小麦对白粉菌过敏性反应与H_2O_2及3种氧化酶活性变化的关系(英文)

[目的]研究小麦对白粉菌过敏性反应与H2O2及3种氧化酶活性变化的关系,为探讨小麦对白粉菌抗性生理机制奠定基础。[方法]以白粉菌菌株17(Bgt17)和菌株6(Bgt6)及小麦品种扬158为试验材料,测定H2O2处理的小麦叶片过敏性细胞数量及POD、PPO、SOD酶活性。POD活性测定参照Hammershmidt方法并加以改进,用0.05mmol/LpH值为5.5的磷酸缓冲液进行提取,以愈创木酚作为底物测定,以1minA470变化0.01为1个酶活力单位;PPO活性测定参照ConstableandRyan方法进行,用0.05mmol/LpH值为5.5的磷酸缓冲液提取,以儿茶酚为底物测定,以1minA525变化0.01为PPO活力单位;SOD活性的测定参照Levine等方法进行,用提取液(50mmol/LMet,0.1mol/LpH值为7.8的磷酸缓冲液,1mmol/L二硫苏糖醇(DDT))提取。测定SOD对氮蓝四唑(NBT)光还原的抑制率,以抑制NBT光还原50%为1个酶活力单位。[结果]乳突在亲和性与非亲和性组合叶表细胞形成的差异不大,非亲和性反应叶表细胞过敏性反应发生早且明显比亲和性反应多;接种Bgt17后不同时间及非亲和性组合中含H2O2的过敏性反应细胞频率明显高于亲和性组合;用5、10、20mmol/L抗坏血酸处理后,小麦叶片中过敏性细胞数量显著减少,15.0与30.0mmol/LH2O2处理后诱导亲和性反应小麦叶片中过敏性细胞数量显著增加;接种白粉菌后,小麦叶片中POD、PPO、SOD活性均呈先上升后下降趋势,且非亲和性组合比亲和性组合3种酶的活性高。[结论]HO、PPO、SOD活性与小麦对白粉菌抗性存在一定相关性。     

核桃仁种皮中多酚的提取工艺研究

核桃仁种皮中含有抗氧化性的多酚类物质,为探讨乙醇浓度、料液比、提取时间、提取温度对核桃仁种皮中多酚提取效果的影响,试验运用正交设计对核桃仁种皮中多酚的提取条件进行了优化。确定最佳工艺条件为:60%的乙醇,料液比为1∶8,提取时间30 min,提取温度55℃,核桃仁种皮的多酚粗提得率为5.71 mg.g-1。     

菜用型甘薯多酚含量的测定及其基因型差异

筛选了7个菜用型甘薯品种(系),采用L9(34)正交试验,研究了Na2CO3量、反应温度、FC添加量与反应时间对多酚反应效率的影响,进而选出最适反应条件,并分析多酚含量的不同基因型差异。结果显示,多酚反应体系最佳组合:6 mL 10%Na2CO3、2 mL Folin-Ciocalten、反应温度35℃、反应时间30 min;确定这4个因素的影响主次顺序为:Na2CO3>FC添加量>反应温度>反应时间。统计分析结果表明,菜用型甘薯叶的多酚含量在基因型间存在一定差异,其中含量较高的福薯7-6与福薯10,分别达4.47和3.96 mg/g FW,与其它品种(系)差异极显著,而与福薯10遗传背景相似的福薯11,多酚含量最低,为1.67 mg/g FW。     

多酚氧化酶基因NtPPOE差异表达的效应分析

为研究烟草中多酚氧化酶基因的功能,本研究从GenBank中挑选一个烟草多酚氧化酶基因Polyphenol oxidase E(基因登录号:NW_015916967.1),命名为NtPPOE。构建NtPPOE基因的CRISPR载体和超表达载体,分别得到NtPPOE基因抑制/过量表达的转化植株。研究表明,基因敲除植株NtPPOE基因相对表达量降低,PPO活性降低,多酚类物质含量升高;超表达植株NtPPOE基因相对表达量升高,PPO活性升高,多酚类物质含量降低。NtPPOE基因表达量与PPO酶活性呈正相关,与多酚类物质含量呈极显著性负相关,NtPPOE基因的表达影响了烟草多酚代谢。本研究通过分析NtPPOE基因差异表达产生的效应,初步验证了NtPPOE基因的功能,并得到了NtPPOE基因抑制/过量表达的突变植株,为PPO基因功能研究和不同PPO活性品种的育种提供材料基础。     

The control of polyphenol oxidase activity in fruits and vegetables

Objective of this research was to find alternative methods for the control of polyphenol oxidase (PPO) activity in fruits and vegetables with the purpose of reducing or eliminating the use of SO₂ for this purpose. Interactions between the use of ascorbic acid, citric acid, EDTA, sodium metabisulphite and heat treatment (70 °C for 2 min) in the control of PPO activity were studied in avocado (var. Fortuna), banana (var. Nanica), apple (var. Ana, Fuji, Gala & Golden), pear (var. D'Agua), peach (var. Real), potato (var. Bintje), eggplant (var. Super F100), mushroom (Agaricus bisporus) and hearts-of-palm (Euterpe edulis Mart.). The results demonstrated that PPO of avocado and eggplant was most resistant to inhibition by the methods used. The least efficient method tested for the control of PPO was the addition of ascorbic acid and EDTA, while the most efficient methods investigated included the use of ascorbic acid, citric acid, sodium metabisulphite and heat treatment. The results indicated that, with the exception of PPO from avocado, the most adequate alternative method to substitute for the use of SO₂ in the control of PPO was a combination of ascorbic acid, citric acid and heat treatment.     

