CYTOTOXICITY AND GENOTOXICITY OF POLYETHYLENIMINE AND NICKEL CHLORIDE IN RED SEA BREAM (Pagrosomus major) FIN CELL LINE RSBF

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Sequences Characterization of Microsatellite DNA Sequences in Pacific Abalone (Haliotis discus hannat)

The microsatellite-enriched library was constructed using magnetic bead hybridization selection method, and the microsatellite DNA sequences were analyzed in Pacific abalone Haliotis discus hannai. Three hundred and fifty white colonies were screened using PCR-based technique, and 84 clones were identified to potentially contain microsatellite repeat motif. The 84 clones were sequenced, and 42 microsatellites and 4 minisatellites with a minimum of five repeats were found (13.1% of white colonies screened). Besides the motif of CA contained in the oligoprobe, we also found other 16 types of microsatellite repeats including a dinucleotide repeat, two tetranucleotide repeats, twelve pentanucleotide repeats and a hexanucleotide repeat. According to Weber(1990), the microsatellite sequences obtained could be categorized structurally into perfect repeats (73.3%), imperfect repeats(13.3%), and compound repeats (13.4%). Among the microsatellite repeats, relatively short arrays (＜ 20 repeats) were most abundant,accounting for 75.0%. The largest length of microsatellites was 48 repeats, and the average number of repeats was 13.4. The data on the composition and length distribution of microsatellites obtained in the present study can be useful for choosing the repeat motifs for microsatetlite isolation in other abalone species.     

CYTOTOXICITY AND GENOTOXICITY OF POLYETHYLENIMINE AND NICKEL CHLORIDE IN RED SEA BREAM（Pagrosomus major）FIN CELL LINE RSBF

A continuous marine fish cell line RSBF(i.e.Red Sea Bream Fin)was utilized to screen the cytotoxicity and genotoxicity of polyethylenimine(PEI)and nickel cholride(NiCl2)in this study on the deleterious effects of aquatic genotoxins on fish.At the 0.01 to 1μgml concentration tested,PEI had acute toxicity to the treated RSBF cells(IC50=1.12,0.92,0.88 and 0.64μg/ml PEI for time 0 h,24 h,48 h and 72 h after treatment,respectively)and markedly inhibited their proliferation in a dose-dependent manner.At the 0.001 to 5 μmol/L concentration tested,NiCl2 posed no acute toxicity but significantly stimulated their growth(107?214?of control).Random amplified polymorphic DNA(RAPD)technique was used to detect the genotoxic effects of PEI and NiCl2 by comparing the RAPD banding patterns of the control and treated cells.RAPD analysis indicated that at the concentrations tested,PEI was more genotoxic than NiCl2 to RSBF cells;that there was a slight dose-dependent response in the genotoxic effect of PEI bue not NiCl2;and that RAPD technique might provide a sensitive,non-specif-ic gentoxic endpoint.And the potent cytotoxicity and genotoxicity of PEI on fish cells showed that we should be cautious in utilizing it as gene vector in fish gene transfer and human gene therapy.     

