高效解磷细菌的筛选及其对玉米苗期生长的促进作用

采用改良后的PVK平板,从石灰性土壤上长势良好的野生植物根表分离到44株解磷细菌,通过NBRIP液体摇瓶实验,培养7 d后发现：K₃菌株培养液中全磷浓度高达643.2 μg·ml^-1,可溶性磷为584.8 μg·ml^-1,约有12.9%的磷酸三钙被溶解出来,为对照(CK)的10.5倍;K₉菌株培养液的全磷浓度为608.5 μg·ml^-1,可溶性磷浓度为606.4 μg·ml^-1.盆栽试验的结果表明：接种解磷细菌的处理玉米株高、茎粗和干质量显著高于CK;将有机肥作为载体和解磷细菌一同混合施入土壤的处理,玉米苗干质量较单施解磷菌显著增加.经初步鉴定, K₃、K₉为假单胞菌属.     

玫烟色拟青霉适温性及抗多菌灵特性的种内变异

在15 ℃～35 ℃的温度范围内比较测定了玫烟色拟青霉(Paecilomyces fumosoroseus) 6株野生菌的菌落生长、产孢量及孢子活力.结果显示,3个指标均以25 ℃左右最好,但同一温度下菌株间或同一菌株在不同温度下的差异显著, 菌株Pfr116和Pfr6206具有相对稳定优良的生长、产孢及活孢率性状.对不同浓度多菌灵对6株野生菌的菌落生长和单孢菌落形成的影响测定表明,菌株间存在较大差异.不同多菌灵浓度对单孢菌落形成的抑制率观察值可用逻辑斯蒂模型较好地拟合(r²≥0.90).用所获参数估计的最低禁菌浓度MIC值显示,Pfr4205和Pfr116对多菌灵表现为低抗性（MIC≤20.0 μg·ml^-1）;Pfr153、Pfr612和Pfr2175属于低水平中抗,MIC值仅略大于20 μg·ml^-1;而Pfr6206的MIC值高达93.5 μg·ml^-1,接近MIC>100 μg·ml^-1的高抗标准.因此,Pfr6206为适温性和抗多菌灵特性俱佳的菌株,可作为抗多菌灵和适应不同季节或地区的害虫生防菌剂的候选菌株.     

青岛近海及其临近海域冬季微微型浮游植物的分布

微微型浮游植物（0.2～2.0 μm） 是海水中最小的自养浮游生物, 在世界各海域广泛分布, 并在海洋有机物质循环中起着非常重要的作用.利用荧光显微技术对青岛近海及其邻近海域冬季微微型浮游植物丰度进行了调查,研究了微微型浮游植物的空间变化和昼夜变化的特征, 并分析了微微型浮游植物丰度与环境因子的相关性. 结果显示, 冬季该海域以富含藻红素（Phycoerythrin-rich, PE）的聚球藻(Synechococcus, Syn)细胞占优势,微微型真核藻类（Picoeukaryote, Euk）次之,而富含藻蓝素（Phycocyanin-rich, PC）的聚球藻细胞数量很低, 未发现原绿球藻（Prochlorococcus, Pro）的存在.Syn 的变化范围为8.97×10³～1.95×10⁵ cells·ml^-1, 平均4.67×10⁴ cells·ml^-1; Euk的变化范围为1.95×10²～1.01×10⁴ cells·ml^-1, 平均2.39×10³ cells·ml^-1. Syn丰度在胶南以南海域出现高值区域, 在即墨海域和崂山东南海域出现低值区域. Euk丰度在日照海域出现高值区域; 崂山海域为低值区域; 各水层Syn和Euk丰度均无明显差异(P﹥0.05). 对胶州湾中部连续站4层水体的24 h昼夜连续变化进行观测发现, Syn、Euk丰度都有明显昼夜波动.相关性分析表明： Syn与温度、 电导率呈正相关, 与溶氧浓度呈显著负相关; Euk与盐度和溶氧浓度呈显著负相关. 微微型浮游植物对总浮游植物生物量的贡献约为20％.     

