DNA条形码在鳞翅目昆虫中的应用

2003年,Hebert等提出DNA条形码后,快速而精确的特点使它在物种鉴定中得到了广泛的应用。鳞翅目是昆虫纲中第二大目,其物种鉴定任务复杂而艰巨,因此DNA条形码具有广阔的应用前景。该文主要针对DNA条形码概况以及近年来它在鳞翅目昆虫中的研究情况予以综述。     

微型DNA条形码在水产类物种掺假检测中的应用

水产类物种掺假导致了商业欺诈、食品安全等问题,建立一种快速高效的水产类物种掺假检测技术迫在眉睫.目前,标准DNA条形码(FDB)在水产类物种掺假检测中已经得到了广泛应用,但是食品加工中的高温、高压、反复冻融和酸碱变化等因素均会使DNA发生降解,导致标准DNA条形码扩增失败.微型DNA条形码(MDB)可以利用相对较短(＜...     

物种丰富度垂直分布格局及影响机制

物种丰富度分布格局是一定地域内物种丰富度沿三维空间的立体分布,包括物种丰富度在经度、纬度和垂直梯度(海拔高度和海水深度)三个维度上的空间分异。近年来物种多样性的垂直分布格局与机制研究得到了生物地理学家和生态学家的重视。物种丰富度的垂直分布格局存在多种类型,但随海拔增加而物种数减少的单调递减模型和中海拔物种丰富度最高的单峰模型较为常见。目前在机制研究中验证较多的是气候稳定性、生物因子(种间相互作用)、能量、生境异质性、干扰、进化时间、物种分化速率、面积、中域效应(mid-domain effect)、生态位保守性(niche conservatism)等假说和机制。物种丰富度的分布格局是多方面因素综合作用的结果;由于地理、地形、气候、地质演化历史、物种库和进化历史、物种分化速率、干扰等差异,在不同地区存在着特别的物种丰富度空间分布格局和机制;处于同一地区的不同类群的物种也因进化扩散历史和生态适应能力不同而呈现多样化的分布格局。因此,对不同地区和类群的物种丰富度格局和机制进行研究应具体分析后才能得到可信结论。     

中国不同地理区域鸟兽物种丰富度的相关性

生物类群之间物种丰富度的相关性研究是当前物种多样性研究中的热点问题之一,目前,中国尚无相关的研究报道。我们收集了中国三种区域类型：动物地理亚区、行政区和保护区的鸟兽名录,分析了行政区与保护区、动物地理区和经纬度带中鸟兽物种数比值及其相关性。 结果表明：不同区域、动物地理区和经纬度带中鸟兽物种数都显著相关。保护区尺度鸟兽物种 数的相关系数为0.818和动物地理区中的华北区为0.768,其他所有区域和地理区域的鸟兽物 种数的相关系数都高于0.850。因此,鸟兽物种数的相关关系在一定程度上具有预测价值。我们发现不同区域鸟兽物种数比值无显著性差异;但是,不同区域间鸟兽物种数 比值差异显著。该比值在中国呈中间低四周高的分布趋势,其中东北地区最高。我们还利用历史累积调查数据与非历史累积调查数据进行了鸟兽物种数比值及其相关性分析,发现利用累积数据计算的相关性低于非累积数据计算的相关性,但利用累积数据计算的鸟兽物 种数比值高于非累积数据计算的比值。最后,探讨了为什么鸟类与兽类的物种数目会相关。我们根据物种－面积公式,S=CA^Z,导出了两个生物类群物种丰富度的相关关 系式。利用全国不同区域数据拟合,得到Z₁/Z₂＝0.913,Z₁/Z₂接近于1。于是 ,C₁/C2可视为近似等于R_am。本研究可推广到其他不同生物类群物种。物种数量的相关关系为快速评估区域的物种多样性提供了一条途径。     

中国芦苇沼泽植物物种丰富度分布格局及其驱动因素

明确大尺度物种丰富度的分布格局及其影响因素一直是生态学及生物地理学领域的核心议题之一,其对于预测全球环境变化背景下生物多样性的响应和制定减少生物多样性丧失的保护方案有着重要意义。本文基于全国芦苇(Phragmites australis)沼泽野外调查数据,结合气候、地理、土壤等环境因子,探讨了中国芦苇沼泽植物物种丰富度的分布格局及其驱动机制。结果表明:(1)中国的芦苇沼泽植物物种丰富度总体表现为亚热带湿润区和温带湿润半湿润区较高,青藏高原区、温带干旱半干旱区和滨海区较低的特点;(2)芦苇沼泽植物物种丰富度与年降水、土壤有机碳、土壤总氮、纬度均呈显著正相关,而与年均温、最冷月最低温、土壤pH值、海拔之间表现为显著负相关;(3)土壤因子特别是土壤pH值是影响芦苇沼泽植物物种丰富度的最主要因素,其次分别为淹水状况、气候因子和地理因子;(4)结构方程模型结果表明土壤因子、淹水状况和气候因子直接影响植物物种丰富度,而地理因子通过调控土壤因子、淹水状况与气候因子间接影响植物物种丰富度。总体而言,中国芦苇沼泽植物物种丰富度具有空间异质性,其分布格局受到多种因素的共同影响,土壤因子是影响植物物种丰富度...     

