天然缺失inlAB的非典型单核细胞增多性李斯特菌生物学特性鉴定

摘要：【目的】 InlA与InlB是单核细胞增多性李斯特菌重要的毒力因子,其介导的黏附作用是细菌建立感染的前提。本研究拟探明天然缺失inlAB基因簇的非典型单增李斯特菌的表型与基因型特征。【方法】针对inlAB天然缺失株S10,进行生化特征、细胞黏附力、小鼠体内毒力、感染相关基因检测、谱系分析等。【结果】 S10株为具有典型单增李斯特菌生化特征的1/2b型菌株,对HeLa细胞的黏附力显著低于其他菌株（p<0.05）,对小鼠毒力较弱。S10缺失inlAB及与其毗邻的lmo0431、lmo0432、lmo0436、lmo0437基因,但具有李斯特菌第一毒力岛中完整的毒力基因构成。S10分布于谱系Ⅰ的进化枝上,与4b型菌株的遗传距离较近。【结论】 S10为单增李斯特菌inlAB天然缺失株代表该类非典型菌株的首次报道。S10具有典型的单增李斯特菌谱系Ⅰ基因背景,inlAB可能通过独立的重组或水平转移事件缺失于基因组。     

药用植物内生放线菌的分离、筛选及活性菌株YIM 61470鉴定

从云南西双版纳热带雨林多种药用植物中分离到272株内生放线菌,活性筛选表明 146株菌的发酵产物具有抗菌活性,其中94株菌具有拮抗病原细菌活性,127株菌具有抑制病原真菌的功能.分离菌株YIM 61470具有广谱抗菌活性,通过形态特征、培养特征、生理生化特征、细胞化学分类特征和基于16S rRNA基因序列的相似性分析等研究,菌株YIM 61470被鉴定为链霉菌属(Streptomyces)氢化链霉菌(S.llydrogenans)的一个菌株.     

葱鳞葡萄孢菌的异核现象

本文报导了葱鳞葡萄孢菌(Botrytis squamosa)的异核现象。采用荧光染色法证实了该菌的分生孢子和菌丝细胞均是多核的。以该菌的自然形态和生长特征为遗传标记,在193个单孢子培养物中获得了三种菌型:菌核型(S)、菌丝型(M)、界于菌核和菌丝之间的菌核菌丝型(SM)。SM型在形态上与原始菌株相同。S型和M型在连续的传代过程中其特征保持稳定,因而被认为是同核型。SM型在连续传代过程中是不稳定的,且一再分离出S型和M型,因此被认为是异核型。用传代稳定的S型和M型重新合成的异核体在特征上相同于原始菌株和SM型。用这个合成的异核体再次进行单孢子分离,三种菌型重新被获得。三种菌型在致病力方面没有表现出显著的差异。     

1株拮抗真菌的生防细菌Xj11的分离与鉴定

从江西省井冈山自然保护区发病毛竹中分离到1株具有明显抑制植物病原真菌活性的生防细菌Xj11。其对小麦赤霉病菌、尖孢镰刀菌、甘蔗节菱孢菌、链格孢菌和炭角菌均有较强的抑制作用。通过形态特征和培养特征观察、生理生化实验及16S rDNA序列系统发育分析,初步鉴定该菌为唐菖蒲伯克霍尔德氏菌,命名为Burkholderia gladioli strain Xj11。     

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。     

内蒙古碱湖中紫色非硫光合细菌的分离和鉴定

从内蒙古碱湖水样中分离得到一株紫色非硫光合细菌,命名为JH1-6.对该菌株进行了形态学观察、生理生化鉴定、活细胞吸收光谱以及16S rDNA序列分析.16S rDNA序列分析结果表明该菌株与沼泽红假单胞菌的16S rDNA序列同源性高达99%,结合形态特征和生理生化特性以及活细胞吸收光谱特征等,确定菌株JH1- 6在分类地位上属于沼泽红假单胞菌(Rhodopseudomonas palustris).     

