A novel and efficient assay for identification and quantification of Acidithiobacillus ferrooxidans in bioleaching samples

Acidithiobacillus ferrooxidans is a Gram-negative, acidophilic, and chemolithotrophic bacterium that is active in bioleaching. The leaching efficacy is directly influenced by the biomass changes of this specie in bioleaching microbial community. In order to perform a simple and sensitive assay on A. ferrooxidans from mixed strains in this process, a novel assay was developed based on sandwich hybridization assay with the aid of S1 nuclease treatment and fluorescent labeling. In the work, a designed DNA probe complementary to the conservative region of its 16S rRNA was synthesized, which showed high accuracy for distinguishing homologous species with the exclusion of even-only two base pairs difference. The specificity of this assay was verified in different systems with mixed strains, and the quantitative result was proved by comparison of microscopic cell counting. The detection sensitivity was about 8 × 10(2) cells/ml and the inter-assay coefficient of variation of three independent assays was from 3.8 to 7.7 %, respectively. In addition, the cycle of assay was about 3-4 h when the cost estimated was less than $0.5 per sample. This assay method might be applied for identifying and monitoring any kind of bacterial strain from a mixed microbial flora in bioleaching or other areas.     

百香果内生细菌多样性及促生长特性

内生菌可为植物提供营养成分,也可以通过代谢产物促进植物生长,目前很少出现关于百香果(Passiflora edulia Sims)内生菌的研究.百香果是西番莲科西番莲属的一种草质藤本植物,主要生长于亚热带与热带地区,对其内生菌进行分离纯化,依据插入序列指纹图谱(IS-PCR)结果对所得菌株聚类,经16S rRNA基因测...     

The complete mitochondrial genome of the damsel bug Alloeorhynchus bakeri (Hemiptera: Nabidae)

The complete sequence of the mitochondrial DNA (mtDNA) of the damsel bug, Alloeorhynchus bakeri, has been completed and annotated in this study. It represents the first sequenced mitochondrial genome of heteropteran family Nabidae. The circular genome is 15, 851 bp in length with an A+T content of 73.5%, contains the typical 37 genes that are arranged in the same order as that of the putative ancestor of hexapods. Nucleotide composition and codon usage are similar to other known heteropteran mitochondrial genomes. All protein-coding genes (PCGs) use standard initiation codons (methionine and isoleucine), except COI, which started with TTG. Canonical TAA and TAG termination codons are found in eight protein-coding genes, the remaining five (COI, COII, COIII, ND5, ND1) have incomplete termination codons (T or TA). PCGs of two strands present opposite CG skew which is also reflected by the nucleotide composition and codon usage. All tRNAs have the typical clover-leaf structure, except the dihydrouridine (DHU) arm of tRNA(Ser (AGN))which forms a simple loop as known in many other metazoa. Secondary structure models of the ribosomal RNA genes of A. bakeri are presented, similar to those proposed for other insect orders. There are six domains and 45 helices and three domains and 27 helices in the secondary structures of rrnL and rrnS, respectively. The major non-coding region (also called control region) between the small ribosomal subunit and the tRNA(Ile )gene includes two special regions. The first region includes four 133 bp tandem repeat units plus a partial copy of the repeat (28 bp of the beginning), and the second region at the end of control region contains 4 potential stem-loop structures. Finally, PCGs sequences were used to perform a phylogenetic study. Both maximum likelihood and Bayesian inference analyses highly support Nabidae as the sister group to Anthocoridae and Miridae.     

基于荧光激活细胞分选技术的超高通量酶活性筛选方法及其应用

基于荧光激活细胞分选(FACS)技术的超高通量酶活性筛选方法是新出现的一类高通量筛选技术.它利用流式细胞仪高灵敏度、高通量的特点,能以极高的速度(108/天)对大容量酶基因文库进行筛选.FACS筛选技术的出现突破了常规筛选方法低效、耗时、费力等瓶颈问题,极大地提升了人类对大容量基因文库的探索能力,因此在新酶基因筛选、酶活性检测、酶定向进化等领域有广泛的应用潜力.综述了FACS超高通量酶活性筛选方法的最新研究进展,着重介绍了其在酶定向进化中的应用.     

乌兰布和沙漠典型灌木群落结构及数量特征

灌木群落是乌兰布和沙漠生态系统中最重要的组成部分,根据野外样地的调查资料,从各群落物种组成、生活型谱、盖度特征、物种多样性、群落相似性等方面对乌兰布和沙漠中典型灌木群落结构和数量特征进行分析,为该区生物多样性保护与植物资源持续利用提供理论依据。结果表明:(1)梭梭、白刺、沙冬青、沙蒿、油蒿、盐爪爪是乌兰布和沙漠中主要灌木群落类型;各典型灌木植被群落中仅出现12科47属共计58种植物,各群落间物种组成差异较大,但均以藜科、菊科、禾本科、蒺藜科、豆科等干旱荒漠区的表征科数量居多;(2)沙漠中分布范围最广的梭梭群落、白刺群落、沙冬青群落和油蒿群落其生活型谱均以一年生植物所占比例最大,与研究区干旱的气候特征相吻合;(3)各群落中灌木层植物所占群落盖度的比重均超过了50%,其贡献率大于草本植物;(4)分布于生境条件较好的群落类型其物种多样性相对较高,从整体看各典型灌木群落的H′多样性指数的变化范围在0.98～3.42之间,而D多样性指数的变化范围在0.415 5～0.879 5之间,其物种多样性指数较低,物种组成比较简单;(5)各典型植物群落的相似性系数在0.068 2～0.604 7之间,均未达到极相似水平,其中白刺群落、梭梭群落以及沙冬青群落之间呈轻度相似水平,而其他各群落之间相似性系数均低于0.25,表明各群落间生境差异较大。     