基于DNA条形码技术鉴定库尔勒机场的蝙蝠物种

随着航空业的发展,野生动物与航空器之间的冲突愈演愈烈,研究机场周边飞行动物对机场动物撞击防范工作具有重要意义。蝙蝠作为世界上唯一的飞行类哺乳动物,也是严重影响夜间飞机飞行安全的隐患之一,但由于蝙蝠体型较小,发生撞击后往往无法发现相对完整的尸体,多为血液和毛发等,因此物种鉴定比较困难。从库尔勒机场防鸟网采集到一具蝙蝠样本,基于DNA条形码技术（DNA barcoding）,通过16S rRNA遗传距离分析,发现棕蝠属（Eptesicus）与库尔勒样本群体间遗传距离为0.031~0.092,属内与库尔勒样本种间差异最大的是南美棕蝠（Eptesicus diminutus）,遗传距离为0.092;差异最小的是大棕蝠（Eptesicus serotinus）,遗传距离为0.031。结合形态学分析并通过解剖证实该个体的性腺尚未发育,确定了库尔勒机场挂网蝙蝠物种为大棕蝠。本研究为机场不易辨认或者保存不完整样本的物种鉴定提供方法依据。在确定物种后,了解其生活史特征,有利于机场动物撞击防范工作的精准实施,从而最大限度降低机场损失。     

次生林不同类型森林边缘的鸟类物种丰富度及个体多度比较

边缘效应对动物的分布及行为会产生一定的影响,在鸟类生态学研究中已证实某些鸟类在森林内部和森林边缘区域存在着物种丰富度和个体多度的差异。于1999至2001年的春夏季,在吉林省左家自然保护区对阔叶林/农田边缘、阔叶林/灌丛边缘及阔叶林/针叶林边缘3种不同类型边缘地带的鸟类物种丰富度及个体多度进行了比较研究。结果表明,不同年间鸟类物种丰富度无显著变化,但个体多度存在着一定的波动。不同类型森林边缘的鸟类物种丰富度存在着一定的差异,阔叶林/灌丛边缘的鸟类物种丰富度最高,而阔叶林/针叶林边缘的鸟类物种丰富度最低。鸟类个体多度的总体趋势在3种不同类型的边缘差异不显著,但存在种间差异,灰椋鸟、灰头啄木鸟和喜鹊在阔叶林/农田边缘的个体多度最高,斑啄木鸟、黄胸、三道眉草和日本树莺在阔叶林/灌丛边缘的个体多度最高,而沼泽山雀、冕柳莺和山在阔叶林/针叶林边缘的个体多度最高。     

中条山油松人工林下物种对群落物种丰富度分布格局的贡献

该研究以中条山油松人工林群落为研究对象,研究林下不同大小的子群落对群落物种丰富度分布格局的贡献,并确定影响该区域群落物种丰富度分布格局的关键种,为区域物种多样性保护提供理论依据.结果 表明:(1)该地区林下物种频度分布格局呈明显右偏,且不同样方物种丰富度存在明显差异.(2)常见种对群落丰富度分布格局的贡献大于稀有种.(...     

横断山区树线以上区域种子植物的标本分布与物种丰富度

植物标本是植物空间分布信息的重要来源, 也是估算物种丰富度的主要数据资料。本文收集了有关数据库和标本馆的标本资料, 以分析横断山区树线以上区域种子植物标本的采集现状和物种丰富度。将树线以上区域(4,100–5,500 m)划分成14个100 m海拔带, 将每号标本的海拔信息记录到相应的海拔带内。共收集8,316号标本信息, 记载种子植物1,820种, 其中横断山区特有种655种。这些标本在物种间的分布极不均匀, 仅有1–2号标本的物种最多, 共974种, 占53.5%。各海拔带内标本总数、种均标本数和物种丰富度随海拔的增加而下降, 但物种稀疏曲线不能很好地描述物种丰富度沿海拔梯度的分布格局。因此, 需要开展更多的样地调查和标本采集工作, 为物种丰富度的估算积累更多的资料。     

