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. 2011 Jul;1(1):11-19.
doi: 10.1007/s13205-010-0001-5. Epub 2011 Jan 18.

Development of ITS sequence-based markers to distinguish Berberis aristata DC. from B. lycium Royle and B. asiatica Roxb

Subramani Paranthaman Balasubramani, Gurinder Singh Goraya, Padma Venkatasubramanian

Development of ITS sequence-based markers to distinguish Berberis aristata DC. from B. lycium Royle and B. asiatica Roxb

Subramani Paranthaman Balasubramani et al. 3 Biotech. 2011 Jul.

Abstract

The stems of Berberis aristata DC. (Berberidaceae) are used in the South Asian traditional medicine as a key ingredient in formulations for eye care, skin diseases, jaundice, rheumatism and also in diabetes. B. lycium Royle and B. asiatica Roxb. are traded as equivalents of B. aristata. Conventional macro-morphology and microscopic examination does not aid in critically distinguishing the three species. DNA markers were developed by amplifying and sequencing the complete internal transcribed spacer region (ITS1, 5.8S rRNA and ITS2) from the genomic DNA, using universal primers. The markers developed are efficient and reliable in authenticating B. aristata, B. asiatica and B. lycium. These are useful as molecular pharmacognostic tool in quality control of raw drugs.

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Figures

Fig. 1
Fig. 1
PCR amplification of complete ITS sequence. Lane M 100 bp DNA ladder; lanes 1–3 B. aristata; lanes 4–6 B. asiatica; lanes 7–9 B. lycium. Approximately, 700 bp amplicon can be observed with all samples of Berberis spp.
Fig. 2
Fig. 2
Complete ITS sequence of B. aristata, B. asiatica and B. lycium. Species-specific 20 mer primers (forward and reverse) for each species is shown underlined in the sequence
Fig. 3
Fig. 3
Multiple alignment of the complete ITS sequence of B. aristata, B. asiatica and B. lycium
Fig. 4
Fig. 4
PCR amplification of genomic DNA from Berberis spp. with AR1F and AR1R. Lane M 100 bp DNA ladder; lanes 1–5 B. aristata; lanes 6–8 B. asiatica; lanes 9–11 B. lycium. A 405 bp amplicon is observed only in B. aristata samples
Fig. 5
Fig. 5
PCR amplification of genomic DNA from Berberis spp. with AS1F and AS1R. Lane M 100 bp DNA ladder; lanes 1–4 B. asiatica; lanes 5–7 B. lycium; lanes 8–10 B. aristata. A 401 bp amplicon is observed only in B. asiatica samples
Fig. 6
Fig. 6
PCR amplification of genomic DNA from Berberis spp. with BL2F and BL2R. Lane M 100 bp DNA ladder; lanes 1–5B. lycium; lanes 6–8 B. aristata; lanes 9–11 B. asiatica. A 232 bp amplicon is observed only in B. lycium samples
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