果梅果实发育过程中有机酸含量及相关代谢酶活性的变化特征

以优选荔波野生梅单株‘荔波-3’和‘荔波-11’为试验材料,采用高效液相色谱法(HPLC)测定了果梅果实在发育过程中有机酸含量及相关代谢酶活性变化特征。结果表明:(1)‘荔波-3’和‘荔波-11’果梅果实中柠檬酸和苹果酸主要在果实发育后期积累,花后140d两品种的柠檬酸积累量达到最大,且果实中总酸的积累量也相应的达到最大值。(2)柠檬酸积累量与柠檬酸合成酶(CS)活性呈极显著正相关关系,‘荔波-11’因CS活性高于‘荔波-3’,使得其果实成熟时的柠檬酸含量高于后者。(3)苹果酸积累量与磷酸烯醇式丙酮酸羧化酶(PEPC)和NAD-苹果酸脱氢酶(NAD-MDH)活性呈极显著正相关关系,但与NADP-苹果酸酶(NADP-ME)活性呈负相关关系;在花后45~100d内,果梅果实中PEPC和NAD-MDH活性迅速升高,而NADP-ME活性相对较低,使苹果酸积累量迅速增加;在花后120~140d,‘荔波-3’果实中PEPC、NADP-MDH活性显著高于‘荔波-11’,而其NADP-ME活性低于‘荔波-11’,导致成熟时‘荔波-11’的苹果酸含量高于‘荔波-3’。可见,果梅果实的有机酸主要在果实发育后期积累,有机酸积累差异受多个酶协同调控;苹果酸的积累差异主要由NAD-MDH、PEPC和NADP-ME活性协同变化引起,柠檬酸积累差异主要受CS活性变化影响。     

不同梨品种果实有机酸含量变化与相关酶活性的研究

以梨品种‘鸭梨’、‘茌梨’和‘八里香’为试材,对不同品种果实有机酸积累及相关酶活性随发育的动态变化特征进行分析比较,探讨果实有机酸含量与相关酶活性的关系。结果表明:(1)梨果实发育过程中有机酸含量呈逐渐下降趋势,果实成熟时的总有机酸含量以‘茌梨’最高,‘鸭梨’次之,‘八里香’最低,且3品种间总酸含量差异显著。(2)果实发育后期,‘鸭梨’、‘茌梨’和‘八里香’的柠檬酸含量几乎相同,而苹果酸的含量差异显著,说明苹果酸是引起品种间总酸含量差异的主要因素。(3)在苹果酸代谢中,不同品种间NADP-ME酶活性在果实发育后期差异显著,而NAD-MDH和PEPC酶活性无显著差异,表明NADP-ME酶是引起‘鸭梨’、‘茌梨’和‘八里香’成熟果实苹果酸含量差异的主要原因。(4)不同品种的柠檬酸代谢酶CS、Cyt-ACO、Mit-ACO和NAD-IDH的活性变化趋势一致,且成熟时无显著差异,表明它们不是引起梨果实酸含量差异的主要原因。(5)梨果实发育过程中有机酸含量与相关代谢酶活性的相关性分析表明,苹果酸和柠檬酸代谢酶共同影响梨果实的酸含量水平。     