中国黑戈壁地区植物物种丰富度格局的水热解释

我国黑戈壁地区自然环境恶劣,植物多样性格局受到极端干旱气候的影响而形成了特殊的分布格局。为了揭示黑戈壁地区极端气候对物种丰富度格局的影响,通过对5000 km样线内174个样方中的植物进行调查,结合气候数据,研究影响中国黑戈壁地区植物物种丰富度格局的气候因素以及不同生活型植物物种丰富度格局与气候关系的差异。结果表明,水热动态假说对物种丰富度格局的解释率为62.3%,未解释部分为37.7%,说明其能够很好的解释黑戈壁地区的植物群落物种丰富度格局;能量对物种丰富格局的单独解释率仅为3.5%,水分的单独解释率为16.4%,两者共同解释率为42.4%,水分和能量共同决定的水资源可利用性主导着物种丰富度格局;不同生活型植物对气候因子的响应存在显著差异,水热综合作用对草本植物丰富度格局的解释率为65.4%,但对灌木仅有37.9%,表明水热动态假说不适用于解释灌木植物的物种丰富度格局,植物对气候的适应特性及种间相互作用对物种丰富度格局有着重要的影响。     

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。     

苹果园鳞翅目夜蛾科DNA条形码鉴定

为了检验DNA条形码在鳞翅目夜蛾科蛾类鉴定中的可行性,本文对采自北京昌平苹果园内的夜蛾科14种71头蛾类标本分别提取了DNA,并扩增了线粒体cox1及核基因28S,利用系统发育树、遗传距离、阈值等方法进行了鉴定和比较分析。同时,检验了目前BOLD系统的鉴定成功率。实验表明,基于cox1基因和BOLD系统的鉴定成功率达到了100%,而基于28S则很低,为64.8%。用不同方法构建的系统发育树,鉴定结果均相同。93%的种内遗传距离小于1%,94%的种间遗传距离为大于3%,种内种间的遗传距离形成明显的3%阈值现象。     

Identification of species within Tetrastigma (Miq.) Planch.(Vitaceae) based on DNA barcoding techniques

Many species of Tetrastigma (Miq.) Planch. (Vitaceae) have long been used as medicinal plants in China, and some are endangered due to overexploitation. Although adulterants are often added to traditional Chinese medicines, there is no reliable or practical method for identifying them. In this study, we used four markers (rbcL, matK, trnH-psbA and internal transcribed spacer [ITS]) as DNA barcodes to test their ability to distinguish species of Tetrastigma. The results indicated that the best barcode was ITS, which showed significant inter-specific genetic variability, and thus its potential as a DNA barcode for identifying Tetrastigma. Multiple loci provided a greater ability to distinguish species than single loci. We recommend using the combined rbcL+matK+ITS barcode for the genus. Phylogenetic trees from each barcode were compared. Analyses using the unweighted pair group method with arithmetic mean discriminated an equal or greater percentage of resolvable species than did neighbor joining, maximum likelihood, or maximum parsimony analyses. Additionally, five medicinal species of Tetrastigma, especially T. Hemsleyanum, could be identified precisely using DNA barcoding.     

DNA barcoding confirms polyphagy in a generalist moth,Homona mermerodes (Lepidoptera: Tortricidae)

Recent DNA barcoding of generalist insect herbivores has revealed complexes of cryptic species within named species. We evaluated the species concept for a common generalist moth occurring in New Guinea and Australia, Homona mermerodes, in light of host plant records and mitochondrial cytochrome c oxidase I haplotype diversity. Genetic divergence among H. mermerodes moths feeding on different host tree species was much lower than among several Homona species. Genetic divergence between haplotypes from New Guinea and Australia was also less than interspecific divergence. Whereas molecular species identification methods may reveal cryptic species in some generalist herbivores, these same methods may confirm polyphagy when identical haplotypes are reared from multiple host plant families. A lectotype for the species is designated, and a summarized bibliography and illustrations including male genitalia are provided for the first time.     

DNA条形码技术在田间常见蓟马种类识别中的应用

蓟马类害虫种类多、 体型小, 传统的形态学鉴定方法难以快速准确识别。本研究利用DNA条形码通用型引物, 以我国田间常见的25种蓟马为靶标扩增其线粒体DNA细胞色素C氧化酶亚基Ⅰ (mitochondrial cytochrome c oxidase subunit Ⅰ gene, mtDNA COⅠ) 基因 (约650 bp), 通过对靶标片段碱基序列的测序及比对分析, 以邻接法 (NJ法) 构建系统发育树, 并以Kimura双参数模型计算种内、 种间遗传距离。结果表明： 聚类分析与形态学鉴定结果一致, 表现为较长的种间分支和较短的种内分支, 每个单系分支对应一个物种, 同一物种不同单倍型的最初分支自展值均为100%。25种蓟马的种内平均遗传距离为0.0027, 种间平均遗传距离为0.2757, 种间遗传距离为种内遗传距离的102.1倍; 而且种内、 种间遗传距离没有重叠区域。结果说明基于COⅠ基因的DNA条形码技术可以用于不同种类蓟马的快速准确鉴别。     

