一株耐辐射考克氏菌的分离与鉴定

从10kGy60Coγ射线辐照处理的新疆戈壁土壤中分离到一株耐辐射微生物I-7R,该菌菌落为橙红色革兰氏阳性球菌。最适生长温度为30℃,(G+C)mol%含量为67.5%,不具有氨苄青霉素、氯霉素、四环素、卡那霉素、潮霉素、利福平及壮观霉素等抗性。UV辐射生存曲线显示,I-7R菌株与大肠杆菌K-12相比,具有较强的紫外辐射抗性。16S rDNA序列比较分析表明I-7R菌株与Kocuria rosea、K.erythromyxa以及K.polaris同源性达到99%。结合I-7R菌株生理生化试验结果,该株菌归属于考克氏菌属(Kocuria),并与Kocuria rosea最为相近,暂命名为K.rosea I-7R。     

耐辐射球菌清除活性氧自由基及对DNA的保护作用

用化学发光法分析了耐辐射球菌 (D .radiodurans)的两个菌株 (R1、KD830 1 )和大肠杆菌对超氧阴离子自由基、羟基自由基和过氧化氢的清除能力。通过羟基自由基诱导的DNA氧化损伤化学发光模型考察了耐辐射球菌细胞提取物对自由基氧化导致DNA氧化损伤的保护作用。结果表明 ,耐辐射球菌提取物能显著清除O-2·、H2 O2 和·OH ,清除能力明显高于大肠杆菌 ,并能有效地防止自由基所致DNA氧化损伤 ,表现出超强的抗氧化和耐辐射能力 ,这归因于耐辐射球菌中具有较高的抗氧化酶活性     

耐辐射动球菌recO和recR基因的克隆表达及功能验证

为研究耐辐射动球菌Rec O和RecR蛋白的抗紫外辐射损伤效应,本研究通过PCR扩增耐辐射动球菌rec O和recR基因的全长序列,构建重组质粒p GEX-2T-rec O和p GEX-2T-recR并转化到大肠杆菌E.coli BL21(DE3)中,IPTG诱导重组蛋白表达,Western Blot检测蛋白表达,测定紫外辐照前后E.coli BL21(DE3)、空载菌株及表达菌株的生存率,并采用实时荧光定量PCR技术检测工程菌中耐辐射动球菌rec O、recR基因及E.coli BL21(DE3)中Rec FOR途径相关基因(rec F、rec O、recR、rec A、rec G、ruv A、ruv B、ruv C、ssb)的表达情况。结果表明,Rec O和RecR融合蛋白在大肠杆菌中成功诱导表达;当紫外辐照剂量大于8 J·m-2,导入p GEX-2T-rec O和p GEX-2T-recR的工程菌存活率均高于对照组,说明耐辐射动球菌RecR和Rec O蛋白能增强E.coli BL21(DE3)对紫外辐射的抵抗能力;Rec O工程菌中rec F、recR、rec A、rec G、ruv A、ruv B、ruv C、ssb的表达明显高于对照组,RecR工程菌中Rec FOR途径其他相关基因的表达与对照组差异均不显著,表明耐辐射动球菌recR和rec O基因可通过不同的调控路径提高E.coli BL21(DE3)的抗紫外辐射能力。本试验结果为耐辐射动球菌的耐辐射机制研究提供了一定的科学依据。     

耐辐射动球菌类胡萝卜素提取物的抗氧化活性研究

耐辐射动球菌(KR)是一种从高放射性污染环境中分离出来的具有抗辐射、抗氧化特性的橘黄色球菌。为探明KR抗氧化特性与菌体内代谢物活性的关系,从KR中分离提取纯化类胡萝卜素,利用高效液相色谱(HPLC)和液质联用(LC-MS)分析提取物的组成和性质,通过体外高铁离子还原能力和DPPH自由基清除试验、体外脂类氧化抑制试验和DNA氧化损伤的抑制试验,比较KR类胡萝卜素提取物与番茄红素、β-胡萝卜素的体外抗氧化能力及生物大分子氧化保护能力。结果表明,KR提取物中,其中一种产物的最大吸收波长在471.9 nm和501.1 nm处,分子式C_(48)H_(66)O_2;另一种产物的最大吸收波长在464.6 nm,分子式C_(24)H_(34)O。KR类胡萝卜素提取物的高铁离子还原能力在浓度为2μg·mL~(-1)时达到1.22;DPPH自由基清除率在浓度为4μg·mL~(-1)时达到94.26%;对脂类氧化的抑制率在浓度为8μg·mL~(-1)时达到54.29%;对DNA氧化损伤的抑制率在浓度为8μg·mL~(-1)时达到84.78%。本研究结果为进一步探究KR抗氧化作用及其机制,以及医疗、保健等领域的抗氧化药物研发奠定了基础。     

