石榴果实发育期果皮褐变及相关酶活性变化

为探明石榴果实发育期果皮褐变及其相关酶活性、总酚和抗氧化能力的变化规律及相互关系,以泰山红和泰山三白甜石榴为试材,研究各发育期果皮褐变度、多酚氧化酶(PPO)活性、过氧化物酶(POD)活性、苯丙氨酸解氨酶(PAL)活性、总酚含量和DPPH自由基清除率的变化。结果表明,2个品种果皮褐变度、PPO活性、POD活性、总酚含量和DPPH自由基清除率随着发育天数的增加逐渐降低,均在7月15日达到最高。2个品种果实发育期果皮PAL活性呈先降低后升高再降低的变化趋势,分别在7月15日和9月13日出现峰值。泰山红抗褐变能力高于泰山三白甜,二者分别在9月23日和9月13日采收能减轻褐变的发生。相关性分析表明,PAL、PPO、POD、总酚和DPPH自由基清除率均与石榴果实酶促褐变密切相关,石榴果实酚类物质含量高低与其抗氧化能力极显著正相关。本研究结果为深入揭示石榴果实褐变机理提供了理论依据。     

小麦抗根腐病突变体抗病机理的探讨

以离体筛选获得的小麦抗根腐病突变体及其亲本为材料,在根腐病菌毒素作用下,研究了过氧化物酶、超氧化物歧化酶和苯丙氨酸解氨酶等防御酶系的变化。结果表明,毒素可使上述酶活性、同工酶谱带发生改变。突变体酶的变化比亲本显著,这些变化与它们的抗病性强弱相一致。可以认为,突变体在毒素作用下具有较强的防御酶活性是产生较强抗性的原因。     

辐照对苦荞种子发芽及苗期细胞膜与保护酶活性的影响

利用不同剂量(100~500Gy)的60Coγ射线辐照处理3个苦荞品种的干种子,研究γ射线对苦荞幼苗的性状、膜透性和保护酶活性的生物学效应。结果表明,3个苦荞品种的发芽率、苗高、根长随着辐照剂量的增加逐渐降低,细胞膜透性、丙二醛含量、SOD酶活性和POD酶活性则逐渐增加,但在0~300Gy剂量辐照范围内的晋荞2号和九江苦荞的SOD酶活性呈现增加-降低-增加的趋势。γ射线对不同品种各性状产生的影响程度也不同,3个品种辐照敏感性依次为九江苦荞>西农1号>晋荞2号。     

辐照鸡肉检测方法研究

研究了辐照对鸡腿肉脂肪酶、过氧化物酶、过氧化氢酶、鸡油的过氧化物值含量的影响 ,并探讨了不同贮藏时间对脂肪和油脂的自氧化作用。结果表明 ,脂肪酶活性随辐照剂量的增加而下降 ,过氧化物酶活性随剂量增加而上升 ,过氧化氢酶活性随剂量的增加呈下降趋势 ,油脂的过氧化物值也随辐照剂量的增加而增加。贮藏时间对过氧化物没明显影响。     

苹果种质资源矮生基因型的研究

本文测定了38个杂交组合的2597个F_1代苹果实生苗新梢伸长区节间皮层或相应部位叶片中的过氧化物酶同工酶酶谱,统计了3种杂交类型(即普通型品种之间、普通型与矮生型品种之间、矮生型与矮生型品种之间)的过氧化物酶同工酶酶9带的比例。研究表明,该比例与杂交后代出现矮生型和普通型实生苗的理论比例相符,表明苹果新梢伸长区节间皮层和相应部位叶片申的过氧化物酶同工酶酶9带是矮生型苹果岭标记,而且可能是受两对互补基因所控制。     