苦瓜叶提取物对美洲斑潜蝇取食和产卵行为的抑制作用

美洲斑潜蝇是危害蔬菜、观赏植物的重大害虫之一.苦瓜叶乙醇提取物(浓度为2000～4000 μg·ml^-1)对美洲斑潜蝇成虫的取食和产卵都具有较强的抑制作用.用环己烷、乙酸乙酯、正丁醇和水依次对乙醇提取物进行萃取,并测试了4种萃取物对美洲斑潜蝇成虫取食和产卵的抑制作用.结果表明: 环己烷、乙酸乙酯、正丁醇和水萃取物在浓度为1000 μg·ml^-1时,处理后2 d对美洲斑潜蝇成虫的拒食率分别是11.08%、34.89%、22.99%和 0,产卵抑制率分别是0、30.91%、6.45%和 0.其中,乙酸乙酯萃取物的活性最强,当其浓度为4000 μg·ml^-1时,处理后2 d对美洲斑潜蝇成虫的拒食率和产卵忌避率分别为70.95％ 和69.49％.乙酸乙酯萃取物经硅胶柱层析分离得到(19S,23E)-5β,19-环氧-19-甲氧葫芦素-6,23-二烯-3β,25-二醇 (化合物1)、(19R,23E)-5β,19-环氧-19-甲氧葫芦素-6,23-二烯-3β,25-二醇(化合物2) 和3β,7β,25-三羟基葫芦素-5,23-二烯-19-醛缩-3-O-β-D-吡喃葡糖苷（化合物3）,3种化合物在供试的浓度（100～400 μg·ml^-1）条件下对美洲斑潜蝇的取食和产卵行为都有明显的抑制作用.在400μg·ml^-1浓度时,化合物1、化合物2和化合物3对美洲斑潜蝇成虫的拒食率分别是66.89%、53.53%和78.02%,产卵抑制率分别是76.32%、58.36%和78.36%.     

青海盐湖嗜盐微生物类群及F16菌株生长特性和抗菌、抗肿瘤活性

从青海盐湖底泥中分离出45株嗜盐微生物.其中,丝状真菌F16的抗菌和抗肿瘤活性最强,其发酵液的乙酸乙酯粗提物能抑制4种细菌的生长,尤其对大肠杆菌的抑制作用最强,具有较强的细胞毒活性,当粗提物浓度为50 μg·ml^-1时对肝癌细胞BEL7402的抑制率可达76.91％.F16菌株的最适生长温度为15 ℃;培养基盐度升高,对F16的抑制性增强,当盐度超过15％时,F16不生长;在pH值为5～9范围内,F16生长良好.     

不同品系萼花臂尾轮虫休眠卵的形态特征

对采自青岛和芜湖两地的萼花臂尾轮虫在3种温度(20 ℃、25 ℃和30 ℃)和2种藻类食物浓度(1.0×10⁶和5.0×10⁶ cells·ml^-1)下所产休眠卵的长径、短径和体积等形态特征进行了显微测量、计算和分析.结果表明,2种食物浓度下,培养温度以及培养温度和品系间的交互作用均对轮虫休眠卵的长径、短径和体积具有显著影响.当食物浓度分别为1.0×10⁶和5.0×10⁶ cells·ml^-1时,轮虫在20 ℃下所产休眠卵的长径、短径和体积均最大;在25 ℃和30 ℃下所产休眠卵的短径和体积均最小.品系对轮虫休眠卵长径、短径和体积的影响也取决于食物浓度.当食物浓度为1.0×10⁶ cells·ml^-1时,芜湖品系轮虫的休眠卵长径、短径和体积(156.00 μm、99.95 μm和12 269.11 μm³)均显著大于青岛品系轮虫的休眠卵(145.13 μm、91.97 μm和10 498.19 μm³);而当食物浓度为5.0×10⁶ cells·ml^-1时,芜湖品系轮虫的休眠卵长径、短径和体积(155.68 μm、100.85 μm和12 348.59 μm³)均与青岛品系轮虫的休眠卵(156.63 μm、98.04 μm和12 054.20 μm³)之间无显著差异.两品系中,仅芜湖品系轮虫休眠卵的长径、短径和体积分别与温度呈曲线相关.同一温度下,两品系轮虫的休眠卵体积均随着食物浓度升高而增大;但30 ℃下芜湖品系轮虫所产休眠卵体积却随着食物浓度的升高而减小.     

