线粒体COⅠ基因在昆虫DNA条形码中的研究与应用

自2003年DNA条形码(DNA barcodes)概念出现以来,DNA条形码技术(DNA barcoding)受到生物分类学领域普遍关注,线粒体细胞色素氧化酶亚基I(mtDNACOⅠ)被用作动物类群的主要条形码序列,基于该基因片段的昆虫条形码研究在国内外广泛开展。本文在概述DNA条形码、条形码技术及已开展的昆虫条形码研究计划的基础上,总结了昆虫mtDNACOⅠ条形码及其技术在发现和描述隐种、种类分子鉴定以及系统发育等方面的研究进展,分析了细胞核线粒体假基因(Numts)对mtDNACOⅠ条形码扩增的影响,提出检测和避免Numts的方法,并对DNA条形码技术的进一步研究和应用进行了讨论和展望。     

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。     

戴云山国家级自然保护区访花昆虫DNA条形码数据集

访花和传粉昆虫对于维持生态系统功能具有重要作用,但我国相关昆虫类群的本底数据非常缺乏。作为基于特定基因序列的物种划分方法, DNA条形码在标本鉴定、新物种发现、生物多样性保护、种群遗传和进化等研究领域具有重要的应用价值。本文报道了福建戴云山国家级自然保护区双翅目、膜翅目和鞘翅目3个类群访花昆虫的815条线粒体COI条形码数据,并详细提供了所获样品的海拔分布信息。该数据集可为地区性昆虫多样性的DNA条形码数据库构建、隐存种发现、海拔梯度物种遗传多样性和生物多样性保护等方面研究提供帮助。     

真菌DNA条形码技术研究进展

DNA条形码(DNA barcoding)技术作为一门新兴的物种鉴定方法以其灵敏、精确、方便和客观的优势,在动植物和微生物的分类鉴定中已经得到广泛应用.真菌鉴定中常用作标准条形码的是核核糖体DNA内转录间隔区(Internal transcribed spacer,ITS),如今也有一些新型条形码被发现和应用到实际操作中,如微条形码、ND6、EF3.本文对DNA条形码技术的产生和发展做出了总结,通过研究其在真菌中应用的实际案例分析了DNA条形码技术的优缺点及发展趋势,并指出DNA条形码技术将以全新的视角来弥补传统分类学的不足,最终实现生物自身的序列变异信息与现有形态分类学的结合.     

DNA条形编码在蚜虫类昆虫中的应用

2003年提出的DNA条形编码技术给生物分类研究带来了空前的繁荣,众多学者对此进行了分析和讨论.蚜虫类昆虫具有多型、转主寄生等复杂的生物学特性,其形态特征多有特化或退化,因此,DNA条形编码在蚜虫类昆虫中的应用必将给蚜虫分类学研究带来巨大的活力.文章总结了国际DNA条形编码技术的研究进展和现状,并展望了DNA条形编码在蚜虫类昆虫研究中应用的方向,该研究技术主要用于对蚜虫物种快速准确的鉴定、解决多型性问题、发现隐存分类单元,探讨蚜虫种间的系统发育关系、蚜虫与寄主植物的关系,解释蚜虫地理分布格局和推测近期分化物种的成因等.     

DNA条形码在生态学研究中的应用与展望

DNA条形码是利用标准的DNA片段对物种进行快速鉴定的技术,已在生物学各相关领域得到广泛应用。随着DNA条形码技术的不断发展和完善,已成功应用于生态学领域的相关研究中。本文综述了DNA条形码在物种快速鉴定和隐存种发现、群落系统发育重建和生态取证、群落内物种间相互关系研究等方面的应用,并介绍了DNAmetabarcoding技术和环境DNA条形码在生物多样性和生态学研究领域中的应用。最后,结合新的测序技术和未来大科学装置的发展,在相关数据库逐渐完善,新分析方法和计算模型不断开发使用的情景下,对DNA条形码在生态学相关领域的应用前景进行了展望。     

