Real-Time PCR Quantification of Chloroplast DNA Supports DNA Barcoding of Plant Species

2016 ◽  
Vol 58 (3) ◽  
pp. 212-219 ◽  
Author(s):  
Hitomi S. Kikkawa ◽  
Kouichiro Tsuge ◽  
Ritsuko Sugita
Keyword(s):  
Chloroplast Dna ◽  
Real Time ◽  
Dna Barcoding ◽  
Plant Species ◽  
Real Time Pcr

Molecular Tracing of the Origin of Six Different Plant Species in Bee Honey Using Real-Time PCR

2017 ◽  
Vol 100 (3) ◽  
pp. 744-752 ◽  
Author(s):  
Yajun Wu ◽  
Yange Yang ◽  
Mingchang Liu ◽  
Bin Wang ◽  
Meige Li ◽  
...  
Keyword(s):  
Real Time ◽  
Plant Species ◽  
Real Time Pcr ◽  
Sensitivity Testing ◽  
Foreign Pollen ◽  
Geographic Regions ◽  
Fair Market ◽  
Chaste Tree ◽  
Parallel Tests

Abstract The quality of honey is significantly influenced by floralorigin. Mislabeling floral species occurs frequently in bee honey products. To protect consumers from economic fraud and maintain a fair market environment, methods to identify floralspecies in honey are necessary. In our study, real-time PCRs were established, targeting six honey types mainly produced in China (canola, Chinese milkvetch, Chinese chaste tree, locust tree, litchi, and longan). Sensitivity testing on DNA fromplant tissues exhibited LODs of about 0.5–5 pg/μL. For DNA extracts of pollen sediments from different honeyspecies, LODs ranged from 13.6 to 403.2 pg/μL. In an experiment to determine the practical LODs of honey in which adulterant honey was spiked in the genuine honey, adulterant honey as low as about 0.1–0.5% was detected in 90–100% in 10 parallel tests. Additionally, pollen was spiked in the honey and stored under various conditions to investigate the migration of pollen DNA into the honey supernatant. Finally, the efficiency of our method was investigated by testing honey samples of unknown compositions from different geographic regions. Of the 159 honey samples that were supposed tobe monofloral that had been collected in five provinces, a small portion were found to be contaminated with foreign pollen(7%). The methods proved to be specific, sensitive, and reliable in identifying the six plant species in honey, which would be a useful tool during the market supervision and QC of honey products.

Molecular identification of dried squid products sold in China using DNA barcoding and SYBR green real time PCR

2020 ◽  
Vol 37 (7) ◽  
pp. 1061-1074
Author(s):  
Rongzhen Shi ◽  
Manhong Huang ◽  
Jing Wang ◽  
Chuhan He ◽  
Xiaoguo Ying ◽  
...  
Keyword(s):  
Real Time ◽  
Dna Barcoding ◽  
Molecular Identification ◽  
Real Time Pcr ◽  
Sybr Green

scholarly journals TaqMan Real-Time PCR Assays To Assess Arbuscular Mycorrhizal Responses to Field Manipulation of Grassland Biodiversity: Effects of Soil Characteristics, Plant Species Richness, and Functional Traits

2010 ◽  
Vol 76 (12) ◽  
pp. 3765-3775 ◽  
Author(s):  
Stephan König ◽  
Tesfaye Wubet ◽  
Carsten F. Dormann ◽  
Stefan Hempel ◽  
Carsten Renker ◽  
...  
Keyword(s):  
Species Richness ◽  
Real Time ◽  
Plant Species ◽  
Real Time Pcr ◽  
Environmental Samples ◽  
Large Scale ◽  
Arbuscular Mycorrhizal ◽  
Sequence Type ◽  
Plant Biodiversity ◽  
Pcr Assays

ABSTRACT Large-scale (temporal and/or spatial) molecular investigations of the diversity and distribution of arbuscular mycorrhizal fungi (AMF) require considerable sampling efforts and high-throughput analysis. To facilitate such efforts, we have developed a TaqMan real-time PCR assay to detect and identify AMF in environmental samples. First, we screened the diversity in clone libraries, generated by nested PCR, of the nuclear ribosomal DNA internal transcribed spacer (ITS) of AMF in environmental samples. We then generated probes and forward primers based on the detected sequences, enabling AMF sequence type-specific detection in TaqMan multiplex real-time PCR assays. In comparisons to conventional clone library screening and Sanger sequencing, the TaqMan assay approach provided similar accuracy but higher sensitivity with cost and time savings. The TaqMan assays were applied to analyze the AMF community composition within plots of a large-scale plant biodiversity manipulation experiment, the Jena Experiment, primarily designed to investigate the interactive effects of plant biodiversity on element cycling and trophic interactions. The results show that environmental variables hierarchically shape AMF communities and that the sequence type spectrum is strongly affected by previous land use and disturbance, which appears to favor disturbance-tolerant members of the genus Glomus. The AMF species richness of disturbance-associated communities can be largely explained by richness of plant species and plant functional groups, while plant productivity and soil parameters appear to have only weak effects on the AMF community.

scholarly journals A TaqMan real-time PCR method based on alternative oxidase genes for detection of plant species in animal feed samples

PLoS ONE ◽  
2018 ◽  
Vol 13 (1) ◽  
pp. e0190668 ◽  
Author(s):  
Maria Doroteia Campos ◽  
Vera Valadas ◽  
Catarina Campos ◽  
Laura Morello ◽  
Luca Braglia ◽  
...  
Keyword(s):  
Real Time ◽  
Plant Species ◽  
Real Time Pcr ◽  
Alternative Oxidase ◽  
Animal Feed ◽  
Pcr Method ◽  
Feed Samples

A real-time PCR assay to differentiate the B and Q biotypes of the Bemisia tabaci complex in Cyprus

2009 ◽  
Vol 99 (6) ◽  
pp. 573-582 ◽  
Author(s):  
L.C. Papayiannis ◽  
J.K. Brown ◽  
N.A. Seraphides ◽  
M. Hadjistylli ◽  
N. Ioannou ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Real Time ◽  
Plant Species ◽  
Bemisia Tabaci ◽  
Dna Sequences ◽  
Real Time Pcr ◽  
Middle Eastern ◽  
Polymorphism Analysis ◽  
Pcr Assay ◽  
The Real

AbstractA real-time PCR assay based on TaqMan® technology was developed and evaluated for the rapid detection of the B and Q biotypes of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). A survey was conducted during 2005–2007 in order to identify the distribution and prevalence of B. tabaci biotypes in Cyprus using the real-time PCR assay. More than 700 adult whiteflies collected from 35 cultivated and weed plant species were individually haplotyped using TaqMan® PCR, and the results of the assay were validated by restriction fragment length polymorphism analysis and DNA sequencing of the mitochondrial cytochrome oxidase I (mtCOI) gene. Two biotypes, B and Q, were identified in the collected plant species on the island. The real-time PCR and RFLP assay consistently yielded the same results, although the real-time assay was more sensitive and less time consuming. Phylogenetic analysis of the mtCOI DNA sequences corroborated the identity of the B and Q biotypes 100% of the time and by phylogenetic analysis the haplotypes grouped, as expected, in the major North African-Mediterranean-Middle Eastern clade of the B. tabaci complex.

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