DNA条形码技术在生物分类学中的应用前景

DNA条形码(DNA barcoding)技术作为近年来发展起来的一门物种鉴定的新兴技术,已引起越来越多的关注。这一技术的主要目的是鉴定已知物种和发现的新物种。DNA条形码提供了可信息化的分类学标准和更加敏感的分子差别模式,相对于传统生物鉴定的优势在于可以揭示隐存种,鉴定缺少形态数据或形态不易区分的种类,为物种鉴定提供简单有效的工具。综述了DNA条形码技术的产生、发展和基本原理以及在生物分类学中的应用及存在的问题。     

Ditylenchus gigas n. sp. parasitizing broad bean: a new stem nematode singled out from the Ditylenchus dipsaci species complex using a polyphasic approach with molecular phylogeny

Morphologial, biochemical, molecular and karyological analyses of different populations and races of the stem and bulb nematode Ditylenchus dipsaci have suggested that it represents a species complex, of which only D. dipsaci sensu stricto and its morphologically larger variant, known as the giant race of the stem and bulb nematode, are plant parasites of economic importance. The present study singles out the giant race from this complex, herein described as a new species named Ditylenchus gigas n. sp., on the basis of morphological and molecular data obtained from several populations collected from broad beans in southern Italy, southern Spain and Lebanon. The new species epithet, which refers to the large body size of the nematode with respect to the normal races, must be considered to be conspecific with the D. dipsaci‘giant race’ from Fabaceae in recent literature. Morphologically, the new species is characterized by a body size 1·5–2 times longer than the ‘normal race’, stylet delicate (11·5–13·0 μm long) with knobs distinctly sloping backwards, and long post‐vulval uterine sac (81–150 μm long). Results of molecular analysis of rDNA sequences including the ITS1‐5.8S‐ITS2 region, the D2–D3 fragment of the 28S gene, the small 18S subunit, the partial mitochondrial gene for cytochrome c oxidase I (mtCOI), and hsp90 gene sequences, support the new taxonomic species status for the former D. dipsaci giant race from Vicia faba, and clearly distinguish D. gigas n. sp. from D. dipsaci sensu stricto.     

Genetic study of Mediterranean and South American populations of tomato leafminer Tuta absoluta (Povolny, 1994) (Lepidoptera: Gelechiidae) using ribosomal and mitochondrial markers

BACKGROUND: Before its introduction into Europe at the end of 2006, Tuta absoluta (Povolny, 1994) was confined solely to South America. Currently, this invasive pest is well established in various European and Mediterranean countries, causing important economic losses to tomato (Lycopersicon esculentum Mill.) crops. In order to study the genetic variability of this pest, 23 Mediterranean and ten native South American populations were analysed with nuclear ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) markers. RESULTS: The internal transcribed spacers 1 (ITS1) and 2 (ITS2) of rDNA and a fragment in the mtDNA gene encoding cytochrome oxidase I (COI) were PCR amplified and sequenced in T. absoluta. Sequence analyses consistently revealed neither intrapopulation nor interpopulation variation in either genomic region. CONCLUSIONS: High genetic homogeneity was detected in T. absoluta populations from the Mediterranean Basin and South America, based on mtCOI and ITS rDNA sequence analysis. A single genetic type was identified in this pest. Copyright © 2011 Society of Chemical Industry     

Genetic variation and mtCOI phylogeny for Bemisia tabaci (Hemiptera, Aleyrodidae) indicate that the ‘B’ biotype predominates in Iran

Despite a large number of investigations on the molecular genetics and population structure of the whitefly Bemisia tabaci (Gennadius) complex, no such study had been conducted in Iran. The genetic variation of B. tabaci was examined using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) for 18 field collections from cucumber, eggplant, and tomato in four provinces of Iran. PCR amplification and restriction digestion with two enzymes detected 388 RFLP fragments, of which 16 fragments showed polymorphisms. Cluster analysis of these data placed all B. tabaci individuals within a single group, and there was no evidence for between- or within-population genetic variation. Phylogenetic (Clustal W) analysis of 42 B. tabaci mtCOI sequences (n = 21 field collections) from Iran, and a comparison with well-studied haplotype or biotype reference sequences available in public sequence databases, revealed that the Iranian B. tabaci populations were most closely related to the B biotype at 0–1.2% nucleotide identity. The B biotype is a well-known member of a sister clade from the Middle East–North African region of the world, owing to its nearly worldwide distribution and invasive characteristics. This report indicates that a single major haplotype of B biotype is prevalent in Iran and that its closest relative is the B biotype. Also, given the extent of known variation in the Middle East and African continent, data indicate somewhat surprisingly that the B. tabaci collections sampled in Iran had limited genetic variation and population substructure. Knowledge that the B biotype of B. tabaci predominates in Iran is important for designing effective pest management strategies given that biotypes of B. tabaci are known to differ greatly with respect to insecticide resistance, host range, virus–vector interactions, and other key biological characteristics.     