叶面喷施钙镁肥对‘妃子笑'荔枝果肉苹果酸积累的影响

为探讨叶面喷施钙镁肥对‘妃子笑''荔枝果肉苹果酸积累的影响,该文对‘妃子笑''荔枝树冠作喷布0.3%氯化钙(Ca)、0.3%氯化镁(Mg)及其二者混合(Ca+Mg)等水溶液处理,以树冠喷布清水为对照(CK),测定不同生长时期果肉水溶性钙和镁、苹果酸等含量及苹果酸代谢相关酶活性的动态变化,并作多元线性相关分析。结果表明:(1)苹果酸含量呈“L”型变化,Mg、Ca和Ca+Mg处理在果实发育前期促进苹果酸积累,Ca处理在后期促进苹果酸积累。(2)果肉水溶性钙含量总体呈上升趋势,水溶性镁含量大致呈“M”的动态变化趋势。(3)CK和Ca处理的苹果酸含量与NADP-ME活性、Ca+Mg处理的苹果酸含量与PEPC和NAD-MDH活性等均呈正相关,CK的苹果酸含量与PEPC活性、MS活性呈负相关。(4)水溶性钙抑制NAD-MDH、NADP-ME等活性,水溶性镁抑制NAD-MDH、MS等活性。综上认为,钙、镁叶面营养通过改变水溶性钙、镁等含量和苹果酸代谢途径不同关键酶活性而影响果肉苹果酸积累,其中Ca处理可能通过积累更多的苹果酸而抑制果肉呼吸作用,进而使果肉减少糖分损失,在生产中可作施肥技术应用。该研究结果为我国荔枝实际生产提供一定的理论参考和技术支持。     

菠萝叶片中有机酸含量、转氨酶和脱氢酶活性的昼夜变化

研究了菠萝叶片中苹果酸、柠檬酸、草酰乙酸、α－酮戊二酸和草酸等有机酸和转氨酶、脱氢酶活性的昼夜变化。结果表明,除了草酸外,其他４种有机酸的含量都呈白天降低晚上增高的趋势。有机酸的含量在上午９∶００时有最高值,下午１８∶００时有最低值。苹果酸、柠檬酸、草酰乙酸、α－酮戊二酸和草酸的昼夜最大差值分别为１４０、６１、３１．８、８．２和６．７ｍｍｏｌ·ｋｇ－１ＦＷ。异柠檬酸脱氢酶和琥珀酸脱氢酶活性的昼夜变化与有机酸的趋势一致,白天降低晚上增高。可溶性蛋白和游离氨基酸含量以及谷草转氨酸、谷丙转氨酶活性在下午１８∶００时有最高值,凝胶电泳显示菠萝叶片中有新的蛋白带出现。     

烤烟主流烟气内源有害成分与烟叶化学成分相关性

研究了不同生态产区烟叶主要化学成分与主流烟气内源有害成分的关联性.结果表明:初烤烟叶内源有害成分和危害性指数(H值)与烟叶部位具有较好的关联性.4-甲基亚硝基-1-(3-吡啶基)-1-丁酮(NNK)、苯并[a]芘、氢氰酸(HCN)、氨和苯酚的释放量表现为上部叶＞中部叶,巴豆醛释放量表现为中部叶＞上部叶,H值表现为上部叶＞中部叶.不同内源有害成分与烟叶含碳、含氮化学成分的相关性复杂,同一类型烟叶化学成分对不同内源有害成分呈现出不同的相关关系.卷烟危害性指数与烟叶中烟碱、蛋白质、总氮、主要多酚和有机酸含量呈显著正相关,而与钾离子、总糖、还原糖、淀粉等含碳类物质含量呈显著负相关.适当降低烟叶含氮化合物含量、增加烟叶钾含量可能会降低烟叶的危害性指数.     

‘鸭梨’ב京白梨’杂交后代果实有机酸积累差异及相关酶活性的研究

该研究以‘鸭梨’ב京白梨’杂交后代高酸个体(GS-Y14)和低酸个体(DS-Y182)为试材,系统分析了果实发育过程中有机酸积累动态及相关酶活性的变化特征。结果表明:(1)GS-Y14属于苹果酸优势型果实,DS-Y182属于柠檬酸优势型果实,且成熟时两者在总酸含量上表现出的差异主要是由于苹果酸含量的差异所致。(2)苹果酸酶(NADP-ME)是苹果酸形成的关键酶,在梨果实发育过程中NADP-ME起分解作用,且该酶活性在两类个体果实发育后期差异显著,即NADP-ME是引起GS-Y14和DS-Y182中苹果酸含量不同的主要原因,进而导致成熟时两类个体果实酸积累的差异。(3)梨果实磷酸烯醇式丙酮酸羧化酶(PEPC)活性的升高有利于其柠檬酸的合成,而柠檬酸合酶(CS)是影响其柠檬酸含量变化的关键酶;细胞质乌头酸酶(Cyt-ACO)和线粒体乌头酸酶(Mit-ACO)早期对果实柠檬酸的含量变化影响较小,后期异柠檬酸脱氢酶(NAD-IDH)活性对柠檬酸在后代个体中的积累有一定影响。     

