Free proline accumulation in sugarcane under water restriction and spittlebug infestation

A cigarrinha-das-raízes Mahanarva fimbriolata (Stål) (Hemiptera: Cercopidae) tornou-se uma praga importante da cultura da cana-de-açúcar na região centro-sul do Brasil. Estratégias de controle têm sido desenvolvidas, mas os danos promovidos por essa praga e sua interação com a cana-de-açúcar ainda são pouco caracterizados. em altos níveis de infestação da praga, os sintomas nas plantas são muito semelhantes à restrição hídrica severa. Este trabalho foi conduzido com o objetivo de determinar se o estresse promovido pela infestação de ninfas de cigarrinha-das-raízes resulta em acúmulo de prolina livre, além de comparar dois genótipos de cana-de-açúcar quanto à tolerância ao déficit hídrico. Dois experimentos foram conduzidos em delineamento inteiramente casualizado, em esquema fatorial 2 x 2 x 4 com duas cultivares (SP80-1816 e RB72454), dois níveis de estresse (controle e dez ninfas por planta ou 50% de restrição hídrica) e quatro épocas. O déficit hídrico promovido por M. fimbriolata não resultou em acúmulo de prolina, ilustrando a existência de mecanismos distintos para a percepção do estresse hídrico promovido pela praga e pela variação no potencial osmótico nas células das raízes. A cultivar RB72454 acumula maiores teores de prolina livre, e o acúmulo de massa seca e o crescimento dos colmos são menos afetados nessa cultivar sob restrição hídrica. Os teores de solutos compatíveis provavelmente não podem ser utilizados para quantificar o estresse promovido por M. fimbriolata em cana-de-açúcar. A cultivar RB72454 é mais tolerante ao déficit hídrico do que a cultivar SP80-1816.

Free amino acids and sugars in rye grain: implications for acrylamide formation

Acrylamide forms from free asparagine and sugars during cooking, and products derived from the grain of cereals, including rye, contribute a large proportion of total dietary intake. In this study, free amino acid and sugar concentrations were measured in the grain of a range of rye varieties grown at locations in Hungary, France, Poland, and the United Kingdom and harvested in 2005, 2006, and 2007. Genetic and environmental (location and harvest year) effects on the levels of acrylamide precursors were assessed. The data showed free asparagine concentration to be the main determinant of acrylamide formation in heated rye flour, as it is in wheat. However, in contrast to wheat, sugar, particularly sucrose, concentration also correlated both with asparagine concentration and with acrylamide formed. Free asparagine concentration was shown to be under genetic (G), environmental (E), and integrated (G × E) control. The same was true for glucose, whereas maltose and fructose were affected mainly by environmental factors and sucrose was largely under genetic control. The ratio of variation due to varieties (genotype) to the total variation (a measure of heritability) for free asparagine concentration in the grain was 23%. Free asparagine concentration was closely associated with bran yield, whereas sugar concentration was associated with low Hagberg falling number. Rye grain was found to contain much higher concentrations of free proline than wheat grain, and less acrylamide formed per unit of asparagine in rye than in wheat flour.

Effects of water availability on free amino acids, sugars, and acrylamide-forming potential in potato

Irrigation is used frequently in potato cultivation to maximize yield, but water availability may also affect the composition of the crop, with implications for processing properties and food safety. Five varieties of potatoes, including drought-tolerant and -sensitive types, which had been grown with and without irrigation, were analyzed to show the effect of water supply on concentrations of free asparagine, other free amino acids, and sugars and on the acrylamide-forming potential of the tubers. Two varieties were also analyzed under more severe drought stress in a glasshouse. Water availability had profound effects on tuber free amino acid and sugar concentrations, and it was concluded that potato farmers should irrigate only if necessary to maintain the health and yield of the crop, because irrigation may increase the acrylamide-forming potential of potatoes. Even mild drought stress caused significant changes in composition, but these differed from those caused by more extreme drought stress. Free proline concentration, for example, increased in the field-grown potatoes of one variety from 7.02 mmol/kg with irrigation to 104.58 mmol/kg without irrigation, whereas free asparagine concentration was not affected significantly in the field but almost doubled from 132.03 to 242.26 mmol/kg in response to more severe drought stress in the glasshouse. Furthermore, the different genotypes were affected in dissimilar fashion by the same treatment, indicating that there is no single, unifying potato tuber drought stress response.