耐辐射奇球菌中辐射/干燥响应基序的研究进展

耐辐射奇球菌是迄今为止被发现抗逆性最强的极端生物之一,是研究DNA损伤应激响应与修复机制的较为理想模式生物.其中的辐射/干燥响应基序是一组相对保守的17-bp长的回文序列,广泛地分布在众多不同功能的DNA损伤响应基因中,与抗电离辐射、氧化损伤及干旱等有关.本文结合该领域以往的研究文献与最新研究,探讨了耐辐射奇球菌中辐射/干燥响应基序的研究进展,阐释了其所涉及的DNA损伤应激响应通路,并且进一步展望了其潜在的生物学功能、意义与应用.     

耐辐射异常球菌磷酸戊糖途径对DNA损伤修复的影响

耐辐射异常球菌(D.radiodurans)超强的辐照抗性归因于其高效的DNA损伤修复系统,磷酸戊糖途径(PPP)作为细胞各种糖类代谢枢纽,是合成核苷酸和核酸所需的核糖以及生成细胞还原力(NADPH)的重要途径。为探究D.radiodurans R1中PPP对DNA损伤修复的影响,本研究敲除PPP中编码限速酶葡萄糖-6-磷酸脱氢酶(G6PD)的基因zwf,比较UV辐照后突变株Δzwf与野生型D.radiodurans的生理生化差异。结果表明,zwf突变导致菌株对UV辐照敏感;UV辐照后Δzwf胞内NADPH、D-核糖、核苷酸合成前体IMP、UMP含量明显降低;葡萄糖酸-δ-内酯及D-核糖能完全和部分恢复Δzwf的UV辐射抗性。综上可知PPP通过提供DNA损伤修复原料及还原力,在DNA损伤修复过程中起着重要作用。本研究结果为DNA损伤修复机理提供参考。     

植物次生代谢响应UV-B辐射胁迫的生态学意义

平流层臭氧的减薄已导致地表中波紫外辐射(UV-B,280～320 nm)增强,使植物体内的次生代谢发生改变,而次生代谢产物与抗紫外辐射、抑制昆虫、防止病菌感染和其他食草动物取食、凋落物分解、他感作用等方面存在复杂联系,进而影响生态系统的种类组成、种间关系以及生物的多样性,并导致生态系统的生产力、物质循环、地球化学循环和能量流动等功能的改变,从而影响生态系统的平衡.本文综述了UV-B辐射增强对植物群落和生态系统的影响以及对次生代谢物影响的生态学意义,并展望了该领域的研究方向.     

耐辐射奇球菌Deinococcus radiodurans中非编码RNA

耐辐射奇球菌(Deinococcus Radiodurans)对电离辐射、紫外线以及强氧化剂等方面具有惊人的抗性。非编码RNA(non-coding RNA,ncRNA)在参与转录调控、RNA的加工与修饰、mRNA的转录、蛋白质的运输与稳定性调节等方面具有非常明显的作用。本文通过概率上下文无关算法(Stochastic ContextFree Grammar,SCFG),对耐辐射奇球菌R1菌株的基因间序列进行相似二级结构预测,结果发现,在耐辐射奇球菌R1基因组的非编码区存在28个非编码RNA家族。其中IRE家族成员最多,其次是Histone3、tRNA和SECIS家族。所发现的DicF与ctRNA_pGA1t、mRNA等家族成员为细菌所特有。与其他生物比较分析结果显示,可以用这些非编码RNA来研究耐辐射奇球菌的生物学功能,并为进一步研究其DNA损伤与修复分子机制提供依据。     

辐射诱变选育栀子新品系

为选育产量高、活性物质含量高、抗逆性强的栀子新品种,选取现有本地栀子种质资源中表现较好的类型的枝条,利用60Co γ射线进行辐射诱变处理,然后将优选的株系通过扦插繁殖、栽培,经过多年的稳定和筛选,选育出了具有树型适中、分枝匀称、冠形圆满、花果期集中、结实量多、果实大、果实中栀子苷与色素含量高、抗寒、适应性强等特点的新品系——天顺栀子。经急性毒性试验,结果表明该新品系与普通栀子一样安全无毒性。     

