基于线粒体COⅠ基因的齿小蠹属昆虫DNA条形码研究

齿小蠹属（鞘翅目： 小蠹科）昆虫是植物检疫中经常截获的类群, 为探讨线粒体细胞色素C氧化酶亚基Ⅰ(COⅠ)基因的特定区段作为DNA条形码快速准确鉴定齿小蠹种类的可行性, 以齿小蠹属昆虫为研究对象, 测定分析了线粒体COⅠ基因462 bp碱基序列。序列分析结果显示： 变异位点为259个, 保守位点203个, 简约信息位点181个, 自裔位点78个。所有位点中, A, G, C和T碱基平均含量分别为30.7%, 16.5%, 17.0%和35.8%。A+T含量较高, 为66.5%, 明显高于G+C含量, 表现明显的A+T碱基偏嗜, 且A与T含量相当, 符合昆虫线粒体基因碱基组成的基本特征。转换与颠换结果显示： 该段序列未达到饱和, 可以得到准确的进化分析。利用Kimura 2-parameter模型分析遗传距离得到, 同物种间的遗传距离介于0.002~0.007之间, 不同种间的遗传距离介于0.056~0.431间, 平均遗传距离为0.199, 说明该段序列能够区分不同物种。基于COⅠ基因序列构建的邻接法系统发育树（NJ树）显示, 同一物种聚为同一小支, 且分支自展值均为100%; 近缘种能聚集在一起, 且置信度很高（≥97%）。结果表明应用基于COⅠ基因片段的DNA条形码进行齿小蠹属昆虫分类鉴定具有可行性。     

基于DNA条形码对北京地区蔷薇科花卉上蚜虫的快速鉴定

蔷薇科花卉植物是北京城市绿化的主要观赏植物,也是多种蚜虫的寄主植物。由于蚜虫体型小,形态特征特化或退化,具有复杂的多型现象,依据传统的形态学特征往往无法实现对物种的准确而快速的鉴定,因此会影响对蚜虫的及时有效防治。本研究应用DNA条形码技术,基于COⅠ基因序列分析,对北京地区主要蔷薇科花卉上的蚜虫进行编码,试图在花卉种植和害虫防治时用于物种的快速、准确鉴定。实验得到9属12种蚜虫的65条COⅠ基因序列,种内平均差异为0.70%,种间平均差异为9.45%,最高可达12.59%。基于COⅠ序列构建了NJ树和MP树,绝大多数物种的样品有效地聚为一支,且支持率达到了90%以上,构建的系统发育树可以很好的显示相同物种样品的聚类。结合遗传距离和系统发育树分析表明,基于COⅠ序列的DNA条形码能有效区分83%的北京地区蔷薇科花卉蚜虫物种。     

基于DNA条形码的四川老君山国家级自然保护区尺蛾科昆虫种类鉴定

为了解四川老君山国家级自然保护区鳞翅目Lepidoptera尺蛾科Geometridae资源,于2019年6—8月和2020年5—6月通过灯诱法采集2 402号标本,运用DNA条形码技术对标本进行分子鉴定。通过BOLD数据库,邻接法构建系统发育树和ABGD在线工具对获得的297条线粒体细胞色素C氧化酶亚基Ⅰ(COⅠ)基因序列进行物种界定,结合BOLD和GenBank中70条相关种序列共367条COⅠ基因序列分析遗传距离。发现297条序列隶属于191种,其中95种鉴定到种,35种鉴定到属;鉴定到属或种水平的序列隶属于92属。计算遗传距离发现,种内遗传距离为0～4.36%,平均种内遗传距离为0.63%,种间遗传距离为3.43%～19.12%,平均种间遗传距离为11.91%。结果表明,保护区尺蛾科昆虫种类丰富,基于COⅠ基因的DNA条形码技术能有效区分尺蛾科物种。     

