scholarly journals The efficiency of universal mitochondrial DNA barcodes for species discrimination of Pomacea canaliculata and Pomacea maculata

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8755
Author(s):  
Adrian Kannan ◽  
Suganiya Rama Rao ◽  
Shyamala Ratnayeke ◽  
Yoon-Yen Yow
Keyword(s):  
16S Rdna ◽  
Dna Barcoding ◽  
Peninsular Malaysia ◽  
Coi Gene ◽  
Diagnostic Tools ◽  
Species Discrimination ◽  
Pomacea Canaliculata ◽  
Agricultural Wetlands ◽  
Barcoding Gap ◽  
Species Specific

Invasive apple snails, Pomacea canaliculata and P. maculata, have a widespread distribution globally and are regarded as devastating pests of agricultural wetlands. The two species are morphologically similar, which hinders species identification via morphological approaches and species-specific management efforts. Advances in molecular genetics may contribute effective diagnostic tools to potentially resolve morphological ambiguity. DNA barcoding has revolutionized the field of taxonomy by providing an alternative, simple approach for species discrimination, where short sections of DNA, the cytochrome c oxidase subunit I (COI) gene in particular, are used as ‘barcodes’ to delineate species boundaries. In our study, we aimed to assess the effectiveness of two mitochondrial markers, the COI and 16S ribosomal deoxyribonucleic acid (16S rDNA) markers for DNA barcoding of P. canaliculata and P. maculata. The COI and 16S rDNA sequences of 40 Pomacea specimens collected from six localities in Peninsular Malaysia were analyzed to assess their barcoding performance using phylogenetic methods and distance-based assessments. The results confirmed both markers were suitable for barcoding P. canaliculata and P. maculata. The phylogenies of the COI and 16S rDNA markers demonstrated species-specific monophyly and were largely congruent with the exception of one individual. The COI marker exhibited a larger barcoding gap (6.06–6.58%) than the 16S rDNA marker (1.54%); however, the magnitude of barcoding gap generated within the barcoding region of the 16S rDNA marker (12-fold) was bigger than the COI counterpart (approximately 9-fold). Both markers were generally successful in identifying P. canaliculata and P. maculata in the similarity-based DNA identifications. The COI + 16S rDNA concatenated dataset successfully recovered monophylies of P. canaliculata and P. maculata but concatenation did not improve individual datasets in distance-based analyses. Overall, although both markers were successful for the identification of apple snails, the COI molecular marker is a better barcoding marker and could be utilized in various population genetic studies of P. canaliculata and P. maculata.

scholarly journals Molecular analysis of the mitochondrial markers COI, 12S rDNA and 16S rDNA for six species of Iranian scorpions

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Parisa Soltan-Alinejad ◽  
Javad Rafinejad ◽  
Farrokh Dabiri ◽  
Piero Onorati ◽  
Olle Terenius ◽  
...  
Keyword(s):  
16S Rdna ◽  
Phylogenetic Analyses ◽  
Coi Gene ◽  
Diagnostic Tools ◽  
Morphological Identification ◽  
Species Determination ◽  
Important Species ◽  
Mitochondrial Markers ◽  
12S Rdna ◽  
Rdna Sequences

Abstract Objectives Annually, 1.2 million humans are stung by scorpions and severely affected by their venom. Some of the scorpion species of medical importance have a similar morphology to species with low toxicity. To establish diagnostic tools for surveying scorpions, the current study was conducted to generate three mitochondrial markers, Cytochrome Oxidase I (COI gene), 12S rDNA and 16S rDNA for six species of medically important Iranian scorpions: Androctonus crassicauda, Hottentotta saulcyi, Mesobuthus caucasicus, M. eupeus, Odontobuthus doriae, and Scorpio maurus. Results Phylogenetic analyses of the obtained sequences corroborated the morphological identification. For the first time, 12S rDNA sequences are reported from Androctonus crassicauda, Hottentotta saulcyi, Mesobuthus caucasicus and M. eupeus and also the 16S rDNA sequence from Hottentotta saulcyi. We conclude that the mitochondrial markers are useful for species determination among these medically important species of scorpions.