‘金冠’苹果及其优系(SGP-1)果实发育期间有机酸代谢特征比较北大核心CSCD

以‘金冠’苹果及其优系‘SGP-1’为试材,测定果实发育期间有机酸组分、含量和苹果酸代谢相关酶活性,分析它们的变化规律及相关关系,以探索苹果有机酸积累的关键时期和关键酶,揭示果实低酸成因。结果表明:(1)苹果果实发育期间,‘SGP-1’的有机酸含量显著低于‘金冠’,成熟时仅为‘金冠’的二分之一,且主要由苹果酸、奎宁酸、酒石酸和柠檬酸组成,幼果期以奎宁酸为主,成熟期以苹果酸为主。(2)‘SGP-1’的苹果酸含量显著低于‘金冠’,其在幼果期和膨大期变化规律与‘金冠’相反,且积累关键时期和快速下降期早于‘金冠’;‘SGP-1’果实其余酸组分含量变化趋势与‘金冠’基本一致,但在幼果期显著高于‘金冠’,在成熟期与‘金冠’差异不显著。(3)‘SGP-1’的苹果酸代谢相关酶活性在幼果期均显著高于‘金冠’,成熟期持平或显著低于‘金冠’;幼果期MDH活性和成熟期ME活性在两材料间变化规律相反。(4)‘SGP-1’的幼果期苹果酸积累与PEPC和VHA活性呈极显著正相关关系,而同期‘金冠’则与MDH、PEPC和VHA活性呈极显著负相关关系;‘SGP-1’膨大期苹果酸积累与MDH、PEPC活性呈极显著负相关关系,与PEPCK和VHA活性呈显著正相关关系,而同期‘金冠’则与PEPC、ME和VHP活性呈极显著或显著负相关关系;二者成熟期苹果酸积累均与MDH、PEPC、ME和VHP活性呈极显著或显著负相关关系。研究发现,‘SGP-1’是以苹果酸为主的低酸型‘金冠’苹果变异优系,对苹果酸积累起主要调控作用的酶种类和活性与‘金冠’不同,导致了‘SGP-1’的低酸品质,该研究结果为深入探索果实低酸形成机理和培育高糖低酸新品种奠定了基础。     

菠萝叶片中有机酸含量,转氨酶和脱氨酶活性的昼夜变化

研究了菠萝叶片中苹果酸,柠檬酸,草酰乙酰,α－酮戊二酸和草酸等有机酸和转氨酶、脱氢酶活性的昼夜变化。结果表明,除了草酸外,其他４种机酸的含量都呈白天降低晚上增高的趋势。     

外源水杨酸对铝胁迫下菊芋根系分泌物的影响

为探究铝胁迫对菊芋根系分泌物的影响以及外源水杨酸(SA)的缓解作用,该文以耐铝型南京菊芋和铝敏感型资阳菊芋为试验材料,采用土培法,设置铝浓度500μmol·L^-1,分析了不同浓度(10、100、1 000μmol·L^-1)SA对铝胁迫下菊芋根系分泌物中有机酸、氨基酸以及根尖相关代谢酶活的影响。结果表明：(1)单铝胁迫会导致菊芋根系分泌物中柠檬酸、草酸、苹果酸浓度升高,且南京菊芋升高幅度大于资阳菊芋;柠檬酸合酶和苹果酸脱氢酶在单铝胁迫下活性增强;脯氨酸含量显著提升,总氨基酸浓度均显著减少。(2)外源SA加入后,南京菊芋根系分泌的柠檬酸、草酸、苹果酸浓度均得到不同程度提高,但经高浓度(1 000μmol·L^-1)SA处理后资阳菊芋根系分泌草酸显著降低,且在各浓度SA处理下苹果酸浓度均无明显变化;柠檬酸合酶活性出现不同程度的增强,但对南京菊芋根尖中苹果酸脱氢酶活性影响不大,且高浓度(1 000μmol·L^-1)SA处理后显著降低了资阳菊芋根尖中苹果酸脱氢酶活性;脯氨酸含量显著下降,从总氨基酸浓度变化来看,...     

