掷孢菌科的研究II．中国叶表掷孢酵母的初步分类及掷孢酵母属的种类

作者多年来从我国禾本科等植物叶表分离了大量掷孢酵母,本文重点报道70株掷孢酵母与类酵母的初步分类,共分为三属,即掷孢酵母属(Sporobolomyces),布勤掷孢酵母属(Bul-lera)与腥掷孢菌属(Tilletiopsis);其中掷孢酵母属占75％。至于53株掷孢酵母,共分为三种,即攻红掷孢酣母(Sporobolomyces roseus)、赭色掷孢酵母(Sporobolomyces salmonicolor)、Sporobolomyces shibatanus);经统计,后者最为常见,占此属的84％。这些种在我国尚未见报道,作为新记录。     

南极乔治王岛土壤微生物的生物学特征及生物活性

从南极乔治王岛土壤分离出77株放线菌和丝状真菌,它们大多数是嗜冷菌或兼性嗜冷菌。它们能在0℃生长,适宜温度为5-15℃、15—20℃或l 5—28℃。用多种条件研究了77株菌的生物活性,发现32％的菌株有抗微生物活性(包括革兰氏阳性和阴性细菌、酵母、青霉等)。其中5株菌对精原细胞、10株对KB细胞有抗肿瘤活性。根据形态培养特征、细胞壁化学组分、放线菌大多数属链霉菌(Streptomyces),其次是诺卡氏菌(Nocardia)和类诺卡氏菌(Nocardioides)。 丝状真菌有5种不同的类型,它们都属金孢霉属(Chysosporium)。     

枝顶孢属真菌的抑菌活性及其代谢产物研究

用组织分离法从采集于广东鼎湖山的腐殖质中分离得到一株枝顶孢属真菌(Acremonium sp. SC0105),其固体发酵物的乙醇提取物对金黄色葡萄球菌(Staphylococcus aureus)有较强的抑制作用。经多种柱层析,从固体发酵物中分离得到5个化合物。通过光谱分析,分别鉴定为姜糖脂B (1)、姜糖脂C (2)、D-甘露醇(3)、酒渣碱(4)、枝顶孢素F(5)。其中酒渣碱是首次从枝顶孢属真菌中分离得到。     

掷孢菌科的研究III．不同掷孢酵母属产掷孢子的能力及担子菌酵母形成特大厚垣孢子的规律

本文以分离自中国的大量掷孢酵母为材料,研究了不同属的掷孢酵母生掷孢子的能力,以及此能力的持久性,发现属间差异很大。此外还报道了142株不同酵母的产厚垣孢子的情况,发现与担子菌有关的4属酵母在接近致死温度时,都会出现特大厚垣孢子如红掷静酵母Sporobolomyces),布勒掷孢酵母(Bullera),红酵母(Rhodotorula)与隐球酵母(Cryp-tococcus),而属于子囊菌的酵母属(Saccharomyces)则无此现象,从而对某些接近担子菌的酵母又增加了新的认识。     

耐热酵母与酿酒酵母原生质体融合的研究

酿酒酵母(saccharomyces cerevisiae)396是利用甘蔗糖蜜生产酒精的生产菌株,假丝酵母(Candida sp·)C₆是我们从云南温泉底泥中筛选到的一株能在45℃生长良好的耐热酵母,我们应用原生质体融合技术进行了两菌株属间融合的研究。通过亚硝基胍(NTG)诱变得到的营养缺陷型菌株396(arg^－)和C6(lys^-),其融合频率为O．91×10^-5。从检出的融合子中挑选6株进行考核、其细胞体积平均为亲株的1．3倍,DNA含量平均为亲株的1．6倍,培养特征、形态、大小和生理生化特征表现不一,特别是糖类的同化试验。除F₂、F₁₄外,其他4株可以排除异核体形成的可能性。比较了在28℃,40℃和45℃培养条件下出发亲株396、C₆、直接亲株396(arg^-)C₆(lys^-)和融合子F₁、F₇、F₁₂,F₁₃的生长曲线、基质利用率和乙醇产率等,得到一株在40℃培养条件下糖的利用率为94．3％、乙醇产量为59．7g／L的属间融合株F₁₃。     