scholarly journals Cytochrome Oxidase I (COI) Divergence Assessment of Family Nemipteridae from Malaysian Waters

2019 ◽  
Vol 1 (1) ◽  
pp. 41-48
Author(s):  
Liew You En ◽  
Salwani Abdullah ◽  
Tan Min Pau ◽  
Mazlan Abd Ghaffar ◽  
Alias Man ◽  
...  
Keyword(s):  
Species Identification ◽  
East Coast ◽  
Peninsular Malaysia ◽  
Coi Gene ◽  
Species Discrimination ◽  
Trawl Survey ◽  
Putative Species ◽  
Commercially Important ◽  
Nucleotide Divergence ◽  
Coi Sequences

DNA Barcoding, primarily focusing on cytochrome c oxidase subunit I (COI) gene has been appraised as an effective tool for species identification. Nonetheless, species identification based on molecular approach is essential for discrimination of look-alike species. In this study, we focused on the marine fishes Family Nemipteridae, one of the commercially important group distributed within the surrounding seas of Malaysia. Some of the samples were collected during the National Demersal Trawl Survey in the Exclusive Economic Zone of East Coast Peninsular Malaysia by the Department of Fishery Malaysia. A 652bp region of COI was sequenced for 74 individuals from nine putative species. Additional 34 COI sequences from GenBank were also included in this study making the total number of samples analysed to 108 individuals. The average Kimura 2-parameter (K2P) nucleotide divergence was 0.34% among individuals within species and 6.97% within genera. All putative species formed monophyletic clades in both Neighbour-joining (NJ) and Maximum-likelihood (ML) trees. However, there was a potential misidentification in specimen identified as Nemipterus tambuloides, as the specimen did not group with their own taxa. It was genetically grouped in Nemipterus thosaporni clade. This study supports the effectiveness of COI gene in species discrimination of Family Nemipteridae.

scholarly journals Identification of CITES-Listed Euphorbia royleana through DNA Barcoding Technology: A New Facet in Wildlife Forensics

2021 ◽  
Vol 3 (2) ◽  
pp. 260-272
Author(s):  
Mukesh Thakar ◽  
Tina Sharma
Keyword(s):  
Genetic Distance ◽  
Dna Barcoding ◽  
Related Species ◽  
Species Discrimination ◽  
Closely Related Species ◽  
Wildlife Forensics ◽  
Interspecific Genetic Distance ◽  
Species Specific ◽  
Endangered Status ◽  
Discrimination Method

Disorganized and chaotic collection of the Euphorbia plant species from the wild is one of the major reasons for its endangered status. According to CITES, the trade in Euphorbia royleana species is prohibited under Appendix II. However, the trade continues unabated as current identification methods do not discriminate between closely related species.  In the present study, a DNA barcoding method has been used to establish inter- and intra-specific divergences of both matK and rbcL regions by using pairwise genetic distance measurement methods for evaluating the maximum barcoding gap. The matk and rbcL yielded a 100% amplification and sequencing success rate to distinguish closely related species of Euphorbia royleana unambiguously. The matk and rbcL showed average interspecific genetic distance divergence values of 0.031and 0.015, respectively. The maximum number of species-specific SNPs was observed in matK sequences at seven consecutive sites, which could distinguish Euphorbia royleana from closely related species.  The best candidate barcoding region to identify Euphorbia royleana was found to be matK with a single-locus barcoding approach. Furthermore, the species discrimination method was developed with the help of species-specific SNPs derived from the matK barcoding region to accurately authenticate Euphorbia royleana, and it provided 100% species resolution

scholarly journals SNP Typing Using Multiplex Real-Time PCR Assay for Species Identification of Forensically Important Blowflies and Fleshflies Collected in South Korea (Diptera: Calliphoridae and Sarcophagidae)