慈竹叶蝉类害虫DNA条形码分析

叶蝉类昆虫形态结构多样,在农林生态系统的物种多样性和植物保护工作中扮演着重要角色,但其物种的准确鉴定一直是农林植保工作中的难点。DNA条形码技术极大促进了农林生态系统物种的快速、准确鉴定。本研究经过连续2年的野外调查采集慈竹Bambusa emeiensis主要叶蝉种类,扩增了广泛分布于中国慈竹的12种主要叶蝉类害虫的线粒体基因COⅠ和16S rRNA序列片段,并进行了遗传距离、系统发育及矢量Klee-diagram图分析。结果显示:慈竹叶蝉昆虫COⅠ基因序列片段(590 bp)种内遗传距离为0.004,种间遗传距离为0.283;16S rRNA基因序列片段(463 bp)种内遗传距离为0.003,种间遗传距离为0.257;不同种间存在明显的条形码间隔。2个基因序列片段的分子系统发育分析结果与形态学研究谱系关系一致。Klee-diagram图分析结果和分子系统发育结果一致。上述结果表明,COⅠ和16S rRNA基因适用于慈竹叶蝉类昆虫的物种鉴定,可为竹林叶蝉类昆虫的准确快速鉴定提供参考方法。     

部分山雀科鸟类的DNA条形码与物种识别

近年来,DNA条形码(DNA barcodes)被认为是鸟类物种识别和分类的有效手段。采用遗传距离和构树这两种方法检验了线粒体基因COⅠ片断作为DNA条形码区分包括长尾山雀在内的山雀科鸟类的识别效果。此次实验分析了92条COⅠ序列,其中50条来自GenBank,代表了该科的30个物种。分析结果表明:尽管山雀科鸟类呈现出较大的种内变异,但是该科鸟类的种间遗传距离还是远大于种内的遗传距离。COⅠ条形码可识别出大部分的山雀科物种,同时该条形码还可鉴定出亚种组、亚种、甚至地理种群。然而,对于近期分化和存在杂交的物种,COⅠ难以鉴定出。在此情况下,探索应用核基因条形码或者采用基于碱基属性的鉴定方法则可以弥补这一缺陷。比较古北界、东洋界与新北界(北美)的山雀类发现,古北界和东洋界的山雀具有较大的种内变异且种群内具有较大的遗传分化,推测这很可能和第四纪冰川的影响有关。     

DNA条形码在鞘翅目昆虫分子系统学研究中的应用

近年来,DNA条形码(DNA Barcoding)技术已经成为生物分类学研究中备受关注的新型技术,并在鞘翅目昆虫系统发育研究中得到广泛应用。本文总结了鞘翅目昆虫DNA条形码研究所用COⅠ基因序列,概述了DNA条形码在鞘翅目昆虫的物种分类鉴定、发现新种和隐存种、系统发育关系研究等方面的应用,并对DNA条形码研究技术新进展和标准序列筛选需要注意的问题进行了讨论。     

水葱新害虫——黑粒脉蜡蝉形态及DNA条形码研究

本文报道发生在贵州省威宁县草海国家级自然保护区水葱Schoenoplectus tabernaemontani上的1种新害虫——黑粒脉蜡蝉Nisia fuliginosa的形态及其DNA条形码。详细描述了黑粒脉蜡蝉卵、若虫及成虫的主要形态特征;同时通过PCR获取黑粒脉蜡蝉的线粒体COⅠ基因序列;运用MEGA 6.0分析其序列组成及遗传距离,以Nisia sp.和Phaconeura sp. 2种粒脉蜡蝉科Meenoplidae昆虫为外群,采用邻接法构建系统发育树。系统发育树显示,黑粒脉蜡蝉6个样品(卵、若虫及成虫)的COⅠ基因序列能较好地聚为一支,且可明显与其他种类区分开来;遗传距离分析结果表明,黑粒脉蜡蝉种内遗传距离为0,3种粒脉蜡蝉科昆虫种间遗传距离为0.166 8～0.236 1,种内遗传距离与种间遗传距离不存在重叠现象。初步证实了COⅠ基因序列可作为DNA条形码片段,从而实现黑粒脉蜡蝉卵、若虫及成虫快速而准确的物种鉴定。此外,本文还报道了黑粒脉蜡蝉在水葱上的危害状况。     

