Abstract：
BACKGROUND: Ocimum tenuiflorum L. is a highly traded medicinal with several therapeutic values. Green Tulsi and purple Tulsi are two subtypes in O. tenuiflorum and both have the same medicinal properties. Recent reports have revealed that purple Tulsi contains higher quantities of methyl eugenol (ME), which is moderately toxic and potentially carcinogenic. Therefore, we developed an allele-specific PCR (AS-PCR) method to distinguish the green and purple Tulsi.
RESULTS: Using the green Tulsi as a reference, 12 single nucleotide polymorphisms (SNPs) and 10 insertions/deletions (InDels) were identified in the chloroplast genome of the purple Tulsi. The C > T SNP at the 1,26,029 position in the ycf1 gene was selected for the development of the AS-PCR method. The primers were designed to amplify 521 bp and 291 bp fragments specific to green and purple Tulsi, respectively. This AS-PCR method was validated in 10 accessions from each subtype and subsequently verified using Sanger sequencing. Subsequently, 30 Tulsi powder samples collected from the market were subjected to molecular identification by AS-PCR. The results showed that 80% of the samples were purple Tulsi, and only 3.5% were green Tulsi. About 10% of the samples were a mixture of both green and purple Tulsi. Two samples (6.5%) did not contain O. tenuiflorum and were identified as O. gratissimum.
CONCLUSIONS: The market samples of Tulsi were predominantly derived from purple Tulsi. The AS-PCR method will be helpful for quality control and market surveillance of Tulsi herbal powders.
RESULTS: Using the green Tulsi as a reference, 12 single nucleotide polymorphisms (SNPs) and 10 insertions/deletions (InDels) were identified in the chloroplast genome of the purple Tulsi. The C > T SNP at the 1,26,029 position in the ycf1 gene was selected for the development of the AS-PCR method. The primers were designed to amplify 521 bp and 291 bp fragments specific to green and purple Tulsi, respectively. This AS-PCR method was validated in 10 accessions from each subtype and subsequently verified using Sanger sequencing. Subsequently, 30 Tulsi powder samples collected from the market were subjected to molecular identification by AS-PCR. The results showed that 80% of the samples were purple Tulsi, and only 3.5% were green Tulsi. About 10% of the samples were a mixture of both green and purple Tulsi. Two samples (6.5%) did not contain O. tenuiflorum and were identified as O. gratissimum.
CONCLUSIONS: The market samples of Tulsi were predominantly derived from purple Tulsi. The AS-PCR method will be helpful for quality control and market surveillance of Tulsi herbal powders.
摘要：
背景：tenuiflorumL.是一种具有多种治疗价值的高度交易的药物。绿色Tulsi和紫色Tulsi是O.tenuiflorum的两个亚型，都具有相同的药用特性。最近的报道显示，紫色的Tulsi含有更多的甲基丁香酚（ME），具有中等毒性和潜在致癌性。因此,我们开发了等位基因特异性PCR（AS-PCR）方法来区分绿色和紫色Tulsi。
结果：使用绿色Tulsi作为参考，在紫色Tulsi的叶绿体基因组中鉴定出12个单核苷酸多态性（SNP）和10个插入/缺失（InDels）。选择ycf1基因中1,26,029位的C>TSNP用于AS-PCR方法的开发。引物设计用于扩增对绿色和紫色Tulsi特异的521bp和291bp片段，分别。这种AS-PCR方法在来自每种亚型的10个种质中进行了验证，随后使用Sanger测序进行了验证。随后，从市场上收集的30个Tulsi粉末样品通过AS-PCR进行分子鉴定。结果表明，80％的样品是紫色的Tulsi，只有3.5%是绿色塔尔西。约10%的样品是绿色和紫色Tulsi的混合物。两个样品(6.5%)不含有0.tenuiflorum,并且被鉴定为0.gratissimum。
结论：Tulsi的市场样品主要来自紫色Tulsi。AS-PCR方法将有助于Tulsi草药粉的质量控制和市场监督。
结果：使用绿色Tulsi作为参考，在紫色Tulsi的叶绿体基因组中鉴定出12个单核苷酸多态性（SNP）和10个插入/缺失（InDels）。选择ycf1基因中1,26,029位的C>TSNP用于AS-PCR方法的开发。引物设计用于扩增对绿色和紫色Tulsi特异的521bp和291bp片段，分别。这种AS-PCR方法在来自每种亚型的10个种质中进行了验证，随后使用Sanger测序进行了验证。随后，从市场上收集的30个Tulsi粉末样品通过AS-PCR进行分子鉴定。结果表明，80％的样品是紫色的Tulsi，只有3.5%是绿色塔尔西。约10%的样品是绿色和紫色Tulsi的混合物。两个样品(6.5%)不含有0.tenuiflorum,并且被鉴定为0.gratissimum。
结论：Tulsi的市场样品主要来自紫色Tulsi。AS-PCR方法将有助于Tulsi草药粉的质量控制和市场监督。
