片突菱纹叶蝉感染枣疯植原体与Wolbachia的检测及系统发育分析

[目的]探讨片突菱纹叶蝉Hishimonus lamellatus Cai et Kuo不同种群中枣疯植原体与沃尔巴克氏体Wolbachia的感染情况和Wolbachia在不同器官组织分布,明确枣园菱纹叶蝉中Wolbachia的感染类型和分类地位,为研究Wolbachia感染对枣疯植原体潜在介体叶蝉生物学及生态学影响奠定基础.[方法]通过枣疯植原体和Wolbachia的基因特异性引物对片突菱纹叶蝉田间自然种群和实验室种群进行分子检测和鉴定.[结果]田间采集的片突菱纹叶蝉成虫植原体感染率在55％-61％之间,而Wolbachia感染率为3％-4％.田间采集的片突菱纹叶蝉自然种群经室内饲养,在1-4龄若虫中检测到Wolbachia,2-5龄若虫中检测到了植原体.片突菱纹叶蝉实验室饲养无植原体种群在其卵巢、卵和若虫中发现感染Wolbachia,在其唾液腺和消化道也检测到了Wolbachia,感染率在58％-100％之间.基于Wolbachia的wsp基因构建系统发育树,发现片突菱纹叶蝉体内的2个Wolbachia株系同属于B大组,但不同于B大组其他株系,属于新株系wLam1和wLam2.[结论]片突菱纹叶蝉成虫采自田间种群可以感染枣疯植原体和Wolbachia,无植原体叶蝉实验室饲养种群成虫感染Wolbachia显著高于田间种群,片突菱纹叶蝉体内2个Wolbachia株系属于B大组.这一研究结果为Wolbachia作为介体叶蝉生物防治剂进一步利用提供了基础信息.     

扁平圆扁螺(Hippeutis complanatus)早期发育的形态观察

在水温18℃左右条件下培养扁平圆扁螺(Hippeutis complanatus),使其产卵,在光镜下观察其早期发育过程,详细描述各发育阶段的形态特征.扁平圆扁螺胚胎发育属典型的包囊幼虫发育类型,其过程共分为6个时期,即卵裂期、囊胚期、原肠期、担轮幼虫期、面盘幼虫期和仔螺形成期.胚胎于面盘幼虫晚期孵化,在水中完成仔螺形成过程,腹足从三角变为长楔形,幼虫肾退化,螺壳平伏形成仔螺.     

快速STR分型在法医DNA分析中的应用研究进展

STR分型是目前世界通用的DNA指纹分析方法.传统的分型方法包括DNA提取、DNA定量、PCR扩增和对扩增后的STR片段进行检测和分析,耗时较长(8¹0 h).一些情况下很难满足案件的实际需求.法医学家们在加快STR分型的方法研究一直不懈努力.现从快速提取DNA、无提取直接PCR、快速PCR、微流控芯片技术在STR快速分型方面的应用四方面,对近年来出现的可进行快速STR分型的新技术、新方法进行综述.     

4种快速PCR检测试剂法医学应用的初步研究

采用快速PCR扩增,探索其法医学应用价值.将AmpFLSTRIdentifiler试剂盒分别与4种不同的快速PCR检测试剂联合构建快速PCR体系,以9947A为模板,采用各自优化的循环参数进行扩增,并将各分型结果与常规方法进行比较,结果表明:联合构建的4种快速PCR体系均可获得与常规方法一致的DNA分型结果,扩增用时最短可减少至22 min.可见应用快速PCR方法进行扩增,可获得与常规方法一致的STR分型,并且显著缩短扩增时间,提高DNA分型速率.     

蛋白质晶体的物理性质及其鉴别方法研究进展

蛋白质晶体由蛋白质分子有序排列的框架组成,其内部富含溶剂（水）分子,外观形貌多种多样,其物理性质与常规固态晶体相比有很大不同。我们针对蛋白质晶体的一些物理性质（包括低密度、机械性能、声学、热学、光学等性质）,进行了研究进展评述,并根据蛋白质晶体的特殊物理性质,评述了人们发展出的从蛋白质结晶液中区分蛋白质晶体的方法。     

肠出血性大肠杆菌O157∶H7 EspA和EspB蛋白的重组表达及其免疫效果

目的：通过DNA重组技术表达肠出血性大肠杆菌（EHEC）0157：H7的EspA和EspB蛋白,并分析它们的免疫保护性。方法：采用PCR技术从EHEC0157：H7基因组中扩增espA和espB基因,连接至pET-22b（4-）载体上,转化至宿主细胞大肠杆菌BL21（DE3）,经IPTG诱导表达,用亲和层析纯化目的蛋白,SDS-PAGE测定其相对分子质量,免疫小鼠分析其免疫保护性。结果：重组espA和espB基因片段的测序结果与GenBank中的相应基因序列完全一致,一致性均为100％;得到了纯度为95％以上的重组EspA和EspB蛋白,免疫小鼠所得到的抗体效价均为10^6。结论：重组EspA和EspB蛋白获得了可溶性表达,表达的蛋白具有良好的免疫保护性,为进一步制备疫苗奠定了基础。     