DNA条形码技术在北京百花山地区夜蛾科物种鉴定中的应用

为了探讨DNA条形码技术在夜蛾物种鉴定中的可行性, 本研究利用条形码通用引物扩增了北京百花山地区43种夜蛾75个样本的线粒体细胞色素C氧化酶亚基I （mitochondrial cytochrome c oxidase subunit I, COI）基因序列, 以Kimura双参数模型进行种内种间遗传距离分析、 使用邻接法（neighbor-joining, NJ）和最大简约法（maximum parsimony, MP）分别构建系统发育树, 并利用分子序列差异阈值对样本进行分子可操作分类单元（molecular defined operational taxonomic units, MOTU）划分。结果表明： 所有夜蛾种类通过系统发育树可以成功区分; 种内平均遗传距离(0.03%)远远小于种间平均遗传距离(11.29%); 采用较为保守的1%的序列差异阈值将75个夜蛾样本分为42个MOTU, 正确率为95%, 除了MOTU04包含2个物种外, 剩余41个MOTU与形态种呈现一一对应的关系。结果显示, 基于COI基因的DNA条形码对于本研究中所涉及的夜蛾具有较好的区分, 可以作为一种有效的工具在夜蛾科昆虫物种鉴定中进行应用。     

基于DNA条形码对桃属植物上蚜虫的快速鉴定

桃属(Amygdalus)植物是我国重要的果树,有些种类是常见的城市绿化观赏树种,也是多种蚜虫的寄主。蚜虫的体型小,有复杂的多型现象,传统的形态学特征往往无法实现对物种的准确而快速的鉴定。本研究应用DNA条形码技术,基于COⅠ基因序列分析,对我国桃属植物上的蚜虫进行编码,为桃属植物上蚜虫的物种快速、准确鉴定提供有力的支持。本研究共编码桃属植物上蚜虫12种,其中蚜亚科Aphidinae 6属10种,毛管蚜亚科Greenideinae 1属1种,毛蚜亚科Chaitophorinae 1属1种。共获得COⅠ基因序列96条,种内平均差异为0.76%,种间为5.7%¹5.5%。基于COⅠ序列构建了NJ树,绝大多数物种的样品有效地聚为一支,且支持率达到了95%以上。结合遗传距离和系统发育树分析表明,基于COⅠ序列的DNA条形码能有效区分99%的桃属植物上的蚜虫物种。     

DNA条形码试剂盒检测技术在大小蠹属种类鉴定中的应用

[目的]DNA条形码技术已成为生物分类鉴定的有力工具.DNA条形码技术的相关问题,如物种种内和种间的遗传距离出现重叠区域,将直接影响到物种鉴定的准确性.我们应用DNA条形码试剂盒检测技术来快速、准确地鉴定口岸截获的检疫性大小蠹属种类.[方法]针对大小蠹昆虫设计引物以提高PCR扩增效率.运用自主研发的基因条码分析软件找出基因片段上区分每个物种的多态位点规律,作为该物种的鉴定特征并建立数据库,应用于物种鉴定.[结果]使用针对大小蠹属昆虫设计的引物成功扩增出325 bp的COI基因片段.将大小蠹属12种昆虫的COI基因片段上的核苷酸诊断位点的组合作为物种的鉴定特征,可以准确地区分近似种.通过比对植物检疫鉴定系统数据库里的鉴定特征,将6个大小蠹属的未知样品成功鉴定到种(核苷酸序列一致性为100％),与形态鉴定结果一致.[结论]结果表明DNA条形码试剂盒检测技术可以准确鉴定大小蠹属的种类.该检测技术可以应用于其他经济重要性有害生物的检测鉴定.     

Uniform hypothesis of cell behavior--movement, contact inhibition of movement, adhesion, chemotaxis, phagocytosis, pinocytosis, division, contact inhibition of division, fusion.

The cell has been represented as a charged liquid drop. Contrary to the DLVO-theory, the effect of the surface potential upon the value of the interfacial tension of the cell membrane has also been taken into consideration. The cell membrane has visco-elastic properties and its constituents may move against each other. Cell movement is caused by the appearance of a small number of the electrically charged constituents of the cell membrane on the leading edge of the cell. This produces a local decrease in the surface tension and the cell membrane expansion. At the moment of contact between two cells proton transfers occur between the strongly negatively charged microvilli of one cell and the body of the other, analogous to a condenser breakdown. This, through the effect on the surface tension, causes contact inhibition of movement. The distribution of the proton dissociable groups modifies the interaction between the cells (differentiation) and between the cell and the substratum (adhesion). Adsorption of the charged compounds at the surface of the cell membrane, decreasing the surface potential and increasing the surface tension, causes the phenomena of chemotaxis, phagocytosis and pinocytosis. Cell division, considered in the terms of the surface energy, requires an adequate supply of considerable quantities of energy inversely proportional to the surface potential value. In case of a reduction of the distance between the cells, their surface potential and the energetic barrier of the cell division processes increases, and causes contact inhibition of cell division. Due to their high charge, division of neoplastic cells is inhibited much later than division of normal cells, or is completely ininhibited due to geometric conditions. Fusion of the cell membrane in the intra-cellular and intercellular processes is a reverse process in relation to the cell division.     