湘西北海拔高度对烤烟常规化学成份含量的影响

通过野外栽培试验、全自动化学流动分析测定法和火焰光度测定法,研究了湘西北慈利县与石门县海拔高度对烤烟常规化学成份含量的影响,主要结果是：1)烤烟总烟碱含量和总氮含量均与海拔高度呈极显著的负相关,而钾含量与海拔高度呈正相关．2)烤烟总糖含量与海拔高度间无明显的相关性,海拔高度对烤烟氯含量无影响．3)高海拔区域种植烤烟有利于降低烟叶的烟碱含量、提高烟叶的钾含量,从而提高烟叶的可用性与安全性、     

New bioactive derivatives of nonactic acid from the marine Streptomyces griseus derived from the plant Salicornia sp.

Three new compounds, butyl homononactate (5), butyl nonactate (6), 8-actyl homononactic acid (7), along with four known compounds homononactic acids (1), nonactic acid (2), homononactyl nonactate (3), homononactyl homononactate (4) were isolated from the marine Streptomyces griseus RSH0407, derived from the plant Salicornia sp., Chenopodiaceae. Their structures were elucidated by extensive spectroscopic techniques and by comparison with data reported in the literature. The absolute configuration of 3 was first reported by using X-ray copper radiation. Compound 5 exhibited cytotoxic activities against the HCT-8, A2780, BGC-823, BEL-7402, and A549 cell lines in vitro, with IC₅₀ values of 2.87 ± 0.20, 4.90 ± 0.30, 2.19 ± 0.32, 5.07 ± 0.23 and 1.78 ± 0.18 μM, respectively, and compounds 4–7 showed weak antibacterial activities against four bacterials respectively.     

Reassessing the role of phospholipase D in the Arabidopsis wounding response

Plants respond to wounding by means of a multitude of reactions, with the purpose of stifling herbivore assault. Phospholipase D (PLD) has previously been implicated in the wounding response. Arabidopsis ( Arabidopsis thaliana ) AtPLD α 1 has been proposed to be activated in intact cells, and the phosphatidic acid (PA) it produces to serve as a precursor for jasmonic acid (JA) synthesis and to be required for wounding-induced gene expression. Independently, PLD activity has been reported to have a bearing on wounding-induced MAPK activation. However, which PLD isoforms are activated, where this activity takes place (in the wounded or non-wounded cells) and what exactly the consequences are is a question that has not been comprehensively addressed. Here, we show that PLD activity during the wounding response is restricted to the ruptured cells using ³²P_i-labelled phospholipid analyses of Arabidopsis pld knock-out mutants and PLD -silenced tomato cell-suspension cultures. pldα1 knock-out lines have reduced wounding-induced PA production, and the remainder is completely eliminated in a pldα1 / δ double knock-out line. Surprisingly, wounding-induced protein kinase activation, AtLOX2 gene expression and JA biosynthesis were not affected in these knock-out lines. Moreover, larvae of the Cabbage White butterfly ( Pieris rapae ) grew equally well on wild-type and the pld knock-out mutants.     