市售冷却牛肉中主要细菌的常规分离与鉴定

利用常用纯培养的方法, 根据细菌的菌落形态、菌落颜色、革兰氏染色等常见特征, 从市售冷却牛肉中, 选取菌落形态差别比较明显的菌株共32株, 其中保鲜膜包装冷却牛肉样品共12株, 未包装冷却牛肉样品共20株; 同时选取两样品中的优势菌株各4株进行进一步的研究(8株细菌编号为：S01~S08, 其中S01~S04为未包装冷却牛肉样品; S05~S08为保鲜膜包装冷却牛肉样品), 通过ARDRA(Amplified ribosomal DNA restriction analysis) 以及16S rDNA 序列等进行分类研究, 确定该细菌的分类地位, 并结合形态、常规生理生化特性进行鉴定, 确定各细菌所属种。实验表明：S01为假单胞菌属中的恶臭假单胞菌(Pseudomonas putida), S02为希瓦氏菌属下的(Shewanella cincia sp.), 而S03和S05为希瓦氏菌属下的腐败希瓦氏菌(Shewanella putrefaciens), S04为窄食单胞菌属中的嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia), S06为嗜冷杆菌(Psychrobacter sp.), S07为葡萄球菌属中的松鼠葡萄球菌(Staphylococcu sciuri), S08为微杆菌属中的产左聚糖微杆菌(Microbacterium laevaniformans)。证明两样品的共有优势菌为希瓦氏菌属。通过对样品可培养微生物情况进行初步的调查分析, 为冷却肉加工工艺提供一定的理论基础。     

基于16S rDNA测序技术分析功能性便秘患者粪便菌群

目的 运用16S rDNA测序技术分析功能性便秘（FC）患者粪便菌群特征,以期寻找FC特征菌,为FC诊治寻找新的治疗靶点。方法 收集福建中医药大学附属第二人民医院60例实验组（FC患者）和30例正常组的粪便,采用16S rDNA测序技术分析粪便菌群结构,并进行基因功能的预测。结果 两组在门水平上,主要由厚壁菌门和拟杆菌门组成;在属水平上,主要优势菌以粪杆菌属和拟杆菌属为主。实验组粪便菌群丰富度指数Chao1、Observed＿OTUs较正常组升高（P<0.05）,多样性指数Shannon指数和Simpson指数虽有上升趋势,但差异无统计学意义（P> 0.05）。布劳特菌属、罗氏菌属、 Ruminococcus＿gnavus＿group、Ruminococcus＿gauvreauii＿group、Eubacterium＿hallii＿group、Anaerostipes、Lachnospiraceae＿ND3007＿group、Dorea、乳球菌属在实验组中丰度均显著降低（P<0.01）。实验组特征菌为小杆菌属,正常组为布劳特菌属和埃希菌-志贺菌属。实验组蔗糖降解Ⅲ（P...     

中国齿状真菌研究5.齿耳属(担子菌门,皱孔菌科)(英文)

对我国木生齿状真菌齿耳属进行了研究,报道了该属3个中国新记录种:阔纤毛齿耳菌Steccherinum ciliolatum、山生齿耳菌S.oreophilum和强壮齿耳菌S.robustius。3个种分别采集自黑龙江、四川和吉林省。从形态学及分子系统学方面确定了该属3个种类的分类地位。阔纤毛齿耳菌主要特征为:子实体平伏,奶油色至浅黄色,短齿,边缘呈绒毛状,孢子椭圆形;山生齿耳菌主要特征为:子实体小,平伏至反卷,菌盖表面光滑,子实层体齿状至孔状,骨架囊状体锥形,孢子窄椭圆形;强壮齿耳菌的主要特征为:子实体平伏至反卷,菌齿浅橘红色至暗黄色,孢子宽椭圆形。分子系统学方面,Steccherinum 3个种类获得了高的支持率,分属不同分支,显著区别于该属的其他种类。根据采集到的标本对它们进行了详细的描述和绘图,并给出了我国该属已知15种的检索表及每种的简要特征。     

一个新的古菌类群———奇古菌门(Thaumarchaeota)

基于16S rRNA基因的系统发育关系,古菌域(Archaea)被分为两个主要类群:广古菌门(Euryarchaeota)和泉古菌门(Crenarchaeota)。近20年来,微生物分子生态学技术的快速发展和应用显示,在中温环境中广泛存在着大量的未培养古菌,而且它们可能在自然界重要元素(N、C)的生物地球化学循环中发挥着重要作用。最初,这些未培养古菌因在16S rRNA基因系统发育上与泉古菌关系较密切而被称作中温泉古菌(non-thermophilic Crenarchaeota)。而近年来,对更多新发现的中温古菌核糖体RNA基因序列和其它分子标记物进行的分析均不支持中温泉古菌由嗜热泉古菌进化而来的假设,而揭示其可能代表古菌域中一个独立的系统发育分支。基因组学、生理生态特征等分析也显示中温泉古菌与泉古菌具有明显不同的特征。因而专家建议将这些古菌(中温泉古菌)划分为一个新的门,成为古菌域的第三个主要类群—Thaumarchaeota(意译为奇古菌门)。这一新古菌门提出后得到其他研究证据的支持和认可。本文对目前已知的奇古菌门的分类地位演化、基因组学、多样性和生理代谢特征等作一简要综述。     