不同利福平浓度压力下耐辐射球菌的自发突变率与突变谱研究

根据耐辐射球菌(Deinococcus radiodurans) rpoB基因的保守性,建立了rpoB/Rifr 突变分析系统。为研究不同利福平(rifampin, Rif)浓度对耐辐射球菌自发突变率和突变谱的影响,分别检测耐不同利福平浓度下辐射球菌的突变率。结果表明：在5μg/ml的利福平浓度下耐辐射球菌的突变率显著高于其在25和50μg/ml浓度下的突变率。随利福平浓度的不同而表现出的突变谱差异,表明在不同利福平浓度引起的活性自由氧(reactive oxygen species, ROS)压力下,耐辐射球菌可能使用不同抗突变的应对策略。在低利福平浓度下,耐辐射球菌重点修复碱基替换突变导致的DNA损伤。这可能是因为ROS的主要作用是导致DNA出现氧化性碱基损伤。     

THE ISOLATION OF NUCLEOSIDE DIPHOSPHATES FROM ALKALINE EXTRACTS OF SOIL

Two pyrimidine nucleoside diphosphates have been isolated from sodium hydroxide extracts of soils. They were initially detected in fractions which had been separated by ion-exchange chromatography, and were purified by a combination of ion-exchange and paper chromatography. Their ultraviolet spectra indicated that one ester contained thymine and the other uracil, and this was confirmed by hydrolysis. The ratio of P to pyrimidine was 2, but their acid stability showed that the esters were not nucleoside-5’ pyrophosphates. Both gave a positive reaction to a colour test for deoxypentose and comparison with reference samples prepared from calf-thymus DNA showed that they were thymidine-3′: 5’ diphosphate and deoxyuridine-3′:5’ diphosphate. The nucleoside diphosphates may be artefacts, derived from polynucleotides by acid and alkaline hydrolysis, but their isolation confirms, for the first time, the existence in soil of characteristic structural units of DNA.     

紫外分光光度法测定农田土壤微生物生物量氮可行性初探

采用氯仿熏蒸浸提-紫外分光光度法和消化法比较测定了田间定位试验不同施肥处理土壤、添加植物残体土壤、添加葡萄糖土壤的微生物生物量碳、氮(SMBC,SMBN)。结果表明,当土壤微生物生物量氮含量较高时(>20 mg kg-1),采用分光光度法与消化法测定的SMBN具有显著正相关关系(P<0.05),但当SMBN量较低时(<20 mg kg-1)时,分光光度法测定与消化法测定的SMBN没有显著相关性。当土壤中添加麦秸和玉米秸时,土壤浸提液颜色较深(黄色),不适合采用分光光度法测定SMBN。因此,熏蒸提取–分光光度法测定SMBN,仅适于土壤浸提液无色透明、且SMBN含量较高的土壤。     

Confirmation of polybrominated biphenyl residues in feeds and dairy products, using an ultraviolet irradiation-gas-liquid chromatographic technique.

An ultraviolet (UV) irradiation-gas-liquid chromatographic (GLC) detection procedure was used to confirm the presence of polybrominated biphenyl (PBB) residues in sample extracts after GLC quantitation. Chromatograms of PBB standard and sample extract solutions showed similar photodecomposition peak patterns dependent on time and intensity of UV irradiation. Confirmation of PBBs in sample extracts by this procedure was possible for any amount detectable by the GLC system employed. Prolonged UV irradiation resulted in complete disappearance of all PBB peaks from the chromatogram, permitting their distinction from background peaks due to extraneous sample material unaffected by UV irradiation.     

土壤溶解性有机碳四种测定方法的对比和转换

针对土壤溶解性有机碳(DOC)不同测定方法之间可比性较差的问题,应用TOC仪法、容量法、紫外分光光度法和比色法分别测定了中亚热带丘陵山地6个土属共46个新鲜土壤样品的DOC含量。结果表明:参照TOC仪法测得的DOC数据,容量法一致性地低估20%~67%;比色法测定DOC含量较低的底土时(200 mg/kg)仅低估7%~27%,但在分析DOC含量较高的表土(600 mg/kg)时最不敏感,低估达53%~93%;紫外分光光度法在DOC含量较高时也存在一定的低估,但在DOC含量较低时高估65%~189%。4种方法测得的DOC数据均呈极显著正相关关系(P0.01)。统计分析获得的线性或指数方程可应用于将容量法、紫外分光光度法和比色法的测定结果向TOC仪法的数据转换,而且有必要针对表土和底土使用不同的转换方程或参数,这些经验转换方程的建立有助于增强不同研究结果之间DOC数据的可比性,也有利于推动土壤DOC测定标准的完善。     

Champagne wine polyphenols protect primary cortical neurons against peroxynitrite-induced injury

White wines are generally low in polyphenol content as compared to red wines. However, Champagne wines have been shown to contain relatively high amounts of phenolic acids that may exert protective cellular actions in vivo. In this study, we have investigated the potential neuroprotective effects of Champagne wine extracts, and individual phenolics present in these extracts, against peroxynitrite-induced injury. Organic and aqueous Champagne wine extracts exhibited potent neuroprotective activity against peroxynitrite-induced injury at low concentrations (0.1 microg/mL). This protection appeared to be in part due to the cellular actions of individual components found in the organic extracts, notably tyrosol, caffeic acid, and gallic acid. These phenolics were observed to exert potent neuroprotection at concentrations between 0.1 and 10 microM. Together, these data suggest that polyphenols present in Champagne wine may induce a neuroprotective effect against oxidative neuronal injury.     

