特定电磁波(TDP)对植物超氧物歧化酶的影响

本文报道了特定电磁波(TDP)处理对水稻、小麦、玉米、番茄等植物种子吸胀、萌发过程中及其幼芽、幼苗中超氧化物歧化酶(SOD)活力及其同工酶的影响,电磁波长为0.25～50μm,处理时间为1～5 h 不等,处理温度控制在30±2℃,SOD 活力测定采用酶抑制氯化硝基四唑兰(NBT)光还原法。试验结果表明,0.25～50μm的电磁波明显提高了上述植物中 SOD 活性,增加幅度一般在11%～100%之间,试验结果亦指出,特定电磁波未改变 SOD 同工酶酶谱。作者认为 TDP 对 SOD 活力的提高有助于提高植物的抗逆性能力。     

Fractionation of soil humic acids by preparative polyacrylamide gel electrophoresis in the presence of concentrated urea

To separate soil humic acids (HAs) into their constituents and characterize them, polyacrylamide gel electrophoresis (PAGE) was carried out in the presence of 7 M urea using a preparative electrophoresis system. Two types of soil HAs were fractionated into nine fractions by PAGE. The dark-colored constituents were recovered from the electrophoretic fractions by precipitation on acidification, and the brown-colored constituents dissolved in the acidic solution of fast-moving fractions were recovered by adsorption onto DAX-8 resin. High-performance size-exclusion chromatography (HPSEC) confirmed that the constituents of the HAs were separated based on their molecular size by PAGE. The dark-colored constituents exhibited higher degrees of humification than did the corresponding unfractionated HAs, except for the constituents remaining in the electrophoretic gels at the end of electrophoresis. Diffuse reflectance infrared spectroscopy revealed that the chemical properties of the dark-colored constituents changed regularly: the content of carboxyl groups decreased and the proportions of proteinous, aliphatic and polysaccharide moieties increased with increasing molecular size. The humification degrees of the constituents adsorbed onto DAX-8 resin were considerably lower than those of the corresponding unfractionated HAs. The chemical properties of the DAX-8-adsorbed constituents were different from those of the dark-colored constituents. Observation of electrophoretic fractions under blue light (470 nm) and HPSEC with fluorescence detection at an excitation wavelength of 460 nm and an emission wavelength of 520 nm showed that green fluorescent substances were largely concentrated in the smallest molecular size fractions and were partitioned into both the dark-colored precipitates and DAX-8-adsorbed fractions. The proportion of organic carbon recovered by precipitation and adsorption onto DAX-8 resin was 45–63%, indicating that substantial parts of the HA constituents were missing. The unrecovered constituents were considered to be acid-soluble, nearly colorless substances. The dissociation of the acid-soluble constituents from the acid-insoluble dark-colored constituents during the preparative PAGE of soil HAs was ascribed to disruption of hydrogen bonding and hydrophobic interactions caused by concentrated urea.     

甜菜SRAP-PCR反应体系的优化

为了建立适宜甜菜的SRAP反应体系,以40个不同类型的甜菜品系为试验材料,研究了PCR反应体系的主要成分对SRAP扩增结果的影响。对SRAP反应体系中的DNA模板浓度、Taq DNA聚合酶浓度、引物浓度以及dNTP浓度进行了探索,确立了适合甜菜的SRAP反应体系为:在20μL反应体系中,模板DNA为40ng,Taq DNA聚合酶1.0U,引物60ng,dNTP(2.5mmol/L)为1.6μL,扩增程序为94℃预变性3min,反应前5个循环在94℃1min,35℃1min,72℃1min的条件下运行;随后的35个循环复性温度提高到50℃,最后72℃延伸10min。并利用该反应体系对甜菜40个不同品系进行SRAP反应,用6%的变性聚丙烯酰胺凝胶电泳检测,不同品系间DNA谱带多态性丰富,证实该体系稳定可靠,可以用于甜菜的分子标记研究。     

活性污泥中细菌对聚丙烯酰胺的生物降解研究

聚丙烯酰胺(Polyaerylamide,PAM)是一类重要的水溶性高分子聚合物,已广泛应用到工农业生产的各个领域和人们的日常生活中.同时,PAM在环境中的残留、迁移和降解对环境的污染也日趋严重,尤其是降解后的单体丙烯酰胺对人类的神经系统有很大的危害.本文从胜利油田的活性污泥中筛选出3株聚丙烯酰胺降解菌,通过比较筛选出一株降解效果较好的菌,命名为AS-2.根据生理生化特性分析,初步鉴定为海球菌属.采用室内培养方法,研究了AS-2对聚丙烯酰胺生物降解的最佳条件.结果表明,当降解时间为5 d,pH=8,温度为40℃,碳源为原油,氮源为NaNO3,原油和NaNO3的含量分别为2.5,1.4g·L-1时,AS-2对聚丙烯酰胺的降解率达到45.23%.通过对聚丙烯酰胺生物降解前后的红外谱图比较,推断出AS-2主要降解了聚丙烯酰胺侧链的酰胺基,将酰胺基降解为羧酸和游离的氨基.用高效液相色谱检测生化后的PAM溶液,未检测出单体丙烯酰胺.     

