产朊假丝酵母细胞壁33 ku蛋白的功能研究

通过胰蛋白酶和枯草杆菌蛋白酶对产朊假丝酵母Candida utilis细胞壁的酶解,发现一种分子质量为33 ku的酵母细胞壁主要结构蛋白. 研究显示,在细胞壁上这种蛋白质与细胞壁绝大多数蛋白质成分不同, 它不被胰蛋白酶水解,但对枯草杆菌蛋白酶的作用敏感.33 ku蛋白存在于酵母菌整个对数生长期的细胞壁中,特别是在对数早期细胞壁中,它是唯一的对胰蛋白酶作用不敏感的蛋白质成分.实验证明,该蛋白质对维系酵母细胞壁骨架成分葡聚糖的相互连接和细胞壁的完整结构,具有重要作用,是一种重要的酵母细胞壁嵌合蛋白.     

Analysis of Candida utilis genomic DNA,homologous to cDNA of chicken A1 (1) collagen gene

Genomic fragments, homologous to chicken A1(1) collagen cDNA encoding triple-helical domain, were revealed by Southern analysis in various fungi. Such a genomic fragment from Candida utilis was cloned and sequenced. Analysis of the obtained DNA sequence revealed the 119 bp segment, which has possibly originated from the 54bp module common for the fibrillar collagen genes of higher eukaryotes.     

Differential location of regulatory and nonregulatory trehalases inCandida utilis cells

The isolation of vacuoles by density gradient centrifugation of protoplast lysates fromCandida utilis cells showed a high specific activity for nonregulatory trehalase in vacuoles whereas the regulatory trehalase activatable by phosphorylation behaves as a cytoplasmic enzyme. The vacuolar trehalase is a glycoprotein that can be precipitated by Con A-Sepharose. Treatment of this enzyme with endo H reduced its reactivity with the lectin without loss of enzyme activity and decreased its apparent molecular weight by gel filtration.Abbreviations cAMP adenosine 3, 5-cyclic monophosphate - Con A Concanavalin A - CP buffer 10mM sodium citrate-phosphate pH 6.8 - endo H endo--N-acetyl glucosaminidase H - PMSF phenyl-methyl-sulphonylfluoride - PNPG p-nitrophenyl--glucoside - PNPP p-nitrophenyl-phosphate     

Continuous cultivation of the yeast Candida utilis at different dilution rates on pineapple cannery effluent

Candida utilis was grown on a pineapple cannery effluent in a chemostat at dilution rates ranging between 0.05 and 0.65 h^–1 to establish optimal conditions for biomass production and chemical oxygen demand (COD) reduction. Sucrose, fructose and glucose were the main sugars in the effluent. Maximum value for cell yield coefficient and productivity were (0.686, g_x/g_s) and (2.96, g_x/l/h) at a dilution rate of 0.425 and 0.475 h^–1, respectively, while maximum COD reduction (98%) was attained at a dilution rate of 0.1 h^–1. The maintenance coefficient attained a value of (0.093, g_s/g_x/h). An increase in dilution rate produced a higher protein content of the biomass.     

Rice ubiquitin promoters: deletion analysis and potential usefulness in plant transformation systems

Strong constitutive promoters form a cornerstone for basic and applied research using transgenic plants. GUS (beta-glucuronidase) expression levels from constructs containing RUBQ1 or RUB2 rice ubiquitin promoters were 8- to 35-fold higher in transgenic rice [Oryza sativa (L.)] plants, respectively, when compared to the 35S promoter. Deletion analysis of the 5'-upstream region of RUBQ2 revealed a putative enhancer region that produced a 2.4-fold increase in transient GUS expression. Southern blot analysis showed that three to seven copies of the GUS gene were stably inserted into R0 and R1 plants and inherited in a monogenic fashion.     

Growth of Candida utilis on ethanol and isopropanol

Candida utilis grew on ehtanol and an ethanol-isopropanol-water (22:2:1 vols) mixture but not on isopropanol alone. Acetone accumulated in all cultures containing isopropranol but its presence in the alcohol mixture did not lower growth rate or yield significantly when compared with growth experiments on ethanol alone. Growth rate and yield declined at ethanol concentrations greater than 1% (v/v) and 0.3% (v/v) respectively. In a 0.3% (v/v) alcohol mixture, acetate was found only during the exponential growth phase. In a 3% (v/v) mixture, acetate and ethyl acetate accumulated during growth whereas acetaldehyde was present only during the exponential growth phase.     