不同处理对水稻病程相关蛋白和过氧化物酶的影响

以稻白叶枯菌 (Xanthomonasoryzaepv.oryzae.XOO)弱毒株 75 1和Ni(NO3) 2 作为诱导因子处理水稻幼苗。表明不同处理均在第 2d表现出最大诱导效果 ,Ni(NO3) 2处理后的相对诱导效果达 50 8% ;可溶性蛋白质IEF和SDS PAGE显示 ,诱导蛋白一般在诱导后 2 4h及 48h出现。Ni(NO3) 2 处理后 48h ,SDS PAGE发现上位叶有新增谱带 (MW分别为 54 95和 3 2 3 6kD) ;IEF显示上位叶出现pI5 1谱带 ,诱导叶出现pI5 2 6谱带 ;POD同工酶诱导叶新增加 4条酶带 (Rf分别为 0 62、0 63、0 64及0 73 ) ,上位叶新增加两条酶带 (Rf为 0 63和 0 64)。不同诱导因子处理后 ,可溶性蛋白质含量增加 ,诱导后 2 4～ 48h增幅最大 ;还显示不同诱导因子对同一植物诱导PRP的效力不同     

栽培大麦辐射敏感性与进化的关系

大麦幼苗组织的过氧化物酶同工酶有3个酶带区,8条酶带。辐照后,在偏负极端出现一个新的酶带区,增加1—3条酶带。辐射剂量效应可用线性回归方程Y=A+BX拟合。大麦品种的过氧化物酶同工酶变化与辐射敏感性有关,与进化类型直接相关。协方差分析表明,裸大麦对辐射的敏感性显著高于皮大麦,四棱和六棱大麦则显著高于二棱大麦。据此可以认为,在进化上属原始类型的品种,辐射敏感性较低。     

土壤类过氧化物酶活性对长期施肥的响应

为探究不同施肥处理对土壤类过氧化物酶活性的影响，以灰漠土和红壤长期定位试验站施肥处理土壤为研究对象，借助纳米矿物颗粒的类酶颜色反应原理，研究了不施肥(CK)、施用化肥(NPK)、施用化肥和有机肥(NPKM)和施用有机肥(M)土壤类过氧化物酶活性的变化。结果表明：与天然辣根过氧化物酶(HRP)相似，土壤也具有经典的催化颜色反应，灰漠土和红壤类过氧化物酶活性范围为0.4～1.0 U/kg。长期施肥显著改变了土壤类过氧化物酶活性，其中以M处理土壤类过氧化物酶活性为最高，可达0.7 U/kg，比CK处理增加了70%以上。不同施肥年限下，CK和NPK处理土壤类过氧化物酶活性并无显著差异，而M和NPKM处理土壤类过氧化物酶活性则随施肥时间线性增加(R²=0.71,P<0.01)。高温灭菌处理后，土壤类过氧化物酶活性大多没有改变，表明其活性是土壤本身固有的氧化还原特性；但M处理红壤类过氧化物酶活性比CK和NPK处理红壤稳定性更高。皮尔森相关性分析表明，类过氧化物酶活性可能影响土壤可溶性矿质元素(如硅和镁)水平。因此，施肥措施可以通过改变土壤固有类过氧化物酶活性而影响养分周...     

水分胁迫下玉米细胞膜伤害及其保护酶活性的变化

以2个不同抗旱性玉米品种为材料,研究了水分胁迫下玉米细胞膜伤害及其保护酶活性的变化。其细胞膜保护酶系统的超氧化物歧化酶(SOD)的活性在水分胁迫初期下降,之后升高,最后持续下降。过氧化物酶(POD)的活性初期变化复杂,后期成倍升高。     

鸡粪对高产冬小麦根系活力和光合性能的影响

在高产条件下 ,设置不同的鸡粪用量梯度 ( 1 5、30、45m3 hm2 ) ,以不施鸡粪肥为对照 ,研究其对小麦根系活力和光合性能的影响。结果表明鸡粪肥处理提高了单株成穗率和单位面积穗数 ,增加了千粒重 ,但最高分蘖数和平均单穗粒数表现出了下降趋势。开花后旗叶过氧化物酶活性随穗灌浆的进行而逐渐增强 ,群体光合速率增加 ;根群的吸收活力增高 ,这种正效应随鸡粪肥用量的增加而增大。     

Characterization of polyphenol oxidase and peroxidase and influence on browning of cold stored strawberry fruit

