scholarly journals Identification of Tinomiscium petiolare from Vietnam using the DNA barcode

2021 ◽  
Vol 182 (2) ◽  
pp. 114-122
Author(s):  
B. B. Thinh ◽  
R. V. Doudkin ◽  
L. D. Chac ◽  
H. V. Chinh ◽  
Q. V. Hoi ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Dna Barcode ◽  
Pcr Amplification ◽  
Rapid Identification ◽  
Medicinal Species ◽  
Cycle Sequencing ◽  
Stage Of Development ◽  
Pcr Products ◽  
Genetic Sequences ◽  
Total Dna

Background. Tinomiscium petiolare Hook.f. & Thomson is a medicinal species of the family Menispermaceae. This species is currently being intensively exploited for therapeutic purposes. Precise and rapid identification of T. petiolare is critical and essential for the classification, propagation, use and conservation of its genetic resources. In recent years, DNA barcoding has been known to be a fast and sensitive method for identifying species at any stage of development, using short DNA sequences. In this study we have performed the identification of T. petiolare specimens in Vietnam based on the sequence analysis of 4 DNA barcode loci: ITS, matK, rbcL and rpoC.Materials and methods. Total DNA was extracted from leaf samples using DNeasy Plant Mini Kit. PCR amplification of the ITS, matK, rbcL and rpoC regions was carried out on the GeneAmp PCR System 9700 with specific primers. The purified PCR products were sequenced on the ABI 3500 Genetic Analyzer system, using BigDye®Terminator v3.1 Cycle Sequencing Kit. These genetic sequences were analyzed and compared, and a phylogenetic tree was constructed using BioEdit, BLAST, and MEGA 6 programs.Results and conclusion. The success rate of amplification and sequencing was 100% for all 4 DNA barcode loci (ITS, matK, rbcL and rpoC) in the studied specimens. The produced sequence sizes of ITS, matK, rbcL and rpoC in the specimens were 574 bp, 810 bp, 527 bp and 488 bp, respectively. Further, we identified that all studied specimens were genetically related to each other and associated with the same species T. petiolare. Overall, the results of the study generated the most complete DNA barcode database of T. petiolare collected in Vietnam, contributing to the taxonomy and identification of this species. 

ISSR-Derived Species-Specific SCAR Marker for Rapid and Accurate Authentication of Ocimum tenuiflorum L.

Planta Medica ◽  
2017 ◽  
Vol 84 (02) ◽  
pp. 117-122 ◽  
Author(s):  
Amit Kumar ◽  
Vereena Rodrigues ◽  
Priyanka Mishra ◽  
Kuppusamy Baskaran ◽  
Ashutosh Shukla ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
High Volume ◽  
Inter Simple Sequence Repeat ◽  
Rapid Identification ◽  
Medicinal Species ◽  
Accurate Identification ◽  
Sequence Characterized Amplified Region ◽  
Medicinal Value ◽  
Ocimum Tenuiflorum ◽  
Species Specific

Abstract Ocimum tenuiflorum has been widely used in traditional medicine and has high medicinal value. High volume trade of this potential medicinal plant species led to unscrupulous adulteration of both crude drugs as well as formulations. Morphology-based authentication is difficult in cases of incomplete or damaged samples and in dried herbal materials. In such cases, PCR-based molecular methods may aid in accurate identification. The present study aimed at developing species-specific DNA marker(s) for the authentication of O. tenuiflorum. A species-specific amplicon (279 bp) generated through an inter-simple sequence repeat marker (UBC 835) in all individuals of O. tenuiflorum was cloned, sequenced, and a primer pair was developed (designated as CIM-OT-835F/CIM-OT-835R). The newly developed sequence characterized amplified region marker was validated through PCR amplification in all available seven species of Ocimum, and its specificity for O. tenuiflorum was confirmed with the consistent generation of an amplicon of 177 bp. The developed marker can be used for accurate and rapid identification of the species for certification purposes and will be useful in quality control of medicinal preparations containing this important medicinal species.

scholarly journals Gene Sequence Analysis of Mitochondrial COI in Genus Saperda (Coleoptera: Cerambycidae: Lamiinae)

2021 ◽  
Author(s):  
Wei XU ◽  
Qinhua Gan ◽  
Jian Pu ◽  
Yingwen Pan ◽  
Bo Cai ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Detection Rate ◽  
Gene Sequence ◽  
Mitochondrial Gene ◽  
Dna Barcode ◽  
Nuclear Gene ◽  
Rapid Identification ◽  
Mitochondrial Coi ◽  
Gene Sequence Analysis ◽  
Total Dna