Association of free amino acids with caries experience and mutans streptococci levels in whole saliva of children with early childhood caries

Objective: The aim of the present study was to identify the free amino acid content in whole saliva of children with (CE) and without early childhood caries (CF) (ECC), correlating these findings with caries experience and mutans streptococci (MS) levels in saliva.Design: Seventy-eight healthy children, both genders, 6-71 months age, were selected to participate in the study. Following examination for dmft scores calculation, unstimulated whole saliva was collected from all 78 participants, stored at -80 degrees C, and used for amino acid analysis, on a Biochem 20 plus amino acid analyzer. Stimulated whole saliva was collected from 52 children, transported, diluted and plated on MSB agar medium for detection of MS in cfu/mL.Results: Forty different free amino acids were identified in whole saliva, with great variation in their concentration. A statistically significant relation was found between caries experience and the presence of free proline and glycine. While proline (p = 0.0182) was more frequently absent in the CF group, the absence of glycine (p = 0.0397) was more often observed in the CE group. In the presence of higher levels of MS, free glycine reduced the risk of experiencing dental caries (p = 0.0419). Conversely, the presence of proline was found to increase the risk of experiencing the disease (p = 0.0492).Conclusions: The presence of free proline and absence of free glycine in children with ECC, highly contaminated with MS, increased the chances of experiencing dental caries in the present population. Further studies are needed to better understand this phenomenon. (C) 2008 Elsevier Ltd. All rights reserved.

Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma Cruzi

Over the past three decades, L-proline has become recognized as an important metabolite for trypanosomatids. It is involved in a number of key processes, including energy metabolism, resistance to oxidative and nutritional stress and osmoregulation. In addition, this amino acid supports critical parasite life cycle processes by acting as an energy source, thus enabling host-cell invasion by the parasite and subsequent parasite differentiation. In this paper, we demonstrate that L-proline is oxidized to Δ(1)-pyrroline-5-carboxylate (P5C) by the enzyme proline dehydrogenase (TcPRODH, E.C. 1.5.99.8) localized in Trypanosoma cruzi mitochondria. When expressed in its active form in Escherichia coli, TcPRODH exhibits a Km of 16.58±1.69 µM and a Vmax of 66±2 nmol/min mg. Furthermore, we demonstrate that TcPRODH is a FAD-dependent dimeric state protein. TcPRODH mRNA and protein expression are strongly upregulated in the intracellular epimastigote, a stage which requires an external supply of proline. In addition, when Saccharomyces cerevisiae null mutants for this gene (PUT1) were complemented with the TcPRODH gene, diminished free intracellular proline levels and an enhanced sensitivity to oxidative stress in comparison to the null mutant were observed, supporting the hypothesis that free proline accumulation constitutes a defense against oxidative imbalance. Finally, we show that proline oxidation increases cytochrome c oxidase activity in mitochondrial vesicles. Overall, these results demonstrate that TcPRODH is involved in proline-dependant cytoprotection during periods of oxidative imbalance and also shed light on the participation of proline in energy metabolism, which drives critical processes of the T. cruzi life cycle.

