ITS-based Identification of Manyleaf Paris Rhizome and Its Adulterants

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[Objective] To discriminate manyleaf paris rhizome and its commonly mixed adulterants by ITS sequence analysis. [Method] The experimental materials included manyleaf paris rhizome samples collected from Yunnan Province and Sichuan Province, commercial manyleaf paris rhizome samples purchased from market and the control Yunan manyleaf paris rhizome. The ITS fragments of these materials were amplified using specific primers for bidirectional sequencing. The sequencing results were multiply aligned by software Clustal W, based on which the mutated sites were analyzed by PAUP 4.0b10 for constructing the phylogenetic tree (most parsimony, MP). [Results] The ITS length of the samples varied between 622-639 bp, of which there were 53 mutated sites in ITS1 sequence, more than that in ITS2 and 5.8 s regions. Phylogenetic analysis showed that the manyleaf paris rhizome samples collected from various origins fell into a different branch. From the phylogenetic tree, there are 14 identification sites among manyleaf paris rhizome and adulterants in sequence alignment plot. [Conclusion] The ITS sequence can distinguish the genuine Paris and its adulterants. But more specific markers for accurate and rapid identification purposes should be exploited, combined with other gene sequences, to distinguish analogue materials with close genetic relationship. [Objective] To discriminate manyleaf paris rhizome and its commonly mixed adulterants by ITS sequence analysis. [Method] The experimental materials included manyleaf paris rhizome samples collected from Yunnan Province and Sichuan Province, commercial manyleaf paris rhizome samples purchased from market and the control Yunan manyleaf The sequencing results were multiplicated by software Clustal W, based on which the mutated sites were analyzed by PAUP 4.0b10 for constructing the phylogenetic tree (most parsimony, MP). [Results] The ITS length of the samples varied between 622-639 bp, of which there were 53 mutated sites in ITS1 sequence, more than that in ITS2 and 5.8 s regions. Phylogenetic analysis showed that the manyleaf paris rhizome samples collected From the various origins fell into a different branch. From the phylogenetic tree, there are 14 identification sit es among manyleaf paris rhizome and adulterants in sequence alignment plot. [Conclusion] The ITS sequence can distinguish the genuine Paris and its adulterants. But more specific markers for accurate and rapid identification purposes should be exploited, combined with other gene sequences, to distinguish analogue materials with close genetic relationship.
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