肉果秤锤树叶提取物的抑菌作用研究

[目的]探讨秤锤树叶的逐级提取物的抑菌作用。[方法]采用试管稀释法测定秤锤树叶的各级提取物一总浸液、正丁醇萃取液、乙酸乙酯提取液对各试验菌的最小抑菌浓度(MIC)。[结果]各级提取物对试验菌分别有不同程度的抑菌作用,其中对金黄色葡萄球菌的抑菌作用最为明显;各提取物的抑菌强度依次为:乙酸乙酯萃取液(对4种实验菌均明显抑制:MIC≤31 mg/ml)、正丁醇萃取液(金黄色葡萄球菌:MIC≤62 mg/ml、其余3种菌的MIC相似:MIC≤125)、总浸液(仅对金黄色葡萄球菌抑制明显:MIC=125 mg/ml)。[结论]秤锤树叶的乙酸乙酯提取物具有广谱抗茵作用,其逐级提取物富集的抑茵活性逐级增强。     

Molybdenum availability, nitrogen limitation, and phytoplankton growth in natural waters

Sulfate inhibits molybdate assimilation by phytoplankton, making molybdate less available in seawater than it is in freshwater. As a result, nitrogen fixation and nitrate assimilation, both processes that require molybdenum, may require a greater expenditure of energy in seawater than in freshwater. This may explain in part why coastal marine ecosystems are usually nitrogen limited whereas lakes usually are not. Experimentally increasing the ratio of sulfate to molybdate (i) inhibits molybdate uptake, (ii) slows nitrogen fixation rates, and (iii) slows the growth of organisms that use nitrate as their nitrogen source.     

Dimethyl sulfoxide protects tightly coupled mitochondria from freezing damage

Dimethyl sulfoxide prevented loss of respiratory control and decrease in efficiency of oxidative phosphorylation when plant mitochondria were stored in liquid nitrogen. Respiration was severely inhibited and was not stimulated by adenosine diphosphate when mitochondria were frozen in liquid nitrogen without dimethyl sulfoxide. Thus, isolated mitochondria provide a model system for the study of the effects of freezing on biological membranes and of the prevention, by dimethyl sulfoxide, of freezing damage.     

Nitrogen fixation by a blue-green epiphyte on pelagic sargassum

Nitrogen fixation by Dichothrix fucicola, an epiphyte on pelagic Sargassum, was measured in May and June 1972 in the western Sargasso Sea and the Gulf Stream. This is the first report of nitrogen fixation by a heterocyst-bearing blue-green alga in the open ocean, and also the first observation of nitrogen fixation in the genus Dichothrix. Cellular carbon/nitrogen ratios suggested that the Dichothrix was nitrogen-starved. In dense aggregations of Sargassum, such as rafts or windrows, the enrichment of surface seawater with combined nitrogen from nitrogen fixation may be pronounced.     

Adenosine 3',5'-monophosphate, adrenocorticotropic hormone, and adrenocortical cytosol protein synthesis

In cultures of mouse adrenocortical tumor cells (Sato's minimal deviation Y-1 clonal strain), the acceleration of steroid biosynthesis after exposure to adrenocorticotropic hormone or cyclic adenosine 3',5'-monophosphate is maximum within 15 to 60 minutes and precedes any significant increase in labeling of protein with [4,5-(3)H]leucine. However, when cytosol proteins are separated by acrylamidegel electrophoresis, rapid changes in the amount and labeling of several protein fractions are evident in less than 30 minutes and are no longer evident within 60 minutes. This finding supports the proposal that the effects of tropic hormones and their intracellular mediators involve rapid selective effects on protein synthesis.     

固氮施氏假单胞菌电子传递复合体rnf基因簇的功能鉴定

能量供应是限制生物固氮效率的重要因素,电子传递复合体是生物固氮过程中能量产生的重要组成部分。基因组分析表明,在联合固氮菌施氏假单胞菌A1501基因组中存在两套编码电子传递复合体rnf,分别为基因簇rnf1和rnf2,其中rnf1位于固氮基因岛上,rnf2基因簇位于核心基因组上。为了明确两套电子传递体在生物固氮过程中的功能,分别构建了rnf1和rnf2基因簇的突变株并测定了相关表型。结果发现,在基本培养基中两个基因簇的突变都不影响菌体生长,可能相互之间存在着功能互补;固氮酶活性测定结果表明,位于固氮基因岛中的rnf1基因簇缺失造成固氮酶活下降80%以上,而rnf2基因簇的缺失对酶活无显著影响。进一步采用启动子-lacZ融合表达策略探究了基因簇rnf1对固氮酶结构基因nifH转录的影响,发现rnf1极性突变株中nifH的启动子转录活性仅为野生型的17.59%,推测rnf1缺失造成了固氮条件下能量产生匮乏,胞内碳氮比失衡,进而造成固氮系统表达受抑制。     

