Refining DNA Barcoding Coupled High Resolution Melting for Discrimination of 12 Closely Related Croton Species

Osathanunkul, Maslin; Suwannapoom, Chatmongkon; Ounjai, Sarawut; Rora, Jantarika A.; Madesis, Panagiotis; Boer, Hugo de

doi:10.1371/journal.pone.0138888

Cited by 33 publications

(23 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…These barcoding systems provide open barcode databases and standard protocols for species identification. In recent years, a lot of quick identification methods based on DNA barcode markers were developed, such as Allele-specific PCR18, Real-time PCR19, High Resolution Melting (HRM) analysis20, PCR-RFLP21, and etc. These methods have simpler test procedures and shorter time-consuming than conventional methods that were more convenient for rapid identification.…”

mentioning

confidence: 99%

Rapidly discriminate commercial medicinal Pulsatilla chinensis (Bge.) Regel from its adulterants using ITS2 barcoding and specific PCR-RFLP assay

Shi

Zhao

Yao

et al. 2017

Sci Rep

View full text Add to dashboard Cite

Pulsatillae radix is a conventional traditional Chinese medicine (TCM) with common name Baitouweng, and has notable effects on inflammation and dysentery. Pulsatilla chinensis (Bge.) Regel is the only source plant of Baitouweng recorded in Chinese Pharmacopoeia, but its adulteration often occurs in the market that possibly affects medicinal efficacy and safety. We have established an internal transcribed spacer 2 (ITS2) barcode library based on 105 plant samples from 12 Pulsatilla species and 10 common adulterants. Results indicate that ITS2 barcoding can accurately distinguish Pulsatilla species from their adulterants. Pulsatilla chinensis can be discriminated from 11 congeneric species by two stable single nucleotide polymorphisms (SNPs) in the ITS2 region. Additionally, a quick specific PCR-RFLP identification assay based on the ITS2 barcode was developed. Using specific primers ITS2/PR1 combined with restriction enzyme Bgl I, Pu. chinensis can rapidly be differentiated from other species via simple and low-cost test procedures. Furthermore, 30 commercial Baitouweng products were tested and only two products were derived from authentic Pu. chinensis. Thus, these two molecular approaches provide practical tools for quick identification of commercial Baitouweng products and can help ensure the safe use of this TCM product.

show abstract

mentioning

confidence: 99%

Rapidly discriminate commercial medicinal Pulsatilla chinensis (Bge.) Regel from its adulterants using ITS2 barcoding and specific PCR-RFLP assay

Shi

Zhao

Yao

et al. 2017

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Fatores como o elevado número de espécies, somados a ampla distribuição geográfica e considerável diversidade morfológica, contribuem para que o gênero Croton seja considerado de alta complexidade taxonômica, sendo estimulada a realização de estudos para elucidação das relações filogenéticas do gênero (RIINA et al, 2009). Estratégias de estudos moleculares associadas a identificação, sequenciamento e comparação de regiões conservadas do genoma (a exemplo da técnica de DNA Barcode) podem contribuir na identificação e diferenciação de espécies de Croton, auxiliando no desenvolvimento de estudos taxonômicos e filogenéticos (OSATHANUNKUL et al, 2015).…”

Section: Resultsunclassified

CONTRIBUIÇÕES DA BIOLOGIA MOLECULAR PARA CARACTERIZAÇÃO, CONSERVAÇÃO E USO DO GÊNERO Croton L. (Euphorbiaceae)

Silva¹,

Freitas²,

Santos³

et al. 2019

Enci Bio

View full text Add to dashboard Cite

The genetic-molecular knowledge of the species that present relevance as a natural resource is fundamental for the elaboration of strategies of conservation and sustainable use of biodiversity. Molecular markers are important tools for obtaining relative genetic data from population estimates to identification of genomic regions associated with phenotypic characteristics. The aim of this study was to present an overview of the available knowledge in the literature on the application of molecular markers for the study of species of the genus Croton, indicating both the main gaps and perspectives regarding the available molecular tools and its application in the understanding of the genus, aiming at the conservation and use of these species. For that, searches were made on databases (indexers) of scientific publications. In summary, genetic studies with the application of molecular markers were restricted to seven species of Croton spp., corresponding to less than 1% of the species of the genus, and most dominant molecular markers were used, a fact that limits the possibility of authors to broaden their discussions and conclusions. The data presented indicate the lack of knowledge regarding the genetic characteristics and the population genetic structure for Croton spp.. Consequently, this reality testifies to a set of opportunities for conducting new research that contributes to the characterization and use of this biodiversity.

show abstract

“…Based on the results above, it was estimated that the ITS would be a suitable barcoding marker for the HRM analyses for the target species. ITS has also been used for plant species identification with excellent results [ 2 , 5 , 19 , 20 ].…”

Section: Resultsmentioning

confidence: 99%

Evaluation of a DNA-based method for spice/herb authentication, so you do not have to worry about what is in your curry, buon appetito!

et al. 2017

Self Cite

View full text Add to dashboard Cite

It is long believed that some spices may help protect against certain chronic conditions. Spices are usually parts of plants that have been powdered into small pieces. Have you ever wondered what the curry powder in your dish is made of? The aim of this work was to develop an appropriate DNA-based method for assessment of spice identity. Selecting the best marker for species recognition in the Zingiberaceae family. Six DNA regions were investigated in silico, including ITS, matK, rbcL, rpoC, trnH-psbA and trnL. Then, only four regions (ITS, matK, rbcL and trnH-psbA) were included in the simulated HRM (High-resolution Melting) analysis as the results from previous analysis showed that rpoC and trnL may not be suitable to be used to identify Zingiberaceae species in HRM analysis based on both the percentage of nucleotide variation and GC content. Simulated HRM analysis was performed to test the feasibility of Bar-HRM. We found that ITS2 is the most effective region to be used for identification of the studied species and thus was used in laboratory HRM analysis. All seven tested Zingiberaceae plants were then able to be distinguished using the ITS2 primers in laboratory HRM. Most importantly the melting curves gained from fresh and dried tissue overlapped, which is a crucial outcome for the applicability of the analysis. The method could be used in an authentication test for dried products. In the authentication test, only one of seven store-sold Zingiberaceae products that were tested contained the species listed on their labels, while we found substitution/contamination of the tested purchased products in the rest.

show abstract

Refining DNA Barcoding Coupled High Resolution Melting for Discrimination of 12 Closely Related Croton Species

Cited by 33 publications

References 59 publications

Rapidly discriminate commercial medicinal Pulsatilla chinensis (Bge.) Regel from its adulterants using ITS2 barcoding and specific PCR-RFLP assay

Rapidly discriminate commercial medicinal Pulsatilla chinensis (Bge.) Regel from its adulterants using ITS2 barcoding and specific PCR-RFLP assay

CONTRIBUIÇÕES DA BIOLOGIA MOLECULAR PARA CARACTERIZAÇÃO, CONSERVAÇÃO E USO DO GÊNERO Croton L. (Euphorbiaceae)

Evaluation of a DNA-based method for spice/herb authentication, so you do not have to worry about what is in your curry, buon appetito!

Contact Info

Product

Resources

About