小麦内生细菌及其对根茎部主要病原真菌的抑制作用

对小麦植株不同生育期、不同器官的内生细菌进行了分离和数量变化分析.结果表明,根、茎、叶及未成熟籽粒等器官中存在大量的内生细菌,鲜组织中平均约含内生细菌5.0×10⁵ CFU·g^-1,其中根系中内生细菌数量达7.8×10⁵ CFU·g^-1,而茎秆、叶片和未成熟籽粒中内生细菌数量分别为4.8×10⁵、3.2×10⁵和2.8×10⁵ CFU·g^-1.内生细菌数量在不同生育期也存在差异,幼苗期平均约为3.1×10⁵ CFU·g^-1、拔节期和灌浆期分别为5.7×10⁵和7.0×10⁵ CFU·g^-1.不同小麦田块之间存在明显差异,长武县一田块植物鲜组织中内生细菌的数量为6.1×10⁵ CFU·g^-1,而大荔县一田块约为3.9×10⁵ CFU·g^-1.试验结果发现,对小麦全蚀病菌具有拮抗作用的内生细菌有51株、对小麦纹枯病菌具有抑制作用的内生细菌有45株.用平板对峙法测定,有71株对两种病原真菌均有拮抗作用,对小麦全蚀病菌抑菌圈直径大于10 mm的有23株,其中来源于根系、茎秆、叶片和籽粒的分别为6株、7株、9株和1株;对小麦纹枯病菌抑菌圈超过10 mm的有20株,其中来源于根系、茎秆、叶片和籽粒的分别为7株、5株、7株和1株,说明从小麦叶片诱捕分离的内生细菌中,对小麦全蚀病菌和纹枯病菌抑菌作用较强的分离株比率最高,其次为茎秆,而根部和未成熟籽粒中比例明显较低.     

刺参对浅海筏式贝类养殖系统的修复潜力

浅海筏式养殖滤食性贝类产生大量的粪便和假粪（总称生物沉积物）,对海水养殖环境产生一系列影响;而沉积食性海参能够有效清除颗粒有机物,在海水养殖系统中扮演“清道夫”的生态角色.为评估刺参在浅海筏式贝类养殖系统中的生物修复潜力,本文在不同季节现场研究了贝 参混养模式下刺参对贝类生物沉积物的摄食及生长和排泄特征.结果表明: 刺参能够在新设计的养殖设施中与滤食性贝类混养,最大生长率达0.34%·d^-1; 并可通过摄食有效清除贝类生物沉积物, 摄食率为0.1746 g·g^-1·d^-1（夏季,21.2 ℃）、0.0989 g·g^-1·d^-1（秋季,19.2 ℃）和0.0050 g·g^-1·d^-1（冬季,7.7 ℃）;刺参主要通过排泄溶解形态的NH₄⁺N和PO₄^3- -P来促进沉积物中营养盐的再生,其排泄率也呈现明显的季节变化.基于现场试验数据,估算了刺参在桑沟湾的生物修复潜力, 刺参与贝类混养可摄食4.5～159.6 kg·hm^-2·d^-1生物沉积物、排泄1 382.5～3 678.1 mmol·hm^-2·d^-1NH₄⁺ -N及74.6～335.7 mmol·hm^-2·d^-1PO₄^3--P.表明刺参对浅海筏式贝类养殖系统具有较大的生物修复潜力,贝-参混养模式不仅能够取得较大的生态效益,而且能显著增加养殖生产的经济效益.     