Nitrogen fixation by floating diatom mats: a source of new nitrogen to oligotrophic ocean waters

Nitrogen fixation, apparently by bacterial endosymbionts, is associated with intertwining chains of two species of the diatom Rhizosolenia. In situ fixation rates were enhanced by incubation in the dark, whereas concurrent shipboard experiments either underestimated or did not detect nitrogen fixation. This is the first example of nitrogen fixation associated with a bacteria-diatom symbiosis in the pelagic zone, and it indicates that these systems may contribute a significant amount of "new" nitrogen to oligotrophic waters.     

生物固氮基因簇结构与进化研究进展

在固氮菌基因组中,固氮相关基因都是作为一个或几个基因簇存在,是细菌生物固氮的遗传基础。这其中包括固氮酶结构基因nifHDK,以及参与电子转运和铁钼辅因子合成及组装的其他相关基因。对不同菌中固氮基因簇的结构和进化进行了简要的比较分析。比较所有已测序固氮菌中的固氮基因簇,发现均至少含有六个保守基因:nifH、nifD、nifK、nifE、nifN和nifB,并且这6个基因在所有已鉴定的系统中都是固氮所必需的。尽管在不同种类的固氮微生物中,固氮基因簇的构成及组织模式有所不同,但是它们在结构、催化机制和系统进化上紧密相关。同时也对固氮基因的研究方向提出了展望。旨在为今后进一步研究生物固氮机制,了解固氮微生物的进化,扩展生物固氮多样性提供理论基础。     

茶叶对烟气脱硫废水的处理研究

[目的]研究利用茶叶处理烟气脱硫废水,为烟气脱硫废水的处理研究提供参考。[方法]对不同品种的茶叶在不同的浸泡次数、浸泡温度及浸泡时间下茶汤中茶多酚的含量进行测定,分析不同条件下茶汤中茶多酚含量的变化。利用茶汤与烟气脱硫废水进行络合反应,得出两者反应完全时的体积比,确定反应的最佳条件。利用ICP-AES法测定烟气脱硫废水及反应沉淀上清液中金属的含量。[结果]结果表明,浸泡水温对茶汤中茶多酚含量影响较大。茶汤处理烟气脱硫废水的最佳条件是:绿茶在75℃的浸泡水温下浸泡1次20m in,此时,所用茶汤量最少,沉淀上清液的金属含量也较少。[结论]找到了较好的处理烟气脱硫废水的方法。     

红豆草根瘤菌的耐盐和共生性状改造

本实验以042B 的 DNA 转化 ROG31细胞,在以0.2 mol/L NaCl 为选择标记的YMA 培养基上,获得37个耐盐的菌落,纯化后又经30代的转管移植,有8株转化子获得了稳定的耐盐性,再将这8株转化子回接红豆草植株,得到1株耐0.4 mol/L NaCl 和固氮能力较好的红豆草根瘤菌 ROt6。供体菌042B 和转化子 ROt1和 ROt6在0.3 mol/L NaCl 培养基中生长时,细胞内积累大量谷氨酸。     

一株色季拉山长鞭红景天根际溶磷菌的分离、鉴定及其低温适应性分析

[目的]从西藏色季拉山长鞭红景天根际土壤中分离筛选出溶磷菌株,为高寒植物根际促生菌的开发利用和丰富溶磷菌种质资源提供理论基础和科学依据.[方法]以长鞭红景天为研究物种,采用溶磷圈法从其根际土壤中分离溶磷菌,以钼锑抗比色法测定溶磷量进行复筛;筛选出溶磷效果最好的菌株,测定其产吲哚乙酸(IAA)、固氮促生特性,结合形态、生理生化特性和16S rRNA序列分析进行分类鉴定.以芽孢杆菌为对照,将芽孢杆菌和分离获得的溶磷菌株分别在4、10、15、20和30 ℃下培养7 d,研究其低温适应性.[结果]从长鞭红景天根际土壤中筛选出21株具有溶磷作用的菌株,其中PSB19菌株在PKO无机磷培养基上的溶磷圈直径/菌落直径(D/d)最高,液体溶磷量达392.15 μg/mL,具有产IAA和固氮特性,其在4和10℃的低温条件下生长良好.通过形态、生理生化鉴定和16S rDNA序列分析,将PSB19菌株归类于粘着箭菌(Ensifer adhaerens).[结论]PSB19菌株有较好的溶磷、产IAA和固氮特性及低温适应性,具有作为生物菌肥在农业生产及植被恢复等方面应用的潜力.     