Polyphenol oxidase and peroxidase were extracted from two different varieties of strawberry fruit (Fragaria x ananassa D, cv. 'Elsanta' and Fragaria vesca L, cv. 'Madame Moutot') and characterized using reliable spectrophotometric methods. In all cases, the enzymes followed Michaelis-Menten kinetics, showing different values of peroxidase kinetics parameters between the two cultivars: Km = 50.68 +/- 2.42 mM ('Elsanta') and 18.18 +/- 8.79 mM ('Madame Moutot') mM and Vmax = 0.14 +/- 0.03 U/g ('Elsanta') and 0.05 +/- 0.01 U/g ('Madame Moutot'). The physiological pH of fruit at the red ripe stage negatively affected the expression of both oxidases, except polyphenol oxidase from 'Madame Moutot' that showed the highest residual activity (68% of the maximum). Peroxidase from both cultivars was much more thermolable as compared with PPO, losing over 60% of relative activity already after 60 min of incubation at 40 degrees C. The POD activation energy was much lower than the PPO activation energy (DeltaE = 97.5 and 57.8 kJ mol-1 for 'Elsanta' and 'Madame Moutot', respectively). Results obtained from d-glucose and d-fructose inhibition tests evidenced a decreasing course of PPO and POD activities from both cultivars as the sugar concentration in the assay medium increased. Changes in CIE L*, a*, b*, chroma, and hue angle values were taken as a browning index of the samples during storage at 4 degrees C. A decrease in L* was evident in both cultivars but more marked in 'Elsanta'. PPO and POD activities from cv. 'Elsanta' were very well-correlated with the parameter L* (r2=0.86 and 0.89, respectively) and hue angle (r2=0.85 and 0.93, respectively). According to these results, the browning of the fruit seemed to be in relation to both oxidase activities.     

Characterization and role of polyphenol oxidase and peroxidase in browning of fresh-cut melon

Polyphenol oxidase (PPO) and peroxidase (POD) were extracted from two different varieties of melon ( Cucumis melo L. cantalupensis cv. Charentais and C. melo L. inodorus cv. Amarillo) and characterized using reliable spectrophotometric methods. In both cases the enzymes followed Michaelis-Menten kinetics, showing different values of kinetics parameters between the two cultivars: K m = 7.18 +/- 0.70 mM ('Charentais') and 6.66 +/- 0.20 mM ('Amarillo') mM; V max = 7.93 +/- 0.35 units/min ('Charentais') and 13.82 +/- 0.37 units/min ('Amarillo'), relative to PPO; K m = 24.0 +/- 2.10 mM ('Charentais') and 5.05 +/- 0.19 mM ('Amarillo') mM; V max = 344.83 +/- 10.32 units/min ('Charentais') and 80.64 +/- 2.01 units/min ('Amarillo'), relative to POD. Optimum pH for PPO was 7.0 for 'Charentais' and 7.5 for 'Amarillo, whereas it was 4.5 for both cultivars relative to POD. Melon PPO had maximum activity at 60 degrees C in both 'Charentais' and 'Amarillo' cultivars, whereas POD maximum activity was found at 45 degrees C in 'Charentais' and at 25 degrees C in 'Amarillo'. POD from both cultivars showed higher thermolability compared with PPO, losing >90% of relative activity after only 5 min of incubation at 70 degrees C. POD's activation energy was much higher than that of PPO (Delta E (#) = 86.3 and 160.6 kJ mol (-1) for 'Charentais' and 'Amarillo', respectively). PPO and POD activities from both cultivars showed a decreasing pattern as sugar concentration in the assay medium increased, except in POD extract from 'Charentais', which maintained its activity in the presence of high d-glucose concentration (up to 5 M). Changes in L*, a*, b*, chroma, and hue angle values were chosen to describe the browning development in the samples during storage at 5 degrees C. A slight decrease in L* value and a more marked reduction of a* value were noted in both cultivars above all at the end of storage period. POD activity during storage at 5 degrees C was highly correlated with changes of parameters a*, b*, chroma, and hue angle ( r (2) from 0.82 to 0.97) for cultivar 'Charentais'. According to these results, only POD activity seemed to be involved in browning of minimally processed melon.     

