γ-亚麻酸产生菌Mucor sp．EIM-10的筛选及分子鉴定

为了获得高产γ-亚麻酸（γ-linolenic acid,GLA）的菌株,利用苏丹黑染色法筛选获得1株GLA产生菌EIM-10,通过摇瓶培养,其生物量可达11．882g／L,菌丝体油脂含量可达18．86％。气相色谱-质谱（GC—MS）分析表明其γ-亚麻酸质量分数（占总脂肪酸）高达27．68％。为进一步鉴定该菌株,克隆测定了该菌18SrRNA基因序列,并对其进行系统进化树分析,结果表明该菌属于毛霉属,与Mucor racemosus、Mucor plumbeus、Mucor ramosissimus与Mucor circinelloides同属一个分支。     

总枝状毛霉对去氢表雄酮的生物转化研究

从土样中筛选到1株能转化甾体的菌株,经形态观察,鉴定为总枝状毛霉(Mucor racemosus)。利用该菌株对去氢表雄酮进行生物转化研究,转化产物分离后,通过红外、质谱和二维核磁波谱分析,确定了化学结构。研究结果表明转化产物是7α-羟基去氢表雄酮和7β-羟基去氢表雄酮。     

尘螨变应原Der f1全序列的原核表达及生物信息学分析

目的 构建尘螨变应原Der f1原核表达体系,并了解其分子特征.方法 提取粉尘螨总RNA,用RT-PCR合成Der f1编码基因,将其克隆至pMD19-T载体,亚克隆至表达载体pET-28a( ),转化至大肠杆菌并用IPTG诱导表达.用生物信息学软件对测序结果进行分析并预测其空间结构.结果 从粉尘螨总RNA中扩增获得Der f1 cDNA片段,成功构建了表达质粒pET-28a( )-Der f1,Western blotting显示原核表达获得成功.测序结果提交GenBank,登陆号为EU095368,该基因长966 bp,与参考序列同源性达99.9%,推测其编码氨基酸321个,属疏水蛋白,位于细胞外,信号肽位于1～18氨基酸处.同源性分析提示Der f1和Eur m1相似率为88%,而Der f1和Der p1的相似率为77%,分子进化树中粉尘螨和梅氏嗜霉螨聚成一簇.Der f1的二级结构由α-螺旋(109 aa,33.96%)、延伸链(55 aa,17.13%)、β-转角(18 aa,5.61%) 和随机卷曲(139 aa,43.30%)组成.结论 尘螨变应原Der f1原核表达获得成功,为进一步生产重组变应原奠定了基础.生物信息学分析表明粉尘螨和梅氏嗜霉螨的亲缘关系可能更近,而与屋尘螨关系稍远,此与现行的形态学分类系统并不符合.     

氧化亚铁硫杆菌的诱变机理研究

目的：通过物理诱变提高氧化亚铁硫杆菌（T.f）的活性。方法：利用紫外线对高氧化亚铁硫杆菌进行诱变,对比紫外线辐照120s、240s和300s后培养的氧化亚铁硫杆菌的活性,得到最佳的辐照时间。结果：紫外线诱变氧化亚铁硫杆菌的最佳时间为240s,超过该值后T.f菌很难培养。结论：经过240s的紫外线辐照后,利用Leathen培养基培养的氧化亚铁硫杆菌与对照组相比,浓度为对照组的1.21倍,pH值低0.28,氧化还原电位的绝对值比值为1.13,在同样时间条件下,培养液中Fe2＋浓度的曲线变化斜率分别为0.683和0.236。     

尘螨变应原Der f1 真核表达载体的构建及转染CHO 细胞

目的:构建尘螨变应原Der f1真核表达载体,转染真核细胞并进行蛋白表达.方法:根据Genebank中Der f1基因的核酸序列(AB034946),设计引物,采用PCR法,从保存的JM109工程菌中扩增Der f1编码基因,克隆到真核表达质粒pcDNA3.1/myc-hisA上,以脂质体法转染CHO细胞,经G418筛选,进行稳定表达细胞株的筛选和鉴定.结果:将目的基因Der f1成功连接到pcDNA3.1/myc-hisA-Derf1并转染CHO细胞,获得稳定表达的CHO细胞株.结论:成功构建了尘螨变应原Der f1真核表达载体,并转染CHO细胞表达蛋白质.     