机载云降水粒子成像仪所测数据中伪粒子的识别

机载云粒子成像仪(Cloud Imaging Probe,简称CIP)和降水粒子成像仪(Precipitation Imaging Probe简称PIP)所测数据中伪粒子的存在会直接导致仪器所测数据质量下降。利用山西Y-12探测飞机在太原地区的三次航测资料对飞机采样期间伪粒子图像的特点及其成因进行了分析和归类,并在此基础上提出了综合利用到达时间间隔阈值和图像处理识别技术的伪粒子识别方法。利用所提的方法对航测的粒子图像资料进行处理、统计和分析,统计结果表明,在一次测量过程中伪粒子的出现概率最高可达45.2%,其平均出现概率分别是36.01%(CIP)和8.64%(PIP);在所有伪粒子成分中,破碎形成的伪粒子的出现概率是最高的,它可以占到伪粒子总数的95%以上;其次是条纹状粒子和并存粒子,相对来说,空白粒子和线状粒子出现的概率是比较低的。研究还发现,机载平台的飞行状态对伪粒子各成分的出现概率也会造成影响。利用所提方法对仪器测量的粒子谱、粒子数浓度和冰水含量值进行订正,订正结果表明伪粒子对仪器量程内的粒子谱、粒子数浓度和冰水含量值均有影响,其中在云粒子谱影响上,伪粒子对粒子谱两端的影响较大,其中对小云粒子谱的影响主要是在400 μm以下,对大云粒子端的影响主要是在1000 μm(CIP)和2000 μm(PIP)以上。所提方法和阈值对于以Y-12飞机为机载探测平台,以CIP和PIP为探测仪器所获取的其它航次云微物理图像资料处理也是有一定的参考使用价值。     

Identification of Kalidium species (Chenopodiaceae) by DNA barcoding

DNA barcoding is an increasingly prevalent molecular biological technology which uses a short and conserved DNA fragment to facilitate rapid and accurate species identification. Kalidium species are distributed in saline soil habitat throughout Southeast Europe and Northwest Asia, and used mainly as forage grass in China. The discrimination of Kalidium species was based only on morphology-based identification systems and limited to recognized species. Here, we tested four DNA candidate loci, one nuclear locus (ITS, internal transcribed spacer) and three plastid loci (rbcL, matK and ycf1b), to select potential DNA barcodes for identifying different Kalidium species. Results showed that the best DNA barcode was ITS locus, which displayed the highest species discrimination rate (100%), followed by matK (33.3%), ycf1b (16.7%), and rbcL (16.7%). Meanwhile, four loci clearly identified the variant species, Kalidium cuspidatum (Ung.-Sternb.) Grub.var. sinicum A. J. Li, as a single species in Kalidium.     

A Practical Method for Accurate Measurement of Trace Level Fluorine in Mg‐ and Fe‐Bearing Minerals and Glasses Using Electron Probe Microanalysis

Fluorine plays an important role in magmatic and hydrothermal processes, but due to its low abundance in geological samples determining F is difficult by electron probe microanalysis. By using a W‐Si multi‐layered pseudocrystal as the diffraction crystal instead of thallium acid phthalate (TAP), count rates were considerably higher, which however introduced spectral interferences between FKα and FeLα and MgKβ lines when normal integral mode is applied. In this study, we developed a protocol using a W‐Si multi‐layered pseudocrystal for measuring accurately trace level F in both minerals and glasses. First, we used differential mode with an optimised PHA (pulse height analysis) setting in signal processing, instead of normal integral mode, which completely eliminated the second‐order MgKβ line. Second, the overlap of the first‐order FeLα on FKα peak, which cannot be filtered by modifying the PHA setting, was calibrated quantitatively using F‐free minerals and silicate glasses. Applying this two‐step method, F was determined in a number of reference glasses, as well as in glasses synthesised from powders of the rock reference materials AC‐E, GS‐N and DR‐N. Our data are consistent within error with F concentrations determined by other methods, demonstrating the reliability of this method.     

玻璃质壳类底栖有孔虫卷转虫属Ammonia spp.的DNA保存方式和提取效能比较研究

对有孔虫的DNA进行有效保存及提取是开展有孔虫分子鉴定工作的前提。本研究以探索适合玻璃质壳类底栖有孔虫的DNA保存及提取方法为目的,以中国近海优势属——玻璃质类底栖有孔虫卷转虫属(Ammonia spp.)为模式生物,考察不同温度(20—120°C,间隔10°C)烘干、冷冻(–20°C、–80°C,有水、无水)以及脱氧胆酸钠裂解液(Na deoxycholate,简称DOC)处理(含和不含EDTA)保存有孔虫DNA方法,同时进行了DOC裂解液法和Guanidine裂解液法对Ammonia spp.DNA提取效能的比较。结果显示,烘干保存法中20°C组和30°C组显著优于40°C组(P0.05);冷冻保存法中无水组显著优于有水组(P0.05);DOC裂解液保存法各组间无显著差异(P0.05)。三种保存方法综合效果表现为冷冻组显著优于烘干组和裂解液组(P0.05);两种DNA提取方法的提取效果都很好,但DOC法更廉价且操作简便。本文给出了一套适用于玻璃质壳类有孔虫的DNA保存和提取的优化方案。     

