柠檬酸和抗坏血酸对蝴蝶兰叶外植体褐变发生的影响

目的:探究柠檬酸和抗坏血酸对蝴蝶兰叶片外植体褐变发生的影响以及对PPO活性变化影响的作用机理.方法:以褐变率和褐变指数为参考数据,分析柠檬酸和抗坏血酸对外植体PPO活性和PPO反应产物积累的影响以及与外植体褐变发生的关系.结果:分别用100mg/L柠檬酸共培养和50mg/L抗坏血酸浸泡处理叶片外植体,经离体培养3d,褐变率分别比对照降低94.9%和54.9%,离体培养6d,褐变指数低于对照的0.53,分别为0.46和0.36,同时PPO活性降低.结论:推测柠檬酸抑制褐变的原因是直接与酶蛋白作用,抗坏血酸则与新生醌类物质结合.     

Proteomic analysis of core breakdown disorder in Conference pears (Pyrus communis L.)

2-DE was applied to study core breakdown disorder in controlled atmosphere stored 'Conference' pears. This physiological disorder is characterized by internal browning of the fruit tissue and the development of cavities. Suitable protein phenol extraction/ammonium acetate-methanol precipitation and 2-DE protocols for a wide pH range were established for pear tissue. The protein expression profiles of healthy, sound (intact tissue of pears with core breakdown) and brown tissue were analyzed with the univariate non-parametric Kolmogorov-Smirnov test and multivariate statistical techniques such as principal component analysis and partial least square discriminant analysis. Both statistical approaches revealed interesting differentially expressed proteins between healthy and disordered pears. LC-ESI-MS/MS identification of differentially expressed proteins between healthy and sound tissue revealed their participation in the energy metabolism, the antioxidant system and ethylene biosynthesis. Up-regulated characteristic proteins in brown tissue were mainly involved in energy metabolism and defense mechanisms. Proteomics coupled to univariate and multivariate statistical techniques seems to be an efficient approach to get a better insight into the different mechanisms and pathways leading to the core breakdown disorder.     

On the specificity of alcoholic acidic silver nitrate reagent for the histochemical localization of ascorbic acid. A reappraisal.

The specificity of the alcoholic acidic silver nitrate staining method for the histochemical localization of ascorbic acid was reappraised. It was found that the method is by and large better suited for the localization of ascorbic acid in both animal and plant tissues due to its greater specificity, which is ensured by employing reagent made in carbon dioxide saturated glass distilled water as well as by carrying out the reaction at a low temperature (0-4 degrees C) and at a pH of 2-2.5.     

Interaction of ascorbic acid and colchicine with sucrose utilization byCunninghamella elegans

The stimulating effect of ascorbic acid, administered to colchicine-treated mycelia ofCunninghamella elegans, on respiration, keto acid production and carbohydrate accumulation was lower than the highly increased rate of sugar absorption. Furthermore, the increasing ascorbic acid concentration in colchicine media favoured lower carbohydrate accumulation, keto acid production, carbon dioxide output and highly stimulated the utilization of the absorbed sugars for the formation of non-carbohydrate compounds. In addition, the vitamin seemed to counteract the inhibitory effects of colchicine on nucleoprotein and soluble nucleotide synthesis by colchicine-treated mycelia particularly when the drug was administered at high levels.     

Control of lethal browning of tissue culture plantlets of Cavendish banana cv. Formosana with ascorbic acid

Cavendish banana cv. Formosana is a high yielding commercial cultivar resistant to race 4 of Fusarium oxysporum f. sp. cubense. Mass micropropagation of this cultivar has a serious problem of high mortality due to lethal browning of plantlets. The mineral contents in leaves and corms of diseased and healthy plantlets were similar. Amendment of culture medium with anion exchange resins, cation exchange resins, polyvinylpyrrolidone or activated charcoal did not reduce the disease incidence. However, addition of ascorbic acid to the surface of culture medium not only prevented the development of lethal browning but also greatly increased the number of plantlets produced. Even at 0.005% ascorbic acid was able to reduce the disease incidence by more than 60% and caused over 8-fold increase in number of plantlets produced. When cultures raised from 12 different Formosana corms were tested, ascorbic acid was able to reduce disease incidence by an average of 83%, and increase the number of plantlets in each test. When diseased plantlets were transferred to culture medium with ascorbic acid, all of them recovered, and resumed normal growth and multiplication, while all control plantlets on culture medium without ascorbic acid died after one month.     