Phenolic compounds and related enzymes are not rate-limiting in browning development of fresh-cut potatoes

The effect of minimal processing on polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia-lyase (PAL), and phenolic compounds was studied in five potato cultivars (Agria, Cara, Liseta, Monalisa, and Spunta). Minimal processing caused an overall increase in PPO, POD, and PAL activities. The isoform pattern of PPO was the same for all of the cultivars before and after processing. No latent PPO was detected. The isoperoxidase pattern was approximately the same among cultivars. An increase in POD activity was related to the specific induction of an acidic isoperoxidase. PAL showed an induction pattern characterized by the presence of a maximum peak of activity after 4 days of processing for all of the cultivars. The sequence of browning susceptibility of potato cultivars was as follows: Monalisa > Spunta > Liseta > Cara > Agria. Browning development was only partially correlated to PAL activity (only during the first 4 days after wounding). However, this correlation could not explain the above sequence of browning susceptibility. Minimal processing caused an increase of chlorogenic acid, whereas tyrosine content remained unchanged. In summary, no significant correlation was found between either rate or degree of browning and any other biochemical and physiological attribute investigated (PPO, POD, hydrogen peroxide, ascorbic acid content, and initial phenolics content as well as total and individual phenolics accumulation).     

Effect of wounding on phenolic enzymes in six minimally processed lettuce cultivars upon storage

The effect of wounding on polyphenol oxidase (PPO), peroxidase (POD), and phenylalanine ammonia-lyase (PAL) was studied in six minimally processed lettuce (Lactuca sativa L.) cultivars upon storage for 7 days at 5 degrees C (Iceberg Mikonos (IM), I. Green Queen (IGQ), I. Asdrúbal (IA), Little Gem Sandra (LGS), Romaine Cazorla (RC), and R. Modelo (RM)). Wounding of lettuce tissue midribs (because of minimal processing) caused an exponential increase in PPO activity due to the activation process from latent to fully active PPO by following first order kinetics in the time range from 3.7 days (LGS) to 6.3 days (RC). However, total PPO activity (active plus latent) remained constant. Isoform pattern of PPO changed upon storage probably because of posttranslational processes. POD activity linearly increased with induction of new POD isoenzymes. PAL activity presented a typical bell-shaped induction pattern in four cultivars. Only IM and IGQ showed a second induction response which has not been previously described in the literature. IM was the cultivar most susceptible to browning and RC was the cultivar least susceptible. However, no clear correlation was observed between browning and any of the biochemical and physiological attributes investigated (PPO, PAL, and POD activities, total and individual phenols accumulation, and ascorbic acid content).     

Polyphenol oxidase and peroxidase expression in four pineapple varieties (Ananas comosus L.) after a chilling injury

Pineapple internal browning (IB) is a chilling injury that produces enzymatic browning associated with flesh translucency. Pineapple biodiversity allowed the investigation of how polyphenol oxidase (PPO) and peroxidase (POD) activities with their different isoforms are involved in the IB mechanism. Fruits of four varieties that expressed IB symptoms differently, Smooth Cayenne (SCay) and the hybrids MD2, Flhoran 41 (Flh 41), and Flhoran 53 (Flh 53), were stressed by cold. The susceptible varieties showed classical brown spots but different patterns of IB, whereas MD2 and controls showed no IB. Enzymatic activities were measured on fruit protein extracts and PPO and POD isoforms separated on mini-gels (PhastSystem). Only PPO activity was significantly enhanced in the presence of IB. Up to six PPO isoforms were identified in the susceptible varieties. PPO was barely detectable in the nonsusceptible variety MD2 and in controls. The number of PPO isoforms and the total PPO activity after chilling are varietal characteristics.     

Partial characterization of peroxidase and polyphenol oxidase activities in blackberry fruits

A partial characterization of peroxidase (POD) and polyphenol oxidase (PPO) activities in blackberry fruits is described. Two cultivars of blackberry (Wild and Thornless) were analyzed for POD and PPO activities. Stable and highly active POD and PPO extracts were obtained using insoluble poly(vinylpyrrolidone) and Triton X-100 in 0.05 M sodium phosphate, pH 7.5, buffer. Blackberry POD and PPO activities have a pH optimum of 6.5, in a reaction mixture of 0.2 M sodium phosphate. Optimal POD activity was found with 3% o-dianisidine. Maximum PPO activity was found with catechol (catecholase activity) followed by 4-methylcatechol. Polyacrylamide gel electrophoresis of blackberry extracts under non-denaturing conditions resolved in various bands. In the POD extracts of Wild fruits, there was only one band with a mobility of 0.12. In the Thornless POD extracts there were three well-resolved bands, with R(f) values of 0.63, 0.36, and 0.09. Both the Wild and Thornless blackberry cultivars produced a single band of PPO, with R(f) values of 0.1 for Wild and 0.06 for Thornless.     

