不同酿酒酵母菌株来源异麦芽糖酶IMA1的克隆、表达及表征（英文）

【目的】异麦芽糖酶IMA1在充分利用含有α-1,6-O-糖苷键的低聚糖中起着关键作用。【方法】在本研究中,对来自4株酿酒酵母菌株(包括3株嗜酸性菌株)来源的异麦芽糖酶IMA1进行克隆、表达、纯化和表征。【结果】研究发现,4种异麦芽糖酶IMA1表现出类似的pH和温度依赖性,但表现出不同的动力学参数和热稳定性。IMA1-A对α-MG(α-甲基葡糖苷)表现出最高的结合亲和力、转换数、催化效率和热稳定性。结构和序列分析表明,2个远离活性位点和底物结合位点的氨基酸的差异对异麦芽糖酶IMA1的动力学参数和热稳定性有重要影响。【结论】本研究结果对进一步研究异麦芽糖酶IMA1的结构-功能关系奠定了基础。     

反刍动物产志贺毒素大肠杆菌的分离鉴定和致病潜力分析

产志贺毒素大肠杆菌（Shiga toxin-producing Escherichia coli,STEC）是重要的食源性病原,而STEC往往以正常菌群的形式存在于牛羊等反刍动物肠道。[目的] 本研究对牛羊粪便样品中的STEC分离和鉴定并对分离株进行致病潜力分析。从江苏、云南和河北等地共分离到羊源STEC菌株11株,牛源STEC菌株1株,另新疆农业大学佟盼盼组馈赠牛源菌株10株。[方法] 通过细菌选择培养及特异性基因stx1和stx2的检测进行分离鉴定;并通过Vero细胞毒性试验、溶血活性试验和毒力因子的检测分析STEC分离株的致病潜力。[结果] 分离到羊源分离株11株,分离率17.5%（11/63）;分离得到牛源分离株1株,分离率0.7%（1/134）;11株羊源分离株中有5株对Vero细胞具有强的毒性,3株有溶血活性;11株牛源分离株中有5株对Vero细胞具有强的毒性,3株有溶血活性。11株羊源STEC分离株eae基因携带率为63.6%（7/11）,而11株牛源STEC分离株eae基因携带率仅为9.0%（1/11）。[结论] 结果表明羊源STEC菌株的分离率和致病潜力高于牛源菌株,所以,除牛外,羊作为STEC菌株宿主也应该得到更多的重视。     

2016-2017年宁夏地区牛源金黄色葡萄球菌的全基因组分析

[目的] 了解宁夏地区奶牛乳腺炎金黄色葡萄球菌（Staphylococcus aureus,SA）代表菌株的基因组序列基本特征,进一步探究其耐药基因型、毒力及进化关系,为兽医临床防治提供理论依据。[方法] 采用纸片法对97株金黄色葡萄球菌临床分离株进行抗菌药物敏感性试验,同时进行葡萄球菌蛋白A（Staphylococcus aureus protein A,spa）分型、多位点序列分型（multilocus sequence typing,MLST）,根据分型结果选取16株代表菌株进行全基因组测序,并对获得的测序序列进行处理分析。[结果] 药敏试验结果显示97株分离株对18种抗菌药物存在不同程度的耐药,其中9株耐甲氧西林金黄色葡萄球菌（methicillin-resistantStaphylococcus aureus,MRSA）对青霉素、氨苄西林、苯唑西林、头孢噻呋、磺胺异噁唑、红霉素、庆大霉素和克林霉素等8种抗菌药物完全耐药,甲氧西林敏感金黄色葡萄球菌（methicillin-sensitiveStaphylococcus aureus,MSSA）菌株对青霉素、氨苄西林、磺胺异噁唑耐药率较高。耐药基因数据库（antibiotic resistance genes database,ARDB）注释分析显示16株代表菌株共携带21种耐药基因,其中norA、tet38、bacA、mepA的携带率较高,与药敏试验结果具有一定的相关性。毒力基因数据库（virulence factors of pathogenic bacteria,VFDB）注释分析显示所有菌株携带多种与粘附、宿主免疫逃逸、分泌、胞外酶编码、铁摄取等疾病相关的毒力基因,MRSA菌株均携带较多毒力因子,MSSA菌株携带毒力因子数目不等。基因岛预测结果显示16株代表菌株存在不同数量的基因岛且MRSA菌株携带基因岛数目及毒力基因岛较多,但耐药基因岛数目与MSSA差异不明显。SNP分析结果显示部分分离株同源性较高,同源性较高的两株MRSA的全基因组基本序列特征差异较小,携带的耐药、毒力基因情况相似。[结论] 宁夏地区牛源SA分离株耐药性情况严重且具有较高的毒力水平,本研究为家畜相关MRSA（livestock-associated MRSA,LA-MRSA）与MSSA基因组序列信息的比较分析及宁夏地区SA感染的临床防控提供参考依据。     