质粒使矿井细菌具有对银离子的抗性

<正> 较早时候发现的银离子抗性细菌与使用含银化合物治疗烧伤病人有密切关系,这种菌还存在于照相胶片再处理工厂的废弃物中。现在,Biogen(瑞士日内瓦)的一个小组在美国犹他州的银矿土壤中发现了抗银离子的微生物。施氏假单孢杆菌银离子抗性菌株的有关基因位于该菌的一个质粒上(该菌有三种质粒)。这种抗性看来是银抗性微生物的一种天然进化现象。人们还不知道这个质粒如何使细菌对银离子产生抗性。这个质粒上的基因可以产生某种化合物,能与银离子形成一种惰性复合物。生产的化合物可能是金属硫因的类似物。已知     

不同食细菌线虫取食密度下线虫对细菌数量、活性及土壤氮素矿化的影响

采用悉生培养微缩体系,探讨了不同食细菌线虫取食密度下线虫(Caenorhabditis elegans) 对细菌(Bacillus subtilis)数量和活性及土壤氮素矿化的影响.结果表明,线虫对细菌的取食,促进了细菌的增殖,并在不同线虫取食密度下对细菌的增殖促进作用总体表现为:接种20条·g^-1＞10条·g^-1＞40条线虫·g^-1处理.线虫在促进细菌增殖的同时,明显提高了土壤呼吸强度和土壤蔗糖酶、脲酶和磷酸酶的活性,但不同取食密度处理间差异不明显.线虫与细菌之间的相互作用显著提高了土壤铵态氮和矿质态氮含量,促进了土壤氮的矿化.不同取食密度处理间,线虫对土壤氮素矿化的促进作用与对细菌的增殖促进作用趋势一致.     

Effect of metal microelements on biochemical indices of probiotic bacteria

The influence of metal-microelements (Zn, Cu, Ag, Au) in colloid and ionic form on the main biochemical parameters (ATP-ase activity, transmembrane potential and respiratory activity) of E. coli G35 N#1-413 and Ent. faecalis G35 N#4-410 probiont-strains has been studied with the goal to create complex metal-bearing probiotic preparations. Monotonous dose-dependent inhibitory influence of all mentioned metals in ionic form on the bacteria functional activity has been established. Metals' colloids had certainly stimulating influence on microorganisms' biochemical parameters in the same concentration limits with the following maximum positive effects: for Zn - 2x10(-7)-2x10(-4) mg/ml, for Cu - 8.4x10(-7)-8.4x10(-6) microg/ml, for Ag - 23x10(-7)-23x10(-4) microg/ml, for Au - 5x10(-6) microg/ml by metals. It has been shown, that cultural characteristics of probiont-bacteria in the presence of both ionic and colloid forms of studied metals remain stable. Certain stimulation of probiont-bacteria' main enzymes and life-support processes by colloidal forms of the studied metals-microelements in determined concentration and particle dimension limits is the evidence of a possibility of metals' colloids application in probiotic preparations' composition in order to increase the resistance and functional activity of probiont-microorganisms.     

Silver accumulation and resistance in Escherichia coli R1

E. coli R1 contains at least 2 large plasmids (83 and 77 kb) while E. coli S1 was previously cured of the 83 kb plasmid. Transformation using artificial competence, high-voltage electroporation, and plasmid mobilization experiments with the mobilizing plasmid RP4, failed to ascertain the 83 kb plasmid was responsible for Ag(+)-resistance. Silver accumulation by an Ag(+)-sensitive E. coli S1 strain was 5-fold higher than an Ag(+)-resistant E. coli R1 strain. The Ag(+)-resistant E. coli R1 strain produced 33% more H2S and 32% more intracellular acid labile SH than the Ag(+)-sensitive E. coli R1 strain when grown in the absence of AgNO3. Hydrophobic interaction chromatography revealed E. coli R1 displayed higher cell surface hydrophobicity than E. coli S1. HPLC protein analysis of cell-free extracts also revealed differences between protein fractions in E. coli R1 and S1 strains.     