植物抗寒基因表达调控研究进展

低温胁迫是植物生长过程中的主要非生物胁迫因子之一,也是影响农作物生产的主要因素之一。研究表明,在植物体内存在着一个复杂的对低温胁迫信号感知及传导的网络系统,该系统中大量的相关基因已有报道,这些基因不仅仅涉及植物激素的应答,还涉及到植物基因的转录调控及转录后的修饰与调控等各个方面。该文就近年来国内外有关植物抗寒基因表达调控的研究进展进行综述。     

烟草磷酸吡哆醛水解酶的分离纯化与表征

采用硫酸铵沉淀、DEAE-Sepharose Fast Flow阴离子交换、Sephadex G-100凝胶过滤和SP Sephadex C-25阳离子交换柱层析等步骤,对烟草磷酸吡哆醛水解酶进行了分离纯化。结果表明:该酶被纯化了119.6倍,得率为28.49%,经凝胶过滤和SDS-PAGE测得该酶的全分子量为49.6kDa,亚基分子量约为25kDa;该酶最适温度为50℃,最适反应pH为5.5;Mg2+、Ca2+、Mn2+等对该酶有激活作用,金属离子螯合剂EDTA对酶有抑制作用,加入Mg2+后抑制作用得到解除;在最适反应条件下,测得反应底物磷酸吡哆醛(PLP)和磷酸吡哆胺(PMP)的Km值分别为0.23mmol/L和0.56mmol/L。     

‘寒富’苹果二倍体及其同源四倍体叶片超微结构和叶绿素荧光参数特征

采用扫描电镜和石蜡切片法,以‘寒富’苹果二倍体及经秋水仙素加倍获得的同源四倍体植株为材料,比较两种倍性植株叶片超微结构、叶绿素含量及叶绿素荧光参数的日变化规律。结果显示:(1)与二倍体植株相比较,其同源四倍体叶片厚度、栅海比、气孔长、气孔宽、分别增加了15.1%、16.1%、70.5%、27.2%,而气孔密度显著减少了58.7%;其同源四倍体叶保卫细胞中叶绿体数和叶绿素含量分别比二倍体植株高出125.3%、37.7%。(2)‘寒富’苹果同源四倍体与其二倍体的叶绿素荧光参数PSⅡ的原初光能转化效率(Fv/Fm)、PSⅡ的潜在光化学效率(Fv/F0)和以吸收光能为基础的光合性能指数(PI)值的日变化趋势相似,但PI平均值比二倍体显著高出38.6%。研究表明,同源四倍体较二倍体叶片在形态上更大、更厚,气孔更大、密度更小,栅海比更大,表现出抗病的叶片结构;同时同源四倍体较二倍体含有更高的叶绿素含量,表现更优良的光合特性。     

刺山柑多酚类物质含量及其抗氧化活性研究

采用Folin-Ciocalteu法,测定南疆地区刺山柑不同时期不同器官多酚类物质含量,并用DPPH法对其抗氧化活性进行比较研究。结果表明:(1)多酚类物质在不同时期刺山柑不同器官中均有分布,刺山柑不同器官中花蕾中多酚类物质含量最高,茎中多酚类物质含量最少;其中5～8月份花蕾中多酚类物质含量随月份的增加而减少,茎中多酚类物质含量随月份的增加而增加,其他器官中多酚类物质含量变化与花蕾基本相似。(2)不同时期刺山柑的不同器官均有清除DPPH自由基的能力,花蕾的抗氧化活性最强,且花蕾的抗氧化活性随月份的增加而减弱,茎的抗氧化活性最弱。     

3×3完全双列杂交F1不同阶段生长特点的分析

为了了解鲤杂交F1不同组合、不同阶段生长性状的变化情况,研究以建鲤、黄河鲤和黑龙江野鲤3个鲤品种双列杂交F1为试验材料,通过组合、组合内性别间、性别、不同时期体重、体增重以及协方差分量的分析,来确定完全双列杂交F1生长性状的变化特点,以及在此过程中起重要作用的影响因素。结果表明:不同时期各个组合体重不同,不同时期组合内性别间体重差异不同;不同时期不同性别间体重差异不同;不同协方差分量,同一组合PIT标记17个月时的体重的最小二乘估计值不同,同一个协方差分量,9个组合中极值估计值出现的组合也不同;除PIT标记17个月后的体厚作为协方差分量外,性别之间体重没有差异,其余分量均是雌鱼体重显著高于雄鱼体重。这些说明选取合适的协方差分量对组合的选择和育种的结果有重要影响。