有淀粉糖化酶活性的耐高温酿酒酵母的构建研究

通过诱导Hu—TY一1(耐高温酿酒酵母)产孢及单孢分离得到的单倍体株Hu—TY—l—A(Sa-ccharamyces cerevisiae)与具有STAl、STA2、STA3(淀粉糖化酶)遗传因子的酵母(S．Dias-taticus)单倍体和纯合二倍体进行相同交配型的种间原生质体融合,融合率为3．3×1O^-6-8．7×1O^-6。融合子的细胞形态、大小、DNA含量、生长和发酵特性等方面均不同于亲株。融合株BA-2、BA-3、CA-18、DA-2、CCA-30、其细胞体积与DNA含量为亲株之和,30℃生长速率与生物量为亲株的2—3倍,40℃生长速率与生物量为亲株HU－TY－1－A的1．3－1．6倍,能够利用淀粉,并且在40℃高温葡萄糖发酵酒精产率比糖化酵母高,比HU－TY－1－A低,是一些有淀粉糖化酶、耐高温的酿酒酵母新菌株。     

L-丝氨酸生产菌的选育及不同碳源对发酵的影响

以一株黄假单胞属(Pseudomonas flava)菌株A3为出发菌株,经过紫外(UV)诱变和硫酸二乙酯(DES)逐级诱变处理和选育,选育出一株能够以甲醇为唯一碳源的兼性甲基营养型菌JW-01(Mth^R、Gly^R)。在含甘氨酸30g/L、甲醇1%的发酵培养基中发酵3d后L-丝氨酸产量为6．2g/L,较出发菌株提高了67．6%。该菌具有较好的传代稳定性。     

四株糖单孢菌分离株的分类学研究

从广西地区的土样中,分离到4株细胞壁Ⅳ型,糖型A, 无枝菌酸的假诺卡氏菌科的放线菌菌株,编号分别为191、221、202、和212。根据4株 菌的形态学特征和细胞化学特征,将其归入糖单孢菌属。与该属5个已知种的7个代表株进行 的rDNA的BamHI酶切片段长度类型分析(Ribotyping)的结果表明：191为青绿色糖单孢菌(S.viridis),202为青灰色糖单孢菌(S.caesia),221和212为相同的与青绿色糖单孢菌(S .viridis)的亲缘关系最近的种,但不同于已知的任何一个种。     

致病酵母菌基因组多态性及亲缘关系的研究

致病酵母是条件致病菌感染中最常见的菌群。其属间、种间及种内的分型具有重要的流行病学及临床意义。以随机扩增多态性(Randomly Amplified Polymorphism DNA markers,RAPD)的方法对48株临床上常见的酵母菌属间、种间及种内基因组型的多态性进行了研究,并以多种引物扩增带型的相似性系数的高低来评价酵母菌之间的亲缘关系。结果表明:RAPD带型可清楚的显示出假丝酵母(Candida)及相关酵母属间、种间及种内的差异,亲缘关系的研究表明假丝酵母属与隐球菌属(Cryptococcus)、丝孢酵母属(Trichosporon)的相似性系数为80％,除季也蒙假丝酵母(C. guilliermondii)外,假丝酵母属中不同种间的相似性系数为82％～87％,同种不同株间的相似性系数>90％。大多数属、种基因组分型的结果和形态学分类结果相符。     