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Hari Jang ◽  
Sang Eon Shin ◽  
Kwang soo Ko ◽  
Seong Hwan Park
Keyword(s):  
South Korea ◽  
Real Time ◽  
Dna Barcoding ◽  
Real Time Pcr ◽  
Coi Gene ◽  
Nucleotide Sequencing ◽  
Morphological Identification ◽  
Growth Data ◽  
Important Species ◽  
Species Specific

Medicolegal entomology—a subfield of forensic entomology—is mainly used in medicolegal investigations to estimate the postmortem interval (PMI). The minimum PMI of a corpse invaded by necrophagous immature insects can be estimated because the PMI is near to or earlier than the oviposition time of the larvae that hatched and fed on the corpse. As the growth speeds of larvae differ depending on temperature and species, species-specific growth data are used to estimate the minimum PMI. While morphological identification of adult necrophagous flies can be done by a well-trained entomologist, identification of larvae is relatively difficult. Larvae can only be identified up to the family level and developmental stage by observing the posterior spiracles. For these reasons, the molecular biology method of DNA barcoding has been developed. DNA barcoding that targets the mitochondrial cytochrome c oxidase subunit I (COI) gene is commonly used. COI sequences are currently acquired using polymerase chain reaction (PCR) and Sanger sequencing, which are too time-consuming and complex for practical use in medicolegal investigations. To compensate for these limitations and facilitate the use of entomology for medicolegal investigation, we designed a multiplex real-time PCR system to identify nineteen forensically important species of Calliphoridae and Sarcophagidae flies collected in South Korea. In contrast to the Sanger nucleotide sequencing process, this technology only requires a one-step real-time PCR with melt curve analysis of amplicons generated by primers targeting species-specific single nucleotide polymorphisms (SNPs). Multiplex real-time PCR was performed for twelve species of Calliphoridae (four reactions) and for seven species of Sarcophagidae (three reactions). This assay is expected to make it easier and faster for investigating authorities to identify major species of necrophagous flies at beginning of investigation and to increase the utilization of entomological evidence in forensic investigations.

DNA barcoding of common soft scales (Hemiptera: Coccoidea: Coccidae) in China

2015 ◽  
Vol 105 (5) ◽  
pp. 545-554 ◽  
Author(s):  
X.-B. Wang ◽  
J. Deng ◽  
J.-T. Zhang ◽  
Q.-S. Zhou ◽  
Y.-Z. Zhang ◽  
...  
Keyword(s):  
Dna Barcoding ◽  
Coi Gene ◽  
Species Discrimination ◽  
Correct Identification ◽  
Success Rates ◽  
Agricultural Pests ◽  
Data Set ◽  
Operational Taxonomic Units ◽  
Ceroplastes Ceriferus ◽  
Level Identification

AbstractThe soft scales (Hemiptera: Coccoidea: Coccidae) are a group of sap-sucking plant parasites, many of which are notorious agricultural pests. The quarantine and economic importance of soft scales necessitates rapid and reliable identification of these taxa. Nucleotide sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene (barcoding region) and 28S rDNA were generated from 340 individuals of 36 common soft scales in China. Distance-based [(best match, Automated Barcode Gap Discovery (ABGD)], tree-based (neighbor-joining, Bayesian inference), Klee diagrams, and general mixed Yule coalescent (GMYC) models were used to evaluate barcoding success rates in the data set. Best match showed that COI and 28S sequences could provide 100 and 95.52% correct identification, respectively. The average interspecific divergences were 19.81% for COI data and 20.38% for 28S data, and mean intraspecific divergences were 0.56 and 0.07%, respectively. For COI data, multiple methods (ABGD, Klee, and tree-based methods) resulted in general congruence with morphological identifications. However, GMYC analysis tended to provide more molecular operational taxonomic units (MOTUs). Twelve MOTUs derived from five morphospecies (Rhodococcus sariuoni, Pulvinaria vitis, Pulvinaria aurantii, Parasaissetia nigra, and Ceroplastes rubens) were observed using the GMYC approach. In addition, tree-based methods showed that 28S sequences could be used for species-level identification (except for Ceroplastes ceriferus – Ceroplastes pseudoceriferus), even with low genetic variation (<1%). This report demonstrates the robustness of DNA barcoding for species discrimination of soft scales with two molecular markers (COI and 28S) and provides a reliable barcode library and rapid diagnostic tool for common soft scales in China.