In this study, the total DNA of nine species of Saperda (Lopezcolonia) octopunctata (Scopoli, 1772), Saperda (Lopezcolonia) scalaris (Linnaeus, 1758), Saperda interrupta Gebler, Saperda Alberti (Plavilstshikov), Saperda (Saperda) similis Laicharting, 1784, Saperda (Compsidia) populnea (Linnaeus, 1758), Saperda (Saperda) carcharias (Linnaeus, 1758), Saperda (Lopezcolonia) perforata Pallas, 1773 and Saperda ohbayashi were extracted. Two partial sequences of mitochondrial gene and one partial sequence of nuclear gene were amplified. Comparing the COI sequence with the DNA barcode data in GenBank can effectively identify the related species of Saperda. It will be applied to the rapid identification of some species of Saperda in imported wood at ports, and improve the detection rate of plant quarantine.

scholarly journals Hepatitis B Transmission: Is Vertical Transmission the Major Route in Intermediate Endemic Areas? A Proof-of-Concept Study Based on Mother–Child Genotypes

2021 ◽  
Author(s):  
Ujjal Poddar ◽  
Mercilena Benjamin ◽  
Rakesh Aggarwal ◽  
Aditya Narayan Sarangi ◽  
Amrita Mathias ◽  
...  
Keyword(s):  
Hepatitis B ◽  
Region Of Interest ◽  
Cycle Sequencing ◽  
Child Pair ◽  
Mother And Child ◽  
Pcr Products ◽  
Major Route ◽  
Genetic Sequences ◽  
Hepatitis B Transmission ◽  
Genotype A

The route of hepatitis B transmission is believed to be horizontal in India, though pediatric studies showed mother as source in the majority of chronic HBV (CHB) cases. We aimed at establishing the fact that mother–child transmission is the main route of acquisition by documenting genotypically identical viruses in mother–child pairs. Blood samples of consecutive children (≤18 years) with CHB and high DNA (>10,000 IU/mL) and their positive mother were collected from January 2013 to December 2015. Polymerase chain reaction (PCR) products of HBV-DNA were amplified and sequenced by using BigDye Terminator Cycle Sequencing Kit v3.1 and aligned with previously described sequences in the region of interest for genotypes A to G by using BioEdit software. Phylogenetic tree was generated using p-distance algorithm in MEGA software version 6. Genotyping of 59 (33 children and 26 mothers) subjects include genotype A in 24 (40.7%) and genotype D in 35 (59.3%). Both mother–child pair genotyping was possible in 25. The median age of 25 children (20 males) was 9 (interquartile range, IQR: 4–11). The distribution of genotypes among mother–child pairs was similar. The concordance between children and their mothers was 24 of 25 (96%). Evolutionary analyses showed significant similarities between mother and child sequences for both genotype A and D, suggesting thereby the same virus. In conclusion, mother–baby transmission seems to be the major route of acquisition of HBV in children in India and near-complete homology in genetic sequences between mother–child pairs is definite proof for that. However, a larger epidemiological study is required to substantiate our findings.

scholarly journals Molecular identification of species of Physalis (Solanaceae) using a candidate DNA barcode: the chloroplast psbA–trnH intergenic region

Genome ◽  
2018 ◽  
Vol 61 (1) ◽  
pp. 15-20 ◽  
Author(s):  
Shangguo Feng ◽  
Kaili Jiao ◽  
Yujia Zhu ◽  
Hongfen Wang ◽  
Mengying Jiang ◽  
...  
Keyword(s):  
Intergenic Region ◽  
Morphological Traits ◽  
Dna Barcode ◽  
Pcr Amplification ◽  
Genetic Distances ◽  
Success Rates ◽  
Medicinal Species ◽  
Distance Methods ◽  
Identification Of Species ◽  
Related Plant

Physalis L., an important genus of the family Solanaceae, includes many commercially important edible and medicinal species. Traditionally, species identification is based on morphological traits; however, the highly similar morphological traits among species of Physalis make this approach difficult. In this study, we evaluated the feasibility of using a popular DNA barcode, the chloroplast psbA–trnH intergenic region, in the identification of species of Physalis. Thirty-six psbA–trnH regions of species of Physalis and of the closely related plant Nicandra physalodes were analyzed. The success rates of PCR amplification and sequencing of the psbA–trnH region were 100%. MEGA V6.0 was utilized to align the psbA–trnH sequences and to compute genetic distances. The results show an apparent barcoding gap between intra- and interspecific variations. Results of both BLAST1 and nearest-distance methods prove that the psbA–trnH regions can be used to identify all species examined in the present study. In addition, phylogenetic analysis using psbA–trnH data revealed a distinct boundary between species. It also confirmed the relationship between species of Physalis and closely related species, as established by previous studies. In conclusion, the psbA–trnH intergenic region can be used as an efficient DNA barcode for the identification of species of Physalis.

