Relationships between molecular bacterial diversity and chemistry of groundwater in the Wairarapa Valley,New Zealand

Groundwater plays an important role in New Zealand water supplies and hence monitoring activities are conducted regularly. Most monitoring programmes aim to evaluate groundwater chemistry and almost completely overlook the microbial component in this ecosystem. In our present study, the bacterial community structure of groundwater in the Wairarapa Valley was examined using the terminal restriction fragment length polymorphism (T-RFLP), and relationships between bacterial community structure and groundwater chemistry, aquifer confinement and groundwater usage were explored. In addition, the results from this study were compared with a previous T-RFLP survey of the same area in an attempt to detect changes in bacterial community structure over time. The data obtained suggested that bacterial community structure was related to groundwater chemistry, especially to redox conditions. Species composition showed minimal variation over time if groundwater chemistry remained unchanged. These findings reflect the potential of using bacterial communities as biological indicators to evaluate the health of groundwater ecosystems. We suggest that it is important to include this type of broad bacterial diversity assessment criteria into regular groundwater monitoring activities.     

条斑紫菜SSU 中内含子分析

根据GenBank中已知的条斑紫菜(Porphyra yezoensis)SSU和LSU r RNA基因序列分别设计引物,分别以混合个体和单个个体的基因组DNA以及c DNA为模板进行扩增,测序分析后推测SSU r RNA基因中的内含子转录后加工可能仍被保留,而在LSU r RNA基因上游序列中没有发现内含子。长511bp的内含子位于SSU r RNA基因序列的上游,其插入位点为TCTGGTG-CCAGCAGCC,内含子5′和3′端的核苷酸序列分别为AAC-和-ATGG。该内含子的剪接位点以及保守区P,Q,R,S与I型内含子是不同的。研究发现,在同种紫菜中,不同个体SSU基因中内含子的有无、分布及结构特征是不同的,说明内含子对于分类是不太适合的。但是内含子的长度以及核苷酸序列在种间又具有一定的特异性,可以为红藻种间以及种内亲缘关系较远的亚种和不同地理群的区分和鉴定提供参考。     

Molecular identification of ascaridoid nematodes from the deep-sea onion-eye grenadier (Macrourus berglax) from the East Greenland Sea

Macrourus berglax from the East Greenland Sea was studied for the presence of ascaridoid nematodes in 2001, 2002 and 2003. The fishes were collected between 278 and 413 m water depth using a benthopelagic net. Based on the amplification of the internal transcribed spacer ITS-1, 5.8S, ITS-2 and flanking sequences (=ITS+), three ascaridoid nematode species were identified. The prevalence of infestation during the 3 years ranged from 42.9% to 62.9% and 22.9% to 40.0% for the anisakids Anisakis simplex (s.s.) and Pseudoterranova decipiens (s.s.), respectively, and from 28.6% to 60.0% for the raphidascarid Hysterothylacium aduncum. A total of 18 specimens, two of each species and examination year, revealed no sibling species, suggesting a limited distribution of other ascaridoid siblings into the deep sea. The ITS-1, 5.8S and ITS-2 sequences of A. simplex (s.s.) from the East Greenland Sea did not differ from previously published sequence data (GenBank) from other regions in the Atlantic and Pacific oceans. The sequences of P. decipiens (s.s.) corresponded most closely to those of specimens from Richardson Bay, western Pacific, and differed in four positions (0.5%). They corresponded least to those of specimens from Japan (1.5%). The sequence data for H. aduncum differed in two positions in the ITS-1 (0.2%) and three positions in the ITS-2 (0.3%) from sequences from Japan. A high genetic similarity between the regions can be explained by (a) extensive final host migration in the case of A. simplex (s.s.), (b) an overlapping distribution of final host populations along the continental shelves for P. decipiens (s.s.) and (c) a low host specificity and large population size in the intermediate and final hosts for H. aduncum. The occurrence of the identified species in the macrourid fish underlines the potential of cosmopolitan ascaridoid nematodes to distribute not only horizontally but also vertically in the deep sea.     