从新疆吉木乃地区冬季泌乳双峰驼中培养的细菌多样性及产酸、产酶活效果

[目的]本研究拟通过可培养获得驼源细菌,分析菌种分布规律,获得产酸、产酶特性的菌株资源,为驼源益生细菌的开发和利用提供资源和技术支撑。[方法]采用稀释法筛选新疆吉木乃地区冬季养殖双峰泌乳骆驼驼乳、唾液及直肠粪便中的细菌,通过16S rRNA基因序列分析对菌株进行鉴定;透明圈法对产酸和产酶的菌株进行初筛,并对菌株产有机酸能力及产淀粉酶、纤维素酶和蛋白酶活力进行复筛。[结果]共培养出63株细菌菌株,经分子鉴定得知,从驼粪中分离的35株菌以嗜冷杆菌属和不动杆菌属为主;驼乳中分离出的21株菌以假单胞菌属和明串珠菌属为优势菌属;唾液中分离纯化出7株,主要为芽孢杆菌属;平板初筛得到产酸菌共有11株,1株粪源肠膜明串珠菌(Leuconostoc mesenteroides) 2F11M乳酸产量最高,达到3.93 mg/mL;1株奶源乳酸明串珠菌(Leuconostoc lactis) 2N5M乙酸产量最高,达到12.73 mg/mL;1株粪源蒙氏肠球菌(Enterococcus mundtii) 2F17M丙酸产量最高,达到10.36 mg/mL;根据初筛产透明圈,产酶菌株主要分离自驼粪和驼奶,其中产淀粉酶的菌株17株,产纤维素酶的菌株10株,产蛋白酶的菌株15株;1株奶源的贝莱斯芽孢杆菌(Bacillus velezensis) Nai1的淀粉酶活性最高,为509.07 μg/(min·mL),1株粪源鲁氏不动杆菌(Acinetobacter lwoffii) 2F5N的纤维素酶活性最高,为156.87 μg/(min·mL),1株奶源枯草芽孢杆菌(Bacillus subtilis) 2N2N产蛋白酶活性最高,为3.59 μmol/(min·mL)。[结论]从新疆泌乳双峰骆驼中筛选出了多种产酸、产酶菌株,且活力都较好,具有制备微生态制剂的潜力。     

精氨酸代谢调控蛋白ArgR调控嗜热链球菌胞外多糖合成

[目的] 研究精氨酸代谢调控蛋白ArgR对嗜热链球菌胞外多糖（EPS）合成的调控作用。[方法] 利用大肠杆菌异源表达嗜热链球菌ArgR蛋白,通过尿素变性-复性和Ni²⁺亲和层析纯化。采用凝胶电泳迁移（EMSA）和生物膜层干涉（BLI）分析ArgR和eps基因簇中P_epsA启动子的相互作用和动力学信息。构建过表达和弱化argR基因菌株,利用苯酚-硫酸法测定其合成EPS差异。[结果] 大肠杆菌异源表达的ArgR为包涵体,使用尿素变性-复性纯化可获得2.95 mg/mL可溶性蛋白;EMSA和BLI结果显示ArgR和启动子P_epsA有特异性结合,且结合因解离水平低而稳定;过表达argR基因可显著降低嗜热链球菌EPS合成,而弱化argR基因则提高EPS合成。[结论] 本研究表明ArgR能特异性结合嗜热链球菌eps基因簇启动子,并负调控EPS生物合成。     