scholarly journals Barcode DNA Edelweis (Anaphalis javanica) Berdasarkan Gen matK

Jurnal MIPA ◽  
2015 ◽  
Vol 4 (2) ◽  
pp. 131
Author(s):  
Muzakir Rahalus ◽  
Maureen Kumaunang ◽  
Audy Wuntu ◽  
Julius Pontoh
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Dna Barcode ◽  
Living Organism ◽  
Chain Reaction ◽  
Base Pairs ◽  
Barcode Dna ◽  
Reverse Primer ◽  
Total Dna ◽  
Polymerase Chain

DNA barcode merupakan metode identifikasi organisme hidup dengan menggunakan urutan DNA pendek (± 500 pasang basa). Tujuan dari penelitian ini adalah memperoleh barcode DNA Edelweis dan menganalisis kemiripan gen matK Edelweis (Anaphalis javanica) dengan kerabat terdekatnya. Isolasi DNA total Edelweis berhasil dilakukan dengan menggunakan  manual prosedur dari InnuPrep Plant DNA Kit yang dimodifikasi. Gen matK parsial telah diisolasi dengan metode Polymerase Chain Reaction (PCR) menggunakan Primer forward matK-1RKIM-f dan Primer Reverse matK-3FKIM-r. Hasil analisis sekuens menghasilkan barcode DNA edelweis berukuran 843 bp. Hasil analisis kemiripan menunjukkan tingkat kekerabatan terdekat dengan A. margaritaceae yaitu 99.86% pada BOLD System dan 100 % pada NCBI.DNA barcode is a method to identify living organism by using several short sequences of DNA (± 500 base pairs). The purpose of this study was to obtain a DNA barcode and analyze the similarity of matK genes of edelweis (Anaphalis javanica) with its closest relatives. Isolation of total DNA of edelweis has been succesfully done by using modified manual procedures of InnuPrep Plant Kit. matK partial gene has been isolated by the method of Polymerase Chain Reaction (PCR) using forward primer MATK-1RKIM-f and reverse primer MATK-3FKIM-r. Analysis of DNA sequences of edelweis confirmed its DNA barcode size was 843 bp. Furthermore, A. javanica showed similarity 99.86% in BOLD system and 100% in NCBI with A. margaritaceae.

scholarly journals Glyceraldehyde-3-Phosphate Dehydrogenase-Encoding Gene as a Useful Taxonomic Tool for Staphylococcusspp.

2000 ◽  
Vol 38 (12) ◽  
pp. 4351-4355 ◽  
Author(s):  
Javier Yugueros ◽  
Alejandro Temprano ◽  
Beatriz Berzal ◽  
Marı́a Sánchez ◽  
Carmen Hernanz ◽  
...  
Keyword(s):  
Staphylococcus Epidermidis ◽  
Fragment Length Polymorphism ◽  
Pcr Amplification ◽  
Rapid Identification ◽  
Pcr Products ◽  
Dna Fragment ◽  
Encoding Gene ◽  
Staphylococcus Hominis ◽  
Staphylococcus Simulans ◽  
Very High

The gap gene of Staphylococcus aureus, encoding glyceraldehyde-3-phosphate dehydrogenase, was used as a target to amplify a 933-bp DNA fragment by PCR with a pair of primers 26 and 25 nucleotides in length. PCR products, detected by agarose gel electrophoresis, were also amplified from 12 Staphylococcusspp. analyzed previously. Hybridization with an internal 279-bp DNA fragment probe was positive in all PCR-positive samples. No PCR products were amplified when other gram-positive and gram-negative bacterial genera were analyzed using the same pair of primers.AluI digestion of PCR-generated products gave 12 different restriction fragment length polymorphism (RFLP) patterns, one for each species analyzed. However, we could detect two intraspecies RFLP patterns in Staphylococcus epidermidis,Staphylococcus hominis, and Staphylococcus simulans which were different from the other species. An identical RFLP pattern was observed for 112 S. aureusisolates from humans, cows, and sheep. The sensitivity of the PCR assays was very high, with a detection limit for S. aureuscells of 20 CFU when cells were suspended in saline. PCR amplification of the gap gene has the potential for rapid identification of at least 12 species belonging to the genusStaphylococcus, as it is highly specific.

