嗜水气单胞菌外膜蛋白（OMP）的原核表达及其对BALB/c小鼠的免疫保护研究

为确定嗜水气单胞菌(Aeromonas hydrophila,Ah)AS1.927株外膜蛋白(OMP)的原核表达产物是否具有抗原性及其对小鼠的免疫保护力.本研究根据GenBank上嗜水气单胞菌外膜蛋白ompA基因序列(GenBank登录号:AF146597)设计1对引物,克隆出ompA全长基因1020 bp(GenBank登录号:EU309491),经Sal Ⅰ和Xhol Ⅰ双酶切后连接pET-30a(+),转化大肠杆菌(Escherichia cDli)BL21,IPTG诱导表达,并进行SDS-PAGE及Westernblot分析鉴定.结果显示重组基因工程菌E.coli[pET-30a-ompA]表达的融合蛋白分子量约为40.7kD,可以被鼠抗嗜水气单胞菌外膜蛋白抗血清识别.将重组基因工程菌Ecoli[pET-30a-om-pA]口服免疫BALB/c小鼠(Mus mussulus);末次免疫1周后用放射免疫分析小鼠肠粘膜分泌型免疫球蛋白A(sIgA)水平,检测结果揭示,该重组基因工程菌能提高sIgA的分泌(P<0.05);同时在小鼠血清中测出特异性抗体IgG(P<0.05).末次免疫2周后用100 LD50的Ah AS1.927(3.3×105> cfu/mL)腹腔注射攻击小鼠,口服重组菌对小鼠的相对免疫保护力(relative percent survival,RPS)为75%,表明重组基因工程菌可诱导小鼠对AhAS1.927产生一定的免疫保护作用.     

Methylene Blue Stainability of Root-Tip Protoplasts as an Indicator of Aluminum Tolerance in a Wide Range of Plant Species,Cultivars and Lines

We previously developed a new simple technique of methylene blue (MB) staining for the discrimination of aluminum (Al)-tolerant protoplasts from 4 plant species (rice, oats, maize and pea). The objectives of the present study were to confirm the applicability of this technique to a wider range of plant species, cultivars and lines, and to identify a common strategy for the early stage of Al tolerance. A total of 10 plant species, two Brachiaria spp., two Oryza spp., buckwheat, maize, pea, triticale, wheat and barley, corresponding to 18 different plant samples (species, cultivars, and lines), were used. Al tolerance (relative net root elongation of the longest root), which was screened at 20 μM AICI₃ in 0.2 mM CaCl₂ (pH 4.9) for 24 h, ranged widely from 10 to 88. Among cultivars and lines within the same species, Al accumulation in the root tip portion was higher in Al-sensitive plants, corresponding to more severe permeabilization of the plasma membranes (PM). Protoplasts isolated mainly from the epidermis, and outer and central parts of the cortex were stained to different degrees by MB, and the blue color was observed both on the surface and inside the protoplasts. Color pictures obtained after staining for 3 min with 0.1% MB were analyzed by Image Hyper II. The ratio of the heavily stained area at threshold 95 to the entire area stained with MB at threshold 125 was defined as MB stainability. MB stainability was negatively correlated with Al tolerance (y= 48.6e^?0.02x , R ²= 0.676**) suggesting the common importance of permeation characteristics of PM, in addition to PM negativity for Al tolerance in a wide range of plant species, cultivars and lines. Analysis of the PM lipid composition was proposed as an important topic for future studies on the negativity and permeation of PM.     

绵羊宿营法防除天然草地灌木杂草研究Ⅲ新鲜羊粪尿对土壤和植物细胞形态结构及生理的影响

采用盆栽试验研究了绵羊新鲜粪尿对植物组织水势、细胞膜透性和细胞形态结构的影响。结果表明 ,在土壤含水量 30 %左右 ,植物无水分胁迫的可能时 ,各处理植物组织水势呈先上升后下降的趋势 ,而对照则呈持续上升趋势。说明绵羊新鲜粪尿加入土壤后 ,土壤溶液浓度升高 ,影响了植物水势。绵羊新鲜粪尿对植物生理影响表明 ,叶片细胞膜透性在 UD处理与对照差异显著 (P<0 .0 5) ,U和 D单处理与对照间差异不显著 (P>0 .0 5)。根细胞膜透性 UD和 U处理与对照差异显著 (P<0 .0 5)。D处理与对照无显著差异 (P>0 .0 5)。植物细胞形态结构显示 ,各处理均无质壁分离现象 ,但 U和 UD处理的皮层细胞出现叶绿体 ,这可能是尿中尿素水解释放的游离氨被植物吸收所致     