中国兰科植物沿经纬度的水平分布格局

对濒危物种在大尺度上地理分布的研究,有助于制定合理的保护规划和保护策略.兰科植物作为一大类急需保护的濒危物种,研究其在中国境内的地理分布格局具有重要的理论和实践意义.通过文献查阅、自然保护区数据整理收集兰科植物在全国范围内的调查数据,利用ArcGIS10.0和SPASS18.0软件对其地理分布进行了分析,结果表明：中国西南地区是兰科植物的分布中心和分化中心;兰科植物丰富度表现出显著的经度和纬度相关性,与经度之间呈单峰关系,在100°E附近出现峰值,但随纬度升高丰富度不断下降.     

基于指数施肥法的一年生油松菌根化容器苗的供N研究

为了探讨一年生菌根化油松容器苗的最适苗木供N量和供N速率,采用4种不同指数施肥量进行了试验.通过测定苗木生物量和N含量,显示总供N量为80 mg·株-1的处理组在生物量积累和苗木N含量上显著优于其它各组（P〈0.05）;在100 d时,该处理可获得0.47 mg·株-1·d-1的N最适添加速率;在该速率下,苗木生物量、N含量分别可以达到845.60 mg·株-1和6.51 mg·株-1.根据生物量和N含量随着供N量增加的回归拟合结果,在107 d的生长过程中,67.96 mg·株-1至84.15 mg·株-1的供N总量可以使得一年生油松幼苗获得较高的生物量积累水平和N含量.     

Granulation of activated sludge in a continuous flow airlift reactor by strong drag force

Most aerobic granule cultivation has been based on the sequencing batch reactor (SBR) and then the factors that affect aerobic granulations were developed in the SBR. However, little work has been done to cultivate aerobic granules in a continuous-flow bioreactor with simple structure that is realistic for engineering. This work is the first to cultivate aerobic granules in a continuous flow airlift fluidized bed reactor (CAFB) possesses a very simple structure and without settling time and starvation time controlling. The configuration of CAFB was the simplest continuous-flow aerobic granular bioreactor reported by now. The majority of granules could be formatted in the CAFB after 12 days cultivation. The effluent COD concentration maintained at 50 ± 10 mg/L for the variable COD loading rate of 3.5 g COD/L/d and 4.8 g COD/L/d, which confirmed that the CAFB performed good anti-shock abilities. CAFB performed good nitrification ability, however, little denitrification was found under the operating conditions of this study. The shear stress acting on the solid phase were hundreds of times stronger in the CAFB than in the SBR at the same aeration strength. It seems CAFB is very efficient for granulation due to the strong shear-force exertion, which is promising for continuous-flow aerobic granular bioreactor. Protein, positive to the hydrophobicity, was predominant in extracellular polymeric substances in the granules, and favored the granules formation in the CAFB combined with the polysaccharides. However, filamentous bulking always happened in 35 days operation of the CAFB, thus further study on the stability of this bioreactor is urgently necessary.     

Fosmid library construction and end sequences analysis of the Pacific oyster,Crassostrea gigas

The Pacific oyster (Crassostrea gigas) is globally distributed and is one of the most commercially and ecologically important marine organisms. However, little is known about the genome of this species. In this study, a C. gigas fosmid library was constructed that contains 459,936 clones with an average insert size of approximately 40 kb, representing 22.34-fold haploid genome equivalents. End sequencing generated 90,240 fosmid end sequences (FESs) with an average length of 384.27 base pairs (bp), covering approximately 2.58% of the Pacific oyster genome. The FESs were subsequently assembled and annotated, resulting in 6332 sequences with predicted open reading frames≥300 and 1,189,100 bp repeats. Furthermore, a total of 3200 microsatellite repeats were identified, and dinucleotide repeats were found to occur most abundantly, with AG and AAT being the most abundant repeat class of dinucleotides and trinucleotides. We also found that the repeat number was generally negatively proportional to the repeat element length. Microsatellites composition between the transcribed sequences and genomic sequences was shown to be different. Point mutations of microsatellite were non-random and underwent strong selection stress. Overall, a comprehensive sequence resource for the Pacific oyster was created, including annotated transposable elements, tandem repeats, protein coding sequences and microsatellites. These initial findings will serve as resources for further in-depth studies of physical mapping, gene discovery, microsatellite marker developing and evolution studies.