Supplemental ascorbic acid does not affect inferred heat loss in broiler chickens exposed to elevated temperature

The provision of supplemental ascorbic acid has been reported to lower the body temperature of chickens maintained at elevated environmental temperatures. Since body temperature is the net effect of heat production and heat loss, it is not known if the reductions in body temperature were due to a lower heat production or an increase in heat loss. The purpose of this work was to determine if supplemental ascorbic acid facilitates heat loss in chickens exposed to an elevated temperature. On day 12 post-hatch broiler chickens were implanted intra-abdominally with a thermo-sensitive radio transmitter. The following day, birds were placed inside an indirect calorimeter maintained at 34 C for 24 h and provided water containing 0 or 400 ppm ascorbic acid. Oxygen consumption, carbon dioxide production, heat production, respiratory exchange ratio, and body core temperature were measured for 3 h; beginning 21 h after the birds were placed inside the calorimeter. No differences were observed in heat production or body core temperature between birds provided or not and 400 ppm ascorbic acid. This suggests that ascorbic acid has no effect on heat loss. Birds provided ascorbic acid did exhibit a significantly lower respiratory exchange ratio suggesting a greater utilization of lipid for energy production. Although lipid has a lower heat increment compared with protein and carbohydrate, the significance of this finding to birds exposed to elevated temperature is not known. In conclusion, under the conditions of this study the provision of supplemental ascorbic acid to broiler chickens maintained at an elevated temperature did not affect heat loss as inferred from measured heat production and body core temperature.     

Role of exogenous ascorbic acid in tissue status of ascorbic acid-2-sulphate in guinea pigs.

Guinea pigs were given ascorbic acid orally in two doses; a low and a high dose. The tissue levels of ascorbic acid-2-sulphate was estimated in these animals after 15 days of feeding and a subsequent deprivation period of 15 days. The specific activity of the enzymes ascorbic acid sulphotransferase and ascorbic acid-2-sulphate sulphohydrolase was studied. During higher ascorbic acid intake, the activity of ascorbic acid sulphotransferase was increased, whereas ascorbic acid-2-sulphate sulphohydrolase showed a decreased activity. But when ascorbic acid intake was lowered or ceased, the activity of the above enzymes showed a reverse pattern. Possible reasons for the lack of antiscorbutic activity of ascorbic acid-2-sulphate in guinea pigs is discussed.     

Ethanolamine Metabolism in Plant Tissues

Ethanolamine is readily metabolized by oat, pea, wheat, apple and carrot tissue preparations. Ethanolamine-1,2 (14)C was incorporated into the lipid fraction, and (14)C activity was distributed in the organic acid, sugar, acid volatile, carbon dioxide and insoluble residue fractions. The distribution varied with the particular tissue. Incorporation into the lipid fraction occurred in tissue homogenates in the absence of ATP by a Ca(++) activated system similar to that reported for animal preparations. The initial step in ethanolamine oxidation involves an amine oxidase. Glycolaldehyde and glyoxylic acid are metabolic intermediates, the former in the conversion of ethanolamine to carbon dioxide. No evidence was obtained for the operation of an ethanolamine transaminase or for the involvement of phosphorylated intermediates in the conversion of ethanolamine to carbon dioxide.     

Decarboxylation of 2-keto fatty acids by brain

An enzyme system consisting of washed acetone powder of pig brain, adenosine triphosphate, nicotinamide adenine dinucleotide, magnesium, fumaric acid, and ascorbic acid was found to catalyze the oxidative decarboxylation of 2-ketostearic acid. The products were carbon dioxide and heptadecanoic acid. Washing the enzyme powder with ethylene-diaminetetraacetate and other chelating agents destroyed the activity. Further properties of the enzyme system are described. It is believed that this enzyme system accounts in part for the 1-carbon degradation route for brain fatty acids.     