四种菊酯类杀虫剂对枸杞蚜虫线粒体膜流动性的影响

为明确菊酯类杀虫剂对昆虫线粒体膜流动性的影响，以1，6-二苯基-1，3，5-己三烯（1，6-diphenyl-1，3，5-hexatriene，DPH）为荧光探剂，采用荧光偏振法研究了4种菊酯类杀虫剂对枸杞蚜虫Aphis sp.线粒体膜流动性的影响。结果表明，4种菊酯类杀虫剂对枸杞蚜虫线粒体膜流动性均有显著影响，其荧光偏振度显著增大，线粒体膜流动性显著减小。当药剂浓度为100 μmol/L时，对照组枸杞蚜虫线粒体膜的荧光偏振度为0.239，高效氯氰菊酯的影响最大，线粒体膜的荧光偏振度为0.315，联苯菊酯的影响最小，线粒体膜的荧光偏振度为0.287。4种菊酯类杀虫剂对线粒体膜流动性的影响都存在一定的浓度效应，当药剂浓度在0~100 μmol/L范围内，浓度越大影响越大。4种菊酯类杀虫剂在不同温度下对枸杞蚜虫线粒体膜流动性的影响不同，高效氯氰菊酯、联苯菊酯和甲氰菊酯对线粒体膜流动性的影响基本不受温度影响，而氯菊酯在高温条件下产生的影响大，在低温条件下产生的影响小，表现为明显的正温度系数。表明DPH可以作为荧光探剂研究枸杞蚜虫线粒体膜的流动性。     

鸡毒支原体特异膜抗原的制备

采用 Ｓ Ｄ Ｓ聚丙烯酰胺凝胶电泳分离纯化鸡毒支原体膜蛋白。选择切取１１０ Ｋ Ｄ、９８ Ｋ Ｄ、６６～６２ Ｋ Ｄ 和５６ Ｋ Ｄ 特异蛋白条带，以 Ｐ Ｂ Ｓ（ｐ Ｈ７．４）浸泡过夜，浸出液置于透析袋封口，透析袋外散布 Ｐ Ｅ Ｇ（ Ｍ ＝ ６ ０００）浓缩膜抗原后获得特异膜抗原，为鸡毒支原体特异诊断方法的建立和单克隆抗体的制备奠定了基础。     

芸苔链格孢菌毒素对白菜细胞膜透性、SOD酶和POD酶活性的影响

 本文以白菜幼苗为材料,研究了芸苔链格孢菌毒素(AB-毒素)对白菜叶片细胞膜透性及叶片内SOD酶、POD酶活性变化的影响。结果表明,白菜叶片用AB-毒素处理后,细胞膜透性增大,且随处理时间的延长和毒素浓度的增加而增高。不同白菜品种用毒素处理后膜透性的改变与病菌接种的发病程度呈正相关,即电导率大的白菜品种,叶片发病也重。试验结果还发现,芸苔链格孢菌及其毒素处理白菜后,体内SOD酶和POD酶活性的变化基本趋于一致,即处理后12 h内急剧下降,然后(12~48 h)活性上升。而对照处理基本变化不大。可见,芸苔链格孢菌毒素表现了与其病原菌一致的致病作用。     

Differential Response of Southern US Rice (Oryza sativa L.) Cultivars to Ultraviolet‐B Radiation

