Al-Qurainy F, Khan S, Tarroum M, Al-Hemaid F M, Ali M A
Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia.
Genet Mol Res. 2011 Nov 10;10(4):2806-16. doi: 10.4238/2011.November.10.3.
Dried parts of different plant species often look alike, especially in powdered form, making them very difficult to identify. Ruta graveolens, sold as a dried medicinal herb, can be adulterated with Euphorbia dracunculoides. The genomic DNA was isolated from the leaf powder (100 mg each) using the modified CTAB method. Internal transcribed spacer sequences of nuclear ribosomal DNA (nrDNA-ITS), and chloroplast spacer sequences (rpoB and rpoC1) are regarded as potential genes for plant DNA barcoding. We amplified and sequenced these spacer sequences and confirmed the sequences with a BLAST search. Sequence alignment was performed using ClustalX to look for differences in the sequences. A DNA marker was developed based on rpoB and rpoC1 of the nrDNA-ITS for the identification of the adulterant E. dracunculoides in samples of R. graveolens that are sold in local herbal markets. Sequence-characterized amplified region markers of 289 and 264 bp for R. graveolens and 424 bp for E. dracunculoides were developed from dissimilar sequences of this nrDNA-ITS to speed up the authentication process. This marker successfully distinguished these species in extracted samples with as little as 5 ng DNA/μL extract.
不同植物物种的干燥部分通常看起来很相似，尤其是呈粉末状时，这使得它们很难鉴别。作为干燥草药出售的芸香，可能会被大戟属植物掺假。使用改良的CTAB方法从叶粉（各100毫克）中分离基因组DNA。核糖体DNA（nrDNA-ITS）的内部转录间隔区序列以及叶绿体间隔区序列（rpoB和rpoC1）被视为植物DNA条形码的潜在基因。我们对这些间隔区序列进行扩增和测序，并通过BLAST搜索确认序列。使用ClustalX进行序列比对以寻找序列差异。基于nrDNA-ITS的rpoB和rpoC1开发了一种DNA标记，用于鉴定在当地草药市场出售的芸香叶样品中的掺假物大戟属植物。从该nrDNA-ITS的不同序列开发出芸香的289和264 bp以及大戟属植物的424 bp的序列特征扩增区域标记，以加快鉴定过程。该标记成功地在DNA提取物低至5 ng/μL的提取样品中区分了这些物种。