Degradation of 4-aminobenzenesulfonate by a two-species bacterial coculture

The mutualistic interactions in a 4-aminobenzenesulfonate (sulfanilate) degrading mixed bacterial culture were studied. This coculture consisted of Hydrogenophaga palleronii strain S1 and Agrobacterium radiobacter strain S2. In this coculture only strain S1 desaminated sulfanilate to catechol-4-sulfonate, which did not accumulate in the medium but served as growth substrate for strain S2. During growth in batch culture with sulfanilate as sole source of carbon, energy, nitrogen and sulfur, the relative cell numbers (colony forming units) of both strains were almost constant. None of the strains reached a cell number which was more than threefold higher than the cell number of the second strain. A mineral medium with sulfanilate was inoculated with different relative cell numbers of both strains (relative number of colony forming units S1:S2 2200:1 to 1:500). In all cases, growth was found and the proportion of both strains moved towards an about equal value of about 3:1 (strain S1:strain S2). In contrast to the coculture, strain S1 did not grow in a mineral medium in axenic culture with 4-aminobenzenesulfonate or any other simple organic compound tested. A sterile culture supernatant from strain S2 enabled strain S1 to grow with 4-aminobenzenesulfonate. The same growth promoting effect was found after the addition of a combination of 4-aminobenzoate, biotin and vitamin B₁₂. Strain S1 grew with 4-aminobenzenesulfonate plus the three vitamins with about the same growth rate as the mixed culture in a mineral medium. When (resting) cells of strain S1 were incubated in a pure mineral medium with sulfanilate, up to 30% of the oxidized sulfanilate accumulated as catechol-4-sulfonate in the culture medium. In contrast, only minor amounts of catechol-4-sulfonate accumulated when strain S1 was grown with 4ABS in the presence of the vitamins.     

Effectiveness of Entomopathogenic Nematodes in an Alginate Gel Formulation Against Lepidopterous Pests

An improved calcium alginate gel formulation was developed and tested as a carrier for entomopathogenic nematodes against Spodoptera littoralis and Helicoverpa armigera larvae. Mortality of 100% was caused in 4th instar larvae of the two insects by feeding them on 1000 infective juveniles (IJ) g -1 of Steinernema carpocapsae (ALL strain) in the gel for 24 h. Exposing 2nd to 5th instars of H. armigera and 3rd to 6th of S. littoralis to 500 IJ g -1 of S. carpocapsae (ALL strain) resulted in 70-100% larval mortality. Mature larvae were less susceptible to the nematodes. Mortality of larvae exposed to 500 IJg -1 of S. carpocapsae (ALL strain) ranged from about 45-55% at 4 h to 90-95% at 48 h. Fourth instar larvae fed for 24 h with 250 IJ g -1 of nematode strains in gel showed in S. littoralis ranges of susceptibility in the following descending order: S. feltiae (IS -7 strain) = S. carpocapsae (DT strain) = S. feltiae (IS-6 strain) > S. carpocapsae (Mexican strain) = S. carpocapsae (ALL strain) = Heterorhabditis bacteriophora (HP-88 strain) = H sp. (IS-5 strain) > S. riobravae (Texas strain); in H. armigera the rating was: S. feltiae (IS-7 strain) = H. bacteriophora (HP88 strain) > S. carpocapsae (ALL strain) = S. feltiae (IS-6 strain ) = Heterorhabditis sp. (IS5 strain) > S. carpocapsae (Mexican strain) > S. riobravae (Texas strain) . The number of nematodes per larval cadaver increased with mortality rates. In greenhouse tests at 28 &#45 2&#176;C and 90% relative humidity, gel discs containing 500 IJ g -1 of nematodes were pinned to leaves of potted plants of cotton ( Gossypium hirsutum ) (Acala SJ2) and the plants were offered to S. littoralis larvae. Larval mortality of 89 &#45 12.7% was caused by S. feltiae (IS-7 strain) and most of the plant leaves were protected against the larvae by the nematodes. In the control, larval mortality was 3.3 &#45 0.05% and the plants were almost completely defoliated. Possibilities of using the gel-nematode formulation to protect sheltered crops against insect pests are discussed     