原产地和入侵地紫茎泽兰对泽兰实蝇的选择性、卵巢蛋白质含量与解毒酶活性的影响

[目的] 紫茎泽兰是我国危害严重的恶性入侵杂草。比较专一性天敌泽兰实蝇对该杂草入侵前后植株的适应性,是揭示外来植物入侵后适应性机制的重要科学问题之一。[方法] 比较泽兰实蝇对原产地和入侵地紫茎泽兰植株的寄主选择性,并测定寄生于2类植株的上泽兰实蝇卵巢蛋白质含量及乙酰胆碱酯酶、羧酸酯酶、谷胱甘肽S-转移酶活性。[结果] 泽兰实蝇对原产地和入侵地紫茎泽兰的选择无显著性差异;寄生在紫茎泽兰入侵地植株上的卵巢蛋白质含量较原产地植株上更高。解毒酶活力比较表明,入侵地紫茎泽兰上泽兰实蝇的羧酸酯酶活性低于原产地上的,但谷胱甘肽S-转移酶（雌虫）活性比较则相反,乙酰胆碱酯酶活性比较均无显著性差异。[结论] 紫茎泽兰入侵后,专一性天敌泽兰实蝇的适应性有所下降,丰富了外来植物入侵机制中天敌逃逸假说的内涵。     

基于MaxEnt模型的草原步甲属潜在地理分布研究

步甲(Carabid beetle)是草原生态系统重要的昆虫类群之一,对草原生态系统环境监测及维护生物多样性起着重要的作用。基于步甲属(雕步甲和长叶步甲)在宁夏草原131个有效调查记录和19个环境因子,利用最大熵模型(Maximum Entropy Model,简称MaxEnt)预测步甲属潜在空间地理分布,运用受试者工作特征曲线(ROC曲线)检测预测模型精度,刀切法(Jackknife)筛选影响步甲属潜在地理分布的主要生态因子。结果表明：(1) MaxEnt模型预测结果 ROC曲线AUC值分别为0.981、0.914,模型预测结果的可信度较高;(2) 19个环境因子中,对步甲属的潜在空间分布预测贡献率较高的环境因子为最冷月份最低气温、年均降水量、最干季均温、年平均气温;(3)雕步甲主要分布在温性荒漠草原及温性草甸草原的北部,长叶步甲主要分布在温性草甸草原。以上研究为步甲调查和保护提供参考依据。     

人生殖道源乳酸杆菌的筛选及在小鼠阴道假丝酵母病模型中的应用评价

【目的】分离筛选人阴道环境中具有益生特性的乳酸杆菌,探索外阴阴道假丝酵母病的益生菌疗法。【方法】利用含1%碳酸钙的de Man,Rogosa and Sharpe (MRS)培养基从无症状育龄女性阴道分泌物中分离乳酸杆菌,采用共培养方法评价其对白色念珠菌(Candida albicans)的抑制作用,通过对乳酸杆菌的耐酸性能、体外聚集特性和黏附能力测试考察其益生特性,并进行乳酸杆菌株功能化组合。通过构建小鼠外阴阴道假丝酵母病模型,初步探索乳酸杆菌株组合对C. albicans的抑制作用。【结果】从53个样品中分离得到19株乳酸杆菌,筛选获得4株乳酸杆菌(Lactobacillus crispatus ZH08、L. fermentumZH09、L. fermentum ZH11和L. crispatus ZH17)具有较强抑制C. albicans生长的能力。4株乳酸杆菌均能耐受低pH环境,能快速降低培养液pH。其中2株L. fermentum具有更强的抑制活性,能在24 h内快速抑制C. albicans生长,抑制率可达到95%以上;另2株L. crispatus具有更强的聚集特性和...     