基于进化树、距离和特征的DNA条形码方法研究——以百花山地区草螟科为例

DNA条形码是一种快捷高效的分子鉴定新技术,近年来在动物分类学领域中得到迅速的发展和应用。在条形码的研究中有基于进化树、距离和特征3种常用的分析方法:第1种方法需要构建系统发育树,分析样本在树上的聚类情况;第2种方法依赖于物种种内和种间的序列差异;第3种则是通过一系列的诊断特征位点来鉴定物种。本研究扩增了北京百花山地区14种草螟科昆虫88个样本的线粒体细胞色素C氧化酶亚基Ⅰ(mitochondrial cytochrome c oxidase subunitⅠ,COⅠ)基因片段,分别基于进化树、距离和特征方法进行了分析,以探讨不同DNA条形码方法在草螟科物种鉴定中的可行性。结果表明:在使用邻接法(neighbor-joining,NJ)构建的系统发育树上,14个草螟物种各自聚成一个单系,均被成功区分。基于Kimura双参数模型计算遗传距离得出,种内和种内有一个明显的"barcoding gap",且ABGD软件对样本的划分完全符合形态鉴定结果。在所有的草螟物种中都找到了诊断核苷酸位点,基于特征来鉴定草螟物种的成功率为100%。结果显示,这3种方法对于本研究中所涉及的草螟都具有较好的区分,基于COⅠ基因的DNA条形码可以作为一种有效的工具在草螟科昆虫的物种鉴定中进行应用。     

利用植物DNA条形码构建亚热带森林群落系统发育关系——以鼎湖山样地为例

在群落水平上重建植物系统发育关系是当前植物系统学研究的一项重要内容;DNA条形码技术的出现为这一工作的开展提供了便利。本文选取国际通用的植物DNA条形码（rbcL,matK和psbA trnH）,对鼎湖山大样地的183个物种（隶属于24目51科110属）进行测序;分别利用两位点和三位点DNA条形码组合构建该样地植物群落的系统发育关系,并比较不同位点组合构建出的群落系统发育关系的拓扑结构和节点支持率;最后选出一个具有最好拓扑结构和最高节点支持率的鼎湖山大样地群落系统发育关系。在目、科和属这三个水平上,三位点条形码片段组合构建的群落系统发育关系与APG系统获得较好匹配;有些进化分支在相应的APG系统位置解决得不好,却在条形码序列构建的系统发育关系中得到了较好解决。表明综合使用不同进化速率的DNA条形码片段并采取三位点超级矩阵的组合策略,在未采用APG系统大框架的情况下,也能快速而又相对准确地构建出鼎湖山南亚热带森林植物群落的系统发育关系。     

利用植物DNA条形码构建亚热带森林群落系统发育关系——以鼎湖山样地为例

在群落水平上重建植物系统发育关系是当前植物系统学研究的一项重要内容;DNA条形码技术的出现为这一工作的开展提供了便利。本文选取国际通用的植物DNA条形码（rbcL,matK和psbA trnH）,对鼎湖山大样地的183个物种（隶属于24目51科110属）进行测序;分别利用两位点和三位点DNA条形码组合构建该样地植物群落的系统发育关系,并比较不同位点组合构建出的群落系统发育关系的拓扑结构和节点支持率;最后选出一个具有最好拓扑结构和最高节点支持率的鼎湖山大样地群落系统发育关系。在目、科和属这三个水平上,三位点条形码片段组合构建的群落系统发育关系与APG系统获得较好匹配;有些进化分支在相应的APG系统位置解决得不好,却在条形码序列构建的系统发育关系中得到了较好解决。表明综合使用不同进化速率的DNA条形码片段并采取三位点超级矩阵的组合策略,在未采用APG系统大框架的情况下,也能快速而又相对准确地构建出鼎湖山南亚热带森林植物群落的系统发育关系。     

DNA条形码分析方法研究进展

DNA条形码是一段短的、标准化的DNA序列,DNA条形码技术通过对DNA条形码序列分析实现物种的有效鉴定.随着生物DNA条形码序列的大量测定,DNA条形码分析方法得到迅速发展,推动了其在生物分子鉴定中的应用.2003年以来,DNA条形码技术已广泛应用于动物、植物和真菌等物种的鉴定,并有力地推动了生物分类学、生物多样性和生态学等学科的发展.本文在综述DNA条形码技术的基础上,总结了5类主要的DNA条形码分析方法,即基于遗传距离的分析、基于遗传相似度的分析、基于系统发育树的分析、基于序列特征的分析和基于统计分类法的分析,并进一步展望了DNA条形码技术的发展与应用.     