DOP–PCR based painting of rye chromosomes in a wheat background

Genome ◽  
2014 ◽  
Vol 57 (9) ◽  
pp. 473-479 ◽  
Author(s):  
Chuanliang Deng ◽  
Lili Bai ◽  
Shufen Li ◽  
Yingxin Zhang ◽  
Xiang Li ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Chromosome Painting ◽  
Pcr Amplification ◽  
Translocation Line ◽  
Addition Line ◽  
Metaphase Chromosomes ◽  
Application Range ◽  
Degenerate Oligonucleotide ◽  
Pcr Products ◽  
Chromosome Fragments

To determine the appropriateness of chromosome painting for identifying genomic elements in rye, we microdissected the 1R and 1RS chromosomes from rye (Secale cereale L. var. King II) and wheat–rye addition line 1RS, respectively. Degenerate oligonucleotide primed – polymerase chain reaction (DOP–PCR) amplification of 1R and 1RS products from dissected chromosomes were used as probes to hybridize to metaphase chromosomes of rye, wheat–rye addition lines 1R and 1RS, translocation line 1RS.1BL, and allohexaploid triticale. The results showed that (i) the hybridization signal distribution patterns on rye chromosomes using 1R-derived DOP–PCR products as the probe were similar to those using 1RS-derived DOP–PCR products as the probe; (ii) 1R and (or) 1RS could not be distinguished from other rye chromosomes solely by the hybridization patterns using 1R- and (or) 1RS-derived DOP–PCR products as the probe; (iii) rye chromosomes and (or) rye chromosome fragments could be clearly identified in wheat–rye hybrids using either 1R- or 1RS-derived DOP–PCR products as the probe and could be more accurate in the nontelomeric region than using genomic in situ hybridization (GISH). Our results suggested that 1R- and (or) 1RS-derived DOP–PCR products contain many repetitive DNA sequences, are similar on different rye chromosomes, are R-genome specific, and can be used to identify rye chromosomes and chromosome fragments in wheat–rye hybrids. Our research widens the application range of chromosome painting in plants.

scholarly journals Barcode DNA Tumbuhan Pangi (Pangium edule R.) Berdasarkan Gen matK

Jurnal MIPA ◽  
2014 ◽  
Vol 3 (2) ◽  
pp. 113
Author(s):  
Irmi Bangol ◽  
Lidya Irma Momuat ◽  
Maureen Kumaunang
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Dna Barcode ◽  
Chain Reaction ◽  
Closely Related Species ◽  
Plant Dna ◽  
Barcode Dna ◽  
Total Dna ◽  
Polymerase Chain ◽  
Forward Primer

DNA barcoding merupakan suatu teknik yang digunakan untuk mempercepat dan mempermudah proses identifikasi organisme dengan menggunakan potongan gen tertentu. Penelitian ini bertujuan untuk menentukan sekuens DNA barcode tumbuhan pangi berdasarkan gen standar matK dan membandingkannya dengan spesies yang berkerabat dekat di GenBank. DNA total daun pangi diisolasi menggunakan Innuprep plant DNA kit dan berhasil diamplifikasi dengan proses Polymerase Chain Reaction (PCR) menggunakan primer berdasarkan gen matK. Hasil sekuensing fragmen DNA yang menunjukkan panjang 720 bp yang teramplifikasi oleh primer forward dan 780 bp untuk yang teramplifikasi oleh primer reverse. Hasil analisis BLASTn menunjukkan tingkat kemiripan tumbuhan pangi sangat tinggi dengan Trichadenia zeylanical, yaitu 99%, dan diikuti spesies lainnya (Kiggelaria africanal, 98%; Guthriea capensis, 96%; Acharia tragodes, 92%; Erythrospermum phytolaccoides, 92%; Hydnocarpus sp. Chase 1301, 90%; Carpotroche longifolia, 89%; Moultonianthus leembruggianus, 89% dan Pimelodendron zoanthogyne, 88%). Analisis komposisi asam amino menunjukkan bahwa matK Pangium edule dan kesembilan spesies tumbuhan lainnya bersifat hidrofobik.DNA barcoding is a technical used to accelerate and simplify the process identification of organism with by using a snipping of specific genes. This study aimed to determine the DNA sequences of plant barcoding standard pangi based gene matK and compare with closely related species in GenBank. Total DNA was isolated using Innuprep pangi leaf plant DNA kit and successfully amplified by the Polymerase Chain Reaction (PCR) using primers based on the gene matK. The results of sequencing long DNA fragments showed7 20 bp are amplified by the forward primer and 780 bp were amplified by the primer for reverse. Blast analysis of the results showed very extremely high the plant pangi degree of similarity with Trichadenia zeylanical, namely99%, and followed by other species (Kiggelaria africanal, 98%; Guthriea capensis, 96%; Acharia tragodes, 92%; Erythrospermum phytolaccoides, 92%; Hydnocarpus sp. Chase 1301, 90%; Carpotroche longifolia, 89%; Moultonianthus leembruggianus, 89% dan Pimelodendron zoanthogyne, 88%). Analysis of aminoacid composition showed that matK Pangium edule and nine other plant species are hydrophobic.