苯唑西林敏感、mecA基因阳性的猪源金黄色葡萄球菌的流行、分子分型及耐药性研究

[目的] 为了解我国猪源苯唑西林敏感-mecA阳性金黄色葡萄球菌（Oxacillin-susceptible,mecA-positive Staphylococcus aureus,OS-MRSA）的流行情况、菌株分子特征及耐药性,本研究对我国中西部4个省份（甘肃、陕西、河南和广西）的9个规模化养猪场进行鼻腔拭子样本采集。[方法] 运用PCR扩增nuc和mecA基因及苯唑西林耐药性检测对OS-MRSA菌株进行分离鉴定。然后对分离所得的OS-MRSA菌株进行26种毒素编码基因、16种抗生素耐药性以及spa、MLST和SCCmec分型检测。[结果] 结果表明,采集的884份样本中,67份样本7.6%（67/884）分离到金黄色葡萄球菌,包括50株甲氧西林敏感菌株（Methicillin-sensitive Staphylococcus aureus,MSSA）、8株苯唑西林耐受-mecA阳性金黄色葡萄球菌（Oxacillin-resistant mecA-positive,OR-MRSA）和9株OS-MRSA菌株。26种被检毒素编码基因中有9种毒素编码基因被检出,其中hla基因检出率最高,其次为hld、hlb、hlg、sei、sem、seg、sen和seo。此外,67株分离株中仅有16株携带肠毒素编码基因,其中OR-MRSA和OS-MRSA菌株分别占37.5%（6/16）和50.0%（8/16）,且携带毒素编码基因的菌株克隆型均为ST9-t899。16种所测试抗生素中,菌株对12种抗生素表现为耐药,其中MSSA、OR-MRSA和OS-MRSA分离株分别主要对1-8、10-12和7-11种抗生素耐药。所有分离株共有4种克隆型ST398-t571、ST9-t899、ST398-t034和t11241,其中ST9-t899为MRSA菌株唯一克隆型和ST398-t571为MSSA优势克隆型。除4株分离株未检测到SCCmec分型外,IV_b（76.5%,13/17）是MRSA分离株的唯一分型。[结论] 结果表明,我国猪源MRSA分离株对苯唑西林药物敏感性发生了改变,出现了较多的苯唑西林敏感菌株。此外,MSSA和MRSA分离株优势克隆型分别为ST398-t571和ST9-IVb-t899。研究还发现,克隆型与毒素编码基因有显著相关性,携带毒素编码基因的菌株克隆型均为ST9-t899。通过了解我国猪源MSSA、OR-MRSA和OS-MRSA的流行、分子特征和耐药性,可以为我国猪源金黄色葡萄球菌的防控提供数据支持。     

基于生态风险评估探究溶藻细菌2-4的抑藻特性

[目的]基于生态安全评估,探索溶藻细菌及其分泌物的抑藻效果与机制。[方法]分离获得溶藻细菌2-4 (Pseudomonassp.),通过正交试验和动力学模型研究了菌株2-4及其分泌物的抑藻特性,并通过急性毒性试验对菌株2-4及其分泌物进行生态风险评估。[结果]菌株2-4在最优条件下接种体积比(V/V)为15%时对铜绿微囊藻的4 d抑制率达92.81%,抑藻反应符合一级动力学模型(t_1/2=126 h)。菌株2-4具有抑藻多样性,首次报道了假单胞菌对斜生四链藻、蛋白核小球藻和丝藻的抑藻效果。菌株2-4分泌的抑藻活性物质分子量小于500 Da,不耐高温和强酸强碱。急性毒性试验结果显示,高于1.5%的菌液对大型溞有毒,高于2%的菌株分泌物对大型溞和稀有鮈鲫有毒,但对明亮发光杆菌无毒。较安全的使用范围内,菌株2-4 (V/V=1.5%)及其分泌物(V/V=10%)对藻华湖泊水样的叶绿素a去除率为4.83%–42.94%和30.62%–68.69%。[结论]本研究客观分析了假单胞菌2-4及其分泌物在生态安全使用范围内的实际抑藻效果,明晰了溶藻细菌生态毒性与抑藻效率的关系,为生物控藻实践提供理论参考。     