紫茎泽兰入侵前后抗虫物质含量差异及其对泽兰实蝇寄生的响应

【目的】紫茎泽兰是我国重要的入侵杂草,对我国生态环境和农、林、牧业造成重大危害。本研究通过比较紫茎泽兰原产地与入侵地种群地上部抗虫物质含量的差异及其对泽兰实蝇寄生的响应,为探明紫茎泽兰入侵种群持续扩张的化学生态机制提供依据。【方法】选取5个紫茎泽兰种群(3个中国云南种群:C1,C2,C3;2个墨西哥种群:M1,M2),比较原产地和入侵地种群地上部分碳氮比、单宁类(单宁酸、儿茶素、鞣花酸)和类黄酮物质(槲皮素、异槲皮素、山奈酚)含量差异,以及泽兰实蝇寄生前后这些抗虫物质的含量变化。【结果】入侵地种群地上部分碳氮比不同程度地低于原产地种群,茎秆部位差异最明显。紫茎泽兰入侵地种群的单宁类和类黄酮物质含量亦低于原产地种群,其中,芽尖部位单宁酸和儿茶素含量差异尤为明显,分别比原产地种群低26.4%和32.3%。泽兰实蝇寄生后,原产地和入侵地种群单宁类和类黄酮物质含量基本呈上升趋势,其中,M1种群寄生植株在虫瘿破膜前,儿茶素含量比未寄生植株升高了163.2%。【结论】紫茎泽兰入侵种群在抗虫特性方面产生了资源再分配的适应性变化,有利于其在新生境天敌缺乏条件下的持续扩张。虽然泽兰实蝇寄生会诱导紫茎泽兰原产地种群和入侵地种群抗虫物质的增加,但原产地种群增加的幅度更大,表明紫茎泽兰入侵后对专食性天敌胁迫的适应性明显下降。     

Solid-substrate fermentation ofKloeckera apiculata andCandida utilis on apple pomace to produce an improved stock-feed

Apple pomace was used in solid-substrate fermentation with the yeastsKloeckera apiculata orCandida utilis Y15. A total crude protein content of 7.5% (w/w) was achieved after 72 h for each yeast. The concentration of essential amino acids in the modified apple pomace was more than twice that in the control, enhancing its nutritive value as a stock-feed supplement. The fermentation of pomace as described can be used to reprocess this waste material into a useful value-added product for the agricultural sector.H. Rahmat was and R.A. Hodge, G.J. Manderson and P.L. Yu are with the Biotechnology Group, Department of Process and Environmental Technology, Massey University, Palmerston North, New Zealand; H. Rahmat is now with the Department of Chemical Engineering, Queen's University of Belfast, Belfast BT9 5AG, Northern Ireland, UK.     

假臭草水浸提液对4种作物的化感作用

【背景】假臭草是近几年来入侵我国南方的外来杂草,目前已成功侵占海南、福建、广东等省沿海的大部分果园、草地、农田等农业生态系统,给当地农业生产造成极大的危害。【方法】采用室内生物测定方法研究不同浓度的假臭草水浸提液(0.010、0.025、0.050 g·m L~(-1))对4种作物(西瓜、水稻、萝卜和辣椒)种子萌发和幼苗生长的化感作用。【结果】各浓度的假臭草水浸提液对4种作物种子萌发和幼苗生长都有较强的化感作用,并且随着浸提液浓度的增大,化感效应显著增强。当浓度为0.050 g·m L~(-1)时,4种受体作物的化感综合效应顺序为西瓜萝卜水稻辣椒。【结论与意义】明确假臭草对不同供试作物的化感作用,能够为该入侵杂草的科学防治提供理论依据。     

濒危种螺蛳对典型附着藻的摄食特性及其在洱海流域恢复的可行性

通过实验室小试研究濒临灭绝物种螺蛳(Margarya melanioides)对舟形藻(Naviculaceae sp.)、菱形藻(Nitzschia sp.)、孟氏颤藻(Planktothrixmougeotii)以及四尾栅藻(Scenedesmus quadricauda)4种典型附着藻类的摄食探讨其摄食生态学特性,调查海菜花(Ottelia acumiuate)养殖塘中藻的群落结构及水体的pH值、光照、溶解氧(DO)、温度等环境,从摄食生态学角度分析,探讨螺蛳在洱海流域恢复的可行性。摄食实验结果显示:螺蛳对实验选取四种藻摄食率关系为舟形藻菱形藻孟氏颤藻四尾栅藻。对舟形藻摄食的影响因素研究结果表明在温度为25℃时其摄食强度最大,摄食率(IR)为(2.19±0.16)mg个~(-1)d~(-1);螺蛳在光照为0-100001x的范围内均有较强的摄食活动,摄食活动最适宜的光照强度区间为1000-20001x,摄食率IR均达到3.0mg个~(-1)d~(-1)以上;溶解氧DO对其摄食活动影响显著,当DO大于3.6mg/L时螺蛳摄食活跃,低于1mg/L时螺蛳基本停止摄食;螺蛳摄食率大小跟螺蛳重量呈显著负相关P0.05。洱海流域海菜花塘水体的调查结果得出海菜花塘的pH值、光照、DO、温度以及藻等环境因子均适合螺蛳生存繁衍,可通过构建海菜花湿地来实现螺蛳的保种与扩增恢复。     

