高白度低PPO活性小麦种质筛选

本试验对142份小麦种质资源的面粉白度和多酚氧化酶（PPO）活性进行测定,结果表明：面粉白度值高于国家一级粉标准的有73份,占样本总数的51.4%;样本PPO活性低于100 AU.min-1.g-1有41份,占供试样本的28.9%;筛选出9份面粉白度值高于80%,且同时具有较低PPO活性（〈100 AU.min-1.g-1）的小麦种质,可作为小麦白度性状改良的遗传材料。     

252份墨西哥CIMMYT小麦多酚氧化酶的活性差异及品种筛选

[目的]为低PPO活性小麦品种的选育提供参考信息。[方法]按Anderson和Morris等方法并作部分修改对引进的252份墨西哥CIMMYT小麦品种的多酚氧化酶活性进行测定。[结果]PPO活性主要受基因型影响;不同品种间PPO活性相差很大。252份墨西哥CIMMYT小麦的多酚氧化酶(PPO)活性的范围为54.00~285.78AU/(min·g)。其中小于100.00AU/(min·g)的品种有46个,200.00AU/(min·g)以上的有37个,其余皆为100.00~200.00AU/(min·g)。品种PPO活性分布在100.00~200.00AU/(min·g)的个数较多,而在<100.00AU/(min·g)和>200.00AU/(min·g)范围则分布较少,基本上呈正态分布。[结论]通过育种途径可改善小麦制品颜色褐变的现象。筛选出PPO活性低于100.00AU/(min·g)的46个的墨西哥CIMMYT小麦品种,可作为小麦品质育种的材料。     

槲皮素和硫脲对野桑蚕多酚氧化酶的抑制类型

多酚氧化酶(Polyphenol oxidase,PPO)在昆虫的变态发育和免疫防御中起着非常重要的作用,为开发以该酶为靶标的新型害虫抑制剂提供理论依据,以野桑蚕(Bombyx mandarina)五龄幼虫为材料提取多酚氧化酶,探讨了槲皮素和硫脲对该酶的抑制作用。结果表明,这两种抑制剂分别属于竞争性和非竞争性抑制剂。     

板栗多酚氧化酶性质的研究

为解决板栗加工褐变的问题,采用分光光度法对板栗果肉中多酚氧化酶的性质进行了研究,分别求出该酶的适宜温度为40～60 C,最适pH值为4.0和7.0,Vmax=0.93吸光度/min,Km=76.15mmol/L.并探讨了该酶的热稳定性以及抑制剂氯化钠、柠檬酸、四硼酸钠、亚硫酸氢钠和抗坏血酸对酶活力的影响.     

IBA处理对杂交榛根蘖育苗生根指标及相关氧化酶活性的影响

以"龙榛1号"当年萌生条为试材,研究了不同IBA浓度处理(500、750、1 000、1 250mg·L^-1)对平欧杂交榛根蘖育苗生根的影响,分析了根蘖育苗生根过程中过氧化物酶(POD)、多酚氧化酶(PPO)活性的变化,并对外源激素处理与相关氧化酶活性与生根的关系进行了相关性分析,以期探讨其生根机理。结果表明:IBA浓度处理能显著地提高生根率,以1 000mg·L^-1的生根效果最佳,生根率和不定根质量均显著高于其它处理。生根过程中,POD和PPO活性呈先上升后下降趋势,POD活性在育苗后20d出现峰值;PPO活性在40d时达到峰值。生根率与POD、PPO活性呈极显著正相关。说明适宜浓度IBA能增加生根过程POD、PPO活性,从而提高生根率。     

Continuous Spectrophotometric Assay and Properties of Ascorbic Acid Oxidising Factors in Wheat

A continuous spectrophotometric assay was developed to measure ascorbic acid oxidation in crude Na₂SO₄ extracts of flour. The rate of ascorbic acid oxidation in flour extracts measured using this method was similar to the rate in flour-water suspensions and 2–4 fold less than the rate in dough measured using an indophenol-xylene extraction method. Flour extracts appeared to contain two ascorbic acid oxidising factors; one with optimal activity at pH 6·3 and 30 °C and the other with optimal activity at pH 10 and 40 °C. The pH 6·3 factor had properties similar to those of ascorbate oxidase (EC 1·10·3·3) in its pH and temperature stability, strong inhibition by NaN₃, KCN and diethyldithiocarbamate, inactivation by proteases, and greater stereospecificity towards -ascorbic acid than -isoascorbic acid. The pH 6·3 factor was most concentrated in the pollard milling fraction of wheat and was lowest in flour. The pH 10 factor had several properties indicating non-enzymic oxidation of ascorbic acid; it was not inactivated by proteases, it was inhibited poorly or not at all by the above ascorbate oxidase inhibitors, and it had low specificity for stereoisomers of ascorbic acid.     