On the relationship between the two branches of the kynurenine pathway in the rat brain in vivo

In the mammalian brain, kynurenine aminotransferase II (KAT II) and kynurenine 3-monooxygenase (KMO), key enzymes of the kynurenine pathway (KP) of tryptophan degradation, form the neuroactive metabolites kynurenic acid (KYNA) and 3-hydroxykynurenine (3-HK), respectively. Although physically segregated, both enzymes use the pivotal KP metabolite l -kynurenine as a substrate. We studied the functional consequences of this cellular compartmentalization in vivo using two specific tools, the KAT II inhibitor BFF 122 and the KMO inhibitor UPF 648. The acute effects of selective KAT II or KMO inhibition were studied using a radiotracing method in which the de novo synthesis of KYNA, and of 3-HK and its downstream metabolite quinolinic acid (QUIN), is monitored following an intrastriatal injection of ³H-kynurenine. In naïve rats, intrastriatal BFF 122 decreased newly formed KYNA by 66%, without influencing 3-HK or QUIN production. Conversely, UPF 648 reduced 3-HK synthesis (by 64%) without affecting KYNA formation. Similar, selective effects of KAT II and KMO inhibition were observed when the inhibitors were applied acutely together with the excitotoxin QUIN, which impairs local KP metabolism. Somewhat different effects of KMO (but not KAT II) inhibition were obtained in rats that had received an intrastriatal QUIN injection 7 days earlier. In these neuron-depleted striata, UPF 648 not only decreased both 3-HK and QUIN production (by 77% and 66%, respectively) but also moderately raised KYNA synthesis (by 27%). These results indicate a remarkable functional segregation of the two pathway branches in the brain, boding well for the development of selective KAT II or KMO inhibitors for cognitive enhancement and neuroprotection, respectively.     

硬枝树花中抑制H_22荷瘤小鼠肿瘤的活性成分研究

研究了从硬枝树花中提取得到的4个单体化合物松萝酸(usnic acid)、去甲环萝酸(evernic acid)、巴尔巴地衣酸(barbatic acid)和水杨嗪酸(salazinic acid)对H22荷瘤小鼠的抑瘤作用,并且对抑瘤率、胸腺指数、脾指数及小鼠白介素-2含量等各个指标的进行检测,以说明此4种化合物对小鼠肿瘤生长的抑制效果。结果表明,松萝酸高、中剂量组,去甲环萝酸高、中剂量组,巴尔巴地衣酸低剂量组,水杨嗪酸高剂量组对小鼠肿瘤有较好的抑制效果,与阴性对照组比较有极显著差异(P0.01),并且这些组的H22荷瘤小鼠血清中白介素-2的含量显著增加,与抑制肿瘤活性具有相关性。     

A simplified method for the estimation of individual amino acid radioactivity in plasma samples

A method is presented for the quantitative estimation of the individual amino acid radioactivity in biological samples. The material is deproteinized with cold acetone, and, after acetone evaporation, is passed through a column containing 1 g of Amberlite XAD-2, then eluted with 10% ethanol. The samples are derivatized with Sanger's reagent (alkaline 1-fluoro-2,4-dinitrobenzene) and passed again through the Amberlite XAD-2 column; the 10% ethanol eluate is now discarded and the DNP-amino acids eluted with acetone. Aliquots are used for TLC chromatography on Silicagel plates; the spots are identified, cut away and their radioactivity estimated. The actual recovery of radioactivity in the spots is about 86-92% of the initial radioactivity. No contamination with radioactive glucose, lactate, pyruvate or glycerol has been observed.     

2D NMR of the Metabolic Antioxidant Dihydrolipoic Acid and its Derivatives

Dihydroplipoate and lipoate are physiological thiols which in addition to their coenzyme functions exhibit antioxidant activity. For NMR investigations of their protective mechanism in biological and model systems it is very important to know the full assignment of proton and carbon spectra of these molecules in water (D₂O). An unambiguous assignment of proton and carbon NMR spectra has been made for dihydrolipoate and its short chain derivatives bisnor-and tetranor-lipoic acid in D₂O and CDCl₃ solutions using 2D NMR methods.

Oxidation of dihydrolipoic acid produces substantial electron density deshielding of the carbons nearest to the SH groups with the largest shift found at the inner SH group (17.79 ppm in D₂O, 16.93 in CDCl₃) and almost no changes in the tail portion of the molecule. However, bisnor-dihydrolipoic acid and especially tetranor-dihydrolipoic acid have more carbon deshielding near the outer SH group of the molecule which correlates with their known diminished ion chelating activity.

