável nos três ciclos de postura, porém em gemas cruas e cozidas. Com o passar dos ciclos (inicial, intermediário e final), as poedeiras mais velhas, apresentaram ovos (gemas) com um teor mais elevado de ferro disponível, porém uma quantidade inferior de proteínas em especial a fosvitina/lipovitelina, comparando-se com os ciclos inicial e intermediário. A disponibilidade de ferro foi mais elevada em gemas que passaram pelo processo de cocção, porém em poedeiras de postura final.The egg is the product of efficient biological transformation made by a laying hen. It has high nutritional value for human consumption. Most of the iron in egg yolk is bound by phosvitin, a major protein present in the egg yolk. Iron deficiency normally occurs due to insufficient intake of dietary iron or its low the bioavailability. The objective of this research was to verify the protein concentration, specially the phosvitin/lipovitelin in egg yolks of raw and cooked eggs in different posture cycles (initial, intermediate, final), and to evaluate the interference of this protein in the availability of the iron micronutrient. The treatments were carried in different periods using raw and cooked eggs. The yolks of raw eggs of laying hens were used in the initial, intermediate and final phases of posture and yolks of cooked eggs in the three posture cycles. Analyses of proximal composition, concentration of total and available iron, protein concentration in egg yolks, and eletrophoresis in poliacrilamida (SDS-PAGE) were performed. There was difference in the protein concentration and in the levels of iron availability in the three posture cycles. After running through the cycle (initial, intermediate, and final), the eggs yolks of the older hens presented an increased amount of available iron and lower amount of proteins, specially the fosvitin/lipovitelin, if compared to the initial and intermediate cycles. The highest iron availability content was found in cooked egg yolks and in hens in the final posture cycle.

Hordeins are the major storage proteins in barley grains and are responsible for their low nutritional quality. Previously, antisense C-hordein barley lines were generated and were shown to contain a more balanced amino acid composition and an altered storage protein profile. In the present study, a proteomic approach that combined two-dimensional gel electrophoresis (2-DE) and mass spectrometry was used to (1) identify the changes in the protein profile of non-storage proteins (salt soluble fraction) in antisense C-hordein barley lines (L1, L2 and L3) and (2) map the differentially expressed proteins compared to the non-transgenic control line (Hordeum vulgare cv. Golden Promise). Moreover, the changes in the proteins were correlated with the more balanced amino acid composition of these lines, with special attention to the lysine content. The results showed that suppression of C-hordein expression does not exclusively affect hordein synthesis and accumulation. The more balanced amino acid composition observed in the transgenic lines L1, L2 and L3 was an indirect result of the profound alterations in the patterns of the non-storage proteins. The observed changes included up-regulated expression of the proteins involved in stress and detoxification (L1), defence (L2 and L3), and storage globulins (L1, L2 and L3). To a lesser extent, the proteins involved in grain metabolism were also changed. Thus, the increased essential amino acids content results from changes in distinct protein sources among the three antisense C-hordein lines analyzed, although the up-regulated expression of lysine-rich proteins was consistently observed in all lines.