施用根瘤菌对土壤微生物氮素类群数量及土壤氮素的影响

为研究施用根瘤菌条件下,大豆不同生育时期土壤微生物数量及土壤氮素含量动态变化,于大豆苗期、花期、结荚期、成熟期分别对大豆根际土壤氨化细菌、硝化细菌、自身固氮菌数量及土壤碱解氮、全氮含量进行测定。结果表明:施用根瘤菌处理与对照相比,在大豆的不同生育时期土壤氨化细菌、硝化细菌、自身固氮菌数量上存在差异,但在大豆的整个生育期根瘤菌有效增加了微生物氮素类群数量。除成熟期土壤中碱解氮含量根瘤菌处理低于对照,各时期土壤中碱解氮含量根瘤菌处理均高于对照,施用根瘤菌在大豆整个生育期增加了土壤全氮的含量。     

液氮超低温在赤霉素产生菌菌种保藏中的应用

目的:探索液氮超低温在赤霉素产生菌菌种保藏中的应用。方法:用15%的甘油制成的菌丝片段悬浮液经匀速降温预冷后,置于液氮所产生的-196℃的超低温环境中保藏,一定保藏周期后取出液氮管做菌株形态观察及上摇床进行效价测定,检验保藏效果。结果:赤霉素产生菌菌种在液氮超低温保藏条件下,其生物活性、生产能力基本无变化。结论:液氮超低温法适用于赤霉素产生菌菌种保藏,效果优于传统的石蜡油保藏法。     

Enhancement of symbiotic dinitrogen fixation by a toxin-releasing plant pathogen

An approximate doubling in plant growth, total plant nitrogen, nodulation, and overall dinitrogen fixation of alfalfa are the consequences of the action of a toxin delivered by a Pseudomonas infesting the alfalfa rhizosphere. The toxin, tabtoxinine-beta-lactam, inactivates selectively one form of glutamine synthetase in the nodules. Thus, normal glutamine synthetase-catalyzed ammonia assimilation is significantly impaired; yet these plants assimilated about twice the normal amount of nitrogen. How plants regulate dinitrogen fixing symbiotic associations is an important and unresolved question; the current results imply that the glutamine synthetase-catalyzed step in ammonia assimilation, a plant function, strongly influences overall dinitrogen fixation in legumes.     

盆栽条件下土壤无机氮浓度对大豆结瘤、固氮和产量的影响

【目的】大豆结瘤固氮和产量对氮肥的反应不同,实际上是由于大豆共生固氮系统及其根系系统对土壤无机氮浓度的感知不同造成的。通过对不同土壤无机氮浓度下大豆结瘤、固氮及产量影响的研究,探索能提高大豆产量、结瘤和固氮的土壤无机氮浓度,即掌握土壤无机氮浓度与大豆共生固氮和产量的数量关系,为调控氮肥施用量及施用时期、预测氮肥对大豆共生固氮能力和产量的影响提供理论依据。【方法】采用盆栽土培试验方法,分别在第一片复叶充分生长（V2期）、始花期（R1期）、始荚期（R3期）和始粒粒（R5期）一次性施用不同量的氮肥,从而形成不同无机氮浓度的土壤。利用获得的不同无机氮浓度土壤为供试土壤,对各生育时期根瘤数量、干重和固氮酶活性及成熟期产量及其构成因子进行调查,明确大豆根瘤固氮和产量对土壤无机氮浓度的响应,掌握土壤无机氮浓度与氮肥及与大豆固氮和产量的数量关系。【结果】不同时期土壤无机氮浓度处理下的根瘤干重、数量和固氮酶活性均随着大豆生育时期的推进在R4期时达到最大值。R6期时大豆平均根瘤干重、数量和固氮酶活性均表现为：V2期＞R5期＞R3期＞R1期,较CK处理根瘤平均干重分别下降15%、18%、17%和32%;根瘤数量下降13%、18%、19%和20%;固氮酶活性下降19%、22%、23%和32%。不同生育时期土壤无机氮浓度与R6期大豆根瘤干重、数量和固氮酶活性间均具有显著的线性负相关关系,即土壤无机氮浓度越大对根瘤干重、数量和固氮酶活性的抑制作用越大。大豆干物质积累量和产量的变化趋势均表现为：R1期＞R3期＞V2期＞R5期。除R5期不同土壤无机氮浓度处理与CK处理间的生物量和产量差异不显著外,V2、R1和R3期不同土壤无机氮浓度处理,均显著地促进大豆生物量和产量的增加。不同生育时期处理均以N3和N4处理的生物量、株高、株荚数、株荚重、株粒重显著高于其它处理。V2期土壤无机氮浓度对大豆固氮能力和产量的影响最大,而R1期土壤无机氮浓度对大豆生长和产量的影响最大。不同生育时期不抑制大豆固氮同时还提高大豆产量的土壤无机氮浓度不同：V2期土壤无机氮浓度达到135.8 mg•kg -1;R1期土壤无机氮浓度为58-91 mg•kg-1;R3期土壤无机氮浓度为29.4-62.8 mg•kg -1;在R5期土壤无机氮浓度达到102.3 mg•kg -1。【结论】大豆对氮肥的反应主要取决于土壤无机氮浓度的大小,而土壤无机氮浓度大小的调节,除了与氮肥施用量有关外还与大豆的生育时期有关系,可以根据农业生产和科学试验的需要进行调节。其中V2期土壤无机氮浓度对大豆根瘤数量、干重和固氮酶活性的影响大于其它生育时期土壤无机氮浓度处理;而R1期土壤无机氮浓度对大豆生物量和产量的影响大于其它生育时期土壤无机氮浓度处理。     