中华钩线虫的DNA条形码研究及分类意义探讨

海洋线虫是海洋底栖生物中数量上最丰富的类群,在海洋生态系统中起着重要的作用。对海洋线虫进行种类鉴定是线虫研究中最重要的工作之一。目前,海洋线虫的鉴定主要采用形态学的分析方法,但是这种方法往往费时费力,对于如此丰富的物种,急需新的鉴定方法。作者以黄海潮间带自由生活线虫优势种——中华钩线虫(Oncholaimus sinensis)为例,在形态学分类的基础上,将DNA条形码技术引入线虫的鉴定中,探讨了线粒体细胞色素C氧化酶第一亚基(mt COI)基因序列、28S r DNA序列的D2D3区以及18S r DNA序列的部分序列作为DNA条形码在中华钩线虫中的适用性。结果表明,18S r DNA序列可作为该种线虫的DNA条形码,为海洋线虫的DNA条形码研究提供了很好的借鉴。目前,利用DNA条形码技术对海洋线虫进行鉴定的报道在国内还属空白,本研究将是海洋线虫分类学研究的很好补充。同时,对于了解该海域海洋线虫多样性及群落分布格局,开展海洋环境监测,进而对海洋的底质环境状况进行健康评价具有十分重要的科学意义。     

甘肃老豆金矿矿石中金属矿物组成特征研究

老豆金矿床是西秦岭夏河—合作矿集区典型金矿床之一。本文通过显微镜下观察和电子探针分析,确认矿石中存在黄铁矿、毒砂、方铅矿、闪锌矿、辉锑矿、辉锑铅矿、脆硫锑铅矿、块硫锑铜矿、方黝锡矿等多种矿物,确定老豆金矿成矿金属元素组合为Au-As-Sb-Cu-Pb-Zn,根据金元素含量特点判断金元素主要以超显微纳米级自然金(Au0)和固溶体金(Au+)的形式附存于闪锌矿等载金矿物中,方黝锡矿等还原性矿物特征指示矿床形成于氧逸度较低,硫逸度较高的还原环境中。     

Evidence of hybridisation within the genus Chrysoblephus and conserved nuclear sequences of South African sparids (Teleostei: Sparidae)

Several hybridisation events have been recorded in marine fish, yet there is little information for southern African species. Here we present evidence for hybridisation within the genus Chrysoblephus (Sparidae), by analysing a fish caught off KwaZulu-Natal, off the east coast of South Africa. We utilise three mitochondrial (16S, COI and Cyt b) and four nuclear markers (1st intron of the S7 ribosomal protein gene, Tmo-4C4, Histone 3 and 18S rRNA) to resolve the hybrid parentage. MtDNA analyses support C. anglicus as the maternal lineage. Analyses of the Tmo-4C4 gene and the first intron of the S7 gene show six and four heterozygous positions for the hybrid fish respectively. The heterozygous positions are shared by C. anglicus and C. puniceus, which suggests C. puniceus to be the paternal lineage, with the hybrid being a first generation cross. Morphologically, the hybrid mostly resembles C. puniceus, with some meristic counts overlapping with other sparid species. Interestingly, only two of the four nuclear markers show variation within the sparids sampled in this study; 1 160 bp of 18S rRNA and Histone 3 were invariable even between the genera Chrysoblephus, Gymnocrotaphus, Petrus and Pachymetopon, suggesting conserved nuclear evolution or a rapid speciation within this group of endemic fishes.