天山大峡谷原始森林黏细菌的分离鉴定及其抗菌活性

【背景】黏细菌是一类具有多细胞群体行为特征的高等原核生物,其对植物病原真菌和细菌的捕食特性使其在植物病害防治方面具有重要的应用潜力。【目的】探究乌鲁木齐天山大峡谷原始森林可培养黏细菌的多样性并分析其抗菌活性,为发掘黏细菌生防菌株奠定基础。【方法】以天山大峡谷原始森林采集的土样和腐木为分离材料,采用兔粪诱导法和被捕食菌诱导法从中分离纯化黏细菌菌株,结合形态学观察、生理生化测定和16S rRNA基因序列分析确定其分类地位,并以6种植物病原真菌[大丽轮枝菌(Verticillium dahliae)、尖孢镰刀菌萎蔫专化型(Fusarium oxysporum f. sp. vasinfectum)、拟轮枝链孢霉(Fusarium verticillioides)、立枯丝核菌(Rhizoctonia solani)、黄色镰刀菌(Fusarium culmorum)、细极链格孢菌(Alternaria tenuissima)]和1种植物病原细菌[梨火疫病菌(Erwinia amylovora)]为靶标菌,通过平板对峙法和菌苔捕食法测定其抗菌活性。【结果】从采集的样品中分离出70株菌株,经纯化后获得36株黏细菌纯培养物。经鉴定隶属于4个属,黏球菌属(Myxococcus) 30株、孢囊杆菌属(Cystobacter) 3株、珊瑚球菌属(Corallococcus) 2株和原囊菌属(Archangium) 1株。抗菌活性分析显示,本研究获得的36株黏细菌至少对2种植物病原真菌有抗菌活性,表现出广谱的抗真菌活性,初步筛选出一株菌株NSE37-1兼具广谱和高效抗真菌活性;供试的15株黏细菌对梨火疫病菌均具有捕食活性,初步筛选出一株对梨火疫病菌具有较强捕食能力的黏细菌菌株NSE25。【结论】天山大峡谷可培养黏细菌资源比较丰富,黏球菌属是该地区可培养黏细菌菌群中的优势菌。分离纯化出的黏细菌菌株均表现出广谱的抗植物病原菌活性,具有进一步研究和开发的潜在价值。     

嗜水气单胞菌ntrC调控的生理功能及其机制研究

[目的] NtrC是一种与DNA结合的转录调控因子,在激活氮同化基因的转录和维持氮源供应中具有重要作用,本研究拟探究其对嗜水气单胞菌生理功能的影响及其作用机理。[方法] 本研究采用同源重组方法构建了嗜水气单胞菌ATCC 7966 ntrC的缺失株,并以野生株为对照,对缺失株的生理表型进行测定和分析,利用定量蛋白质组学技术比较野生株和ntrC缺失株的蛋白表达差异。[结果] 发现敲除ntrC基因后,嗜水气单胞菌在缺氮、渗透压、重金属离子、氧化以及不同抗生素胁迫下的耐受性都发生显著变化,且这些表型在其补救菌株中均能得到恢复。定量蛋白质组学分析发现,野生株和ntrC缺失株的差异表达蛋白可能参与氨基酸生物合成、抗坏血酸和醛糖酸盐等代谢通路的调控。[结论] 本研究阐明了ntrC在嗜水气单胞菌中的重要作用及其对细菌生物学功能的影响,探讨了ntrC直接或间接调控的蛋白与生理表型之间的联系,研究结果可为未来水产致病菌的防治提供理论支持。     