Enhanced microbial biomass assay using mutant luciferase resistant to benzalkonium chloride

In a biomass assay based on adenosine 5(')-triphosphate (ATP) bioluminescence, extracellular ATP is removed; then intracellular ATP is extracted from the microorganism by an ATP extractant and subsequently reacted with luciferase. To provide a highly sensitive assay, the concentration of benzalkonium chloride (BAC) in the ATP extractant was optimized by using a mutant luciferase resistant to BAC. The use of 0.2% BAC, which was acceptable for the luciferase, simultaneously achieved the maximum extraction of intracellular ATP from microorganisms and the inactivation of the ATP-eliminating enzymes for removal of extracellular ATP. The detection limit (blank+3 SD) for ATP was 1.8x10(-14)M (1.8x10(-18)mol/assay) in the presence of the ATP extractant with coefficients of variation of 0.7 to 6.3%. The reagent system coupled with the ATP-eliminating enzymes allowed for the detection of 93 colony-forming units (CFU)/ml of Escherichia coli ATCC 25922, 170CFU/ml of Pseudomonas aeruginosa ATCC 27853, 170CFU/ml of Proteus mirabilis ATCC 29906, 68CFU/ml of Staphylococcus aureus ATCC 25923, and 7.7CFU/ml of Bacillus subtilis ATCC 6051. The yeast cell of Saccharomyces cerevisiae IFO 10217 could be detected at 1CFU/ml. With 54 kinds of microorganisms, the average ATP extraction efficiency compared to the trichloroacetic acid extraction method was 81.0% in 24 strains among gram-negative bacteria, 99.4% in 13 strains among gram-positive bacteria, and 97.0% in 17 strains among yeast. The ATP contents of the gram-negative bacteria, gram-positive bacteria, and yeasts ranged from 0.40 to 2.70x10(-18)mol/CFU (mean=1.5x10(-18)mol/CFU), from 0.41 to 16.7x10(-18)mol/CFU (mean=5.5x10(-18)mol/CFU), and from 0.714 to 54.6x10(-16)mol/CFU (mean=8.00x10(-16)mol/CFU), respectively.     

Thin film trichloroethylene electrochemical sensor

Pt-Ti and Pb-Pt-Ti thin films were deposited on alumina substrates by sputtering in Ar gas. In this study, an electrodeposited Pb-modified Pt-Ti thin film working electrode was prepared. Optimal sensing conditions were found to be -2.10 V (versus Ag/Ag+ with 0.1 M tetrabutylammonium perchlorate (TBAP) in acetonitrile (AN) solution) sensing potential, 250 rpm agitation rate. At room temperature, the response time was 15 s (90% response time). The correlation of sensing response current, id, and trichloroethylene (TCE) concentration, CL, is id = 2.86CL in the range from 100 to 700 ppm TCE. Additionally, the rate constant of (TCE) cathodic reduction was found to be 2.434 x 10(-3) cm(-1) s(-1).     

Rate of plasmid transfer among Escherichia coli strains isolated from natural populations.

The conjugative plasmid R1 was introduced into ten strains of Escherichia coli isolated from natural populations. Spontaneous nalidixic-acid-resistant mutants of the ten strains served as recipients. The ten donor and recipient strains were mated in all combinations and the rate at which R1 transferred between the strains was determined. The rate of transfer ranged from 5.2 x 10(-11)-1.1 x 10(-18) ml per cell h-1, and averaged 1.3 x 10(-15) ml per cell h-1. The results of these experiments suggest that the rates of conjugative transfer are far too low for plasmids to be maintained as parasites in their host populations. Infectious transfer is insufficient; plasmids must confer a selective advantage to their host to be maintained.     

Elaboration of an electroporation protocol for Bacillus cereus ATCC 14579

An electro-transformation procedure was established for Bacillus cereus ATCC 14579. Using early growth-stage culture and high electric field, the ectroporation efficiency was up to 2 x 10(9) cfu microg(-1) ml(-1) with pC194 plasmid DNA. The procedure was tested with three other plasmids, of various sizes, replication mechanisms and selection markers, and the transformation efficiencies ranged between 2 x 10(6) and 1 x 10(8) cfu microg(-1) ml(-)(1). The effects of two wall-weakening agents on electroporation rates were also evaluated. The transformation rate that was reached with our procedure is 10(3) times higher than that previously obtained with members of the Bacillus genus with similar plasmids, and 10(6) times superior than that achieved with available protocols for B. cereus. The proposed method is quick, simple, efficient with small rolling circle plasmids and large theta replicating plasmids with low copy number per cell, and suitable for many genetic manipulations that are not possible without high-efficiency transformation protocols.     

