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DNA条形码技术是植物比较基因组学中的一种新方法。

[DNA barcoding is a new approach in comparative genomics of plants].

作者信息

Shneer V S

出版信息

Genetika. 2009 Nov;45(11):1436-48.

Abstract

DNA barcoding was proposed as a method for recognition and identification of eukaryotic species through comparison of sequences of a standard short DNA fragment--DNA barcode--from an unknown specimen to a library of reference sequences from known species. This allows identifying an organism at any stage of development from a very small tissue sample, fresh or conserved many years ago. Molecular identification of plant samples can be used in various scientific and applied fields. It would also help to find new species, which is particularly important for cryptogamic plants. An optimal DNA barcode region is a small fragment present in all species of a major taxonomic group, having invariable nucleotide sequence in all members of the same species, but with sufficient variation to discriminate among the species. This fragment should be flanked by low-variable regions for use of universal primers in PCR for amplification and sequencing. The DNA barcode that is well established in animals is a sequence of a fragment of the mitochondrial cytochrome c oxidase gene CO1. However, searching for DNA barcode in plants proved to be a more challenging task. No DNA region universally suitable for all plants and meeting all of the necessary criteria has been found. Apparently, a multilocus or two-stage approach should be applied for this purpose. Several fragments of the chloroplast genome (trnH-psbA, matK, rpoC, rpoB, rbcL) in combinations of two or three regions were suggested as candidate regions with highest potential, but more representative samples should be examined to choose the best candidate. The possibility is discussed to use as DNA barcode internal transcribed spacers (ITS) of nuclear rRNA genes, which are highly variable, widely employed in molecular phylogenetic studies at the species level, but also have some limitations.

摘要

DNA条形码技术被提出作为一种通过将未知标本的标准短DNA片段(DNA条形码)序列与已知物种的参考序列库进行比较来识别和鉴定真核生物物种的方法。这使得能够从非常小的新鲜或多年保存的组织样本中识别处于任何发育阶段的生物体。植物样本的分子鉴定可用于各种科学和应用领域。它还有助于发现新物种,这对隐花植物尤为重要。最佳的DNA条形码区域是一个存在于主要分类群所有物种中的小片段,在同一物种的所有成员中具有不变的核苷酸序列,但具有足够的变异性以区分不同物种。该片段两侧应是低变异性区域,以便在PCR中使用通用引物进行扩增和测序。在动物中已确立的DNA条形码是线粒体细胞色素c氧化酶基因CO1的一个片段的序列。然而,在植物中寻找DNA条形码被证明是一项更具挑战性的任务。尚未发现普遍适用于所有植物并满足所有必要标准的DNA区域。显然,为此应采用多位点或两阶段方法。叶绿体基因组的几个片段(trnH-psbA、matK、rpoC、rpoB、rbcL)以两个或三个区域的组合被建议作为最具潜力的候选区域,但应检查更多代表性样本以选择最佳候选者。还讨论了使用核rRNA基因的内部转录间隔区(ITS)作为DNA条形码的可能性,ITS高度可变,在物种水平的分子系统发育研究中广泛应用,但也有一些局限性。

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