DNA barcoding to map the microbial communities: current advances and future directions

During the last two decades, the DNA barcode development towards microbial community has increased dramatically. DNA barcode development is related to error-free and quick species identification which aid in understanding the microbial biodiversity, as well as the diseases related to microbial species. Here, we seek to evaluate the so-called barcoding initiatives for the microbial communities and the emerging trends in this field. In this paper, we describe the development of DNA marker-based DNA barcoding system, comparison between routine species identification and DNA barcode, and microbial biodiversity and DNA barcode for microbial communities. Two major topics, such as the molecular diversity of viruses and barcode for viruses have been discussed at the same time. We demonstrate the current status and the maker of DNA barcode for bacteria, algae, fungi, and protozoa. Furthermore, we argue about the promises, limitations, and present and future challenges of microbial barcode development.     

DNA Barcoding of Japanese Click Beetles (Coleoptera,Elateridae)

Click beetles (Coleoptera: Elateridae) represent one of the largest groups of beetle insects. Some click beetles in larval form, known as wireworms, are destructive agricultural pests. Morphological identification of click beetles is generally difficult and requires taxonomic expertise. This study reports on the DNA barcoding of Japanese click beetles to enable their rapid and accurate identification. We collected and assembled 762 cytochrome oxidase subunit I barcode sequences from 275 species, which cover approximately 75% of the common species found on the Japanese main island, Honshu. This barcode library also contains 20 out of the 21 potential pest species recorded in Japan. Our analysis shows that most morphologically identified species form distinct phylogenetic clusters separated from each other by large molecular distances. This supports the general usefulness of the DNA barcoding approach for quick and reliable identification of Japanese elaterid species for environmental impact assessment, agricultural pest control, and biodiversity analysis. On the other hand, the taxonomic boundary in dozens of species did not agree with the boundary of barcode index numbers (a criterion for sequence-based species delimitation). These findings urge taxonomic reinvestigation of these mismatched taxa.     

A test of seven candidate barcode regions from the plastome in Picea (Pinaceae)

DNA barcoding, as a tool for species discrimination, has been used efficiently in animals, algae and fungi, but there are still debates on which DNA region(s) can be used as the standard barcode(s) for land plants. Gymnosperms, especially conifers, are important components of forests, and there is an urgent need for them to be identified through DNA barcoding because of their high frequency of collection in the field. However, the feasibility of DNA barcoding in gymnosperms has not been examined based on a dense species sampling. Here we selected seven candidate DNA barcodes from the plastome (matK, rbcL, rpoB, rpoC1, atpF-atpH, psbA-trnH, and psbK-psbI) to evaluate their suitability in Picea (spruce). The results showed that none of them or their different combinations has sufficient resolution for spruce species, although matK+rbcL might be used as a two-locus barcode. The low efficiency of these candidate barcodes in Picea might be caused by the paternal inheritance of the chloroplast genome, long generation time, recent radiation, and frequent inter-specific hybridization aided by wind pollination. Some of these factors could also be responsible for the difficulties in barcoding other plant groups. Furthermore, the potential of the nuclear LEAFY gene as a land plant barcode was discussed.     

DNA barcode goes two-dimensions: DNA QR code web server

The DNA barcoding technology uses a standard region of DNA sequence for species identification and discovery. At present, "DNA barcode" actually refers to DNA sequences, which are not amenable to information storage, recognition, and retrieval. Our aim is to identify the best symbology that can represent DNA barcode sequences in practical applications. A comprehensive set of sequences for five DNA barcode markers ITS2, rbcL, matK, psbA-trnH, and CO1 was used as the test data. Fifty-three different types of one-dimensional and ten two-dimensional barcode symbologies were compared based on different criteria, such as coding capacity, compression efficiency, and error detection ability. The quick response (QR) code was found to have the largest coding capacity and relatively high compression ratio. To facilitate the further usage of QR code-based DNA barcodes, a web server was developed and is accessible at http://qrfordna.dnsalias.org. The web server allows users to retrieve the QR code for a species of interests, convert a DNA sequence to and from a QR code, and perform species identification based on local and global sequence similarities. In summary, the first comprehensive evaluation of various barcode symbologies has been carried out. The QR code has been found to be the most appropriate symbology for DNA barcode sequences. A web server has also been constructed to allow biologists to utilize QR codes in practical DNA barcoding applications.     