Foram aplicados tratamentos térmicos em laranja "Valência", tangor "Murcott" e lima ácida "Tahiti" armazenadas em baixa temperatura e avaliados os efeitos na redução de injúrias pelo frio e sobre outras características bioquímicas. Os seguintes tratamentos foram aplicados: T1: frutas armazenadas a 1°C (Controle); T2: aquecimento rápido das frutas em água quente a 53°C, durante 3 minutos, sendo, em seguida, armazenadas a 1°C; T3: aquecimento lento das frutas em câmara regulada para 37°C, por 2 dias, com posterior armazenamento a 1°C; e T4: aquecimento intermitente em ciclos de 6 dias a 1°C + 1 dia a 25°C. As frutas foram armazenadas durante 90 dias a 1°C e 90-95% de Umidade Relativa (UR), sendo avaliadas a cada 15 dias. Além da incidência dos danos, foram avaliadas também as atividades das enzimas antioxidativas (catalase, glutationa redutase e ascorbato peroxidase). A lima ácida "Tahiti" e o tangor "Murcott" suportaram até 90 dias de armazenamento a 1°C com aquecimento intermitente, não apresentando danos pelo frio. No tratamento controle (armazenamento contínuo a 1°C), os danos pelo frio surgiram aos 30 dias de armazenamento para a lima "Tahiti" e aos 45 dias para o tangor "Murcott". Em laranjas "Valência", as injúrias pelo frio surgiram aos 45 dias de armazenamento, sendo significativamente menores no condicionamento térmico. Os efeitos dos tratamentos térmicos no aumento da resistência das frutas ao frio podem estar relacionados com a atividade das enzimas antioxidativas. Para a laranja "Valência", os tratamentos térmicos, aplicados na forma de condicionamento térmico ou aquecimento intermitente, reduzem as injúrias pelo frio e podem prolongar a conservação das frutas a 1°C e 90-95% UR. Para tangor "Murcott" e lima "Tahiti", o aquecimento intermitente é o tratamento mais eficiente para a redução de injúrias pelo frio e para o aumento na capacidade de armazenamento. In the present study thermal treatments on,'Valência' orange, 'Murcott' tangor and 'Tahiti' (temperature conditioning and intermittent warming) were applied lime. The following treatments were applied: T1: Fruits stored at 1°C (Controls); T2: Fast heating of the fruits in hot water at 53°C for 3 minutes and stored at 1°C; T3: Slow heating of the fruits in camera regulated at 37°C for 2 days, with subsequent storage at 1°C; and T4: Intermittent heating in cycles of 6 days at 1°C + 1 day at 25°C. Fruits were stored under low temperature (1°C) and 90-95% relative humidity (RH) during 90 days. Chilling injury incidence and changes in physical-chemical characteristics were evaluated every 15 days. Activity of catalase, glutathione reductase and ascorbate peroxidase was also determined. Intermittent warming was more efficient than temperature conditioning to reduce chilling injury. 'Tahiti' lime and 'Murcott' tangor supported up to 90 days of intermittent warming stored at 1°C, while control fruits showed chilling injury after 30 days ('Tahiti' lime) and 45 days ('Murcott' tangor). 'Valência' orange was affected by chilling injury after 45 days of cold storage, and temperature conditioning has caused lower incidence of this disorder. The effect of thermal treatments can be related to the activity of antioxidant enzymes. For ´Valência" orange, thermal treatments, applied as thermal conditioning or intermittent heating, reduce injuries caused by cold and can lengthen the conservation of fruits at 1°C and 90-95% RH. The intermittent heating is the most efficient treatment to reduce injuries caused by cold and increase at storage capacity of the 'Murcott' tangor and 'Tahiti' lime.

Lysine is an essential amino acid normally present in very low concentration in cereal seeds. In previous reports we have studied the metabolism of lysine in several distinct high-lysine maize mutants and observed drastic variations in the activity of saccharopine dehydrogenase (SDH), a key enzyme involved in lysine degradation. We have now analyzed the activity of SDH using non-denaturing polyacrylamide gel electrophoresis (PAGE) to identify possible isoenzymes that could explain the patterns of activity previously observed. The results indicated the presence of at least two SDH isoenzymes, one contributing to approximately 90% of the total enzyme activity and a minor form only present in the wild type lines and the opaque-1 mutant. The results suggest that the differences in total SDH activity among the genotypes tested are due to alterations in the predominant SDH isoenzymic form, which is likely to be the bifunctional polypeptide containing lysine 2-oxoglutarate reductase.

Two high lysine maize endosperm mutations, opaque-5 (o5) and opaque-7 (o7), were biochemically characterized for endosperm protein synthesis and lysine metabolism in immature seeds. Albumins, globulins, and glutelins, which have a high content of lysine, were shown to be increased in the mutants, whereas zeins, which contain trace concentrations of lysine, were reduced in relation to the wild-type lines B77xB79+ and B37+. These alterations in the storage protein fraction distribution possibly explain the increased concentration of lysine in the two mutants. Using two-dimensional polyacrylamide gel electrophoresis of proteins of mature grains, variable amounts of zein polypeptides were detected and considerable differences were noted between the four lines studied. The analysis of the enzymes involved in lysine metabolism indicated that both mutants have reduced lysine catabolism when compared to their respective wild types, thus allowing more lysine to be available for storage protein synthesis.