植物蛋白质组研究方法

蛋白质组研究是后基因组时代的热点领域,介绍了蛋白质组概念提出的背景,蛋白质组与基因组的联系与区别,蛋白质纽研究的内容,重点综述了植物蛋白质纽研究的技术．双向凝胶电泳仍为蛋白质分离的核心技术,该技术在某些方面作了改进和发展;生物质谱是蛋白质鉴定的关键技术,进一步发展了生物传感芯片质谱及蛋白质芯片技术;对蛋白质纽研究的信息处理依赖于生物信息学的发展,介绍了国内外相关的蛋白质数据库及建立数据库主要应用的图像软件。     

玉米种子清蛋白 PAGE 谱带稳定性及玉米杂交种纯度检验

利用改进的聚丙烯酰胺凝胶电泳（ＰＡＧＥ）技术对１１个不同玉米（Ｚｅａ ｍａｙｓ Ｌ．）自交系和杂交种种子清蛋白（ａｌｂｕｍｉｎ）进行了电泳分析。结果表明,改进的聚丙烯酰胺凝胶电泳技术对玉米种子清蛋白具有较高的分辨率;玉米种子清蛋白存在丰富的多态性,各自交系和杂交种的电泳图均具有鲜明特征,并构成其独特的生化“指纹”;电泳谱带具有相对稳定性,基本不受产地和收获早晚的影响。     

The characterization of Nigerian varieties of pepper, Capsicum annuum and Capsicum frutescens by SDS- polyacrylamide gel electrophoresis of seed proteins

The possibility of using electrophoresis to characterize varieties of pepper, Capsicum annuum and Capsicum frutescens cultivated in Nigeria was investigated. The SDS- polyacrylamide gel electropherogram of extracted total seed proteins of 10 breeding lines in each of the 6 varieties investigated, revealed a pattern in which 12 polypeptide bands with apparent molecular weight range of 22 to 98 kilodaltons could be distinguished. The result showed that the six varieties could be characterized on the basis of presence/absence and staining intensities of 7 polypeptide bands. It is suggested that SDS- polyacrylamide gel electrophoresis of seed proteins provides a useful analytical technique for the characterization of varieties of pepper and there may be genotype duplicates in the collection of Nigerian Capsicum germplasm.     

Ontogenic changes in the digestive enzyme patterns and characterization of proteases in Indian major carp Cirrhinus mrigala

Digestive enzymes of Cirrhinus mrigala (Ham.) were studied during ontogenic development. Specific amylase activity was detected in first feeding fish. The enzyme activity decreased up to day‐18 and then it increased with the age of fish to reach the highest level on day‐34. Protease activity was 28.61 ± 8.90 mU mg protein^?1 min^?1 on day‐4 and increased with the age throughout the study period. Trypsin activity was 31.86 ± 1.12 mU mg protein^?1 min^?1 on day‐4. The activity decreased up to day‐10 and from day‐12 onwards increased up to day‐26. Chymotrypsin activity was 14.56 ± 2.74 mU mg protein^?1 min^?1on day‐4 and constantly increased up to day‐26. A significant increase in lipase activity was observed between days‐24 and 34. SDS‐PAGE and substrate SDS‐PAGE showed the diversity of protein (17.4–127.8 kDa) and protease activity bands (16.6–88.8 kDa) during ontogenesis. Soybean trypsin inhibitor, phenyl methyl sulphonyl fluoride, N‐α‐p‐tosyl‐l ‐lysine chloromethylketone and N‐tosyl‐l ‐phenylalanine chloromethylketone inhibited the protease activity up to 79.72–97.21, 65.55–94.83, 45.41–75.31 and 40.78–64.72%, respectively. Inhibition study in substrate SDS‐PAGE revealed the abundance of serine proteases and the presence of isoforms of trypsin and chymotrypsin. Ethylenediamine‐tetraacetate showed 5.56–22.78% inhibition of metal ion‐specific enzyme activity.     

基于相对迁移率的节节麦醇溶蛋白组成分析

为准确揭示节节麦醇溶蛋白的组成情况,对不同地理分布的212份节节麦的种子蛋白进行酸性聚丙烯酰胺凝胶电泳(A-PAGE)分析。由于麦醇溶蛋白现有的"数字字母命名"系统并不具备广泛的普适性,将普通小麦——‘中国春’(对照)第1条谱带的迁移率作为标准,其他供试材料的谱带与它的比值作为相对迁移率来确定每条醇溶蛋白条带的位置。结果显示,212份节节麦材料中共存在422种不同迁移率的谱带。所有条带可分为ω、γ、β和α4个区,α区的多态性条带最多,达34.70%。多态性条带品种间遗传相似系数(GS)的变幅为0～0.58,遗传变异较大。聚类分析结果表明,节节麦可能是以伊朗为中心向四周各地扩散传播。中国境内的节节麦至少存在3种类型。基于相对迁移率的表示方法,无需除‘中国春’以外的其他对照材料,即可有效区分A-PAGE胶上距离较近的蛋白条带,为不同麦类作物醇溶蛋白的研究提供统一标准。