DNA barcoding of Gaultheria L.in China (Ericaceae: Vaccinioideae)

Four DNA barcoding loci,chloroplast loci rbcL,matK,trnH-psbA,and nuclear locus internal transcribed spacer (ITS),were tested for the accurate discrimination of the Chinese species of Gaultheria by using intraspecific and interspecific pairwise P-distance,Wilcoxon signed rank test,and tree-based analyses.This study included 186 individuals from 89 populations representing 30 species.For all individuals,single locus markers showed high levels of sequencing universality but were ineffective for species resolvability.Polymerase chain reaction amplification and sequencing were successful for all four loci.Both ITS and matK showed significantly higher levels of interspecific species delimitation than rbcL and trnH-psbA.A combination ofmatK and ITS was the most efficient DNA barcode among all studied regions,however,they do not represent an appropriate candidate barcode for Chinese Gaultheria,by which only 11 out of 30 species can be separated.Loci rbcL,matK,and trnH-psbA,which were recently proposed as universal plant barcodes,have a very poor capacity for species separation for Chinese Gaultheria.DNA barcodes may be reliable tools to identify the evolutionary units of this group,so further studies are needed to develop more efficient DNA barcodes for Gaultheria and other genera with complicated evolutionary histories.     

Identifying Chinese species of Gammarus （Crustacea： Amphipoda） using DNA barcoding

Using a standard cytochrome c oxidase I sequence, DNA barcoding has been shown to be effective to distinguish known species and to discover cryptic species. Here we assessed the efficiency of DNA barcoding for the amphipod genus Gammarus from China. The maximum intraspecific divergence for widespread species, Gammarus lacustris, was 3.5%, and mean interspecific divergence reached 21.9%. We presented a conservative benchmark for determining provisional species using maximum intraspecific divergence of Gammarus lacustris. Thirty-one species possessed distinct barcode clusters. Two species were comprised of highly divergent clades with strong neighbor-joining bootstrap values, and likely indicated the presence of cryptic species. Although DNA barcoding is effective, future identification of species of Gammarus should incorporate DNA barcoding and morphological detection[Current Zoology 55(2):158-164,2009].     

DNA barcoding of Gaultheria L. in China (Ericaceae: Vaccinioideae)

Abstract Four DNA barcoding loci, chloroplast loci rbcL, matK, trnH‐psbA, and nuclear locus internal transcribed spacer (ITS), were tested for the accurate discrimination of the Chinese species of Gaultheria by using intraspecific and interspecific pairwise P‐distance, Wilcoxon signed rank test, and tree‐based analyses. This study included 186 individuals from 89 populations representing 30 species. For all individuals, single locus markers showed high levels of sequencing universality but were ineffective for species resolvability. Polymerase chain reaction amplification and sequencing were successful for all four loci. Both ITS and matK showed significantly higher levels of interspecific species delimitation than rbcL and trnH‐psbA. A combination of matK and ITS was the most efficient DNA barcode among all studied regions, however, they do not represent an appropriate candidate barcode for Chinese Gaultheria, by which only 11 out of 30 species can be separated. Loci rbcL, matK, and trnH‐psbA, which were recently proposed as universal plant barcodes, have a very poor capacity for species separation for Chinese Gaultheria. DNA barcodes may be reliable tools to identify the evolutionary units of this group, so further studies are needed to develop more efficient DNA barcodes for Gaultheria and other genera with complicated evolutionary histories.     

DNA barcoding Central Asian butterflies: increasing geographical dimension does not significantly reduce the success of species identification

DNA barcoding employs short, standardized gene regions (5' segment of mitochondrial cytochrome oxidase subunit I for animals) as an internal tag to enable species identification. Prior studies have indicated that it performs this task well, because interspecific variation at cytochrome oxidase subunit I is typically much greater than intraspecific variation. However, most previous studies have focused on local faunas only, and critics have suggested two reasons why barcoding should be less effective in species identification when the geographical coverage is expanded. They suggested that many recently diverged taxa will be excluded from local analyses because they are allopatric. Second, intraspecific variation may be seriously underestimated by local studies, because geographical variation in the barcode region is not considered. In this paper, we analyse how adding a geographical dimension affects barcode resolution, examining 353 butterfly species from Central Asia. Despite predictions, we found that geographically separated and recently diverged allopatric species did not show, on average, less sequence differentiation than recently diverged sympatric taxa. Although expanded geographical coverage did substantially increase intraspecific variation reducing the barcoding gap between species, this did not decrease species identification using neighbour-joining clustering. The inclusion of additional populations increased the number of paraphyletic entities, but did not impede species-level identification, because paraphyletic species were separated from their monophyletic relatives by substantial sequence divergence. Thus, this study demonstrates that DNA barcoding remains an effective identification tool even when taxa are sampled from a large geographical area.