遗址木构件有害真菌的快速测定方法

介绍了1种快速测定遗址木构件有害真菌的方法:用孢子菌丝悬浮液直接接种于供试木块上,培养4d,即可确定其侵染性;培养8d,即可确定其侵染力。应用该方法成功地测定了灰绿曲霉(Aspergillus glau-cus)、黑曲霉(Aspergillus niger)、枝孢霉(Cladosporiumsp.)、顶青霉(Penicillium corylophilum)、柑桔青霉(Penicilliumcitrinum)、团青霉(Penicillium commune)、黄曲霉(Aspergillus flavus)、微紫青霉(Penicilliumjanthinellum)、总状毛霉(Mucor racemosus)、绿木霉(Trichoderma viride)等10种真菌的侵染性和侵染力。     

纳木措春季水体可培养丝状真菌优势种生态位分析

为探明纳木措春季水体可培养丝状真菌的群落特征,研究在分离纯化丝状真菌的基础上,结合经典分类法、rRNA-ITS转录间隔区测序法确定丝状真菌的群落组成及丰度;同时对水体理化因子进行分析,揭示丝状真菌优势类群生态分化与理化因子的相关性。结果在纳木措水体中共分离纯化出921株丝状真菌,归为20属62种,真菌资源较为丰富。其中,主要优势种(Y>0.02)包括:普通青霉Penicillium commune、酒色青霉Penicillium vinaceum、波兰青霉Penicillium polonicum、青霉菌Penicillium goetzii、灰玫瑰青霉Penicillium griseoroseum、冻土毛霉Mucor hiemalis、总状毛霉Mucor racemosus、格孢腔菌Pleosporales sp.2和壳青霉Penicillium crustosum。优势种生态位指数显示,优势种间生态响应速率之和为负数,低度重叠的种对占比较大(56.94%),总体上呈正关联,但未达到显著水平(χ²>3.841),表明群落正处于衰退阶段。优势种间竞争...     

Developmental changes in functional expression and β-adrenergic regulation of If in the heart of mouse embryo

The hyperpolarization-activated current (If) plays an important role in determining the spontaneousrate of cardiac pacemaker cells. The automatic rhythmicity also exists in working cells of embryonic heart,therefore we studied developmental changes in functional expression and β-adrenergic regulation of If inembryonic mouse heart. The expression of If is high in early developmental stage (EDS) (10.5 d after coitus)ventricular myocytes, low in intermediate developmental stage (IDS) (13.5 d) atrial or ventricular myocytesand even lower in late developmental stage (LDS) (16.5 d) atrial or ventricular myocytes, indicating thatthese cells of the EDS embryonic heart have some properties of pacemaker cells. β-adrenergic agonistisoproterenol (ISO) stimulates If in LDS but not in EDS cardiomyocytes, indicating that theβ-adrenergicregulation of If is not mature in EDS embryonic heart. But forskolin (a direct activator of adenylate cyclase)and 8-Br-cAMP (a membrane-permeable analogue of cAMP) increase the amplitude of If in EDS cells,indicating that adenylate cyclase and cAMP function fairly well at early stage of development. Furthermore,the results demonstrate that If is modulated by phosphorylation via cAMP dependent PKA both in EDSand LDS cells.     

半夏内生真菌的初步研究

从健康无病害的半夏(Pinellia ternate)根、茎、叶和花组织中分离获得内生真菌共计61株,对其进行形态学鉴定后,针对不产孢的菌株,测定了ITSrDNA序列以进行分子鉴定。分离到的内生真菌分别来自11个属,以半知菌为主要群落,其中Hyphomycetes,Zygomycetes和Coelomycetes的比例分别为62.2%,18.1%和8.2%;镰刀菌属(Fusarium spp.)是半夏植物中分离率最高(6%)的属,其余主要优势属为Alternaria,Mucor,Epicoccum,Mortierella和Plectosphaerella。研究表明:从半夏球茎组织中分离到的内生真菌数量多于其他组织。     