104份甘肃小麦品种脂肪氧化酶和多酚氧化酶活性基因等位变异的检测

面粉和面制品的色泽是评价小麦品质的重要指标。小麦脂肪氧化酶(LOX)和多酚氧化酶(PPO)活性对面粉白度及面制品的色泽具有重要影响。为给甘肃省小麦品质育种提供参考依据,以104份甘肃省育成的小麦品种为材料,利用功能标记LOX16、LOX18、PPO18、PPO16和PPO29检测TaLox-B1、PpoA1及Ppo-D1位点的等位变异,分析甘肃小麦品种资源中LOX和PPO活性基因的组成和分布特点。结果表明,在甘肃小麦中,等位变异TaLox-B1a和TaLox-B1b的频率分别为22.12%和77.88%;其中,甘肃冬小麦品种高LOX活性等位变异TaLox-B1a的分布频率(30.56%)高于春小麦(3.13%)。等位变异Ppo-A1a、PpoA1b、Ppo-D1a和Ppo-D1b的频率分别为49.04%、50.96%、50.96%和49.04%;两个PPO基因的等位变异组合Ppo-A1a/Ppo-D1b、Ppo-A1a/Ppo-D1a、Ppo-A1b/Ppo-D1b和Ppo-A1b/Ppo-D1a的分布频率依次为28.85%、20.19%、20.19%和30.77%。说明在甘肃小麦品种中,低LOX活性等位变异(TaLox-B1b)品种比例较高;低PPO活性等位变异(Ppo-A1b、Ppo-D1a)品种分布比例略高于高PPO活性类型;其中,32份小麦品种在TaLox-B1、Ppo-A1和Ppo-D1三个位点同时含有高LOX活性和低PPO活性的等位变异。     

甘蔗基因表达定量PCR分析中内参基因的选择

以甘蔗接种黑穗病菌后0、6、12、24、48、60和72 h时间点的材料为研究对象,应用实时荧光定量PCR(real-time quantitative PCR Real-time qPCR)技术,探讨25S rRNA、GAPDH、β-actin和β-tubulin4个内参基因mRNA水平的表达情况.经geNorm程序统计学分析,4种内参基因的表达稳定性各异,25S rRNA>GAPDH>β-actin>-tubulin,其中以25S rRNA表达稳定性最好,且根据该基因设计的两对定量PCR引物都是可行的.同时,甘蔗PPO基因表达特性的定量PCR分析也显示,甘蔗PPO基因与植物的抗病性相关.结果显示,研究中所筛选的内参基因是合适的.     

Ascorbic acid status as affected by dietary treatment in the Siberian sturgeon (Acipenser baeri Brandt): tissue concentration,mobilisation and L-gulonolactone oxidase activity

A study was conducted to evaluate tissue storage and mobilisation of L-ascorbic acid (AA) in the Siberian sturgeon (Acipenser baeri) fed three different experimental diets. The three treatments consisted of a diet devoid of vitamin C (diet A0) and two diets supplemented with equivalent of 300 mg AA kg^–1 in the form of either silicone-coated ascorbic acid (diet SC) or of ascorbyl-2-polyphosphate (diet AP). During the first phase (4 months) of the trial, six batches of 130 Siberian sturgeon (initial body weight: 25.5±0.5 g) each were fed one of the three diets in duplicate. During the second phase (3 months), fish from groups SC and AP were switched to diet A0 and those fed diet A0 during the first phase were switched to diet SC. Irrespective of the dietary treatment, growth rates were not significantly different from each other. At the end of phase I, in all tissues studied, total ascorbic acid (TAA) concentrations were higher in Siberian sturgeon fed diet AP than in the other two groups. During phase II, tissue ascorbate depletion was also higher in the AP group than in the other two groups. Transfer of the AA-free diet fed group onto a diet supplemented with 300 mg AA kg^–1 (diet SC) led to a slight increase in the TAA concentrations in all tissues. Blood plasma tyrosine concentrations were not significantly different between the three groups. Whole-body collagen levels were affected by dietary AA levels or forms at the end of phase I; the differences were not significant at the end of phase II. Muscle collagen levels were slightly affected. L-Gulonolactone oxidase activity was found in the kidney of Siberian sturgeon, but not in the liver. The ascorbyl-2-polyphosphate appears to be either better utilised by Siberian sturgeon, like in many other teleosts, or more stable than the silicone-coated AA during food processing and storage. Presence of L-gulonolactone oxidase activity in Siberian sturgeon kidney combined with the absence of gross scorbutic signs in AA-free diet fed groups expressing very good growth rates suggested no need of dietary AA byA. baeri.     

茶叶多酚氧化酶的序列分析与结构预测

调用生物信息学方法对已在国际互联网上的生物数据库(DDBJ、Uni-Prot)注册的茶树多酚氧化酶基因的核酸及氨基酸序列进行分析,并对其组成成分、信号肽、跨膜拓扑结构域、疏水性/亲水性、蛋白质二级及三级结构进行预测与推断。结果表明:茶多酚氧化酶不具有信号肽,存在卷曲螺旋,为位于细胞质中的无跨膜的亲水性不稳定蛋白,无规则卷曲是其蛋白质结构的主要结构元件,α-螺旋与β-折叠分散于整个多肽链中,并存在2个结构功能域。