代表性差异分析比较两株来自不同地区的猪源Lactobacillus菌株

[目的]对分离自猪肠道的乳酸杆菌S1菌株进行鉴定,并比较该菌株与同种的001T菌株的基因差异.[方法]对S1菌株进行16S rRNA基因序列分析和种特异PCR检测,并且对S1菌株和Lactobacillus sobrius 001T进行代表性差异分析(Representational difference analysis,RDA).[结果]16S rRNA基因序列分析表明,与S1菌株最相似的已知菌为L.sobrius.变性梯度凝胶电泳分析显示,仔猪空、回肠细菌图谱中有一与S1菌株有相同迁移位置的优势条带,克隆、测序鉴定表明,与该条带相匹配的16S rRNA基因克隆(Clone S)的最相似已知菌也为L.sobrius.16S rRNA基因系统进化分析表明,S1菌株与Clone S和L.sobrius 16S rRNA基因序列同源性分别为99.8%和99.6%.L.sobrius特异性引物也可以扩增S1株菌的16S rRNA基因的特定片段.因此S1菌株可被确定为Lsobrius.RDA对菌株S1和同种的猪源L.sobrius 001T菌株的基因差异进行分析,未发现这两株菌的基因组差异.[结论]猪肠道乳杆酸菌S1菌株属于L.sobrius,其与猪源L sobrius 001T菌株为相似菌株.     

Cross protection between a strain of Staphylococcus epidermidis and eight other species of coagulase-negative staphylococci

Passive protective activity of rabbit antiserum prepared by a representative capsular type II strain of Staphylococcus epidermidis in mice was absorbed out with homologous capsular type strains of S. simulans, S. cohnii, S. xylosus, S. hominis, S. capitis, S. hyicus, S. haemolyticus, and S. saprophyticus in addition to the homologous strain. The minimum amount of the strains required for absorption differed greatly, depending upon the strain. No absorption of the activity was shown with a strain of capsular type I and III of S. epidermidis, S. simulans, and S. cohnii, and a strain of capsular type III of S. hominis. These results suggest a possible capsular type specificity in the cross protection between strains of S. epidermidis and other species of coagulase-negative staphylococci.     

1株抗根结线虫红树林放线菌的筛选与鉴定

从红树林底泥中分离筛选具有抗线虫活性的放线菌,并对活性菌株进行分类鉴定。采用稀释平板法分离放线菌,采用24孔板液体筛选模型筛选具有抗线虫活性的菌株,并对活性菌株进行形态学和生理生化特征研究,测定其16S rDNA序列并进行系统发育分析。筛选得到1株具有杀线虫活性的菌株,编号为HA11090,该菌株发酵液在稀释20倍和40倍后,抗根结线虫校正死亡率分别为70.5%和65.0%。菌株HA11090的16S rDNA序列与Saccharopolyspora hirsuta subsp.kobensis和Saccharopolyspora jiangxiensis的相似性最高,分别为99.35%和99.27%,三者在系统发育树上聚为一个分支,形态和生理生化特征分析显示,菌株HA11090与Saccharopolyspora jiangxiensis基本一致,而与S.hirsuta subsp.kobensis差异较大。鉴定菌株HA11090为Saccharopolyspora jiangxiensis,其发酵液具有较强的抗根结线虫活性,值得进一步研究。     

宋内氏福氏2a双价志贺氏菌芳香族氨基酸营养缺陷型减毒株的研究Ⅱ:生物学特性的研究

进一步对FS54—2、—3、—4、—5、—6、—7及—7b等双价减毒株的侵袭力、毒力、免疫原性与免疫保护性及遗传稳定性等进行了研究,证明双价减毒株仍保持了入侵肠上皮细胞的能力;小动物实验安全低毒;有很好的免疫原性,对小鼠有很好的双重免疫保护性。     