Products of Proline Catabolism Can Induce Osmotically Regulated Genes in Rice1

Many plants accumulate high levels of free proline (Pro) in response to osmotic stress. This imino acid is widely believed to function as a protector or stabilizer of enzymes or membrane structures that are sensitive to dehydration or ionically induced damage. The present study provides evidence that the synthesis of Pro may have an additional effect. We found that intermediates in Pro biosynthesis and catabolism such as glutamine and Δ1-pyrroline-5-carboxylic acid (P5C) can increase the expression of several osmotically regulated genes in rice (Oryza sativa L.), including salT and dhn4. One millimolar P5C or its analog, 3,4-dehydroproline, produced a greater effect on gene expression than 1 mm l-Pro or 75 mm NaCl. These chemicals did not induce hsp70, S-adenosylmethionine synthetase, or another osmotically induced gene, Em, to any significant extent. Unlike NaCl, gene induction by P5C did not depend on the normal levels of either de novo protein synthesis or respiration, and did not raise abscisic acid levels significantly. P5C- and 3,4-dehydroproline-treated plants consumed less O2, had reduced NADPH levels, had increased NADH levels, and accumulated many osmolytes associated with osmotically stressed rice. These experiments indicate that osmotically induced increases in the concentrations of one or more intermediates in Pro metabolism could be influencing some of the characteristic responses to osmotic stress.

Environmental and developmental signals modulate proline homeostasis: evidence for a negative transcriptional regulator.

In many plants, osmotic stress induces a rapid accumulation of proline through de novo synthesis from glutamate. This response is thought to play a pivotal role in osmotic stress tolerance [Kishor, P. B. K., Hong, Z., Miao, G.-H., Hu, C.-A. A. and Verma, D. P. S. (1995) Plant Physiol. 108, 1387-1394]. During recovery from osmotic stress, accumulated proline is rapidly oxidized to glutamate and the first step of this process is catalyzed by proline oxidase. We have isolated a full-length cDNA from Arabidopsis thaliana, At-POX, which maps to a single locus on chromosome 3 and that encodes a predicted polypeptide of 499 amino acids showing significant similarity with proline oxidase sequences from Drosophila and Saccharomyces cerevisiae (55.5% and 45.1%, respectively). The predicted location of the encoded polypeptide is the inner mitochondrial membrane. RNA gel blot analysis revealed that At-POX mRNA levels declined rapidly upon osmotic stress and this decline preceded proline accumulation. On the other hand, At-POX mRNA levels rapidly increased during recovery. Free proline, exogenously added to plants, was found to be an effective inducer of At-POX expression; indeed, At-POX was highly expressed in flowers and mature seeds where the proline level is higher relative to other organs of Arabidopsis. Our results indicate that stress- and developmentally derived signals interact to determine proline homeostasis in Arabidopsis.

Biochemical and physiological responses of sugarcane cultivars to soil water deficiencies

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influência da disponibilidade hídrica e da acidez do solo no teor de prolina livre de guandu

O objetivo deste trabalho foi avaliar a influência da disponibilidade hídrica e da acidez do solo, no teor de prolina livre da parte aérea das cultivares de guandu, IAPAR 43-Aratã e IAC Fava Larga. O delineamento experimental utilizado foi o inteiramente casualizado, em arranjo fatorial 2x3x4 (cultivar x disponibilidade hídrica x acidez), com quatro repetições. em condições de restrição hídrica severa, a cultivar IAC Fava Larga pode ser considerada tolerante, em virtude do maior acúmulo de prolina livre. A presença de alumínio no solo, associada à restrição hídrica severa, apresentou efeito sinergístico sobre os teores de prolina livre.

Interação da deficiência hídrica e da toxicidade do alumínio em guandu cultivado em hidroponia

