不溶性草酸氧化酶的包埋及草酸快速检测方法研究

通过研究不溶性草酸氧化酶包埋方法和草酸快速检测方法,建立氧电极法测定草酸的便捷体系。以大麦麸皮为原料制备不溶性草酸氧化酶,采用5%的琼脂糖包埋,检测包埋酶的稳定性和抗离子干扰能力;以包埋酶为工具酶,通过氧电极法检测草酸含量。试验结果表明:包埋酶连续反应2 h酶活性下降低于5%,贮存30 d活性下降低于8%;草酸检测线性范围0.1～0.8mmol/L,外标法草酸回收率为83%～97%。用氧电极法与显色法测定小麦草酸含量,两种方法的测定值无显著性差异。但氧电极法测定耗时短,反应体系简单,易于推广应用。     

Phylogeny of Limenitidinae Butterflies （Lepidoptera： Nymphalidae） Inferred from Mitochondrial Cytochrome Oxidase I Gene Sequences

The phylogenetic analyses of the subfamily Limenitidinae are performed based on 1471 bp of mtDNA cytochrome oxidase subunit I (C01) gene sequence data which were obtained from 21 individuals spanning 9 genera, along with those of 17species obtained from GenBank, using Apatura iris, Aglais urticae, and Polyura dolon as outgroup species. Although the transitions at the third codon positions of the COI data set were highly saturated, they were still retained for analysis as they contain the majority of the phylogenetic information, and thus, the maximum pasimony (MP) under different weighting schemes and maximum likelihood (ML) trees were reconstructed in this study. The results showed that within this subfamily, the results based on the COI gene sequences are approximately identical to the traditional classification results. However, the clustering of Lexias pardalis and Tanaecia julii within the genus Euthalia as well as the clustering of Phaedyma aspasia within the genus Neptis with weak support are different from that of the current classification scheme made by Chinese scholars. The genus Limenitis is splited into two subelusters in the trees constructed by using MP and ML methods. These results support one of the strongest hypotheses for the tribe relationships within Limenitidinae.     

Response dynamics of three defense related enzymes in cotton leaves to the interactive stress of Helicoverpa armigera (Hübner) herbivory and omethoate application

In order to explore the response dynamics of the activities of defense related enzymes in cotton leaves towards the interactive stress of Helicoverpa armigera herbivory and omethoate application, the activities of phenylalanine ammonia-lyase(PAL), lipoxygenase(LOX), and polyphenol oxidase(PPO) were examined from 6 to 126 h after cotton leaves were treated 12 h of H. armigera herbivory, and then sprayed with 800 mg L–1 omethoate. The results showed that the changes in the activities of PAL, LOX and PPO that occured under the interactive stress of H. armigera herbivory and omethoate application reflected the interactive effects of the two stresses on cotton defense. The similarity between the response dynamics of PAL, LOX, and PPO activities in cotton leaves under the interactive stress and that under H. armigera herbivory treatment alone showed that the induction of H. armigera herbivory on the activities of PAL, LOX and PPO in cotton leaves played a leading role in the interactive effects, and the effect of omethoate application played only a minor role. A joint factor analysis was performed according to a method which has been used to analyze the joint toxicity of pesticides; this analysis sought to clarify if there was a synergistic, antagonistic, or additive effect on PAL, LOX, and PPO activity in cotton leaves resulting from the interactive H. armigera herbivory and omethoate treatment. In the interactive effect on the response of PAL activity in cotton leaves, antagonistic effects of the omethoate application towards H. armigera herbivory were observed at 6 and 12 h. Synergistic effects were then observed at 18 and 30 h. Antagonistic effects were observed from 54 to 78 h and synergistic effects were finally observed at 126 h. The correlation between H. armigera herbivory and omethoate application in the interactive effect on cotton defense responses of LOX activity also fluctuated from synergism to antagonism during the time course. In the interactive effect on PPO activity, only antagonism was observed between H. armigera herbivory and omethoate application. In the interactive stress of H. armigera herbivory and omethoate application on cotton defense responses, omethoate affected the defense responses of cotton to H. armigera herbivory by producing antagonistic and synergistic effects. These results will be useful to understand the relationship between host plant and herbivorous pest.     

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。     

复合生物保鲜剂对腐败希瓦氏菌的抑菌机理

为了研究复合生物保鲜剂对冷藏带鱼优势腐败菌(腐败希瓦氏菌)的抑菌作用机理,通过牛津杯法测定了复合生物保鲜剂对腐败希瓦氏菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC),并测定了复合生物保鲜剂对腐败希瓦氏菌的抑菌活力、细菌生长曲线、细胞膜完整性和细胞的稳定性,对细菌超微结构进行了观察。结果显示:复合生物保鲜剂对腐败希瓦氏菌的MIC与MBC分别为1.6 mg/ml与3.3 mg/ml,且随着作用时间的延长,复合生物保鲜剂对腐败希瓦氏菌的生长有明显抑制作用;尤其经2倍最小杀菌浓度的复合生物保鲜剂处理后,细菌未出现对数生长期;菌液在260 nm处的吸光值明显升高,菌体细胞膜完整性受到破坏;菌体细胞中的碱性磷酸酶升高,菌体细胞壁通透性增大;菌悬液的电导率值明显增大,细胞膜稳定性受损。细菌超微结构观察发现,复合生物保鲜剂作用于菌体细胞后,菌体细胞开始出现皱缩、扭曲变形、表面粗糙和布满泡状物及细胞壁塌陷等现象。表明复合生物保鲜剂可以破坏细菌的细胞内环境和细胞膜稳定性,影响了细菌的正常生长;并且通过影响细菌对营养物质的吸收和代谢物的排出,最终导致菌体死亡。     

Influence of Dietary Isoflavonoids on Stability of Egg Yolk Lipids

An experiment was conducted to investigate the influence of dietary tea polyphenol (TP) and daidzein (DA) on lipid stability of egg yolk. Two-hundred and seventy 27-week-old Hisex Brown laying hens were divided into nine groups, fed a diet supplemented with 0, 5, 10, 20, 40 mg kg-1 TP and 5, 10, 20, 40 mg kg- 1 DA respectively for a period of 8 weeks ad libitum. The dietary addition of TP and DA both could reduce egg yolk LPO content significantly at the end of 4-week experiment (P ＜ 0.05) and keep egg yolk lipid stable at the supplemental dose of 10 mg kg- 1 diet respectively. A significant liner reduction in egg yolk LPO content was found with the increasing of dietary TP and DA. The egg yolk LPO content was decreased by 23.1 and 27.0% (P＜ 0.05) with the supplementation of TP and DA respectively at 40 mg kg-1 diet. The LPO contents in liver, plasma and egg yolk were decreased significantly with the dietary addition of TP or DA respectively ( P ＜ 0.05). There was no significant difference between the effects of TP and DA on improvement of layers‘ antioxidative capacity. The dietary TP and DA both could improve the bird‘s antioxidative level directly irrelevant to SOD and GSH-Px actiVities.     

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。