Elevated ultraviolet‐B (UV‐B; between 290 and 320 nm) radiation, because of depletion of the stratospheric ozone layer, is one of the major environmental factors influencing plant metabolic processes and yield. The southern US rice cultivars contribute greatly towards US rice production, but the effects of elevated UV‐B radiation on these cultivars are not well known. The objective of this study was to determine the effects of elevated UV‐B radiation on leaf photosynthetic rate (P_n), membrane stability, pollen viability, phenolic concentration and yield of eight commercially popular southern US rice cultivars (five inbred cultivars and three hybrids). Plants were grown in a temperature‐controlled greenhouse in Beaumont, TX, USA, and were exposed to UV‐B radiation of 0, 8 or 16 kJ m^?2 day^?1 for 90 days. For most of the cultivars, plants grown under 8 or 16 kJ UV‐B radiation showed significant decreases in P_n, membrane stability, pollen viability, and yield compared with the plants grown under an UV‐B‐free environment, whereas there was a significant increase in leaf phenolic concentration under 16 kJ UV‐B radiation. The hybrid ‘Clearfield XL729’ performed best among the selected southern US rice cultivars under 16 kJ UV‐B radiation.     

Effects of Iron Deficiency on Thylakoid Membrane Structure and Composition in the Alga Dunaliella salina

Abstract

The unicellular green alga Dunaliella was previously proposed as a model photosynthetic organism for adaptation to iron deficiency. The purpose of this study was to find out how iron limitation affects the structure and composition of the photosynthetic system of Dunaliella salina. Iron deprivation did not retard proliferation of D. salina cells, but was associated with a decrease in cell volume and chlorophyll content, and with a four‐fold reduction in iron content and a two‐fold increase in Cu content. Electron microscopic analysis revealed shrinkage of the chloroplast and decrease in stacked thylakoid membranes. Measurements of chlorophyll fluorescence induction in the presence of DCMU and of 77 K chlorophyll fluorescence emission spectra indicated gross changes in the photosynthetic efficiency of reaction centers and in the organization of their associated light harvesting antenna. Differential analysis of protein composition led to the identification of a major thylakoid membrane protein (Tidi), that was specifically induced under iron deprivation. Partial sequencing suggests that Tidi is a novel type of a chlorophyll a/b binding protein. These results clearly show that iron limitation is associated with extensive reorganization of the photosynthetic system in Dunaliella.     

不同钼水平对山羊红细胞膜自由基代谢及其酶活性的影响

本研究旨在观察不同钼水平对山羊红细胞膜自由基代谢及其酶活性的影响。选用27头健康山羊随机分成3组，选用七钼酸铵（[（NH4）6Mo7O24·4H2O]）作为本试验口服钼源，采用（NH4）6Mo7O24·4H2O水溶液对实验山羊每d1次的方式进行灌服。对照组灌服相应剂量蒸馏水，高钼组山羊按每公斤体质量15mg和45mg（以钼计）灌服（NH4）6Mo7O24·4H2O水溶液（15mg Mo·kg^-1·BW组（Ⅱ组）和45mgMo·kg^-1·BW组（Ⅲ组））。试验为期50d，并于第0、10、20、30、40、50天采血，测定红细胞膜中自由基含量和酶活性变化。结果显示，组内比较，Ⅱ组、Ⅲ组GSH—Px、T—AOC、CAT、GST、T—AOC、ATPase和AKP活性在试验前期（0～30d）差异不显著（P〉0．05），试验后期（40～50d）与前期比较下降，差异显著（P〈0．05）；MDA含量、XOD和LDH活性在实验前期（0～30d）差异不显著（P〉0．05），后期（40～50d）与前期（0～30d）比较升高，差异显著（P〈0．05）；组间比较，Ⅱ组、Ⅲ组GSH—Px、T—AOC、CAT、GST、T—AOC、ATPase和AKP活性在实验前期（0～30d）与对照组比较差异不显著（P〉0．05），后期（40～50d）与对照组比较降低，差异显著（P〈0．05）；MDA含量、XOD和LDH活性前期（0-30d）与对照组比较差异不显著（P〉0．05），后期（40～50d）升高，差异显著（P〈0．05）。由此可见，摄入过量的钼会导致山羊红细胞膜抗氧化功能降低、自由基含量升高和能量代谢障碍。     

乳状液膜在金属离子测定中的分离富集作用

简要介绍乳状液膜技术在金属离子分析测定中的分离富集作用以及新的研究和应用进展状况。