Challenges to barcoding an entire flora

DNA barcodes are species‐specific genetic markers that allow taxonomic identification of biological samples. The promise of DNA barcoding as a rapid molecular tool for conducting biodiversity inventories has catalysed renewed efforts to document and catalogue the diversity of life, parallel to the large‐scale sampling conducted by Victorian naturalists. The unique contribution of DNA barcode data is in its ability to identify biotic material that would be impossible to classify using traditional taxonomic keys. However, the utility of DNA barcoding relies upon the construction of accurate barcode libraries that provide a reference database to match to unidentified samples. Whilst there has been much debate in the literature over the choice and efficacy of barcode markers, there has been little consideration of the practicalities of generating comprehensive barcode reference libraries for species‐rich floras. Here, we discuss several challenges to the generation of such libraries and present a case study from a regional biodiversity hotspot in southern Quebec. We suggest that the key challenges include (i) collection of specimens for rare or ephemeral species, (ii) limited access to taxonomic expertise necessary for reliable identification of reference specimens and (iii) molecular challenges in amplifying and matching barcode data. To be most effective, we recommend that sampling must be both flexible and opportunistic and conducted across the entire growing season by expert taxonomists. We emphasize that the success of the global barcoding initiative will depend upon the close collaboration of taxonomists, plant collectors, and molecular biologists.     

Choosing and using a plant DNA barcode

The main aim of DNA barcoding is to establish a shared community resource of DNA sequences that can be used for organismal identification and taxonomic clarification. This approach was successfully pioneered in animals using a portion of the cytochrome oxidase 1 (CO1) mitochondrial gene. In plants, establishing a standardized DNA barcoding system has been more challenging. In this paper, we review the process of selecting and refining a plant barcode; evaluate the factors which influence the discriminatory power of the approach; describe some early applications of plant barcoding and summarise major emerging projects; and outline tool development that will be necessary for plant DNA barcoding to advance.     

Current progress in DNA barcoding and future implications for entomology

DNA barcoding is a technique for identifying organisms based on a short, standardized fragment of genomic DNA. The standardized sequence region is called a DNA barcode because it is like a barcode tag for each taxon. Since the proposition of this concept and the launch of a large project named the Barcode of Life, this simple technique has attracted attention from taxonomists, ecologists, conservation biologists, agriculturists, plant‐quarantine officers and others, and the number of studies using the DNA barcode has rapidly increased. The extreme diversity of insects and their economical, epidemiological and agricultural importance have made this group a major target of DNA barcoding. However, there is some controversy about the utility of DNA barcoding. In this review, we present an overview of DNA barcoding and its application to entomology. We also introduce current advances and future implications of this promising technique.     

Pay Attention to the Overlooked Cryptic Diversity in Existing Barcoding Data: the Case of Mollusca with Character-Based DNA Barcoding

With the global biodiversity crisis, DNA barcoding aims for fast species identification and cryptic species diversity revelation. For more than 10 years, large amounts of DNA barcode data have been accumulating in publicly available databases, most of which were conducted by distance or tree-building methods that have often been argued, especially for cryptic species revelation. In this context, overlooked cryptic diversity may exist in the available barcoding data. The character-based DNA barcoding, however, has a good chance for detecting the overlooked cryptic diversity. In this study, marine mollusk was as the ideal case for detecting the overlooked potential cryptic species from existing cytochrome c oxidase I (COI) sequences with character-based DNA barcode. A total of 1081 COI sequences of mollusks, belonging to 176 species of 25 families of Gastropoda, Cephalopoda, and Lamellibranchia, were conducted by character analysis. As a whole, the character-based barcoding results were consistent with previous distance and tree-building analysis for species discrimination. More importantly, quite a number of species analyzed were divided into distinct clades with unique diagnostical characters. Based on the concept of cryptic species revelation of character-based barcoding, these species divided into separate taxonomic groups might be potential cryptic species. The detection of the overlooked potential cryptic diversity proves that the character-based barcoding mode possesses more advantages of revealing cryptic biodiversity. With the development of DNA barcoding, making the best use of barcoding data is worthy of our attention for species conservation.