Moreover, the proton triplet at position 2 of lipoic acid has strong pH dependence (pK = 4.58) due to the close proximity to the carboxylic group and this feature may be used for monitoring pH.     

D-焦谷氨酸的研制

研究了Ｄ－焦谷氨酸的制备,提纯精制以及分析化验等方面的有关问题。     

CMP-N-acetylneuraminic acid: Is it synthesized in the nucleus

RadioactiveN-acetylmannosamine was injected intravenously into rats to labelN-acetylneuraminic acid (NeuAc) and CMP-NeuAc. Nuclei were isolated from the livers using a non-aqueous technique to prevent leakage of polar metabolites. A preparation was obtained, which was eight times enriched in nuclei based on the ratio DNA/RNA. Free NeuAc and CMP-NeuAc were isolated from this nuclear fraction and from whole liver, and the specific radioactivities were determined. It appeared that at all time points studied, i.e. 1.5, 9.5, and 18 min after injection, the specific radioactivities of free NeuAc as well as of CMP-NeuAc in the nuclear preparation were lower than those in whole liver. Also no significant differences were found between free NeuAc and CMP-NeuAc in the ratio of specific radioactivities in the nuclear fraction/whole liver. Furthermore, no enzyme involved in the synthesis of NeuAc was enriched in the nuclear preparation as compared to various other cytosolic and non-cytosolic enzymes.Because newly synthesized NeuAc is channelled into a special pool and used for activation to CMP-NeuAc [Ferwerda W, Blok CM, van Rinsum J (1983) Biochem J 216:87–92], these results point to a site of activation of NeuAc to CMP-NeuAc other than the nuclear compartment. This might indicate that the nuclear-localized enzyme, CMP-NeuAc synthase, leaves the nucleus before exerting its action.Abbreviations ManNAc kinase (EC 2.7.1.60) ATP:2-acetamido-2-deoxy-d-mannose 6-phosphotransferase - GlcNAc kinase (EC 2.7.1.59) ATP:2-acetamido-2-deoxy-d-glucose 6-phosphotransferase - NeuAc 9-phosphatase (EC 3.1.3.29) N-acetylneuraminate 9-phosphate phosphohydrolase - CMP-NeuAc synthase (EC 2.7.7.43) CTP:N-acetylneuraminic acid cytidylyltransferase - glucose 6-phosphatase (EC 3.1.3.9) d-glucose 6-phosphate phosphohydrolase - p-nitrophenylphosphatase (EC 3.1.3.1/2) orthophosphoric monoester phosphohydrolase - LDH (EC 1.1.1.27) l-lactate:NAD oxidoreductase - (1-4)-galactsyltransferase (EC 2.4.1.38) -N-acetylglucosaminide (1-4)-galactosyltransferase     

Identification and Distribution of m- and p-Hydroxyphenylacetic Acids in the Brain of the Rat

The m and p isomers of hydroxyphenylacetic acid have been identified and quantitated in whole rat brain and in several regions using a capillary column high resolution gas chromatography–mass spectrometry procedure. Their concentrations were: for m-hydroxyphenylacetic acid (mean ± S.E., number of determinations in parentheses)—whole brain, 2.3 ± 0.3 ng/g (7); hypothalamus, 1.2 ± 0.3 ng/g (5); caudate nucleus, 5.5 ± 0.6 ng/g (5); brain stem, 1.8 ± 0.1 ngig (5); cerebellum, 1.2 ± 0.1 ng/g (5) and the “rest,” 1.7 ± 0.1 ng/g (5); and for p-hydroxyphenylacetic acid–whole brain, 10.6 ± 0.7 ng/g (7); hypothalamus, 4.5 ± 0.1 ng/g (4); caudate nucleus, 28.3 ±1.6 ng/g (5); brain stem, 8.6 ± 0.6 ng/g (5); cerebellum, 8.1 ± 0.4 ng/g (9, and the “rest,” 5.3 ± 0.5 ng/g (5). This heterogeneous distribution parallels closely that exhibited by their respective precursor amines, m- and p-tyramine.