Umdale Suraj D, Kshirsagar Parthraj R, Lekhak Manoj M, Gaikwad Nikhil B
Department of Botany, Cytogenetics and Plant Breeding Laboratory, Shivaji University, Kolhapur, Maharashtra, India.
Department of Botany, Yashwantrao Chavan Institute of Science, Satara, Maharashtra, India.
Pharmacogn Mag. 2017 Jul;13(Suppl 2):S224-S229. doi: 10.4103/pm.pm_499_16. Epub 2017 Jul 11.
Sm. is an annual herb widely used in Indian traditional medical practice and commonly known as "Lakshman booti" in Sanskrit. Morphological resemblance among the species of genus . leads to inaccurate identification and adulteration. This causes inconsistent therapeutic effects and also affects the quality of herbal medicine.
This study aimed to generate potential barcode for authentication of and its adulterants through DNA barcoding technique.
Genomic DNA extracted from and its adulterants was used as templates for polymerase chain reaction amplification of the barcoding regions. The amplicons were directed for sequencing, and species identification was conducted using BLASTn and unweighted pair-group method with arithmetic mean trees. In addition, the secondary structures of internal transcribed spacer (ITS) 2 region were predicted.
The nucleotide sequence of ITS provides species-specific single nucleotide polymorphisms and sequence divergence (22%) than A-H (10.9%) and L (3.1%) sequences. The ITS barcode indicates that and are closely related compared to other species.
ITS is the most applicable barcode for molecular authentication of , and further chloroplast barcodes should be tested for phylogenetic analysis of genus Smithia.
The present investigation is the first effort of utilization of DNA barcode for molecular authentication of and its adulterants. Also, this study expanded the application of the ITS2 sequence data in the authentication. The ITS has been proved as a potential and reliable candidate barcode for the authentication of . BLASTn: Basic Local Alignment Search Tool for Nucleotide; MEGA: Molecular Evolutionary Genetic Analysis; EMBL: European Molecular Biology Laboratory; A-H: Photosystem II protein D1- stuctural RNA: His tRNA gene; rbcL: Ribulose 1,5 bi-phosphate carboxylase/oxygenase large subunit gene.
小花远志是一种一年生草本植物，在印度传统医学中广泛使用，在梵语中通常被称为“Lakshman booti”。远志属物种之间的形态相似性导致鉴定不准确和掺假。这会导致治疗效果不一致，也会影响草药的质量。
本研究旨在通过DNA条形码技术生成用于小花远志及其掺假品鉴定的潜在条形码。
从小花远志及其掺假品中提取的基因组DNA用作条形码区域聚合酶链反应扩增的模板。扩增产物用于测序，并使用BLASTn和算术平均法的非加权配对组方法构建系统发育树进行物种鉴定。此外，还预测了内部转录间隔区（ITS）2区域的二级结构。
ITS的核苷酸序列提供了物种特异性的单核苷酸多态性，与A-H序列（10.9%）和L序列（3.1%）相比，序列差异为22%。ITS条形码表明，与其他物种相比，小花远志和[某物种]关系密切。
ITS是小花远志分子鉴定中最适用的条形码，进一步的叶绿体条形码应进行测试，以用于远志属的系统发育分析。
本研究是首次利用DNA条形码对小花远志及其掺假品进行分子鉴定。此外，本研究扩展了ITS2序列数据在鉴定中的应用。ITS已被证明是小花远志鉴定的一种潜在且可靠的候选条形码。BLASTn：核苷酸基本局部比对搜索工具；MEGA：分子进化遗传分析；EMBL：欧洲分子生物学实验室；A-H：光系统II蛋白D1 - 结构RNA：组氨酸转运RNA基因；rbcL：1,5 - 二磷酸核酮糖羧化酶/加氧酶大亚基基因。