Barley (Hordeum vulgare L.) has had an important impact on human nutrition. Hull-less barley is a genetically improved type that has been widely used in recent years. Six Brazilian hull-less barley varieties (IAC-IBON 214-82; IAC 8612-421; IAC 8501-31; IAC 8501-12; IAPAR 39-ACUMAI; IAC 8501-22) were analyzed for storage protein constituents, amino acid contents, and similarity among the hull-less barley varieties. Albumins, globulins, prolamins I and II, and glutelins were extracted and separated by SDS-PAGE. The total protein amino acid contents of the flour were also determined for each variety by TLC and HPLC. Variations in intensity and appearance and disappearance of protein bands were observed among the varieties suggesting genetic variability. However, the amino acid profile did not indicate any major variations in the amino acid concentrations. The high lysine and threonine total concentrations detected in the seeds of the hull-less barley varieties encouraged an investigation into the re...

The capacity of two maize opaque endosperm mutants (o1 and o2) and two floury (fl1 and fl2) to accumulate lysine in the seed in relation to their wild type counterparts Oh43+ was examined. The highest total lysine content was 3.78% in the o2 mutant and the lowest 1.87% in fl1, as compared with the wild type (1.49%). For soluble lysine, o2 exhibited over a 700% increase, whilst for fl3 a 28% decrease was encountered, as compared with the wild type. In order to understand the mechanisms causing these large variations in both total and soluble lysine content, a quantitative and qualitative study of the N constituents of the endosperm has been carried out and data obtained for the total protein, nonprotein N, soluble amino acids, albumins/globulins, zeins and glutelins present in the seed of the mutants. Following two-dimensional PAGE separation, a total of 35 different forms of zein polypeptides were detected and considerable differences were noted between the five different lines. In addition, two enzymes of the aspartate biosynthetic pathway, aspartate kinase and homoserine dehydrogenase were analyzed with respect to feedback inhibition by lysine and threonine. The activities of the enzymes lysine 2-oxoglutate reductase and saccharopine dehydrogenase, both involved in lysine degradation in the maize endosperm were also determined and shown to be reduced several fold with the introduction of the o2, fl1 and fl2 mutations in the Oh43+ inbred line, whereas wild-type activity levels were verified in the Oh43o1 mutant.

Lysine, threonine, methionine and isoleucine are synthesized through the aspartate metabolic pathway. The concentrations of soluble lysine and threonine in cereal seeds are very low. Coix lacryma-jobi (coix) is a maize-related grass and the enzymological aspects of the aspartate metabolic pathway are completely unknown. In order to obtain information on lysine metabolism in this plant species, two enzymes involved in the biosynthesis of these amino acids (aspartate kinase AK, EC 2.7.2.4 and homoserine dehydrogenase HSDH, EC 1.1.1.3) and two enzymes involved in lysine degradation (lysine 2-oxoglutarate reductase LOR, EC 1.5.1.8 and saccharopine dehydrogenase SDH, EC 1.5.1.9) were isolated and partially characterized in coix seeds. AK activity was inhibited by threonine and lysine separately, suggesting the presence of two isoenzymes, one sensitive to lysine and the other sensitive to threonine, with the latter corresponding to approximately 60% of the total AK activity. In contrast to previous results from other plant species, the threonine-sensitive AK eluted from an ion exchange chromatography column at higher KCl concentration than the lysine-sensitive form. The HSDH activity extracted from the seeds was partially inhibited by threonine, indicating the presence of threonine-sensitive and threonine-resistant isoenzymes. LOR and SDH activities were detected only in the endosperm tissue and co-purified on an anion exchange chromatography column, suggesting that the two activities may be linked on a single bifunctional polypeptide, as observed for other plant species. One single SDH activity band was observed on non-denaturing PAGE gels. The Km for saccharopine of SDH was determined as 0.143 mM and the Km for NAD as 0.531 mM. Although SDH activity was shown to be stable, LOR, AK and HSDH were extremely unstable, under all buffer systems tested.