scholarly journals DNA Barcoding of Endangered Paphiopedilum species (Orchidaceae) of Peninsular Malaysia

Phytotaxa ◽  
2019 ◽  
Vol 387 (2) ◽  
pp. 94-104 ◽  
Author(s):  
MICHEAL C. RAJARAM ◽  
CHRISTINA S.Y. YONG ◽  
JUALANG A. GANSAU ◽  
RUSEA GO
Keyword(s):  
Phylogenetic Tree ◽  
Dna Barcoding ◽  
Dna Markers ◽  
High Accuracy ◽  
Peninsular Malaysia ◽  
Dna Barcodes ◽  
Accurate Identification ◽  
Barcoding Gap ◽  
Preliminary Identification ◽  
Sequence Quality

In this study, the efficacy of four DNA markers and their combinations (rbcL, matK, ITS, trnH-psbA) as barcode markers were tested across the endangered Paphiopedilum species from Peninsular Malaysia. Four species of Paphiopedilum were sampled and barcoded. The DNA barcodes reliabilities were evaluated using NCBI BLASTn program, phylogenetic tree via Neighbour-Joining method with 1000 bootstrap replicates in MEGA 6 and barcoding gap assessment. matK is the most promising barcode with high sequence quality (100%), high accuracy in BLASTn (100%), clear resolution of species in Neighbour-Joining phylogenetic tree (100%) and a distinct barcoding gap followed by ITS, trnH-psbA and rbcL. The combination of barcode regions revealed the lack of variation in rbcL and trnH-psbA but they are still useful for preliminary identification followed up by matK for accurate identification.

scholarly journals Evaluating five different loci (rbcL, rpoB, rpoC1, matK, and ITS) for DNA barcoding of Indian orchids

Genome ◽  
2017 ◽  
Vol 60 (8) ◽  
pp. 665-671 ◽  
Author(s):  
Iffat Parveen ◽  
Hemant K. Singh ◽  
Saloni Malik ◽  
Saurabh Raghuvanshi ◽  
Shashi B. Babbar
Keyword(s):  
Dna Barcoding ◽  
Habitat Destruction ◽  
Species Discrimination ◽  
Medicinal Species ◽  
Orchid Species ◽  
Illicit Trade ◽  
Discrimination Capability ◽  
Discrimination Rate ◽  
Identification Tags ◽  
Species Specific

Orchidaceae, one of the largest families of angiosperms, is represented in India by 1600 species distributed in diverse habitats. Orchids are in high demand owing to their beautiful flowers and therapeutic properties. Overexploitation and habitat destruction have made many orchid species endangered. In the absence of effective identification methods, illicit trade of orchids continues unabated. Considering DNA barcoding as a potential identification tool, species discrimination capability of five loci, ITS, matK, rbcL, rpoB, and rpoC1, was tested in 393 accessions of 94 Indian orchid species belonging to 47 genera, including one listed in Appendix I of CITES and 26 medicinal species. ITS provided the highest species discrimination rate of 94.9%. While, among the chloroplast loci, matK provided the highest species discrimination rate of 85.7%. None of the tested loci individually discriminated 100% of the species. Therefore, multi-locus combinations of up to five loci were tested for their species resolution capability. Among two-locus combinations, the maximum species resolution (86.7%) was provided by ITS+matK. ITS and matK sequences of the medicinal orchids were species specific, thus providing unique molecular identification tags for their identification and detection. These observations emphasize the need for the inclusion of ITS in the core barcode for plants, whenever required and available.