金针菇交配型B第三个亚位点的发现与遗传

本研究在分析金针菇减数分裂重组热点时发现B交配型位点可能存在第三个亚位点,为此对同核菌株‘6-3’和‘6-21’的B交配型位点进行全基因组注释,获得8对同源信息素受体基因（pheromone receptor gene,pr）和3对同源信息素前体基因（pheromone precursor gene,pp）,组成了2个结构完整、相距326kb的亚位点α和β;‘6-21’菌株还发现1个‘6-3’菌株不具有的pr,组成第3个亚位点γ,它与亚位点α和β没有连锁。‘6-3’和‘6-21’配对形成异核体后出菇,分离获得142个单孢菌株,其B交配型有8种基因型,通过亲和性试验得到证实,同时还验证了第3个亚位点γ具有与α、β相同的功能。     

Systematic significance of mature embryo of bamboos

The mature embryo of seven species belonging to five genera of Indian bamboos is described. In all these the basic pattern of embryo organisation is same: the scutellar and coleoptilar bundles are not separated by an internode, the epiblast is absent, the lower portion of the scutellum and the coleorhiza are separated by a cleft and the margins of embryonic leaves overlap. The features unique to fleshy fruited bamboos are: presence of a massive scutellum, the juxtaposition of plumule and radicle and the occurrence of a bud in the axil of the coleoptile. The fleshy fruit bearing bamboos should be classified into one group, the tribeMelocanneae. Evidence is provided to recognise additional groups in the subfamilyBambusoideae.     

Taxonomic status of the species of the green algal genusNeochloris

Komárek has recently reviewed the various species assigned to the green algal genusNeochloris Starr (Chlorococcales, Chlorococcaceae) and removed those with uninucleate vegetative cells to a new genus,Ettlia. Watanabe & Floyd, unaware ofKomárek's work, also reviewed the species ofNeochloris and distributed them among three genera—Neochloris, Chlorococcopsis gen. nov., andParietochloris gen. nov.—on the basis of details of the covering of the zoospore and the arrangement of the basal bodies of the flagellar apparatus. This paper reconciles these two treatments and makes additional recommendations at the ranks of genus, family, order, and class.     

Phylogeny of cardinalfishes (Teleostei: Gobiiformes: Apogonidae) and the evolution of visceral bioluminescence

The cardinalfishes (Apogonidae) are a diverse clade of small, mostly reef-dwelling fishes, for which a variety of morphological data have not yielded a consistent phylogeny. We use DNA sequence to hypothesize phylogenetic relationships within Apogonidae and among apogonids and other acanthomorph families, to examine patterns of evolution including the distribution of a visceral bioluminescence system. In conformance with previous studies, Apogonidae is placed in a clade with Pempheridae, Kurtidae, Leiognathidae, and Gobioidei. The apogonid genus Pseudamia is recovered outside the remainder of the family, not as sister to the superficially similar genus Gymnapogon. Species sampled from the Caribbean and Western Atlantic (Phaeoptyx, Astrapogon, and some Apogon species) form a clade, as do the larger-bodied Glossamia and Cheilodipterus. Incidence of visceral bioluminescence is found scattered throughout the phylogeny, independently for each group in which it is present. Examination of the fine structure of the visceral bioluminescence system through histology shows that light organs exhibit a range of morphologies, with some composed of complex masses of tubules (Siphamia, Pempheris, Parapriacanthus) and others lacking tubules but containing chambers formed by folds of the visceral epithelium (Acropoma, Archamia, Jaydia, and Rhabdamia). Light organs in Siphamia, Acropoma, Pempheris and Parapriacanthus are distinct from but connected to the gut; those in Archamia, Jaydia, and Rhabdamia are simply portions of the intestinal tract, and are little differentiated from the surrounding tissues. The presence or absence of symbiotic luminescent bacteria does not correlate with light organ structure; the tubular light organs of Siphamia and chambered tubes of Acropoma house bacteria, those in Pempheridae and the other Apogonidae do not.     