Lysine biosynthesis has been extensively studied and the regulatory enzymes characterized in some of the most important crop plants, however, much less is known about the lysine degradation pathway. Lysine 2-oxoglutarate reductase (LOR) and saccharopine dehydrogenase (SDH) have recently been partially purified and characterized from plants, and have been shown to exist as a single bifunctional polypeptide. We have further characterized these enzymes from rice endosperm in relation to Ca2+ and ionic strength modulation. Optimum pH values of 7.0 and 8.0 were obtained for LOR and SDH, respectively. The LOR domain of the polypeptide was modulated by Ca2+ and ionic strength, whereas the SDH domain was not. It would appear that the modulation by Ca2+ and ionic strength of LOR is a common feature among plant LOR enzymes. S-adenosylmethionine (SAM) did not produce any significant effect on either enzyme activity, indicating that it only plays a role in the regulation of lysine biosynthesis. The effect of S-2-aminoethyl- l-cysteine (AEC) as both a substrate and an inhibitor of LOR activity was also tested. AEC was shown to partially substitute for lysine as a substrate for LOR, but was also able to inhibit LOR activity, possibly competing with lysine at the active site. The higher Km for AEC compared to lysine may reflect a lower binding affinity for AEC.

Summary Aspartate kinase (AK, EC 2.7.2.4) and homoserine dehydrogenase (HSDH, EC 1.1.1.3) have been isolated and partially purified from immature rice (Oriza sativa L.) seeds. Three methods of protein separation (ammonium sulphate precipitation, anion-exchange chromatography and gel filtration chromatography) were used in order to purify and identify the isoenzymes of AK and HSDH. Two peaks of AK activity were eluted from the anion-exchange chromatography column (FFQ-Sepharose) with 230 and 286 mmol/L KCl. The first peak in elution order was inhibited by threonine, while the second peak was strongly inhibited by lysine. An optimum pH of 7.4 was determined for total AK activity. The optimum temperature for AK action was about 35 °C, whereas a combination of 12.5 mmol/L magnesium sulphate and 20 mmol/L ATP produced the highest level of activity. After the ammonium sulphate step, only one form of HSDH that was completely inhibited by threonine, was observed. The threonine-sensitive forms of HSDH and AK co-purified, independent of the protein purification method used exhibiting a molecular mass of 186 kDa. The lysine-sensitive AK had a molecular mass estimated to be 167 kDa. These results confirm the hypothesis that in higher plants, threonine and lysine-sensitive forms of AK are present and suggest that a bifunctional polypetide containing threonine-sensitive AK and HSDH is present is rice seeds.

In plant, the catabolism of lysine has only been studied in some detail in maize. The enzymes lysine 2-oxoglutarate reductase (also known as lysine alpha-ketoglutarate reductase; LOR) and saccharopine dehydrogenase (SDH), which convert lysine into saccharopine, and saccharopine into glutamic acid and 2-aminoadipate 6-semialdehyde, respectively, were isolated from immature rice seeds and partially purified through a three-step purification procedure involving ammonium sulphate precipitation, and anion-exchange and gel-filtration chromatographies, leading to a final yield of 30% for LOR and 24% for SDH. The molecular masses estimated by gel-filtration chromatography on a Sephacryl S200 column and by native non-denaturing PAGE using Ferguson plots were 203 kDa for both enzymes by gel-filtration and 202 kDa for both enzymes by native non-denaturing PAGE. A second band of LOR and SDH activities on native gels was observed for both enzymes with an estimated molecular mass of 396 kDa, which indicated a multimeric structure. Kinetic studies were consistent with an ordered sequence mechanism for LOR, where 2-oxoglutarate is the first substrate and saccharopine is the last product. The results observed for the LOR/SDH activity ratios during purification, the copurification in all three steps, the molecular masses, the relative mobilities on native non-denaturing gels and the pI estimated for LOR and SDH suggest the existence of a bifunctional polypeptide containing LOR and SDH activities.