scholarly journals Analysis of delta-globin gene mutations in Greek cypriots

Blood ◽  
1993 ◽  
Vol 82 (5) ◽  
pp. 1647-1651
Author(s):  
P Trifillis ◽  
A Kyrri ◽  
E Kalogirou ◽  
A Kokkofitou ◽  
P Ioannou ◽  
...  
Keyword(s):  
Globin Gene ◽  
Gene Mutations ◽  
Pcr Amplification ◽  
Rapid Identification ◽  
Restriction Enzyme Digestion ◽  
Globin Genes ◽  
Pcr Products ◽  
Direct Mutation ◽  
In Cis ◽  
Greek Cypriots

We recently described four delta-globin gene mutations in Greek Cypriots studied by polymerase chain reaction (PCR) amplification and automated fluorescence-based DNA sequence analysis (Blood 78:3298, 1991). Selective restriction enzyme digestion of PCR products facilitated direct mutation detection. Twenty-eight additional samples from unrelated Cypriots with Hb A2 levels ranging from 0.6% to 3.6% were studied by PCR and showed the following: twelve had the delta 27 (ala-->ser) mutation, one was heterozygous for the delta IVS-2 AG-->GG change, and none had either the delta 116 (arg-->cys) or delta 141 (leu- ->pro) mutations. The remaining samples were divided into two groups: 11 with borderline normal Hb A2 values that were not pursued; and four with abnormal Hb A2 values. The delta-globin genes from these four samples were sequenced and the same four changes identified in each: a C-->T at -199, a C-->T at codon 4 (thr-->ile), a silent C-->T at codon 97, and an AT deletion at position 722 in IVS-2. The codon 4 change abolishes a Ple I site whereas the codon 97 creates an Nla III site, thus facilitating rapid identification. All four changes are in cis position, suggesting that the -199 C-->T, the C-->T at codon 97, and the AT deletion in IVS-2 are neutral polymorphisms present on the codon 4 (thr-->ile) chromosome. DNA haplotype analysis suggests all five delta-globin gene mutant alleles arose independently on different chromosomal backgrounds.

scholarly journals A Simple PCR Method for Rapid Genotype Analysis ofMycobacterium ulcerans

2000 ◽  
Vol 38 (4) ◽  
pp. 1482-1487 ◽  
Author(s):  
Timothy Stinear ◽  
John K. Davies ◽  
Grant A. Jenkin ◽  
Françoise Portaels ◽  
Bruce C. Ross ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Geographic Region ◽  
Mycobacterium Ulcerans ◽  
Geographic Origins ◽  
Pcr Products ◽  
Total Dna ◽  
Pcr Method ◽  
Clonal Population Structure ◽  
Tissue Specimens ◽  
First Time

Two high-copy-number insertion sequences, IS2404 and IS2606, were recently identified in Mycobacterium ulcerans and were shown by Southern hybridization to possess restriction fragment length polymorphism between strains from different geographic origins. We have designed a simple genotyping method that captures these differences by PCR amplification of the region between adjacent copies of IS2404 and IS2606. We have called this system 2426 PCR. The method is rapid, reproducible, sensitive, and specific for M. ulcerans, and it has confirmed previous studies suggesting a clonal population structure ofM. ulcerans within a geographic region. M. ulcerans isolates from Australia, Papua New Guinea, Malaysia, Surinam, Mexico, Japan, China, and several countries in Africa were easily differentiated based on an array of 4 to 14 PCR products ranging in size from 200 to 900 bp. Numerical analysis of the banding patterns suggested a close evolutionary link between M. ulceransisolates from Africa and southeast Asia. The application of 2426 PCR to total DNA, extracted directly from M. ulcerans-infected tissue specimens without culture, demonstrated the sensitivity and specificity of this method and confirmed for the first time that both animal and human isolates from areas of endemicity in southeast Australia have the same genotype.

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