西藏拉鲁湿地水体冬季可培养丝状真菌多样性及其与理化因子的关系

【背景】拉鲁湿地是我国海拔最高、面积最大的城市天然湿地。开展拉鲁湿地微生物群落结构分析,可为青藏高原生物资源的开发利用和保护提供理论依据,并为湿地生态系统的微生物多样性研究奠定基础。【目的】开展拉鲁湿地水体丝状真菌的多样性研究,探究影响该环境丝状真菌群落分布特征的主要理化因子。【方法】从拉鲁湿地11个水样中分离丝状真菌,应用nr DNA ITS序列分析,并结合经典分类学方法对获得的菌株进行鉴定,运用SPSS和CANOCO软件分析丝状真菌群落结构及其与环境因子的关系。【结果】从拉鲁湿地水体分离的丝状真菌菌株分属于6属13种。拉鲁湿地冬季水体丝状真菌优势属为Mucor、Cladosporium和Galactomyces,优势种为M.hiemalis、M.racemosus和G.geotrichum。总氮与拉鲁湿地水体丝状真菌数量呈显著负相关(P0.05),总氮和总磷对于丝状真菌的分布具有较明显的影响,这2种理化因子与M.racemosus的多度呈正相关性,而与M.hiemalis呈较强的负相关性。【结论】环境因子是影响高原湿地生态系统微生物群落结构的主要因素之一,探讨西藏典型脆弱生态环境中微生物多样性与环境因子的关系具有重要意义。     

Synthesis of 7alpha-hydroxy-dehydroepiandrosterone and 7beta-hydroxy-dehydroepiandrosterone

The fermentation of dehydroepiandrosterone synthesized from the starting material diosgenin using Mucor racemosus produced 7alpha-hydroxy-dehydroepiandrosterone and 7beta-hydroxy-dehydroepiandrosterone. The bioactivity of the microbial metabolites is also discussed. The species M. racemosus was isolated by screening among stains from soil samples collected from various parts of China.     

Einfluß von Epicoccum nigrum,Alternaria alternata,Mucor racemosus und Ulocladium chartarum auf die Protozoenpopulation im Pansen des Rindes (in vitro)

The influence of moulded hay (Alternaria alternata, Epicoccum nigrum, Mucor racemosus, Ulocladium chartarum) and the efficiency of Vitamin B1 substitution to cope these effects on rumen protozoa was investigated using the longterm rumen simulation technique (RUSITEC) for about 25 days. Moulded hay affected medium-sized protozoa to a different extent (Alternaria alternata: ?16 %, Epicoccum nigrum: ?27 %, Mucor racemosus: ?9 %, Ulocladium chartarum: +2 %). The vitamin B1 substitution had positive effects during the feeding of Mucor racemosus and Ulocladium chartarum.     

Detection of extracellular protease in Mucor species

The fungi are characterized by their abilities to produce and secrete enzymes to the external environment. The species of genus Mucor are a group of fungal microbes with important biotechnological potential, which are responsible for production of industrial enzymes. This work evaluated the ability of protease production in twelve species of genus Mucor. The strains were kept for 120 h under the incubation temperature of 28 degrees C on a shaker at 120 rpm. The detection of proteolytic activity was evaluated in all species, the higher activity was detected in Mucor racemosus Fres. f. chibinensis (Neophytova) Schipper.     

大赖草总DNA转化小麦的分子证据

用来自大麦组的４个高度重复序列克隆了ｐＨｖ７１６１,ｐＨｖ７１７８９,ｐＨｖ７１９１、ｐＨｖ７２９３,经地高辛和同位素２种方法标记后作为探针,对新疆大赖草（供体）、春麦７６１（受体）以及用大赖草总ＤＮＡ通过花粉管通道转化成功的大穗转化株基因组在高度严谨条件下进行了分子杂交。结果表明,这４个探针可以探查出基因内一种具有主串产重复单位的散在重复序列。比较受共体和转化体的杂交图谱,发现在转化株中出现了     

利用花粉辐射诱发普通小麦与大赖草染色体易位的研究

将小麦－大赖草Ｌｒ．２和Ｌｒ．１４染色体二体异附加系９４Ｇ１５和９４Ｇ４５即将成熟花粉经＾６０Ｃｏ－γ射线辐射处理,然后分别给普通小麦品种扬麦５号和绵阳１１授粉。辐射Ｍ１的ＰＭＣ ＭＩ期染色体Ｇｉｅｍｓａ Ｃ－分带和基因组ＤＮＡ荧光原位杂交处理后进行染色体配对分析,从１７株Ｍ１单株中筛选到５株ＰＭＣ ＭＩ期大赖草染色体与小麦配对的个体。根据这５株单株自交Ｍ２种子根尖细胞有丝分裂中期染色体Ｃ－分带和     

