广东流溪河水库盔型溞(Daphnia galeata)休眠种群与现生种群的单倍型多样性和遗传分化

休眠卵库作为淡水枝角类生物与遗传信息的储藏库,从沉积物休眠卵库中萌发的枝角类对现生种群的数量与种群遗传结构有着直接的影响.分别采集流溪河水库盔型溞的现生种群和沉积物表层(0~10 cm)的休眠卵,扩增现生种群与休眠卵的线粒体细胞色素C氧化酶亚基I基因,构建了单倍型网络.休眠种群相比现生种群有着较高的单倍型多样性和核酸多样性,初期现生种群分别为0.562、0.00104,末期现生种群分别为0.726、0.00331,休眠种群分别为0.815、0.00761.流溪河盔型溞现生种群与休眠种群存在双向基因流,现生种群到休眠种群的有效迁移率为490.9,休眠种群到现生种群的有效迁移率为527.5.通过构建贝叶斯系统树验证了休眠种群和现有种群中并不存在隐种或者亚种的分化,休眠种群与现生种群之间没有出现较大的遗传分化,现生种群遗传多样性来自于休眠种群,水库的休眠种群更能反映种群真实的遗传多样性.休眠种群与现生种群之间的基因流与休眠卵库大小无关,与休眠卵的萌发有关.     

基于DNA条形码技术的浮游动物休眠卵种类鉴定:以洞庭湖流域常德柳叶湖为例

以洞庭湖流域的典型城市湖泊常德柳叶湖表层沉积物中浮游动物休眠卵为研究对象,采用DNA条形码技术进行种类鉴定,从191个休眠卵中成功获得101条有效序列,鉴定成功率约为53%.根据NCBI数据库比对成功鉴定休眠卵9科12属11种,另有6个样品鉴定到属或科;-3个类群的种间遗传距离平均为种内遗传距离的68倍,表明可以利用线粒体细胞色素c氧化酶第一亚基编码基因(COⅠ)对休眠卵进行有效物种鉴定.通过Neighbor-Joining树进行系统发育分析,发现所鉴定的物种与其参比序列聚类为一支,所有物种分别聚为独立的一个支系,不同物种可有效区分.研究结果均表明,COⅠ基因作为DNA条形码可以实现沉积物中浮游动物休眠卵的物种鉴定.     

两种不同DEM处理方法提取的水系比较

水系不但是重要的自然地理要素,而且对防洪减灾、资源开发等有着重要意义。基于DEM的坡面径流法是最为常用的水系提取方法,其提取精度直接取决于DEM中洼地的处理方法。为了比较不同DEM处理方法对水系提取精度的影响,本文以江西省潦河流域为例,利用基于DEM数据的J＆D算法及基于辅助数据的AGREE方法来处理实验区DEM,然后利用坡面径流法提取水系。结果表明,经两种方法后均可以提取出连续的水系,且提取水系的等级、数量及长度相差不大;在提取精度上,在受人类活动影响较小的山地丘陵地区两种方法提取的水系比较吻合,而在平坦地区或受人类活动影响较大的地区提取的水系差异较大,且AGREE方法处理后提取的水系精度较高。     

大型溞(Daphnia magna)线粒体基因组的测定与序列分析

利用Long-PCR及普通PCR技术,结合引物步移法测序,经拼接、组装后获得中国淮河大型溞(Daphnia magna)线粒体基因组全序列,并对其基因组成、结构特点等进行初步分析.大型溞线粒体基因组全长为14948 bp,A、T、G、C各碱基含量分别为32.37%、34.73%、15.58%、17.31%,表现出明显的AT偏倚.13个蛋白质编码基因、22个t RNA基因、2个rRNA基因和1个D-loop区(控制区)的排列方式与典型的节肢动物线粒体基因组的排列方式略有不同.除COI基因以CTG、ATP8基因以GTG、ND3基因以ATC以及ND6基因以ATT为起始密码子以外,其余9个蛋白质编码基因均以ATG作为起始密码子;9个蛋白质编码基因具有典型的完全终止子TAG或TAA,COI、COII、ND4和ND5等基因采用不完全终止密码子T.基因组包含9个基因间隔区,共81 bp,长度1~62 bp;13个基因重叠区,共77 bp,重叠碱基数在1~27 bp之间,最大重叠在16 S rRNA和tRNA~(Val)基因之间.在预测的22个tRNA基因的二级结构中,发现只有tRNA~(Ser1)基因未能形成完整的二级结构,其他21个基因均可形成典型的三叶草结构.16 S rRNA和12 S rRNA基因长度分别为1373 bp和752 bp,D-loop区全长289 bp.本研究结果为探讨大型溞在溞属中的系统学地位及其与其他种间的系统发生关系等问题提供了数据资源.     