Ascorbic acid in buffalo ovary in relation to oestrous cycle.

Concentration of ascorbic acid was determined in different parts of buffalo ovary at four different stages of oestrous cycle viz. early luteal, mid luteal, late luteal and follicular. The stages were decided from the physical and morphological examinations of corpora lutea. The ovary was dissected in three components viz. corpus luteum, follicular fluid and ovarian stromal tissue for ascorbic acid assay. Corpus luteum showed significant change in concentration of ascorbic acid with the advancement of oestrous cycle, value being highest in late- luteal stage. Follicular fluid and ovarian stromal tissue did not show significant changes in ascorbic acid at any stage of the oestrous cycle. Small follicles, irrespective of the stage of oestrous cycle had, however, significantly higher ascorbic acid content than large follicles.     

Ascorbic acid status of mullet, Mugil cephalus Linn., exposed to cadmium

The effects of acute and chronic cadmium exposures on ascorbic acid concentrations and cadmium accumulation in the tissues of juvenile mullet, Mugil cephalus , are described. Hepatic ascorbic acid reserves were maintained in untreated mullet fed a diet containing ascorbate, but decreased 60% in individuals exposed to 10 mg Cd 1^-1 for six weeks. Ascorbic acid depletion was accompanied by a massive accumulation of cadmium in the liver. Similar changes were found in gill tissue. In contrast, ascorbic acid concentrations were largely unaffected in kidney tissue which had a limited capacity to accumulate the metal. There were marked fluctuations of ascorbic acid levels in the brain. Only small amounts of cadmium were accumulated by this tissue. These results indicate that in mullet liver, gill and brain ascorbic acid stores may be depleted during chronic cadmium poisoning.     

Ascorbic acid and hydroxyproline levels in the serum and granulation tissue of rats with aseptic and infected wounds

Hydroxyproline and ascorbic acid were measured in wound tissues and ascorbic acid was determined in the blood sera of 230 mature male Wistar rats with aseptic and infected surface wounds on days 1-10 (daily), 12, and 15. The principal morphologic characteristics of the granulation tissue were assessed by semiquantitative methods. An infected wound is characterized by increased hydroxyproline levels in the granulation tissue and elevated ascorbic acid concentration in the blood serum. The granulation tissue ascorbic acid level augments during the first nine days in case of an aseptic wound and during twelve days in case of an infected wound. Morphologic and biochemical correlations are indicative of a relationship between the ascorbic acid level on the one hand and granulation tissue vascularization and fibroblast proliferation on the other.     

Impact of paper bridges, activated charcoal, and antioxidants on growth and development of protocorm-like bodies of hybrid Cymbidium

This study aimed to establish whether protocorm-like bodies (PLBs) of hybrid Cymbidium Twilight Moon ‘Day Light’ could be cultured on paper bridges to remove tissue and medium browning and enhance growth. In addition, the effects of two antioxidants (ascorbic acid [AA] and α-tocopherol [AT]) and activated charcoal (AC) on tissue and medium browning were evaluated. Half-PLBs were cultured on Whatman No. 1 paper bridges in 10 ml of medium containing AC, AA, or AT. The production and development of new PLBs was inferior to that on standard agar-based solid medium when half-PLBs were used. The addition of 1 g/l AC, 10 mg/l AA, or 25 mg/l AT did not affect the formation of new PLBs from half-PLBs, but tissue or medium browning was also not observed. Encapsulated PLBs developed from half-PLBs germinated on medium containing AC or antioxidants. This research provides a simple but effective chemical means, through the use of AC or antioxidants, of avoiding tissue or medium browning without negatively impacting the growth and productivity of orchid PLBs. The use of paper bridges, however, significantly reduced PLB-related growth and development parameters relative to agar-based medium.     