产壳聚糖酶内生真菌的筛选、鉴定及酶活力初步研究

植物内生真菌是挖掘不同类型壳聚糖酶及发现新酶的资源宝库。该研究从122株柑橘和血散薯内生真菌中筛选能产生壳聚糖酶的菌株,对其进行鉴定,初步研究酶活力影响因素,为后期其酶学性质及产壳聚糖酶内生真菌与宿主植物病害防御互作关系的研究奠定基础。通过透明圈法初筛结合液体发酵法进行复筛,得到2株可产生壳聚糖酶的内生真菌Stdif9和Stdif9-4,并发现Stdif9-4最高酶活力(0.968 U·mL^-1)显著高于Stdif9(0.780 U·mL^-1)。采用形态学和分子生物学结合的方法将菌株Stdif9-4鉴定为青霉属菌株,即Penicillium sp. Stdif9-4。通过DNS试剂法初步研究影响该菌株产壳聚糖酶活力的因素,发现不同培养时间对菌株壳聚糖酶活力具有显著影响,在培养96 h时,壳聚糖酶活力达到最大值。9种金属离子对菌株的酶活力具有不同影响,其中Mn²⁺和Ca²⁺对壳聚糖酶活力具有明显的激活作用; Ag⁺、Zn²⁺、Cd²⁺、Ba²⁺和Fe³⁺对壳聚糖酶活力具有不同程度的抑制作用,并且Ag⁺的抑制作用最为显著; K⁺和Na⁺对壳聚糖酶活力无显著影响。不同培养代数菌株产酶活力无显著差异,说明其产酶活力稳定。     

陕南早寒武世早期Quadrapyrgites再研究

处于"寒武纪生命大爆发"序幕阶段的梅树村期,生物类群大规模辐射,身体构型快速革新,与前寒武纪生物群面貌明显不同.最近在陕南宁强宽川铺地区梅树村期地层中发现了大量五辐射Punctatus及部分四辐射四方塔形壳属Quadrapyrgites,其中包括1个新种Quadrnpyrgites undulatuscostalis sp.nov..在此基础上对Quadrapyrgites进行了属征补充.双胚层腔肠动物的出现标志着地球生命史的真后生动物演化开端,在生物起源演化历程上占据着关键的位置.本文为研究真后生动物起源演化、生物辐射、体型构建提供了重要实证.     

Description of by-product inhibiton effects on biodesulfurization of dibenzothiophene in biphasic media

As several authors have reported previously, the Biodesulfurization of hydrodesulfurization recalcitrants, such as dibenzothiophene, is not yet commercially viable because mass transfer limitations and feedback inhibition effects are produced during the conversion. This work has been focused to investigate the inhibition process in aqueous and oil-water systems with two different aerobic biocatalysts types, Rhodococcus erythropolis IGTS8 and Pseudomonas putida CECT 5279. The results obtained have proven that global DBT desulfurization process using CECT 5279 was not clearly deactivated due to final product accumulation, under the experimental conditions assayed. Consistently, the desulfurization pattern has been described with the Michaelis-Menten equation, determining the kinetic parameters. On other hand, the assays have shown that important mass transfer limitations produced the decrease of the yields obtained with this Gram(-) strain in biphasic media. With strain IGTS8 it was observed lower mass transfer problems, but contrary the reaction was severely affected by the final product accumulation, in both aqueous and biphasic systems. Therefore it has been proposed an enzymatic kinetic model with competitive inhibition to describe the BDS evolution pattern when this Gram(+) strain was used.     

Biocidal activity of some Mannich base cationic derivatives

A novel series of cationic surfactants was prepared based on Mannich base (produced from the condensation of piperidine and/or morpholine as secondary amine and paraformaldehyde in the presence of 8-hydroxyquinoline). The chemical structures of the synthesized cationic surfactants were confirmed using elemental analyses, FTIR spectroscopy and 1H NMR. Surface activities of the prepared surfactants were measured including: surface tension (gamma), critical micelle concentration (CMC), effectiveness (pi(CMC)), efficiency (Pc20), maximum surface excess (Gamma(max)), minimum surface area (A(min)), interfacial tension (gamma(IT)), emulsification power and foaming power at 25 degrees C. The structural influences on their surface activities and adsorption free energy were discussed. The synthesized cationic surfactants were evaluated for their biocidal activity towards Gram +ve bacteria (Staph. Cocu., Bacillus), Gram -ve bacteria (Salmonella, E. coli), fungi (A. terrus., A. flav.) and yeast (Candida) at 1.0, 2.5 and 5.0mg/mL, respectively. The target compounds showed good inhibition towards Gram +ve bacteria, Gram -ve bacteria and yeast. Meanwhile, excellent fungicidal results were obtained against the various types of fungi under investigation.     