scholarly journals A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 199
Author(s):  
Xiaochun Zhang ◽  
Huan Yu ◽  
Qi Yang ◽  
Ziwei Wang ◽  
Ruocheng Xia ◽  
...  
Keyword(s):  
High Resolution ◽  
Dna Barcoding ◽  
High Resolution Melting ◽  
Its Region ◽  
Its Sequencing ◽  
Melting Temperatures ◽  
Hrm Analysis ◽  
National Drug ◽  
Species Specific ◽  
Its1 And Its2

In recent years, trafficking and abuse of hallucinogenic mushrooms have become a serious social problem. It is therefore imperative to identify hallucinogenic mushrooms of the genus Psilocybe for national drug control legislation. An internal transcribed spacer (ITS) is a DNA barcoding tool utilized for species identification. Many methods have been used to discriminate the ITS region, but they are often limited by having a low resolution. In this study, we sought to analyze the ITS and its fragments, ITS1 and ITS2, by using high-resolution melting (HRM) analysis, which is a rapid and sensitive method for evaluating sequence variation within PCR amplicons. The ITS HRM assay was tested for specificity, reproducibility, sensitivity, and the capacity to analyze mixture samples. It was shown that the melting temperatures of the ITS, ITS1, and ITS2 of Psilocybe cubensis were 83.72 ± 0.01, 80.98 ± 0.06, and 83.46 ± 0.08 °C, and for other species, we also obtained species-specific results. Finally, we performed ITS sequencing to validate the presumptive taxonomic identity of our samples, and the sequencing output significantly supported our HRM data. Taken together, these results indicate that the HRM method can quickly distinguish the DNA barcoding of Psilocybe cubensis and other fungi, which can be utilized for drug trafficking cases and forensic science.

Temporal Detection Limits of Remnant Larval Bloodmeals in Nymphal Ixodes scapularis (Say, Ixodida: Ixodidae) Using Two Next-Generation Sequencing DNA Barcoding Assays

2020 ◽  
Author(s):  
Genevieve A M Lumsden ◽  
Evgeny V Zakharov ◽  
Sarah Dolynskyj ◽  
J Scott Weese ◽  
L Robbin Lindsay ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Dna Barcoding ◽  
Ixodes Scapularis ◽  
Life Stage ◽  
Next Generation ◽  
Host Detection ◽  
Host Identification ◽  
Species Specific ◽  
Generation Sequencing ◽  
Assay Detection

Abstract Using next-generation sequencing DNA barcoding, we aimed to determine: 1) if the larval bloodmeal can be detected in Ixodes scapularis nymphs and 2) the post-moult temporal window for detection of the larval bloodmeal. Subsets of 30 nymphs fed on a domestic rabbit (Oryctolagus cuniculus Linnaeus, Lagomorphia: Leporidae) as larvae were reared and frozen at 11 time points post-moult, up to 150 d. Vertebrate DNA was amplified using novel universal (UP) and species-specific primers (SSP) and sequenced for comparison against cytochrome c oxidase subunit I barcodes to infer host identification. Detectable bloodmeals decreased as time since moult increased for both assays. For the SSP assay, detection of bloodmeals decreased from 96.7% (n = 29/30) in day 0 nymphs to 3.3% (n = 1/30) and 6.7% (n = 2/30) at 4- and 5-mo post-moult, respectively. A shorter temporal detection period was achieved with the UP assay, declining from 16.7% (n = 5/30) in day 0 nymphs to 0/30 in 3-d-old nymphs. Bloodmeal detection was nonexistent for the remaining cohorts, with the exception of 1/30 nymphs at 2-mo post-moult. Host detection was significantly more likely using the SSP assay compared to the UP assay in the first three time cohorts (day 0: χ 2 = 39.1, P &lt; 0.005; day 2: χ 2 = 19.2, P &lt; 0.005; day 3: χ 2 = 23.3, P &lt; 0.005). Regardless of the primer set used, the next-generation sequencing DNA barcoding assay was able to detect host DNA from a larval bloodmeal in the nymphal life stage; however, a short window with a high proportion of detection post-moult was achieved.

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