Fermentation characteristics of some assamica clones and process optimization of black tea manufacturing

Changes in the specific activities of polyphenol oxidase (PPO), peroxidase (POD), and protease and in the relative amounts of flavan-3-ols for eight genetically derived cultivated teas at various stages of leaf maturity and in four succescive seasons were examined. A series of investigations were carried out to study the cross-reactivity of complex polyphenols and PPO-generated orange-yellow theaflavins, as well as of POD oxidized substrates, producing brown so-called thearubigins during fermented tea processing. From the estimation of five major catechins, PPO activities in young shoots, and theaflavin and thearubigin contents of crushed, torn, and curled (CTC) black teas, the superior variety and flavorful flush characteristics were refined. Notable protein hydrolysis by endogenous protease as measured from free amino acids and formation of tannin-protein complex (browning products) was obtained for cultivar character and product quality. Results showed that process optimization with respect to time, temperature, moisture, and pH maximizes PPO-catalyzed desirable theaflavin pigments, whereas POD-mediated chemical reaction produces dull color.     

Effect of wash water temperature and chlorination on phenolic metabolism and browning of stored iceberg lettuce photosynthetic and vascular tissues

Cut tissues from distinct anatomical locations in iceberg lettuce (Lactuca sativa L.) were subjected to washing in cold (4 degrees C) and warm (47 degrees C) water with or without chlorine to assess their propensity to discoloration during storage. Total protein (Bradford method) and phenolic (TPH; Folin-Ciocalteu method) contents and polyphenol oxidase (PPO; spectrophotometric method using catechol as a substrate), peroxidase (POD; guaiacol substrate), and phenylalanine ammonia-lyase (PAL; phenylalanine substrate) activities were determined in photosynthetic and vascular tissue from outer and inner leaves. Unprocessed photosynthetic and inner leaf tissues had significantly higher (P < 0.05) levels of protein and TPH and PPO and POD activities than vascular and outer leaf tissues. PAL activities (on a fresh weight basis) were similar in all tissues. Changes in browning (light reflectance measurement) and phenolic metabolism in all four tissue types were observed during aerobic storage at 5 degrees C over 10 days. PAL activity increased in all tissues after 1-2 days of storage and then gradually decreased. POD activity also increased steadily for the storage duration. Protein content and PPO activity remained constant. Edge browning (measured with a Minolta Chroma Meter) and TPH increased in all tissues, especially in outer vascular tissue. Cut photosynthetic and vascular tissues washed at 4 and 47 degrees C with and without 100 microg mL(-1) chlorine for 3 min were analyzed during 7 days in storage at 5 degrees C. Enzyme activities and accumulation of phenolics in all tissues washed at 47 degrees C were significantly (P < 0.05) lower compared to controls or tissues washed at 4 degrees C. Chlorine had no additional effect at 47 degrees C but significantly (P < 0.05) reduced browning and accumulation of phenolics in lettuce washed at 4 degrees C. These results showed that inherent differences between tissues affect phenolic metabolism and browning in stored, fresh-cut lettuce.     

减压贮藏对去壳黄甜竹笋的保鲜效果及其生理和分子机制

为了探讨减压贮藏延缓采后去壳黄甜竹笋木质化和褐变的效果及其作用机制,本试验以黄甜竹笋为试验材料,笋剥壳后于温度6 ±1℃、相对湿度80%~85%、55 kPa减压环境下贮藏10 d,测定其品质指标、竹笋木质化和褐变关键酶的活性和相关基因的相对表达量。结果表明,减压贮藏(中后期)显著抑制了黄甜竹笋基部切面的褐变(P<0.05),显著延缓了黄甜竹笋的呼吸速率、电导率、硬度,以及丙二醛、纤维素和木质素含量的上升(P<0.05),同时也显著抑制了苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、肉桂醇脱氢酶(CAD)、多酚氧化酶(PPO)的活性及其编码基因的相对表达量(P<0.05)。本研究结果为减压贮藏在黄甜竹笋采后保鲜的应用提供了理论依据。