Characterisation of the yeast Pichia membranifaciens and its possible use in the biological control of Botrytis cinerea, causing the grey mould disease of grapevine

Pichia membranifaciens strain FY-101, isolated from grape skins, was found to be antagonistic to Botrytis cinerea, the causal organism of the grey mould disease of the grapevine. When grown together on solid as well as liquid media, the yeast brings about the inhibition of this parasitic fungus, coagulation and leakage of its cytoplasm, and suppression of its ability to produce the characteristic grey mould symptoms on the grapevine plantlets. In vitro experiments confirm that this yeast can be used as a biological control organism against B. cinerea. An account of the molecular characterisation of P. membranifaciens (complete sequence of the ITS region of its ribosomal DNA, GenBank accession No. AF 270935), as well as the interaction between B. cinerea and the yeast, are given here.     

TORC1 signaling inhibition by rapamycin and caffeine affect lifespan,global gene expression,and cell proliferation of fission yeast

Target of rapamycin complex 1 (TORC1) is implicated in growth control and aging from yeast to humans. Fission yeast is emerging as a popular model organism to study TOR signaling, although rapamycin has been thought to not affect cell growth in this organism. Here, we analyzed the effects of rapamycin and caffeine, singly and combined, on multiple cellular processes in fission yeast. The two drugs led to diverse and specific phenotypes that depended on TORC1 inhibition, including prolonged chronological lifespan, inhibition of global translation, inhibition of cell growth and division, and reprograming of global gene expression mimicking nitrogen starvation. Rapamycin and caffeine differentially affected these various TORC1‐dependent processes. Combined drug treatment augmented most phenotypes and effectively blocked cell growth. Rapamycin showed a much more subtle effect on global translation than did caffeine, while both drugs were effective in prolonging chronological lifespan. Rapamycin and caffeine did not affect the lifespan via the pH of the growth media. Rapamycin prolonged the lifespan of nongrowing cells only when applied during the growth phase but not when applied after cells had stopped proliferation. The doses of rapamycin and caffeine strongly correlated with growth inhibition and with lifespan extension. This comprehensive analysis will inform future studies into TORC1 function and cellular aging in fission yeast and beyond.     

成人牙周炎龈下套氧菌分离鉴定及药敏试验结果分析

分离并鉴定了329例成人牙周炎龈下优势厌氧菌群,并对不同病程中的菌群变迁、厌氧菌的药物敏感性进行了分析.成人牙周炎龈下标本中厌氧菌阳性检出率为97.9%,其中以牙龈紫质单胞菌检出率最高(38.5%),具核梭杆菌次之(18.9%).随着牙周病变程度的加重,牙龈紫质单胞菌、具核梭杆菌、产黑色素普氏菌、星群厌氧链球菌、厌氧消化链球菌的检出率增高(P＜0.05),小韦荣球菌的检出率下降(P＜0.01),表明前5种厌氧菌在AP发病过程中有重要作用,小韦荣球菌与之无关.替硝唑、甲硝唑和克林霉素对438株革兰氏阴性厌氧菌的MIC90分别为1～8,2～8和4～16 mg/L,对278株革兰氏阳性厌氧菌的MIC90分别为16～32,16～64和4～16 mg/L,表明替硝唑和甲硝唑体外抗革兰氏阴性厌氧菌效果优于克林霉素,抗革兰氏阳性厌氧菌作用不如克林霉素.     

IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF A PEPTIDOGLYCAN RECOGNITION PROTEIN FROM THE COTTON BOLLWORM,Helicoverpa armigera

Peptidoglycan recognition proteins (PGRPs) specifically bind to peptidoglycans, and play crucial roles as pattern recognition receptors (PRRs) in mediating innate immune responses. In this study, we identified and characterized a PGRP (HaPGRP‐D) from the cotton bollworm, Helicoverpa armigera. Sequence analysis indicated that HaPGRP‐D is an amidase‐type PGRP. Expression of HaPGRP‐D was upregulated in the hemocytes of H. armigera larvae after injecting Gram‐negative Escherichia coli, Gram‐positive Staphylococcus aureus, or chromatography beads. To test the biological activity of HaPGRP‐D, purified recombinant protein was prepared. Subsequent analysis showed that rHaPGRP‐D (i) could bind and agglutinate Gram‐negative E. coli and Gram‐positive S. aureus in a zinc‐dependent manner, (ii) functioned as an amidase to degrade peptidoglycans in the presence of Zn²⁺, (iii) strongly inhibited the growth of E. coli and S. aureus in the presence of Zn²⁺, (iv) could bind to the surface of hemocytes, (v) increased the phagocytosis of E. coli cells by hemocytes in vitro, and (vi) promoted hemocyte encapsulation on chromatography beads in vitro. These results suggest that HaPGRP‐D plays important roles as PRR, amidase, and opsonin in H. armigera humoral and cellular immune responses.     

Genome Analysis of the Anaerobic Thermohalophilic Bacterium Halothermothrix orenii

Halothermothirx orenii is a strictly anaerobic thermohalophilic bacterium isolated from sediment of a Tunisian salt lake. It belongs to the order Halanaerobiales in the phylum Firmicutes. The complete sequence revealed that the genome consists of one circular chromosome of 2578146 bps encoding 2451 predicted genes. This is the first genome sequence of an organism belonging to the Haloanaerobiales. Features of both Gram positive and Gram negative bacteria were identified with the presence of both a sporulating mechanism typical of Firmicutes and a characteristic Gram negative lipopolysaccharide being the most prominent. Protein sequence analyses and metabolic reconstruction reveal a unique combination of strategies for thermophilic and halophilic adaptation. H. orenii can serve as a model organism for the study of the evolution of the Gram negative phenotype as well as the adaptation under thermohalophilic conditions and the development of biotechnological applications under conditions that require high temperatures and high salt concentrations.     

Inhibition of yeast growth by molybdenum-hydroxylamido complexes correlates with their presence in media at differing pH values

The effects of Mo-hydroxylamido complexes on cell growth were determined in Saccharomyces cerevisiae to investigate the biological effects of four different Mo complexes as a function of pH. Studies with yeast, an eukaryotic cell, are particularly suited to examine growth at different pH values because this organism grows well from pH 3 to 6.5. Studies can therefore be performed both in the presence of intact complexes and when the complexes have hydrolyzed to ligand and free metal ion. One of the complexes we examined was structurally characterized by X-ray crystallography. Yeast growth was inhibited in media solutions containing added Mo-dialkylhydroxylamido complexes at pH 3-7. When combining the yeast growth studies with a systematic study of the Mo-hydroxylamido complexes' stability as a function of pH and an examination of their speciation in yeast media, the effects of intact complexes can be distinguished from that of ligand and metal. This is possible because different effects are observed with complex present than when ligand or metal alone is present. At pH 3, the growth inhibition is attributed to the forms of molybdate ion that exist in solution because most of the complexes have hydrolyzed to oxomolybdate and ligand. The monoalkylhydroxylamine ligand inhibited yeast growth at pH 5, 6 and 7, while the dialkylhydroxylamine ligands had little effect on yeast growth. Growth inhibition of the Mo-dialkylhydroxylamido complexes is observed when a complex exists in the media. A complex that is inert to ligand exchange is not effective even at pH 3 where other Mo-hydroxylamido complexes show growth inhibition as molybdate. These results show that the formation of some Mo complexes can protect yeast from the growth inhibition observed when either the ligand or Mo salt alone are present.     

健康动物肠道正常菌群的分离与鉴定

本文采用多种选择培养基,从健康动物猪、鸿、兔、鱼肠道中分离培养出２２株正常菌群,经过菌落形状、革兰染色、美兰染色镜检及生化反应,初步鉴定为链球菌８株、孔杆菌２株、双歧杆菌４株、芽胞杆菌５林、酵母菌３株。此为优选菌种、制备微囊及生产生态制剂等奠定了良好的基础．