变孢毛霉的一个新变种及对Mucor luteus Linnemann和M.variosporus Schipper的合格化(英文)

根据国际植物命名法规,对两个不合格发表的毛霉种名：Ｍｕｃｏｒ　ｌｕｔｅｕｓ　Ｌｉｎｎｅｍａｎｎ和ＭｕｃｏｒｖａｒｉｏｓｐｏｒｕｓＳｃｈｉｐｐｅｒ进行了合格化。对变孢毛霉（Ｍｉｃｏｒｖａｒｉｏｓｐｏｒｕｓ）的一个新变种进行了描述。     

Zygomycota associated with traditional meju, a fermented soybean starting material for soy sauce and soybean paste

Various zygomycota species were detected during a study of the mycobiota of meju, a brick of dried fermented soybeans, used in Korean cuisine. Two hundred and sixty-eight strains were isolated from 98 finished meju products collected in various regions of Korea from 2009 to 2011, and 96 strains were isolated from in-process meju on various farms from 2010 to 2011. The isolated zygomycota were identified using phenotypic characteristics combined with DNA sequences of the internal transcribed spacer regions of ribosomal DNA and the D1/D2 nuclear ribosomal large subunit. Of 364 zygomycota strains, 108 were identified as Mucor circinelloides, 96 as M. racemosus, 60 as Lichtheimia ramosa, 22 as Rhizopus stolonifer, 16 as Lichtheimia corymbifera, and the other 62 strains comprised 10 other species. The psychrotrophic species, Mucor circinelloides and M. racemosus were predominantly present during low temperature fermentation (LTF) and the thermotolerant species Lichtheimia ramosa and Rhizomucor species were predominant during high temperature fermentation (HTF). The results suggest that temperature has a large influence on the zygomycota composition during the fermentation process of meju.     

应用反向线点杂交技术鉴定临床常见曲霉属和毛霉目真菌

目的应用反向线点杂交技术(reverse line blot hybridization,RLB)快速鉴定临床常见的曲霉属和毛霉目真菌。方法收集我院真菌和真菌病研究中心保存的5种曲霉菌(烟曲霉、黄曲霉、黑曲霉、土曲霉、构巢曲霉)和7种毛霉目真菌(冻土毛霉菌、总状毛霉菌、卷枝毛霉菌、少根根霉、小孢根霉、微小根毛霉、伞状犁头霉),共计98株菌株。利用真菌通用引物ITS1和ITS4对菌株进行PCR扩增,用12个真菌种特异性探针与扩增后产物进行反向线点杂交。将RLB结果与真菌传统形态学鉴定结果、ITS区DNA测序结果进行比较。结果 RLB可以正确鉴定98株实验菌株,与形态学方法和ITS区测序方法鉴定结果100%一致,种特异性探针之间未见交叉杂交,显示出该方法的高度敏感性和特异性。8株阴性对照菌株(白念珠菌、茄病镰刀菌、尖端赛多孢、马尔尼菲青霉、疣状瓶霉、棒曲霉、日本曲霉以及雅致小克银汉霉),使用RLB方法无法鉴定。通过烟曲霉基因组DNA浓度10倍倍比稀释法验证RLB的敏感性为1.8×10-3 ng/μL。结论 RLB技术为实验室早期快速诊断、鉴定临床常见的曲霉属和毛霉目真菌提供参考。     

Physiological control of phosphorylation ribosomal protein S6 in Mucor racemosus.

The level of phosphorylation of ribosomal protein S6 increased with accelerating rates of growth and protein synthesis in Mucor racemosus. Lowered levels of phosphorylation were seen under conditions of metabolic shift-down or the onset of stationary phase, and no phosphorylation was detected in sporangiospores. Changing metabolic states, changing intracellular levels of adenosine triphosphatase, and the level of phosphorylation of protein S6 were correlated in M. racemosus.