Peran Mini-Barcode Internal Transcribed Spacer 2 (ITS2) untuk Identifikasi Molekuler Spesies Anggrek

Mukhamad Su’udi, Zakiyah Ramadany, S. Rohimah, Asyifa Yasmin Arum, Dwi Setyati, F. Ulum
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Abstract

AbstractMolecular identification using short orthological DNA sequences (DNA barcoding) has been applied for the classification of orchid species which is a major step in biodiversity management, conservation, breeding, authenticating components of herbal products, and tracking the adulteration of orchid species. One of the most widely used loci for phylogenetic inference at the generic and infrageneric levels in plants was ITS located between 18S rDNA, 5.8S rDNA, and 26S rDNA. The ITS or ITS2 region has been suggested as a plant barcode in some previous studies. However, DNA barcoding using the entire ITS genome is considered less effective and efficient in the process of PCR amplification and sequencing. Besides, complete DNA barcodes are difficult to obtain from herbarium samples and herbal products because some DNA sequences have been degraded. DNA mini-barcodes were developed over the past ten years to overcome issues related to DNA barcoding. DNA mini-barcodes use shorter DNA segments for PCR amplification, so they can identify species effectively and efficiently compared to the regular DNA barcoding. Specific primers that encode the ITS1 and ITS2 regions need to be designed for the PCR amplification. DNA mini-barcoding ITS has proven useful in species identification, classification studies, and authentication of specific orchid species. Therefore, a new rapid identification method based on the ITS mini-barcode is expected to be established, especially for orchid species. Keywords : DNA mini-barcode, Internal Transcribed Spacer, Orchid, Primer Design
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用于分子鉴定兰花品种的内部条形码2 (ITS2)的作用
摘要利用短同源DNA序列进行分子鉴定(DNA条形码)已被应用于兰科植物的分类,这是生物多样性管理、保护、育种、草药产品成分鉴定和兰科植物掺假追踪的重要步骤。ITS位点位于18S rDNA、5.8S rDNA和26S rDNA之间,是在植物属和非属水平上应用最广泛的系统发育推断位点之一。在以往的一些研究中,已提出ITS或ITS2区域作为植物的条形码。然而,在PCR扩增和测序过程中,使用整个ITS基因组的DNA条形码被认为效率较低。此外,由于一些DNA序列已被降解,因此很难从植物标本和草药产品中获得完整的DNA条形码。DNA微型条形码是在过去十年中发展起来的,以克服与DNA条形码相关的问题。DNA微型条形码使用较短的DNA片段进行PCR扩增,因此与常规DNA条形码相比,它们可以更有效地识别物种。需要设计编码ITS1和ITS2区域的特异性引物进行PCR扩增。DNA迷你条形码ITS已被证明是有用的物种鉴定,分类研究和鉴定特定兰花物种。因此,一种新的基于ITS微型条形码的快速鉴定方法有望建立起来,尤其是兰花品种。关键词:DNA微型条形码,内部转录间隔,兰花,引物设计
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