Six S5 lines of maize, with differences for lysine and oil contents in grains, were used to carry out a complete series of diallelic crosses. The resulting 15 hybrids were grown in a field at two nitrogen levels (10 and 130 kg N ha-1). The general combining ability (GCA) and specific combining ability (SCA) were obtained by using the method 4, model I of Griffing for grain yield, and grain lysine and oil contents. Significant (p < 0.001) interaction was observed between GCA and N levels for grain yield, indicating the selection of different lines for each N level. This interaction was not significant for lysine content, but there were significant effects of GCA for this trait at both N levels (p < 0.1). Significant effects were not observed for GCA or SCA for oil content, however a positive correlation was observed between lysine and oil contents in the hybrids, in the lines and even in the control cultivars. The results indicate the effectiveness of selection for lysine content, irrespective of N level, in the studied non-opaque maize lines, and the possibility of achieving both high lysine and oil content in grains.

Selenium (Se) is an essential element for humans and animals that is required for key antioxidant reactions, but can be toxic at high concentrations. We have investigated the effect of Se in the form of selenite on coffee cell suspension cultures over a 12-day period. The antioxidant defence systems were induced in coffee cells grown in the presence of 0.05 and 0.5 mm sodium selenite (Na2SeO3). Lipid peroxidation and alterations in antioxidant enzymes were the main responses observed, including a severe reduction in ascorbate peroxidase activity, even at 0.05 mm sodium selenite. Ten superoxide dismutase (SOD) isoenzymes were detected and the two major Mn-SOD isoenzymes (bands V and VI) responded more to 0.05 mm selenite. SOD band V exhibited a general decrease in activity after 12 h of treatment with 0.05 mm selenite, whereas band VI exhibited the opposite behavior and increased in activity. An extra isoenzyme of glutathione reductase (GR) was induced in the presence of selenite, which confirmed our previous results obtained with Cd and Ni indicating that this GR isoenzyme may have the potential to be a marker for oxidative stress in coffee.

Lysine is an essential amino acid synthesized in plants via the aspartic acid pathway. The catabolism of lysine is performed by the action of two consecutive enzymes, lysine 2-oxoglutarate reductase (LOR, EC 1.5.1.8) and saccharopine dehydrogenase (SDH, EC 1.5.1.9). The final soluble lysine concentration in cereal seeds is controlled by both synthesis and catabolism rates. The production and characterization of high-lysine plants species depends on knowledge of the regulatory aspects of lysine metabolism and manipulation of the key enzymes. We have for the first time isolated, partially purified, and characterized LOR and SDH from developing sorghum seeds, which exhibited low levels of activity. LOR and SDH were only located in the endosperm and were very unstable during the isolation and purification procedures. LOR and SDH exhibited some distinct properties when compared to the enzymes isolated from other plant species, including a low salt concentration required to elute the enzymes during anion-exchange chromatography and the presence of multimeric forms with distinct molecular masses.

Lysine is catabolyzed by the bifunctional enzyme lysine 2-oxoglutarate reductase-saccharopine dehydrogenase (LOR-SDH) in both animals and plants. LOR condenses lysine and 2-oxoglutarate into saccharopine, using NADPH as cofactor and SDH converts saccharopine into alpha-aminoadipate delta-semialdehyde and glutamic acid, using NAD as cofactor. The distribution pattern of LOR and SDH among different tissues of Phaseolus vulgaris was determined. The hypocotyl contained the highest specific activity, whereas in seeds the activities of LOR and SDH were below the limit of detection. Precipitation of hypocotyl proteins with increasing concentrations of PEG 8000 revealed one broad peak of SDH activity, indicating that two isoforms may be present, a bifunctional LOR-SDH and possibly a monofunctional SDH. During the purification of the hypocotyl enzyme, the LOR activity proved to be very unstable, following ion-exchange chromatography. Depending on the purification procedure, the protein eluted as a monomer of 91-94 kDa containing only SDH activity, or as a dimer of 190 kDa with both, LOR and SDH activities, eluting together.

