Background

Bunium persicum commonly called as Kala zeera, a very high value herbaceous spice used for medicinal purposes is often adulterated with Cuminum cyminum or Safed zeera, a closely related species. Lack of distinctive morphological features makes the identification of genuine kala zeera from its adulterant difficult, the problem is even exaggerated in case of powdered material.

Methodology

Genomic DNA was extracted from all the plant materials by using CTAB-SDS method (Möller et al., 1992) with slight modifications. On the basis of reproducibility and high amplification ability, four universal barcoding loci viz. ITS2, rbcL-a, mat K and psbA-trnH and a specific locus Cum were used in the present study. The amplified PCR products were sequenced bidirectionally and assembled to obtain contigs. The sequences thus obtained were aligned using MUSCLE algorithm (Edgar, 2004) and information pertaining to conserved/ variable/ parsimony informative sites, number of transitions, transversions and Indels was obtained after analyzing the sequences.

Results and conclusion

Among the tested barcoding loci, psbA-trnH has proven to be best barcode in authentication of kala zeera as its amplification and sequencing success was high and it showed the presence of polymorphic sites to detect interspecific variation. This barcode could differentiate between safed zeera and kala zeera in a single reaction, simultaneously.

中文翻译：

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DNA条形码特定和灵敏检测孜然芹掺假在Bunium仙客来

背景

波斯菊通常被称为Kala zeera，一种用于药用目的的非常高价值的草本香料，通常会与Cuminum cyminum或Safed zeera（一种密切相关的物种）掺假。缺乏独特的形态特征使得很难从掺假品中鉴别出真正的卡拉zeera，如果是粉末状材料，问题甚至会更加严重。

方法

通过使用CTAB-SDS方法（Möller等，1992），从植物的所有材料中提取基因组DNA，并稍加修改。在重复性和高放大能力，四个通用条形码位点的基础即。在本研究中使用了ITS2，rbcL-a，mat K和psbA-trnH以及一个特定的基因座Cum。扩增的PCR产物被双向测序并组装以获得重叠群。使用MUSCLE算法（Edgar，2004）对由此获得的序列进行比对，并在分析序列后获得与保守/可变/简约信息位点，转变数，转化和Indels有关的信息。

结果与结论

在测试的条形码基因座中，psbA-trnH已被证明是对卡拉zeera进行身份验证的最佳条形码，因为其扩增和测序成功率很高，并且它表明存在多态性位点以检测种间变异。该条形码可以在单个反应中同时区分安全的玉米和卡拉的玉米。