微囊藻群体总RNA提取方法的比较

微囊藻群体富含多糖类物质是影响微囊藻RNA提取的关键因素.为了获得高质量的微囊藻群体总RNA,对比分析4种针对多糖含量较高的方法——方法 1 PGTX-bead法、方法 2 CTAB-bead法、方法 3 Fast RNAPro Blue Kit和方法 4RNeasy Mini Kit对微囊藻群体总RNA的提取效果.采用琼脂糖凝胶电泳检测微囊藻群体RNA的完整性,Nanodrop ND1000分光光度计检测RNA纯度及浓度,并采用q PCR检测DNA污染情况.结果表明,4种方法都能从微囊藻群体中提取获得RNA,并在去除DNA后都可以进行RT-PCR等后续实验.方法 1 PGTX-bead提取的RNA产量最高,纯度好,DNA污染小,成本低,适合从微囊藻群体中大量提取RNA;方法 2 CTAB-bead提取的RNA样品产量也较高,但DNA污染严重,适合需要同时提取DNA和RNA的样本;方法 3 Fast RNAPro Blue Kit和方法 4 RNeasy Mini Kit提取的RNA产量都较低,但方法 4操作简单,耗时短,所检测目的基因的相对表达量较高,更适合从少量的微囊藻群体中提取总RNA.     

离散型湖泊水体提取方法精度对比分析

基于卫星遥感的陆地水体提取方法多种多样,并且应用广泛.对于水体分布支离破碎的枯水期湖泊,准确的水体提取方法尚不明晰,直接影响湖泊水域面积的提取精度.以鄱阳湖湖区为研究对象,利用ALOS遥感影像,以2.5 m高分辨率全色波段融合影像非监督分类(ISODATA)得到的水体面积为参考值,分别使用归一化水体指数(NDWI)法、NDWIISODATA法和基于近红外(NIR)的ISODATA法提取了10 m分辨率的水体分布,分析了不同方法提取结果之间的差异性及产生原因.结果表明:3种方法均可以较好地提取出水体,但利用ISODATA法提取的水体细部信息更为明显,面积值较NDWI法更大;相对于近红外单波段而言,基于NDWI图像的ISODATA法提取水体的精度更高.纵观3种方法,基于NDWI图像的ISODATA法提取的水体精度最高,基于近红外波段的ISODATA法提取结果次之,NDWI阈值法的提取效果最差.研究结果对于离散型湖泊水体提取方法及数据源的选择等具有重要的借鉴和参考意义.     

基于高分六号数据提取太湖蓝藻方法的对比及适用性分析

以往对太湖蓝藻提取的研究中,较少将国内高分系列卫星影像作为数据源.为此本文基于2019年太湖不同蓝藻暴发时期的高分六号数据,采用随机森林(random forest, RF)法、归一化植被指数(normalized differential vegetation index,NDVI)阈值法和多光谱绿潮指数(multispectral green tide index,MGTI)阈值法对太湖蓝藻进行提取以寻求提取太湖蓝藻的最佳方法,并在此基础之上探讨了RF不同输入变量的适用性.结果表明,2019年太湖南岸和梅梁湖的蓝藻水华富营养化较为严重,NDVI阈值法明显将部分轻度蓝藻漏提,MGTI阈值法对中、重度蓝藻的识别能力不如NDVI法,而RF法能够有效地提取太湖蓝藻.其中将归一化植被指数和归一化水体指数作为输入变量的RF法提取中型暴发期的太湖蓝藻精度最高,总体分类精度和Kappa系数分别为99%和0.97.研究太湖蓝藻的提取方法对太湖的环境治理提供了技术支持,也为其他湖泊蓝藻的遥感监测提供了科学依据.     

五个湖泊中华刺鳅(Mastacembelus sinensis)种群线粒体细胞色素b遗传多样性分析

采用特异性引物对中华刺鳅基因组DNA进行扩增和测序,获得了江苏省5个湖泊的中华刺鳅种群线粒体cyt b基因767 bp的序列.分析表明,cyt b基因中A+T含量略高于G+C含量,144个中华刺鳅共获得cyt b基因单倍型28个,变异位点38个,其中单倍型Hap1分布最为广泛,可能为祖先型单倍型.平均单倍型多样性和平均核苷酸多样性指数分别为0.58751和0.00214,遗传多样性较低.群体间遗传分化系数为0.0356,绝大多数的遗传变异都来自群体内部,而群体间的遗传分化极小.构建的系统进化树未出现明显的以单个湖泊群体的聚簇,5个湖泊的种群间基因交流极其强烈.研究结果表明,当前5个湖泊的中华刺鳅遗传多样性水平比较低,亟待加强保护.     