O objetivo deste trabalho foi avaliar o efeito da interação da deficiência hídrica e da toxicidade do alumínio no crescimento inicial e teores de prolina livre em duas cultivares de guandu, IAPAR 43-Aratã e IAC Fava Larga, cultivadas em hidroponia. As plântulas foram submetidas aos estresses em solução nutritiva (pH 3,8), nos potenciais osmóticos de 0, -0,004, -0,006, -0,008 e -0,010 MPa, com 0, 0,25, 0,50, 0,75 e 1 mmol dm-3 de Al3+. O experimento foi conduzido em sala de crescimento, sob luminária com irradiância média de 190 mmol m-2 s-1 , fotoperíodo de 12 horas e temperatura de 25+1ºC. O delineamento experimental foi inteiramente casualizado, em arranjo fatorial 2x5x5 (duas cultivares de guandu, cinco potenciais osmóticos e cinco níveis de alumínio), com quatro repetições. Os dados foram submetidos às análises de regressão polinomial, agrupamento e componentes principais. A deficiência hídrica causa redução do crescimento da parte aérea do guandu, e a toxicidade do alumínio provoca diminuição do crescimento radicular. Houve aumento nos teores de prolina livre nas duas cultivares sob deficiência hídrica, e apenas na IAC Fava Larga sob toxicidade de alumínio. Na análise multivariada, foi observada alta correlação no crescimento e no acúmulo de prolina na cultivar IAC Fava Larga, o que evidencia provável tolerância aos estresses associados.

METABOLISMO do NITROGÊNIO ASSOCIADO À DEFICIÊNCIA HÍDRICA E SUA RECUPERAÇÃO em GENÓTIPOS DE MILHO

O estresse hídrico afeta profundamente o metabolismo celular vegetal. Neste trabalho, objetivou-se quantificar os efeitos da deficiência hídrica e sua recuperação sobre a atividade das enzimas do metabolismo do nitrogênio: redutase do nitrato (RN), glutamina sintetase (GS) e glutamato sintase (GOGAT) e sobre o acúmulo de prolina em plantas dos genótipos de milho BR 2121 e BR 205. O experimento foi conduzido em casa de vegetação, sob o delineamento inteiramente casualizado, com quatro repetições, utilizando-se vasos que continham 14,3kg de solo. Os tratamentos consistiram da combinação dos dois genótipos e quatro intervalos entre irrigações (1, 3, 5 e 7 dias). No dia da avaliação (49 dias após emergência), os tratamentos com intervalos entre 3 e 7 dias, haviam sido irrigados no dia anterior, caracterizando-se portanto como recuperação da deficiência hídrica leve e severa, respectivamente. As extrações e análises foram realizadas utilizando-se a terceira folha basípeta completamente expandida. As atividades das enzimas estudadas não diferiram entre os tratamentos de estresse hídrico, controle e recuperação do estresse moderado, entretanto as plantas sob recuperação do estresse severo apresentaram atividade enzimática superior à das plantas controle. O acúmulo de prolina livre nas folhas aumentou com o estresse hídrico e respondeu à recuperação do estresse apresentando redução. de modo geral, a atividade enzimática e o acúmulo de prolina apresentaram respostas inversas dentro dos tratamentos.

Atividade da enzima redutase do nitrato e teores de prolina livre em raízes de cana-de-açúcar sob os estresses hídrico e ácido no solo

The aim of this work was evaluate the physiological indicators of association between water deficiency and soil acidity, by determining the nitrate reductase activity, the levels of free proline and initial growth of the root system of seedlings of sugarcane cv. IAC91-5155. After 30 days, the seedlings were transferred to plastic pots with 12 dm3 of dystrophic alic Red Latosol (Oxisol) and submitted to association of three treatments of water availability: no stress (70%), moderate stress (55%) and severe stress (40%), in according with field capacity and three acidity treatments: no stress (55%), moderate stress (33%) and severe stress (23%), considering the base saturation. The experimental design was that of random blocks under factorial scheme of 3x3, with four replicates. After 60 days under the stress association, the levels of free proline, the nitrate reductase activity and the growth of the sugarcane roots system were evaluated in seedlings of sugarcane. The nitrate reductase enzyme activity can be considered a physiological indicator of the effect of the association of acid and water stress in moderate conditions in soil, while the free proline can be considered physiological indicator to both stress in severe conditions. Water deficiency increasing reduced growth of sugarcane roots.

Quantificação da trealose e da prolina livre em cana-de-açúcar sob efeito da disponibilidade hídrica do solo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Atividade da redutase do nitrato e acúmulo de prolina livre em raízes de cana-de-açúcar submetidas aos estresses hídrico e ácido no solo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)