Immunomagnetic separation of scum-forming bacteria using polyclonal antibody that recognizes mycolic acids

Mycolic acid-containing bacteria (mycolata) are thought to be involved in scum formation in aeration basins of activated sludge plants due to their ability to produce biosurfactants and their cell surface hydrophobicity. To isolate these bacteria, immunomagnetic separation (IMS) using an anti-mycolic acid polyclonal antibody was investigated. IMS that targeted Gordonia amarae SC1 exhibited a 100% recovery at 5x10(3) CFU ml(-1). At cell concentration of 7.8x10(6) CFU ml(-1), the recovery was lowered, but 80% of cells were still captured. Effect of bead concentrations on the recovery of SC1 at 10(6) CFU ml(-1) was examined. The results showed that addition of more than 6-7x10(6) beads for 1x10(6) CFU reached a maximum recovery (83%). Furthermore, the IMS procedure optimized with SC1 cells was tested with another mycolata. The results suggested that variation of the recovery for each mycolata is dependent on the specificity of the polyclonal antibody and that mycolata which are recognized by the antibody can be recovered by this procedure.     

Transformation of intact yeast cells treated with alkali cations

Intact yeast cells treated with alkali cations took up plasmid DNA. Li+, Cs+, Rb+, K+, and Na+ were effective in inducing competence. Conditions for the transformation of Saccharomyces cerevisiae D13-1A with plasmid YRp7 were studied in detail with CsCl. The optimum incubation time was 1 h, and the optimum cell concentration was 5 x 10(7) cells per ml. The optimum concentration of Cs+ was 1.0 M. Transformation efficiency increased with increasing concentrations of plasmid DNA. Polyethylene glycol was absolutely required. Heat pulse and various polyamines or basic proteins stimulated the uptake of plasmid DNA. Besides circular DNA, linear plasmid DNA was also taken up by Cs+-treated yeast cells, although the uptake efficiency was considerably reduced. The transformation efficiency with Cs+ or Li+ was comparable with that of conventional protoplast methods for a plasmid containing ars1, although not for plasmids containing a 2 microns origin replication.     

Improved use of organic phosphate by Skeletonema costatum through regulation of Zn2 + concentrations

The maximum growth rate (1.4-2 x 10(5) cells ml(-1) d(-1)), cell final yields (2.6-5.2 x 10(5) cells ml(-1)) and extracellular alkaline phosphatase activity (2.4-10.6 microg phosphate released ml(-1) h(-1)) of the red tide alga, Skeletonema costatum, increased when Zn2+ was increased from 0 to 24 pM, but decreased with 66 pM Zn2+ in growth medium with glycerophosphate as the sole phosphorus source. Extracellular carbonic anhydrase activity and the affinity for HCO3- and CO2 uptake increased when Zn2+ was increased from 0 to 12 pM, but then decreased at higher concentrations. The results suggested that utilization of organic phosphate required more Zn2+ than the uptake of inorganic carbon did, while utilization of dissolved inorganic carbon by Skeletonema costatum was very sensitive to Zn2+ concentration variations.     

马缨丹总岩茨烯对小菜蛾和斜纹夜蛾幼虫的拒食作用

从马缨丹叶片中提取总岩茨烯,测定总岩茨烯对小菜蛾2龄幼虫和斜纹夜蛾1、2龄幼虫的拒食作用.结果表明,在非选择性试验中,1.6 mg·ml^-1的总岩茨烯对小菜蛾2龄幼虫和斜纹夜蛾1龄幼虫有拒食作用,8 h的拒食率分别为62.%和31.1%;在选择性试验中,0.4 mg·ml^-1的总岩茨烯即对小菜蛾2龄幼虫有拒食作用,0.4、0.8和1.6 mg·ml^-1浓度处理下的拒食率分别为52.7%、55.5%和78.9%;总岩茨烯只有在高浓度(1.6 mg·ml^-1)时,才对斜纹夜蛾1龄幼虫有拒食作用,拒食率仅为33.0%.在选择性和非选择性试验中,总岩茨烯对斜纹夜蛾2龄幼虫均无拒食作用.