Detection of Carica papaya Adulteration in Piper nigrum Using Chloroplast DNA Marker-Based PCR Assays

Travadi, Tasnim; Shah, Abhi P.; Pandit, Ramesh; Sharma, Satyawati; Joshi, Chaitanya; Joshi, Madhvi

doi:10.1007/s12161-022-02395-z

Cited by 9 publications

(3 citation statements)

References 33 publications

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“…Following this optimization process, the annealing temperature was set to 60 • C, consistent with singleplex PCR. Many previous studies have chosen 40 cycles to detect short amplicons, approximately 250 bp in length, in chloroplast or mitochondrial DNA [20][21][22]. This number of cycles was determined to be optimal, maximizing yield while minimizing non-specific amplification, and is essential for detecting low-abundance targets in processed foods.…”

Section: Optimization Of Multiplex Pcrmentioning

confidence: 99%

Rapid and Simultaneous Authentication of Six Laver Species Using Capillary Electrophoresis-Based Multiplex PCR

Yang,

Kim,

Kim

et al. 2024

Foods

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Lavers are typically consumed in dried or seasoned forms. However, commercially processed lavers can lead to seafood fraud because it is impossible to authenticate the original species based on morphological characteristics alone. In this study, we developed a capillary electrophoresis-based multiplex polymerase chain reaction (PCR) to authenticate six different laver species. The species-specific primer sets to target the chloroplast rbcL or rbcS genes were newly designed. We successfully established both singleplex and multiplex conditions, which resulted in specific amplicons for each species (N. dentata, 274 bp; N. yezoensis, 211 bp; N. seriata, 195 bp; N. tenera, 169 bp; N. haitanensis, 127 bp; P. suborbiculata, 117 bp). Moreover, the assays were sensitive enough to detect DNA ranging from 10 to 0.1 pg of DNA. The optimized capillary electrophoresis-based multiplex PCR was successfully applied to 40 commercial laver products. In addition to detecting the laver species as stated on the commercial label, the assay discovered cases where less expensive species were mixed in. With its advantageous properties, such as short amplicon size, high specificity, and superior sensitivity, this assay could be used for the authentication of the six laver species.

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Section: Optimization Of Multiplex Pcrmentioning

confidence: 99%

Rapid and Simultaneous Authentication of Six Laver Species Using Capillary Electrophoresis-Based Multiplex PCR

Yang,

Kim,

Kim

et al. 2024

Foods

View full text Add to dashboard Cite

show abstract

“…There are 17 potential barcode regions (matK, rbcL, ITS, ITS2, psbA-trnH, atpF-atpH, ycf5, psbKI, psbM-trnD, coxI, nad1, trnL-F, rpoB, rpoC1, atpF-atpH, and rps16) for plants, having different degrees of universality, specificity, and taxa resolution power that extensively used in the authentication and identification of medicinal plants 4,7 . However, DNA barcoding 8 and species-specific assays [9][10][11] cannot resolve presence of multiple plant species in a single sample 12 . which can be overcome by DNA metabarcoding.…”

Section: Introductionmentioning

confidence: 99%

DNA metabarcoding using newrbcLandITS2metabarcodes collectively enhance detection efficiency of medicinal plants in single and polyherbal formulations

Travadi

Shah

Pandit

et al. 2023

Preprint

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With the widespread adoption of barcoding and next-generation sequencing, metabarcoding is emerging as a potential tool for detecting labelled and unlabelled plant species in herbal products. In this study, newly designed rbcL and ITS2 metabarcode primers were validated for metabarcoding using in-house mock controls of medicinal plant gDNA pools and biomass pools. The applicability of the multi-barcode sequencing approach was evaluated on 17 single drugs and 15 polyherbal formulations procured from the Indian market. The rbcL metabarcode demonstrated detection efficiencies of 86.7% and 71.7% in gDNA plant pools and biomass mock controls, respectively, while the ITS2 metabarcode demonstrated 82.2% and 69.4%. In the gDNA plant pool and biomass pool mock controls, the cumulative detection efficiency increased by 100% and 90%, respectively. A total of 79% cumulative detection efficiency of both metabarcodes was observed in single drugs, while 76.3% was observed in polyherbal formulations. An average fidelity of 83.6% was observed for targeted plant species present within mock controls as well as in herbal formulations. Our results demonstrated the applicability of multilocus strategies in metabarcoding as a potential tool for detecting labelled and unlabelled plant species in herbal formulations.

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“…There are 17 potential barcode regions ( matK, rbcL, ITS, ITS2, psbA-trnH, atpF-atpH, ycf5, psbKI, psbM-trnD, coxI, nad1, trnL-F, rpoB, rpoC1, atpF-atpH , and rps16 ) for plants, having different degrees of universality, specificity, and taxa resolution power that extensively used in the authentication and identification of medicinal plants ( Parveen et al., 2016 ; Kress, 2017 ). However, DNA barcoding ( Vassou et al., 2016 ) and species-specific assays ( Sharma and Shrivastava, 2016 ; Travadi et al., 2022a ; Travadi et al., 2022b ) cannot resolve presence of multiple plant species in a single sample ( Mishra et al., 2016 ), that limitation could be overcome by DNA metabarcoding.…”

Section: Introductionmentioning

confidence: 99%

A combined approach of DNA metabarcoding collectively enhances the detection efficiency of medicinal plants in single and polyherbal formulations

et al. 2023

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IntroductionEmpirical research has refined traditional herbal medicinal systems. The traditional market is expanding globally, but inadequate regulatory guidelines, taxonomic knowledge, and resources are causing herbal product adulteration. With the widespread adoption of barcoding and next-generation sequencing, metabarcoding is emerging as a potential tool for detecting labeled and unlabeled plant species in herbal products.MethodsThis study validated newly designed rbcL and ITS2 metabarcode primers for metabarcoding using in-house mock controls of medicinal plant gDNA pools and biomass pools. The applicability of the multi-barcode sequencing approach was evaluated on 17 single drugs and 15 polyherbal formulations procured from the Indian market.ResultsThe rbcL metabarcode demonstrated 86.7% and 71.7% detection efficiencies in gDNA plant pools and biomass mock controls, respectively, while the ITS2 metabarcode demonstrated 82.2% and 69.4%. In the gDNA plant pool and biomass pool mock controls, the cumulative detection efficiency increased by 100% and 90%, respectively. A 79% cumulative detection efficiency of both metabarcodes was observed in single drugs, while 76.3% was observed in polyherbal formulations. An average fidelity of 83.6% was observed for targeted plant species present within mock controls and in herbal formulations.DiscussionIn the present study, we achieved increasing cumulative detection efficiency by combining the high universality of the rbcL locus with the high-resolution power of the ITS2 locus in medicinal plants, which shows applicability of multilocus strategies in metabarcoding as a potential tool for the Pharmacovigilance of labeled and unlabeled plant species in herbal formulations.

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Detection of Carica papaya Adulteration in Piper nigrum Using Chloroplast DNA Marker-Based PCR Assays

Cited by 9 publications

References 33 publications

Rapid and Simultaneous Authentication of Six Laver Species Using Capillary Electrophoresis-Based Multiplex PCR

Rapid and Simultaneous Authentication of Six Laver Species Using Capillary Electrophoresis-Based Multiplex PCR

DNA metabarcoding using newrbcLandITS2metabarcodes collectively enhance detection efficiency of medicinal plants in single and polyherbal formulations

A combined approach of DNA metabarcoding collectively enhances the detection efficiency of medicinal plants in single and polyherbal formulations

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