5个湖泊河川沙塘鳢(Odontobutis potamophila)种群线粒体细胞色素b基因的遗传变异分析

通过对5个湖泊的河川沙塘鳢种群的线粒体DNA细胞色素b基因进行PCR扩增、测序,获得1141 bp的序列全长.序列分析显示,cyt b基因序列中A+T含量(55.8%)略高于G+C含量(44.2%),共检测到806个多态位点,115个样本得到87个单倍型,平均单倍型多样性为0.969±0.012,核苷酸多样性为0.20081±0.00742,遗传多样性表现高度多样性.太湖种群与大纵湖种群间的遗传距离最近,为0.137,巢湖种群和大纵湖种群之间遗传距离最远,为0.424.分子方差分析表明,群体间遗传分化系数Fst为0.531,变异来自群体内及群体间.cyt b基因序列构建的UPGMA系统进化树显示,5个种群分化成不同的分支系谱,种群间存在的基因交流较少.     

长江中下游5个湖泊黄颡鱼(Pelteobagrus fulvidraco)种群线粒体细胞色素b基因的遗传变异分析

对长江中下游5个湖泊的黄颡鱼种群的线粒体DNA细胞色素b基因进行了PCR扩增、测序,获得了955 bp的序列.分析显示,cyt b序列中A+T含量略高于G+C含量,共检测到54个变异位点,60个样本得到37个单倍型,平均单倍型多样性为0.945±0.018,核苷酸多样性为0.00419±0.00043,遗传多样性表现中等.太湖种群与滆湖种群间的遗传距离最远为0.00651,鄱阳湖种群和巢湖种群之间遗传距离最近为0.00375.分子方差分析(AMOVA)表明,群体间遗传分化系数F_st为0.0684,几乎所有变异都来自群体内,群体间遗传分化极小.cyt b序列构建UPGMA系统进化树显示,5个种群没有分化成不同的分枝谱系,种群间存在广泛的基因交流.     

Sample preparation methods for quantitative detection of DNA by molecular assays and marine biosensors

The need for quantitative molecular methods is growing in environmental, food, and medical fields but is hindered by low and variable DNA extraction and by co-extraction of PCR inhibitors. DNA extracts from Enterococcus faecium, seawater, and seawater spiked with E. faecium and Vibrio parahaemolyticus were tested by qPCR for target recovery and inhibition. Conventional and novel methods were tested, including Synchronous Coefficient of Drag Alteration (SCODA) and lysis and purification systems used on an automated genetic sensor (the Environmental Sample Processor, ESP). Variable qPCR target recovery and inhibition were measured, significantly affecting target quantification. An aggressive lysis method that utilized chemical, enzymatic, and mechanical disruption enhanced target recovery compared to commercial kit protocols. SCODA purification did not show marked improvement over commercial spin columns. Overall, data suggested a general need to improve sample preparation and to accurately assess and account for DNA recovery and inhibition in qPCR applications.     

Modulation of longevity in Daphnia magna by food quality and simultaneous exposure to dissolved humic substances

Various abiotic and biotic factors determine the natural fluctuations of Daphnia spec. populations; food quality and dissolved humic substances (HSs) being among these factors. In this contribution, we try to disentangle the relative impact of food quality and simultaneous HSs exposure on the fertility and longevity of D. magna. It is understood that HS-mediated stress leads to reduced fecundity in well-fed D. magna females; hence, it was expected that poor food, as a second stressor, would aggravate the HS-mediated effects. Three diets were tested: the green algae Pseudokirchneriella subcapitata, baker’s yeast alone, and baker’s yeast plus dissolved ascorbic acid, and exposed D. magna to a HS preparation which has been shown effective in previous bioassays. It was hypothesized that the lifespan and fertility of D. magna would be best when fed green algae, and worst when fed only baker’s yeast. However, contrary to these expectations, any addition of HSs reduced the stress caused by poor food quality and increased lifespan and fecundity. In the yeast series, asexually produced diapausing eggs occurred via a so far unknown pathway. With yeast diet, the expanded lifespans were slightly above, whereas the increased offspring numbers lay below, the corresponding data of the algae-fed individuals. The potential of HSs as an additional food source and as a means to extend the lifespan is discussed. These findings open the innovative perspective that under low quality food conditions, additional stressors at certain intensities may even be beneficial to individuals and populations.     

The research of a comprehensive program for earthquake prediction with seismological methods

In this paper, the MYCIN inexact inference method in Expert System is applied to comprehensive earthquake prediction. And it is proposed that the methods of determining various certainty factors, correcting correlation between anomalous evidences and computing comprehensive certainty factor of occurrence of some moderate or strong earthquake. By use of these methods, 18 earthquake cases since 1966 in North China is tested with seismological anomalies in different seismogenic stage, and the comprehensive certainty factors of occurrence of some moderate or strong earthquake are computed. At last some problems in application are discussed. The Chinese version of this paper appeared in the Chinese edition ofActa Seismologica Sinica,13, 328–337, 1991.