宋内氏痢疾菌无毒株S7表达Vi抗原工程菌的构建

将含有编码Ｖｉ抗原ＶｉａＢ基因片断的质粒转导进入宋内氏痢疾菌无毒株Ｓ７中,组建了重组菌株Ｓ７Ｖｉ。质粒电泳图谱显示重组菌株Ｓ７Ｖｉ中存在被转入的外源质粒带。重组株的生化特性没有改变。菌体凝集及Ｖｉ抗血清标记的ＳＰＡ菌液凝集反应证明在重组株的菌体表面,同时表达了Ｖｉ抗原和宋内氏毒菌的Ｏ抗原。以５×１０~８ＣＦＵ、１０×１０~８ＣＦＵ的重组株免疫近交系的ＬＩＢＰ小鼠,免疫小鼠对宋内氏毒株Ｓ６３攻击的保护率为６０％至９０％,对伤寒毒株Ｔｙ２攻击的保护率为２５％至４０％。     

A conditional lethal mutant of Escherichia coli which affects the processing of ribosomal RNA.

A temperature-sensitive mutant strain was isolated from an RNase III-(rnc) strain of Escherichia coli. At the permissive temperature it behaves like the parental strain, but at the nonpermissive temperature it fails to produce normal levels of 23 S and 5 S rRNA, while instead the 25 S rRNA species becomes very prominent. (The 25 S molecule appears in rnc cells and contains 23 S rRNA sequences). When an rnc+ mutation was introduced to such a strain, or when the rnc mutation was replaced by an rnc+ allele, the strain remained temperature-sensitive. At the permissive temperature such strains synthesized rRNA like any other E. coli strain, but at the nonpermissive temperature they remained unable to synthesize normal levels of 5 S rRNA, and instead a larger molecule was accumulated. The simplest interpretation of theses findings is that the mutant strain contains a temperature-sensitive processing endoribonuclease, RNase E, which normally introduces a cut in the growing rRNA chain somewhere between the 23 S and the 5 S rRNA cistrons. These findings help also to explain the nature and origin of the various rRNA species observed in RNase III- cells and add to our understanding of processing of ribosomal RNA in normal cells of Escherichia coli.     

棉蚜迁飞型和居留型及其杂交后代飞行特性的比较研究

为了明确棉蚜种群飞行能力发生分化的遗传基础,本文对春季木槿上有翅棉蚜中的起飞个体和不起飞个体分别进行连续4代次和2代次的起飞特性选择。结果表明,棉蚜的起飞比率和起飞角度特性表现出了明显的选择响应,并筛选出了棉蚜的迁飞型(M)和居留型(S)。通过对棉蚜性蚜的诱导,摸索出了棉蚜性蚜交配及卵孵化的条件。对迁飞型(M)、居留型(S)、迁飞型和居留型的正反交后代（M♀×S♂,S♀×M♂）、夏季木槿上的滞留蚜（H）等5类棉蚜的起飞特性进行了比较研究。结果表明,迁飞型和居留型的正反交后代的起飞比率显著高于居留型,且稍高于迁飞型。迁飞型雌蚜与居留型雄蚜的杂交后代的起飞角度显著高于居留型和夏季木槿上的滞留蚜。居留型产生有翅蚜的比率低于迁飞型及其与居留型的正反交后代。棉蚜的起飞角度、起飞比率及有翅蚜产生能力表现出了较强的遗传效应。平均起飞时间虽然表现出迁飞型雌蚜与居留型雄蚜的杂交后代显著短于迁飞型雄蚜与居留型雌蚜的杂交后代及夏季木槿上的滞留蚜,但与迁飞型和居留型之间却无显著差异,表现出棉蚜起飞时间特性的遗传效应不明显。     

Identification of Escherichia coli through analysis of 16S rRNA and 16S-23S rRNA internal transcribed spacer region sequences

A bacterial strain, designated BzDS03 was isolated from water sample, collected from Dal Lake Srinagar. The strain was characterized by using 16S ribosomal RNA gene and 16S-23S rRNA internal transcribed spacer region sequences. Phylogenetic analysis showed that 16S rRNA sequence of the isolate formed a monophyletic clade with genera Escherichia. The closest phylogenetic relative was Escherichia coli with 99% 16S rRNA gene sequence similarity. The result of Ribosomal database project's classifier tool revealed that the strain BzDS03 belongs to genera Escherichia.16S rRNA sequence of isolate was deposited in GenBank with accession number FJ961336. Further analysis of 16S-23S rRNA sequence of isolate confirms that the identified strain BzDS03 be assigned as the type strain of Escherichia coli with 98% 16S-23S rRNA sequence similarity. The GenBank accession number allotted for 16S-23S rRNA intergenic spacer sequence of isolate is FJ961337.