RNA干扰对烟草NtPPO8基因沉默的效应分析

多酚氧化酶（PPO）是发生酶促褐变的关键酶,PPO活性过高会使烟叶变褐。为明确烟草NtPPO8基因对PPO活性的影响,建立降低NtPPO8基因表达量的病毒诱导的基因沉默（virus induced gene silencing,VIGS）体系,探究使用RNA干扰技术对烟叶NtPPO8基因表达量及PPO活性的影响。以中烟100和K326为材料,分析在移栽后50d进行VIGS载体侵染的植株,并在侵染后10、20、30、40d分别测定NtPPO8基因表达量及PPO活性。结果表明,接种VIGS载体的转化植株,其中在侵染后20d时NtPPO8基因的沉默效率最高,为78.46%;随着侵染时间推移,NtPPO8基因的沉默效率逐渐降低,在接种后40d内有效表达;且转化植株中的病毒载体会随着植株的生长而向上传导,随着接种部位的上移,沉默效率逐渐降低。因此,研究建立VIGS体系能够有效沉默NtPPO8基因的表达,并且有效降低PPO活性,为提高烟叶耐烤性及降低烟叶褐变比例提供了新的思路和方法。     

马尾松毛虫多功能氧化酶测定正交试验优化研究

应用L16(43)正交试验设计对影响马尾松毛虫多功能氧化酶细胞色素P450测定的因素PMSF浓度、酶原稀释比和离心速度进行优化研究。结果表明供试的16组复配组合中,组合A1B1D1(选用0.2 mol/L PMSF浓度、酶原稀释比为2∶1,离心速度1为5 000 r/min)最优,按照该组合的条件进行马尾松毛虫MFO酶细胞色素P450测定,效果最佳。     

松油烯-4-醇对粘虫几种代谢酶及酚氧化酶的影响

【目的】探讨杀虫植物砂地柏Sabina vulgaris精油中主要杀虫成分松油烯-4-醇的作用机理。【方法】以点滴法处理粘虫5龄幼虫后,测定兴奋期、痉挛期、麻痹期及复苏期等4个中毒阶段试虫体内4种代谢酶及酚氧化酶（PPO）的活性。【结果】松油烯-4-醇仅在麻痹期对酸性磷酸酯酶（ACP）具有一定的抑制作用,而在其它时期均可明显激活ACP及碱性磷酸酯酶（AKP）;对谷胱甘肽S转移酶（GSTs）则表现出先抑制后恢复的趋势;可显著抑制细胞色素P450 O-脱甲基活性,各中毒阶段酶抑制率分别为26.27%、46.03%、80.24%和90.22%;离体和活体条件下,均可显著抑制PPO活性。【结论】松油烯-4-醇对粘虫代谢酶系细胞色素P450、GSTs及PPO的抑制与其毒杀活性有关,而中毒试虫的复苏可能与GSTs活性的恢复有关。该化合物可开发为新型农药或增效剂,在害虫抗性治理中具有潜在的应用价值。     

Antidiabetic and Antioxidant Effects of Polyphenols in Brown Alga <Emphasis Type="Italic">Ecklonia stolonifera</Emphasis> in Genetically Diabetic KK-A<Superscript>y</Superscript> Mice

The dietary intake and control of blood glucose levels are very important in hyperglycemic patients and α-glucosidase inhibitors are a cost-effective means to preventing the progression of diabetes. In search of a natural inhibitor from food materials, α-glucosidase inhibitory activity and the anti-hyperglycemic effects of a brown alga, Ecklonia stolonifera, were investigated using non-insulin dependent diabetic mice. Methanolic extract of E. stolonifera (MEE), which contains a high content of polyphenols, showed strong inhibition of α-glucosidase in vitro. Male KK-A^y mice, a genetically non-insulin dependent diabetic model, showed hyperglycemia with aging, but the ingestion of MEE suppressed the increase in plasma glucose and lipid peroxidation levels in unfasted KK-A^y mice dose dependently. In KK-A^y mice, which were fed the MEE diet for 4 weeks, MEE moderated the elevation of plasma glucose levels after the oral administration of maltose. The polyphenols in MEE were estimated to be phlorotannins by HPLC-PDA and LC/MS analyses. These results demonstrate that E. stolonifera, seaweed typically used as a health food, has strong antidiabetic and antioxidant effects in vivo, thus, it may have beneficial properties in the prevention of diabetes and could be useful in the development of an antidiabetic pharmaceutical and functional food.