Apoptotic Regulation of Caspase Activity

Intracellular cysteine aspartate-specific proteases (caspases) play both signaling and effector roles in realizing the program of cell death. Caspases function as proteolytic cascades unique for each cell type and signal triggering apoptosis. All parts of the proteolytic cascades are duplicated and controlled by feedback signals. Amplification cycles between pairs of caspases (the third and the sixth, the ninth and the third, the twelfth and the sixth, and others) help multiply the initial apoptotic signal. The presence of physiological inhibitors of apoptosis that directly interact with caspases creates a multilevel regulatory network of apoptosis in cell. The caspase proteolytic cascades are also regulated by sphingolipid secondary messengers, among them ceramide, sphingosine, and their phosphates. Moreover, an association of the caspase signaling with ubiquitin-dependent proteolysis is shown in cells. In particular, the use of extracellular activators and inhibitors of caspases allows irreversible activation of apoptosis in tumor cells or the prevention of apoptosis in cortical neurons under neurodegenerative diseases.     

Analysis of twin of eyeless regulation during early embryogenesis in Drosophila melanogaster

The Drosophila Pax6 homolog twin of eyeless (toy) is so far the first zygotically expressed gene involved in eye morphogenesis in Drosophila. The study of its expression during embryogenesis is therefore informative of the initial events of eye development in Drosophila. We have analyzed how the initial expression domain of toy at cellular blastoderm is regulated. We show that the three maternal patterning systems active in the cephalic region (the anterior, terminal and dorsal-ventral systems) cooperate with zygotically activated gap genes to shape the initial expression domain of toy. Whereas Bicoid, Dorsal and Torso signaling synergistically act as activators, Hunchback, Knirps and Decapentaplegic act as repressors.     

New species of Peruvian Orchidaceae III

Sixteen new species are proposed in the generaAckermania, Dressleria, Epidendrum, Maxillaria, Oncidium, Rodriguezia, Sigmatostalix, andTrigonidium. All new species are illustrated.Maxillaria vittariifolia L. O. Williams is newly recorded for Peru. A key is provided forTrigonidium of Peru.Trigonidium loretoense Schltr. andT. peruvianum Schltr. are lectotypified.     

Host plants of someRhinanthoideae (Scrophulariaceae) of eastern North America

Twenty three species in 11 genera were examined in the field to determine hosts. OnlyStriga asiatica andSeymeria cassioides have a narrow host range being restricted to grasses and pines, respectively. These are the only species which cause pronounced and sometimes serious host damage. The other species attach to a great diversity of hosts.     

广西青藓科分类学修订

通过对采自广西24个县(市)的1 147份青藓科植物标本的逐一鉴定及相关文献的查阅,确认有广西青藓科植物11属、44种,其中包括广西青藓科植物新记录属1属,即拟异叶藓属(Pseudokindbergia),新记录种7种,分别为匐枝青藓(Brachythecium procumbens)、阔叶尖喙藓(Oxyrrhynchium latifolium)、泛生尖喙藓(O. vagans)、拟异叶藓(Pseudokindbergia dumosa)、华东细喙藓(Rhynchostegiella sinensis)、长肋拟青藓(Sciurohypnum populeum)和弯叶拟青藓(S. reflexum)。该文提供了修订后的广西青藓科植物名录,并对其中的新记录属、种的主要形态学识别特征、生境和地理分布等进行了详细描述。     

In vitro appraisal of the probiotic value of intestinal lactobacilli

Lactobacilli play a distinctive role in the microbial balance of the chicken gut. In experiments simulating the chicken crop, the antagonism of lactobacilli against Enterobacteriaceae and Salmonella typhimurium was demonstrated and was attributed to lactic acid production. Moreover, adhesion to the crop epithelium was a common characteristic of intestinal lactobacilli. As opposed to salmonellas, lactobacilli were sensitive to deconjugated bile salts at 2.5mm. This sensitivity could lower their chance of proliferation in the small bowel of the chicken tract.The authors are at the Laboratory of Microbial Ecology, Faculty of Agricultural Sciences, State University of Gent, Coupure L., 653, B-9000 Gent, Belgium.