分子生物学技术在赤潮毒素分析监测中的应用

有毒赤潮事件的频繁爆发,不仅对海洋生物及自然资源造成了极大的危害,还严重威胁到公众健康。因此,各国都在积极寻求快速灵敏的检测方法,加强对赤潮毒素的分析及监测。现代分子生物学技术的发展为新型快速检测方法的建立提供了可能。对利用分子生物学技术对赤潮毒素进行检测的几种方法——全细胞PCR法、DNA探针法及信使基因细胞受体法进行了讨论。这些新技术旨在检测出环境中的痕量赤潮毒素和有害生物,杜绝赤潮毒素或有毒藻进入到食物链,从而最大限度保护公众健康、水环境及自然资源。     

基于cox1条形码的鱼肚DNA分子鉴定

为鉴定鱼肚的正品来源,运用一对特异性引物扩增并测定10 个不同价格（700～1 500 元/kg）鱼肚样本的线粒体DNA 细胞色素c 氧化酶Ⅰ（cox1）约660 bp 的序列.通过BLASTN 比对DNA 序列同源度,判定鱼肚所属种类分别为鲈形目石首鱼科（Sciaenidae）、马鲅科（Eleutheronema）、笛鲷科（Lutjanidae）、带鱼科（Trichiuridae）.cox1 条码序列获取便捷,基于此条码序列的物种鉴定技术可用于鱼肚种类鉴别.     

Phaeocystis globosa与Phaeocystis antarctica叶绿体psbA基因的比较

测定了2株球形棕囊藻Phaeocystis globosa P1、P2的psbA基因序列。发现得到的2个序列完全相同。以P2序列对比分析了P．globosa和P．antarctica的psbA基因在DNA序列、氮基酸序列和RNA二级结构上的差异性，发现2种棕囊藻psbA基因DNA序列和氨基酸序列非常保守，无插入／缺失，其核苷酸和氨基酸变异率分别为1．88％和1．13％。与核基因核苷酸的碱基替换不同，psbA基因核苷酸的碱基替换主要发生在密码子的第1位上。且不引起氨基酸的变化，引起氨基酸变化的碱基替换都发生在密码子的第2位和第3位上。在RNA二级结构上两序列的1～4茎环结构完全相同，表现出明显的棕囊藻属的特异性，其它结构区域差异较大。种间差异表现明显。由于psbA基因DNA序列和氨基酸序列非常保守，可能不适宜棕囊藻属的系统发育分析。但其RNA二级结构可能对于棕囊藻的分子分类有一定的参考价值。     

Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum

The dinoflagellate Alexandrium minutum is often associated with harmful algal blooms (HABs). This species consists of many strains that differ in their ability to produce toxins but have similar morphology, making identification difficult. In this study, species-specific rRNA probes were designed for whole-cell fluorescence in situ hybridization (FISH) to distinguish A. minutum from two phylogenetic clades. We acquired the complete SSU to LSU rDNA sequences (GenBank accession numbers JF906989-JF906999) of 11 Alexandrium strains and used these to design rRNA targeted oligonucleotide probes. Three ribotype-specific probes, M-GC-1, M-PC-2, and M-PC-3, were designed. The former is specific for the GC clade ("Global clade") of A. minutum, the majority of which have been found non-toxic, and the latter two are specific for the PSP (paralytic shellfish poisoning)-producing PC clade ("Pacific clade"). The specificity of these three probes was confirmed by FISH. All cells in observed fields of view were fluorescently labeled when probes and target species were incubated under optimized FISH conditions. However, the accessibility of rRNA molecules in ribosomes varied among the probe binding positions. Thus, there was variation in the distribution of positive signals in labeled cells within nucleolus and cytosol (M-GC-1, M-PC-3), or just nucleolus (M-PC-2). Our results provide a methodological basis for studying the biogeography and population dynamics of A. minutum, and providing an early warning of toxic HABs.     

压力传感器的自动标定技术

针对压力测量设备生产和维护过程中,对关键部件压力传感器标定处于手工操作状态、效率低、操作人员不易掌控、标定结果处置复杂的现状,开发了针对压力传感器的自动标定技术。该技术将计算机控制、智能化、曲线拟合和误差分析引入到压力传感器的标定过程中,大幅度提高了标定的效率和可靠性。经此技术标定过的压力探头,在使用中具有互换性,减轻了数据后处理工作的难度。     

Sensitive and rapid detection of two toxic microalgae Alexandrium by loop-mediated isothermal amplification

A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid detection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (PSP). Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer (ITS) of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection limits of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction (PCR) method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.