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低拷贝核基因区域颗粒结合型淀粉合成酶(GBSS1)作为一种新型微型DNA条形码用于鉴别不同鼠尾草属(鼠尾草)物种。

The Low Copy Nuclear Gene Region, Granule Bound Starch Synthase (GBSS1), as a Novel Mini-DNA Barcode for the Identification of Different Sage (Salvia) Species.

作者信息

Parveen Iffat, Techen Natascha, Handy Sara M, Li Jing, Wu Charles, Chittiboyina Amar G, Khan Ikhlas A

机构信息

National Center for Natural Products Research; School of Pharmacy, University of Mississippi, University, MS, USA.

Center for Food Safety and Applied Nutrition, Office of Regulatory Science, U. S. Food and Drug Administration, College Park, MD, USA.

出版信息

Planta Med. 2022 Oct;88(12):985-993. doi: 10.1055/a-1618-6496. Epub 2021 Sep 20.

Abstract

Morphological similarity within species makes the identification and authentication of species challenging, especially in dietary supplements that contain processed root or leaf powder of different sage species. In the present study, the species discriminatory power of 2 potential DNA barcode regions from the nuclear genome was evaluated in 7 medicinally important species from the family Lamiaceae. The nuclear internal transcribed spacer 2 and the exon 9 - 14 region of low copy nuclear gene coding for granule-bound starch synthase 1 were tested for their species discrimination ability using distance, phylogenetic, and BLAST-based methods. A novel 2-step PCR method with 2 different annealing temperatures was developed to achieve maximum amplification from genomic DNA. The granule-bound starch synthase 1 region showed higher amplification and sequencing success rates, higher interspecific distances, and a perfect barcode gap for the tested species compared to the nuclear internal transcribed spacer 2. Hence, these novel mini-barcodes generated from low copy nuclear gene regions (granule-bound starch synthase) that were proven to be effective barcodes for identifying 7 species have potential for identification and authentication of other species.

摘要

物种内部的形态相似性使得物种的鉴定和认证具有挑战性,特别是在含有不同鼠尾草属植物加工根粉或叶粉的膳食补充剂中。在本研究中,评估了来自核基因组的2个潜在DNA条形码区域对唇形科7种重要药用植物的物种鉴别能力。使用距离法、系统发育法和基于BLAST的方法,对核内转录间隔区2以及编码颗粒结合淀粉合酶1的低拷贝核基因的外显子9-14区域的物种鉴别能力进行了测试。开发了一种具有2种不同退火温度的新型两步PCR方法,以实现从基因组DNA的最大扩增。与核内转录间隔区2相比,颗粒结合淀粉合酶1区域显示出更高的扩增和测序成功率、更高的种间距离以及测试物种的完美条形码间隙。因此,这些从低拷贝核基因区域(颗粒结合淀粉合酶)产生的新型微型条形码被证明是鉴定7个物种的有效条形码,具有鉴定和认证其他物种的潜力。

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