Homogenous PCR of Heterogeneous DNA from Phenolic Rich Barks of Terminalia Species for DNA Based Adulteration Detection

Sharma, Satyawati; Sharma, Poojadevi; Yadav, Sheetal; Purohit, Indira; Srivastava, Anshu; Varma, Astha; Shrivastava, Neeta

doi:10.1007/s40011-015-0576-z

Cited by 3 publications

(3 citation statements)

References 26 publications

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“…ITS region was amplified in all the species after DNA isolation from the collected bark samples. [ 15 ] The amplified ITS regions resulted in approximately 700 bp amplicon which was subsequently sequenced. Assembly of raw reads of sequence produced contigs of 667 bp (GeneBank: KF925432.1), 681 bp (GeneBank: KT187391), 720 bp (GeneBank: KC602394.1), and 701 bp (GenBank: KC750922.1) for T. arjuna (Pune, vouchered BVPPERD/PP/0812/11), T. tomentosa (Udaipur, vouchered BVPPERD/PP/0113/02), T. bellirica (Anand, vouchered BVPPERD/PP/0511/13), and T. chebula (Anand, vouchered BVPPERD/PP/0511/14), respectively.…”

Section: Resultsmentioning

confidence: 99%

“…DNA isolated from bark samples was used for amplification of internal transcribed spacer (ITS) locus with the help of universal primers[ 14 ] using method previously reported by Sharma et al . [ 15 ] The amplified products were sequenced, edited, and assembled with the help of Sequence Analysis version 5.2 (Applied Biosystems, CA, USA) and Codon Code Aligner (Trial version, Codon Code, Dedham, MA, USA). Basic Local Alignment Search Tool (BLAST) was performed for verification of assembled contigs and sequence boundaries were determined comparing the sequences with the database.…”

Section: Methodsmentioning

confidence: 99%

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DNA-based simultaneous identification of three Terminalia species targeting adulteration

Sharma

Shrivastava

2016

Phcog Mag

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Background:Various parts of three Terminalia species, namely, Terminalia arjuna (stem bark), Terminalia bellirica (fruit), and Terminalia chebula (fruit) are widely known for their therapeutic principles and other commercial values. However, stem bark of T. bellirica and T. chebula along with Terminalia tomentosa are reported as adulterants of T. arjuna. Correct botanical identification is very critical for safe and effective herbal drugs. DNA-based identification approaches are advancing the conventional methods and sometime proved more beneficial.Objective:The purpose of the study was to develop polymerase chain reaction (PCR) method using internal transcribed spacer (ITS) region to ascertain the identity of T. arjuna herbal material as well as detection of mixing of other three Terminalia species.Materials and Methods:DNA from stem barks samples were isolated and subjected to ITS region amplification and sequencing. Sequences were compared for polymorphic nucleotides determination to develop species-specific primers. Final primers were selected on the basis of in silico analysis and experimentally validated. PCR assays for botanical identification of Terminalia species were developed. Sensitivity testing and assay validation were also performed.Results:The PCR assays developed for Terminalia species were resulted in definite amplicons of the corresponding species. No cross-reactivity of the primers was detected. Sensitivity was found enough to amplify as low as 2 ng of DNA. Mixing of DNA in various concentrations for validation also proved the sensitivity of assay to detect original botanicals in the mixture. The developed methods proved very specific and sensitive to authenticate Arjuna bark to develop evidence-based herbal medicines.SUMMARY Internal transcribed spacer-based species-specific polymerase chain reaction.(PCR) assays were developed to authenticate Terminalia arjuna stem bark and to identify substitution/adulteration of Terminalia bellirica and Terminalia chebula in the genuine starting materialDefinite amplicons were obtained specific to particular species and the assay was found of profound sensitivity to amplify as low as 2 ng of DNAResults of method validation proved that the assay can identify adulterant Terminalia species even when present in lower amountsThe DNA barcodes and PCR methods can also be used to identify Terminalia bellirica and T. chebula related herbal medicinal material. Abbreviations used: ITS: Internal transcribed spacer, BSA: Bovine serum albumin, DMSO: Dimethyl sulfoxide.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

DNA-based simultaneous identification of three Terminalia species targeting adulteration

Sharma

Shrivastava

2016

Phcog Mag

Self Cite

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show abstract

“…The criteria considered, such as high copy number of chloroplast DNA, short amplicon length, specificity, and sensitivity, have significantly contributed to the effectiveness of these primers in accurately identifying and differentiating the target botanicals within herbal products. In the context of lower taxonomic level identification, the utilization of Species-specific Primers derived from the Internal Transcribed Spacer (ITS) region has proven to be a more effective alternative for detections (Hsieh et al, 2021;Sharma et al, 2017). British pharmacies have been pioneers in adopting trnH-psbA-derived species-specific primers for the authentication of Ocimum tenuiflorum (Cartwright, 2016) (Hsieh et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Advancing Herbal Product Authentication: A Comprehensive Review Of DNA-Based Approach For Quality Control And Safety Assurance

Travadi,

Sharma,

Pandit

et al. 2024

eatp

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Overcoming Challenges in DNA Extractions from Triphala Ingredients: A Way Forward for Optimization of Conventional and Digital PCR Assays for Molecular Authentication

Travadi,

Sharma,

Pandit

et al. 2024

Food Anal. Methods

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Homogenous PCR of Heterogeneous DNA from Phenolic Rich Barks of Terminalia Species for DNA Based Adulteration Detection

Cited by 3 publications

References 26 publications

DNA-based simultaneous identification of three Terminalia species targeting adulteration

DNA-based simultaneous identification of three Terminalia species targeting adulteration

Advancing Herbal Product Authentication: A Comprehensive Review Of DNA-Based Approach For Quality Control And Safety Assurance

Overcoming Challenges in DNA Extractions from Triphala Ingredients: A Way Forward for Optimization of Conventional and Digital PCR Assays for Molecular Authentication

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