Changes in hot water soil extracts brought about by nitrogen immobilization and mineralization processes during incubation of amended soils

Summary During incubation of an acid cambisol and an alkaline fluvisol, amended with glucose and nitrate, hot water soil extracts were analysed for N content, ultraviolet absorption, and fluorescence. Humic substances in the hot water extracts and in a neutral sodium pyrophosphate extract were fractionated on polyvinylpyrrolidone and measured spectroscopically. Changes in the hot water and pyrophosphate extract compositions were related to changes in microbial biomass, as estimated by substrate-induced respiration, and the hexosamine content of soil hydrolysates. During the incubation, the microbial population in each type of soil developed quite differently, according to the soil pH. Microbial growth and death in the alkaline soil sample induced a maximum of hot-water-extractable ultraviolet-absorptive non-fluorescent substances. The fluorescence of the hot water soil extract increased steadily with incubation time even after the microbial activity was reduced. A similar increase in fluorescence, in accord with the ultraviolet absorption, was found during incubation of the acid soil samples. After 95 days of incubation, the hot-water-extractable fluorescent and ultraviolet-absorptive substances were reduced. N immobilization induced an increase, and N mineralization a decrease, in dissolved organic N. The relative increase in humic substances in the hot water soil extract was much higher than in the pyrophosphate extract. Therefore, humic material, produced by microbial growth and death, is obviously extractable with hot water.     

THE PERCOLATION OF RAINWATER THROUGH SOIL DEMONSTRATED BY FLUORESCENT DYES

Twelve fluorescent dyes were tested in the laboratory for their stability under conditions of alternate wetting and drying, and persistence of fluorescence in the presence of an acid soil. The strongly fluorescing Pyranine cone, was most suitable for tracing water percolating through soils. In the field this dye could be detected in the soil after several months and couldbe photographed in ultraviolet light. The greater percolation of water against tree trunks and lateral movement in the humus layer were demonstrated.     

THE MOBILIZATION OF IRON BY EXTRACTS OF EUCALYPTUS LEAF LITTER

When aqueous extracts of leaf litter from four closely related Eucalyptus species were reacted with soil material under aerobic conditions their iron mobilizing activity was found to be inversely related to the productivity of the sites on which the species grew. The activity of litter extracts of the four species grown in similar soils in the field was found to increase in the order E. regnans, E. obliqua, E. radiata, E. sieberiana. The results indicate that species-soil interactions could be as important as inherent species characteristics in determining whether or not a species is a ‘podzol former’. The effect on the activity of the extracts of altering their pH before reaction with either soil or with prepared iron oxides suggests that, whereas organic acids could be mainly responsible for mobilizing iron from soil and from anhydrous ferric oxide, polyphenols could be more important than organic acids in mobilizing iron from hydrous ferric oxide. The pH and E_h curves obtained when litter extracts were titrated with a ferric chloride solution showed that iron from this source was strongly reduced by the extracts at low pH.     

星形孢菌素产生菌的选育研究

用紫外线和微波复合诱变星形孢菌素产生菌(Stretomyces sp.4138),以硫酸链霉素抗性偶联进行筛选。结果表明:紫外线处理50s后,在含硫酸链霉素0.8μg/ml的分离培养基上筛选到突变株HZ-09,比原始菌株H-0效价提高了30.46%;再以HZ-09为出发菌株进行微波处理30s后,硫酸链霉素抗性筛选获得突变株HW-25,比原始菌株H-0效价提高了284.20%。菌株HW-25经多次传代,遗传性状稳定。     

辐射对果树花粉发芽率及远缘杂交亲和性的影响

本文研究了不同剂量的60Coγ射线和紫外线辐照对帚型山桃、凯特杏、黑宝石李3种果树花粉后其发芽率及远缘杂交坐果率的影响。结果表明,经60Coγ射线和紫外线照射后,3种果树花粉的发芽率与对照相比都明显降低。3种果树花粉的辐射敏感性有明显差异,其中黑宝石李花粉最敏感,花粉发芽率降低最明显;帚型山桃次之;凯特杏的花粉最不敏感。辐照后的低花粉发芽率降低了远缘杂交的坐果率。