基于4种生态位模型的金钱松潜在适生区预测

金钱松（Pseudolarix amabilis）是我国特有孑遗植物,为国家II级保护植物。基于4种生态位模型（GARP、Bioclim、Domain和Maxent）预测金钱松潜在适生区,采用受试者工作特征曲线（Receiver Operating Characteristic,ROC）和Kappa统计量检验模型的预测效果。预测结果表明金钱松在浙江西北部、安徽南部、湖北南部、湖南北部以及江西北部表现为高度适生,并以这些地带为中心向外延伸至北纬24.43°-33.35°和东经106.41°-123.42°之间,4种模型预测结果的受试者工作特征曲线下面积（Area under recriver operating characteristic curve,AUC）平均值均大于0.9,Kappa平均值亦大于0.75,精度较高。通过"刀切法"分析得出年均温是预测金钱松潜在适生区的关键影响因子,可能为当前金钱松分布格局形成的决定因素。模拟金钱松在末次盛冰期和2070年气候条件下的分布,结果表明其分布格局随气候变化由"南扩北缩"变为"南缩北扩",未来分布面积将大幅减小,气候变化是导致其"南缩北扩"的主要驱动因子。建议在当前金钱松高适分布区域内（江西铜鼓县、湖南张家界和衡阳）建立自然保护区或种子园,并在未来气候条件下高适分布区域内（如安徽北部、河南南部、湖北东南部等地）通过人工引种辅助金钱松的北向迁移。     

Application of a two-stage temperature control strategy for enhanced glutathione production in the batch fermentation by Candida utilis

In batch culture for glutathione production with Candida utilis, a higher temperature (30 °C) was required to hasten cell growth while a lower temperature (26 °C) was needed to increase the production of glutathione. A two-stage temperature control strategy was used to enhance both the yield and the productivity of glutathione. As a result, glutathione production was increased by 5% and 23% of that at 26 °C and 30 °C, respectively, and the intracellular glutathione content reached 2.5% (w/w).     

Continuous culture of Candida utilis: influence of medium nitrogen concentration

When Candida utilis was grown in continuous culture, decreasing the concentration of N in the medium affected cell composition, biomass yield, biomass productivity, maximal growth rate and cell morphology. When the dilution rate was low (0.1 h^-1), reducing N from 1100 to 100 mg/l led to a 40% decrease in RNA content of the cells. Nitrogen-limited growth, which occurred when N<420 mg/l, was associated with significant changes in cell-wall carbohydrates and a significant reduction in the glycogen content of the cells. A set of culture conditions was established which permitted maximal consumption of the main nutrients in the medium and the production of yeast biomass suitable as a source of single-cell protein.     

Antimutagenic activity of soybeans fermented with basidiomycetes in Ames/Salmonella test

Soybeans fermented with either Phellinus igniarius or Agrocybe cylindracea inhibited the mutagenicity of the directly-acting mutagens: 4-nitro-o-phenylenediamine on Salmonella typhimurium strain TA 98 and NaN₃ on S. typhimurium strain TA 100; and indirectly-acting mutagens, 2-aminofluorene using strain TA 98 and benzo[a]pyrene using strain TA 100, in the presence of a supernatant solution from mammalian hepatic microsomes.     

Virulence of Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis in reconstituted human tissue models

Candida parapsilosis is an increasingly important human pathogen. To study the interactions of C. parapsilosis with human tissues, we evaluated the effects of the CBS 604 type strain and three different clinical isolates on reconstituted human oral epithelial and epidermal tissues. The newly described species Candida orthopsilosis and Candida metapsilosis were also examined in these models. Microscopy of reconstituted tissues infected with yeast cells revealed severe attenuation, morphological changes and cellular damage. C. orthopsilosis caused damage similar to C. parapsilosis isolates, whereas C. metapsilosis was less virulent. To further quantitate tissue damage, we measured lactate dehydrogenase (LDH) in the culture supernatant. The relative LDH measurements correlated with our histopathological observations. We also examined the effect of the lipase inhibitor Ebelactone B and proteinase inhibitor Pepstatin A, to establish the utility of this model for studying factors of C. parapsilosis virulence. Both Ebelactone B and Pepstatin A reduced the destruction of epidermal and epithelial tissues. Our data show that reconstituted human tissues are extremely useful for modeling host interactions with C. parapsilosis and for studying fungal virulence factors.