The Opaque-2 (O2) gene encodes a transcriptional activator specifically expressed for grain development of maize. o2 mutants have an opaque and chalky kernel, with a decrease in zein storage protein content, and an increase in the proportions of lysine and tryptophan. In this review, we present recent results investigating genetic properties of the O2 network, using transcriptome and proteome approaches, associated with measurements of activities of enzymes of the aspartate pathway and lysine degradation. The structural polymorphism at the O2 locus was investigated by RFLP in a collection of 51 maize inbred lines. Most polymorphic sites were found outside the coding regions. We then searched for relationships between RFLP polymorphism and (i) mRNA abundance of O2 and of known or suspected target genes, (ii) activity of SDH and (iii) amount of zein isoforms. Polymorphic restriction sites in the 5' upstream regions of the O2 gene were found associated with O2 mRNA abundance (three sites) and the amount of two 19 kDa alpha-zein isoforms (two sites). One restriction site on the 3' side of the O2 gene was found associated with Lor/Sdh mRNA abundance. Our results indicate relationships between polymorphism at the O2 locus and the expression of some of its target genes. Evidence of these associations has to be confirmed on larger samples, and the analysis of the O2 gene sequence should allow more precise testing of the actual involvement of O2 polymorphism in its own transcriptional expression, and in the expression of its target genes. O gene Opaco-2 (O2), expresso especificamente no grão de milho, transcreve para um fator de transcrição da família "leucine-zipper". Mutantes o2 apresentam grãos opacos, redução na quantidade de zeínas e aumento na proporção de lisina e triptofano. Genes cuja expressão é controlada diretamente pelo O2 são conhecidos (alfa-zeínas de 22 kDa, beta-zeínas de 14 kDa, b-32 e cyPpdk1). Nesta revisão, nós apresentamos resultados da caracterização genética de genes relacionados com o O2, através de abordagens de transcritoma, proteoma e de atividades enzimáticas da via metabólica do aspartato e da degradação da lisina. O polimorfismo do locus O2 foi avaliado utilisando-se a técnica de RFLP em 51 linhagens de milho. A maioria dos polimorfismos foi observada nas regiões não codificadoras da proteína. Análises de correlação foram realizadas entre os polimorfismos de RFLP e (i) quantidade de RNAm do O2, cyPpdk, Lor/Sdh e Ahas (ii) quantidade de isoformas de zeínas e (iii) atividade da enzima SDH. Sítios polimórficos foram correlacionados com a quantidade de RNAm do próprio O2, do gene Lor/Sdh e com a quantidade de duas isoformas de a-zeinas de 19 kDa. Nossos resultados indicam a presença de relações entre o polimorfismo do locus O2 e o nível de expressão de genes sob o seu controle. A utilização de um maior número de linhagens e o uso de dados de seqüência do O2 permitirá uma análise precisa da conseqüência do polimorfismo deste fator de transcrição sobre o controle do seu próprio nível de expressão e dos genes por ele controlados.

The nutrition value of a protein is directly related to its amino acid composition. Some of these amino acids, termed essential amino acids, cannot be synthesized by humans and therefore must be supplied in the diet for adults and in particular for infants and children. Lysine is an essential amino acid synthesized via the aspartic acid metabolic pathway, in which threonine, methionine and isoleucine are also endproducts. Moreover, lysine is the first limiting amino acid in all cereal grains. For over 30 years, the aspartic acid metabolic pathway has been studied in higher plants with the aim of identifying and characterizing the key regulatory points controlling the biosynthetic pathway. Two clear distinct time periods, one begining with the development of tissue culture techniques (1970-80's) and the second with the development of plant transformation techniques (90's), has encouraged the production of biochemical mutants and transgenic plants with specific alterations in key enzymes of the pathway, leading to the overproduction and accumulation of threonine in all plant tissues. However, the accumulation of lysine in seeds has been particularly difficult to achieve. Such an observation, associated with the recent biochemical studies on lysine degradation in cereal and legume plant species, has indicated that the manipulation of lysine degradation is as important as the manipulation of lysine synthesis, if the goal of accumulating this amino acid in cereal seeds is to be achieved. In maize, the study and use of other mutants such as the opaque-2 and QPM (Quality Protein Maize) varieties, has contributed significantly to our understanding of the regulatory aspects of the aspartate pathway. The strategies of obtaining cereals rich in lysine and their relevance to the manipulation of other amino acids have been discussed.