Browning and decomposed products of model orange juice

A model solution of orange juice containing sugars, ascorbic acid, and citric acid was prepared and its browning during storage was examined. The solution gradually turned brown. Ascorbic acid (AsA) most contributed to the browning. Citric acid and such amino acids as Arg and Pro promoted the browning. DTPA, a strong chelator, inhibited the browning. 3-Hydroxy-2-pyrone (3OH2P), 5-hydroxymethylfurfural (HMF), furfural, 5-hydroxymaltol, and 2-furoic acid were identified as decomposed products in the stored solution. When 3OH2P was stored, the solution turned slightly brown. Furfural solution added with amino acids turned yellow. 3OH2P showed a positive relation with the browning of retail orange juice during storage.     

High polyphenol oxidase activity and low titratable acidity in browning bamboo tissue culture

Summary Tissue browning that frequently results in the early death of bamboo shoots in vitro correlated directly with polyphenol oxidase (PPO, EC 1.10.3.1) activity and inversely with titratable acidity. It was unrelated to the level of endogenous phenols. During the course of culture, timing of PPO activity paralleled that of explant browning. Browning was highest among shoots cultured in a medium of pH 8, which was consistent with the pH optinum of the bamboo enzyme. The pH optimum was first determined with the crude enzyme, then verified with two purified isozymes. Stability of the bamboo PPO was also highest at pH 10. PPO activities of the severely browning Dendrocalamus latiflorus, the moderately browning Phyllostachys nigra, and the relatively non-browning Bambusa oldhamii were inhibited strongly by ascorbic acid, cysteine, sodium diethyldithiocarbamate, and sodium sulfite. But characterization of bamboo PPO according to enzyme inhibitors was not possible because enzyme extracts of the three species gave varied responses to the traditional substances. Nutrient medium addenda of some PPO inhibitors, namely ascorbic acid, cysteine, kojic acid, and thiourea, mainly enhanced browning. However, ferulic acid at 3 mM and lower concentrations reduced the number of brown shoots per culture, although not the percentage of cultures that browned. Polyvinylpyrrolidone failed completely to suppress browning. The two purified isozymes showed different temperature optima for PPO activity: 60°C and 65°C. The purified isozymes displayed a substrate preference for dopamine, or a cathecol oxidase characteristics.     

Aortic ascorbic acid, trace elements, and superoxide dismutase activity in human aneurysmal and occlusive disease

Altered trace elements and ascorbic acid metabolism have been implicated in the pathogenesis of atherosclerotic cardiovascular disease. However, their role in the disease process, or the effect of atherosclerosis on their tissue levels within plaque, is poorly understood. The present study analyzes the concentrations of Fe, Cu, Zn, and Mn, and ascorbic acid and superoxide dismutase (SOD) activity in tissue samples from 29 patients with abdominal aortic aneurysms (AAA) and 14 patients with atherosclerotic occlusive disease (AOD). It was observed that the Fe and Mn concentrations in AAA and AOD tissue were higher than the levels in nondiseased control aorta, whereas Cu and Zn levels in AAA and AOD tissue were similar to the levels in controls. The Zn:Cu ratio was significantly lower in the AAA tissue in comparison to both AOD and control tissue. In addition, AAA and AOD tissue had low ascorbic acid levels and low Cu,Zn-SOD activity with Cu,Zn-SOD:Mn-SOD ratios of 0.27 and 0.19, respectively, compared to a ratio of 3.20 in control aorta. These data indicate that aorta affected by aneurysms and occlusive disease have altered trace element and ascorbic acid concentrations, as well as low Cu,Zn-SOD activity. Although these observations do not directly support the hypothesis that AAA is associated with aortic Cu deficiency they do suggest a role for oxygen radicals or increased lipid peroxidation in occlusive and aneurysmal disease of the aorta.     