我国小麦落选短体线虫群体遗传多样性分析

 为了明确我国落选短体线虫群体的遗传多样性,利用mtCOI基因标记对落选短体线虫9个地理群体的遗传结构及遗传分化进行分析。结果显示,9个地理群体中共得到101条mtCOI序列,发现28个碱基变异位点,形成14个单倍型。其中,H1单倍型最常见,为7个地理群体的59个个体共有,推测其可能为祖先单倍型。全部地理群体在物种水平呈现中等遗传多样性(H_T = 0.706±0.131),聚类分析显示其可分为类群Ⅰ与类群Ⅱ两个类群,AMOVA分析揭示落选短体线虫整体水平的遗传分化主要来源于种群间。Mantel检验表明落选短体线虫群体的遗传距离与地理距离存在正相关性,但是不同种群之间的遗传分化程度与地理距离没有显著关系。中性检验和错配分布检验均揭示落选短体线虫在整体水平和两个类群上的群体历史动态都处于相对稳定的状态。     

基于DNA条形码技术对江门沿岸海域夏季鱼卵的鉴定

以江门沿岸海域夏季采集到的鱼卵研究对象,利用DNA条形码技术分析鉴定鱼卵种类。本研究获得鱼卵个体有效线粒体COI序列信息217个,测序成功率为68.5%。经BOLD数据库比对分析,成功鉴定鱼卵5目14科19属20种(未知种2种);种内遗传距离为0~0.006,平均遗传距离为0.002,种间遗传距离为0.149~0.325,平均遗传距离为0.255,种间遗传距离为种内遗传的128倍;鱼卵样品以鲈形目数量最多,占51.8%;鲱形目次之,占24.8%;其中鱼卵优势种为黄斑鲾(Photopectoralis bindus)、日本鳀(Engraulis japonicus)、大甲鲹(Megalaspis cordyla)、粗鳞鮻(Chelon subviridis)、金钱鱼(Scatophagus argus)、龙头鱼(Harpadon nehereus)、亚洲(Sillago asiatica)。本次调查川山群岛东部海域获得鱼卵数量、鱼卵种类数均最多,是江门沿岸海域日本鳀、龙头鱼的主要产卵场;黄茅海、镇海湾西部水域均未获得鱼卵,可能与水域环境变化及陆源污染导致产卵环境破坏、亲体量减少有关。该研究结果揭示了江门沿岸海域夏季鱼卵分布格局,为其制定有效的产卵场保护措施提供依据,同时表明DNA条形码技术能有效地对鱼卵进行种类鉴定,可被广泛地应用于我国沿海海域鱼卵鉴定工作。     

广东省半翅目桑树害虫种类与为害调查

【目的】半翅目（Hemiptera）害虫是广东桑树种植区主要为害桑树的害虫种类,每年都会对蚕桑 生产造成较大的经济损失 , 但目前尚未有对广东省半翅目桑树害虫种类系统鉴定与为害调查的相关研究。因此, 针对广东省半翅目桑树害虫进行系统调查和研究,从而为进一步制定出有效的防治措施及开展相关的研究提供 理论依据有着重要的意义。【方法】通过对桑园鳞翅目害虫的调查与采集,利用形态和分子（细胞色素氧化酶 亚基Ⅰ基因 mtCO Ⅰ）鉴定方法对其进行系统鉴定。【结果】共鉴定出 7 种半翅目害虫,分别为双条拂粉蚧（Ferrisia virgata Cockerell）、桑木虱（Anomoneura mori Schwarz）、桑斑叶蝉（Erythroneura mori Matsumura）、尖胸沫蝉 （Aphrophora sp.）、稻绿蝽（Nezara viridula Linnaeus）、瘤缘蝽（Acanthocoris scaber Linnaeus）、离斑棉红蝽（Dysdercus cingulatus Fabricius）。【结论】在鉴定的 7 种半翅目害虫中,双条拂粉蚧、尖胸沫蝉、离斑棉红蝽为广东省首 次报道为害桑树的害虫。