化学固氮的研究

本文讨论了在温和条件下从空气进行化学固氮的问题。在比较了生物固氮和化学固氮的基础上,认识到其间的一些共同特点,这就是为使氮分子活化必须要有过渡金属。随之讨论了有关这方面的机理。为了模拟生物固氮,对固氮酶进行了一系列的研究。本文试就固氮催化剂的化学模型进一步提出一些看法。     

Role of chemotaxis in establishing a specific nitrogen-fixing cyanobacterial-bacterial association

A specific association with the cyanobacterium Anabaena oscillarioides was established by positive bacterial (pseudomonad) chemotaxis to Anabaena oscillarioides heterocysts. This association enhanced nitrogen fixation in A. oscillarioides, and positive chemotaxis was particularly strong during periods of active nitrogen fixation. Addition of compounds known to elicit positive chemotaxis in pseudomonads interfered with the establishment of the association, while removal of these compounds led to reestablishment of the association. Anabaena oscillarioides excretion products, some of which are exuded from heterocyst-vegetative cell junctions, are likely to be responsible for positive chemotactic responses. Chemotaxis-controlled associations such as this one explain in part why aquatic bacterial-algal and bacterial-particle associations occur sporadically and are heterogeneously distributed in time and space.     

Quantitative imaging of nitrogen fixation by individual bacteria within animal cells

Biological nitrogen fixation, the conversion of atmospheric nitrogen to ammonia for biosynthesis, is exclusively performed by a few bacteria and archaea. Despite the essential importance of biological nitrogen fixation, it has been impossible to quantify the incorporation of nitrogen by individual bacteria or to map the fate of fixed nitrogen in host cells. In this study, with multi-isotope imaging mass spectrometry we directly imaged and measured nitrogen fixation by individual bacteria within eukaryotic host cells and demonstrated that fixed nitrogen is used for host metabolism. This approach introduces a powerful way to study microbes and global nutrient cycles.     

Suppression of nitrogen fixation by blue-green algae in a eutrophic lake with trace additions of copper

Nitrogen fixation by blue-green algae in highly eutrophic Clear Lake, California, was severely inhibited by trace amounts of copper. The chelation capacity of the lake is probably saturated by indigenous copper. Additions were only 1/200 of those normally used in algal control. Since nitrogen fixation provides half of the lake's annual nitrogen budget, economical eutrophication control appears possible.     

Cryopreservation, culture, and transplantation of human fetal mesencephalic tissue into monkeys

Studies in animals suggest that fetal neural grafts might restore lost neurological function in Parkinson's disease. In monkeys, such grafts survive for many months and reverse signs of parkinsonism, without attendant graft rejection. The successful and reliable application of a similar transplantation procedure to human patients, however, will require neural tissue obtained from human fetal cadavers, with demonstrated cellular identity, viability, and biological safety. In this report, human fetal neural tissue was successfully grafted into the brains of monkeys. Neural tissue was collected from human fetal cadavers after 9 to 12 weeks of gestation and cryopreserved in liquid nitrogen. Viability after up to 2 months of storage was demonstrated by cell culture and by transplantation into monkeys. Cryopreservation and storage of human fetal neural tissue would allow formation of a tissue bank. The stored cells could then be specifically tested to assure their cellular identity, viability, and bacteriological and virological safety before clinical use. The capacity to collect and maintain viable human fetal neural tissue would also facilitate research efforts to understand the development and function of the human brain and provide opportunities to study neurological diseases.