杉木愈伤组织培养中的褐化控制研究

以杉木茎尖诱导形成的愈伤组织为试验材料,研究维生素C、柠檬酸、半胱氨酸、硫代硫酸钠4种抗氧化剂和活性炭对杉木愈伤组织培养中褐化的影响。结果表明：维生素C和半胱氨酸能有效地控制褐化的发生,且对愈伤组织生长没有明显的抑制作用,是杉木愈伤组织培养较为适宜的抗褐变剂。     

Simultaneous determination of two antioxidants, uric and ascorbic acid, in animal tissue by high-performance liquid chromatography.

A rapid and simple method for the simultaneous analysis of uric and ascorbic acid in extracts of animal tissue is described. The method uses reversed-phase ion-pair chromatography with ultraviolet detection. The technique allows efficient separation of both acids while showing high selectivity, recovery, reproducibility, and sample stability. Calculated levels of both substances in mouse liver tissue were 1.00 +/- 0.05 mumol ascorbic acid/g and 130 +/- 5 nmol uric acid/g.     

Chromium and chronic ascorbic acid depletion effects on tissue ascorbate,manganese, and14C retention from14C-ascorbate in guinea pigs

Chromium (Cr) potentiates the effects of insulin and a role for insulin in ascorbic acid transport has been reported. Therefore, the effects of Cr and ascorbate depletion on tissue ascorbic acid and¹⁴C distribution and excretion after a¹⁴C ascorbate dose were investigated in guinea pigs. As utilization of dietary Cr is affected by interaction with other minerals, tissue manganese (Mn), zinc (Zn), copper (Cu), and iron (Fe) were examined. For 20 wk, 40 weanling animals were fed either a Cr-deficient (<0.06 μg Cr/g diet, ?Cr) or a Cr-adequate (2 μg Cr from CrCl₃/g diet, +Cr) casein-based diet and were given 1 mg ascorbate/d (?C) or 10 mg ascorbate/d (+C) for 20 wk. Animals fed the Cr-depleted diet had decreased weight at 20 wk (p<0.01). Six hours before necropsy, animals were dosed by micropipette with 1.8 μCi ofl-[carboxyl-¹⁴C] ascorbic acid and placed in metabolic cages. Ascorbate supplementation increased Fe concentrations in most analyzed tissues, hepatic¹⁴C, tissue ascorbate and Mn concentration in the adrenal and testes, but decreased the concentrations of Cu in the kidney and Mn in the spleen. Liver Mn concentration was higher and kidney Mn concentration was lower in +Cr animals. Interactions between Cr and ascorbic acid affected Mn concentrations in bone and brain. These results indicate that ascorbate and Cr may affect Mn distribution. Chromium supplementation decreased plasma cortisol, brain¹⁴C and the amount of¹⁴C expired as carbon dioxide. These findings suggest that dietary Cr may affect ascorbic acid metabolism and the metabolic response to stress.     

Ascorbic Acid in Mesencephalic Cultures: Effects on Dopaminergic Neuron Development

Ascorbic acid exists in high intracellular concentrations in fetal rat brain. In mesencephalic cultures the cellular ascorbic acid content drops sharply to undetectable levels when no ascorbic acid is added to the medium, thus creating a model of scorbutic neuronal tissue and affording the study of ascorbic acid's effects on mesencephalic cell development and function. Cultures treated with 0.2 mM ascorbic acid were compared with controls (scorbutic cultures) by using morphological and biochemical indices. Ascorbic acid cultures at 7 and 14 days in vitro showed a marked increase in glial proliferation on glial fibrillary acidic protein staining and increased neurite growth and number on tyrosine hydroxylase staining. Significantly higher dopamine uptake and levels of dopamine and 3,4-dihydroxyphenylacetic acid were also observed after 7 and 14 days of ascorbic acid treatment. The capacity to accumulate ascorbic acid and the ability to retain the intracellular ascorbic acid developed gradually as the cultures matured. Ascorbic acid reached the embryonal levels by day 14 in vitro. We conclude that although neuronal cultures can survive and grow in the absence of detectable levels of ascorbic acid, its presence exerts a broad effect on dopamine neuron morphology and biochemical functioning either directly or through increased glial proliferation, or possibly both.