The Effect of Age and Gender on Al, B, Ba, Ca, Cu, Fe, K, Li, Mg, Mn, Na, P, S, Sr, V, and Zn Contents in Rib Bone of Healthy Humans

The effect of age and gender on major, minor, and trace element contents in the intact rib bone of 80 relatively healthy 15–55-year-old women and men was investigated. Contents or upper limit of contents of 16 chemical elements in the rib bone were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES). Mean values (M?±?SΕΜ) for the mass fraction of Ba, Ca, Cu, Fe, K, Li, Mg, Na, P, S, Sr, and Zn (milligram per kilogram of dry bone) were as follows: 2.54?±?0.16, 171,400?±?4,050, 1.35?±?0.22, 140?±?11, 1,874?±?71, 0.049?±?0.011, 2,139?±?38, 5,378?±?88, 75,140?±?1,660, 1,881?±?51, 291?±?20, and 92.8?±?1.5, respectively. The upper limits of contents of Al, B, Mn, and V were <7.20, <0.65, <0.36, and <0.03, respectively. Statistically significant tendency for the Ca, Mg, and P content to decrease with age was found in the human rib bone, regardless of gender. The mass fraction of Fe in the male rib bone increases with age. It was shown that higher Ca, Mg, Na, P, and Sr mass fractions as well as lower Fe content were typical of female ribs as compared to those in male ribs.     

Distribution of N,P, K,Ca, Mg,S, Zn,Fe, Mn and Cu in groundnut

Summary Concentration of N, P, K, Ca, Mg and S in summer groundnut crop was higher than in kharif while Zn, Fe, Mn and Cu contents were higher in summer crop. Kernel's N, P and Zn; Leaflet's Ca and Mn; Stem's K and Fe; Root's S and Cu and Petiole's Mg contents were highest. Shell's N, P, K, Mg, S, Zn and Cu; Kernel's Ca, Fe and Mn contents were the least. N, P, K, S, Zn and Cu concentrations decreased linearly as the crop grew. Ca, Mg, Fe and Mn concentrations did not display any distinct pattern. Ca concentration was positively correlated with pod yield in both the seasons.     

The effect of malathion,diazinon, and various concentrations of zinc,copper, nickel,lead, iron,and mercury on fish

Acute and chronic toxicity tests for malathion, diazinon, copper (Cu), mercury (Hg), lead (Pb), zinc (Zn), nickel (Ni), and iron (Fe) were conducted. Mortalities ofBarilius vagra andCyprinus carpio (common carp) were variable but LC50-96 hr were similar for pesticides. AdultB. vagra seem to be more sensitive to malathion than juvenile carp. Both juvenile carp and adultB. vagra were extremely sensitive to diazinon. Long-term exposure to pesticides modified morphology and behavior. The LC50-96 values for Cu, Hg, and Pb were 0.3, 0.16, and 0.44, respectively, for smaller fish and 1.0, 0.77, and 1.33, respectively, for larger fish. Replicate LC50 values for Zn, Ni, and Fe were somewhat variable, and for these metals, the size of the fish seemed to affect response because LC50 values increased as fish size increased. Cooper, Pb, Zn, and Fe residues following exposure to sublethal concentrations of these metals for 15 d were significantly greater in whole juvenile common carp than in controls.     

Synthesis,tautomerism, and antimicrobial,anti-HCV,anti-SSPE,antioxidant, and antitumor activities of arylazobenzosuberones

2-Dimethylaminomethylene-1-benzosuberone 1 was coupled with diazotized aniline derivatives to afford a series of the hitherto unreported 2-arylazo-1-benzosuberones 3a–i. The tautomeric structure and the effect of substituents on the tautomeric form (s) of the products 3a–i were discussed. Similar coupling of the enaminone 1 with diazonium salts of heterocyclic amines gave the respective fused azolotriazino-benzosuberones. Some of the newly synthesized compounds showed potent antimicrobial, anti-HCV, antioxidant, antitumor (as topoisomerase I inhibitors), and antimicrobial activities.