scholarly journals Optimization of heterologous microsatellites in Piracanjuba

2015 ◽  
Vol 50 (12) ◽  
pp. 1236-1239 ◽  
Author(s):  
Fausto Moreira da Silva Carmo ◽  
Érico Macedo Polo ◽  
Marcos Antônio da Silva ◽  
Gabriel de Menezes Yazbeck
Keyword(s):  
Microsatellite Markers ◽  
Product Formation ◽  
Pcr Product ◽  
First Time ◽  
Pcr Conditions

Abstract: The objective of this work was to evaluate 41 microsatellite markers for heterologous amplifications in piracanjuba (Brycon orbignyanus). Some markers were tested for the first time. Loci were optimized for PCR conditions and applied to a sample of 49 individuals. Thirty-one loci resulted in PCR product formation, whereas ten loci yielded intelligible polymorphic patterns in the evaluated sample and can be used for amplifications in this species. From the evaluated markers, four loci (BoM1, BoM13, Bh6, and Bh16) are valid to be applied in the study of piracanjuba.

A molecular diagnostic method for detecting Nacobbus in soil and in potato tubers

Nematology ◽  
2005 ◽  
Vol 7 (2) ◽  
pp. 193-202 ◽  
Author(s):  
Brian Kerry ◽  
Belkis Peteira ◽  
Simon Atkins ◽  
Javier Franco ◽  
Rosa Manzanilla-López
Keyword(s):  
Molecular Diagnostic ◽  
Potato Tubers ◽  
Molecular Approaches ◽  
Band Size ◽  
Pcr Product ◽  
Pcr Products ◽  
And Control ◽  
First Time ◽  
Pcr Conditions ◽  
Its Primers

AbstractSpecies of the genus Nacobbus have the potential to reduce yields of major food crops such as potato, sugar beet and tomato in many parts of the world, thus warranting a quarantine effort to avoid their introduction. Molecular studies offer a new method for routine quarantine diagnostics for this nematode that will be faster and more sensitive than previous methods. A primer set was designed from Nacobbus ITS sequences and their specificity confirmed. DNA was extracted from nematodes, soil and potato tubers for use in PCR. Optimised PCR conditions were established and the PCR products were separated on 2% agarose gels, showing that specific ITS primers for the detection of Nacobbus generated a single PCR product, although band size varied slightly between species and soil isolates. The product was generated from DNA extracted from all the Nacobbus samples but not from other nematodes tested (Pratylenchus, Radopholus, Meloidogyne, Globodera, Heterodera). No bands were generated from the uninfested control soil and control tuber DNA samples, thus demonstrating the specificity of the primers. For the first time, Nacobbus was detected in soil and tuber samples using molecular approaches. These results have important applications not only in analysing advisory samples but also in the screening of material for quarantine purposes.

Isolation, Characterization, and Mapping of Genomic Microsatellite Markers for the First Time in Sea-Island Cotton (Gossypium barbadense)

2009 ◽  
Vol 35 (6) ◽  
pp. 1013-1020 ◽  
Author(s):  
Pei-Pei ZHANG ◽  
Xia-Qing WANG ◽  
Yang YU ◽  
Yu YU ◽  
Zhong-Xu LIN ◽  
...  
Keyword(s):  
Microsatellite Markers ◽  
Gossypium Barbadense ◽  
First Time

Identifying the potential of pulsed LED irradiation in synthesis: copper-photocatalysed C–F functionalisation

2018 ◽  
Vol 54 (36) ◽  
pp. 4589-4592 ◽  
Author(s):  
Thomas P. Nicholls ◽  
Johnathon C. Robertson ◽  
Michael G. Gardiner ◽  
Alex C. Bissember
Keyword(s):  
Visible Light ◽  
Product Formation ◽  
Proof Of Concept ◽  
Concept Study ◽  
Catalysed Reaction ◽  
First Time ◽  
Led Irradiation

The results of a proof-of-concept study demonstrate for the first time that pulsed LED irradiation enhances the rate of product formation and the yield of a visible light-mediated photoredox-catalysed reaction.

scholarly journals First Report of Tobacco mild green mosaic virus in Capsicum chinense in Venezuela

Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1108-1108 ◽  
Author(s):  
C. Córdoba ◽  
A. García-Rández ◽  
N. Montaño ◽  
C. Jordá
Keyword(s):  
Mosaic Virus ◽  
Seed Transmission ◽  
Tomato Mosaic Virus ◽  
Capsicum Chinense ◽  
Rt Pcr ◽  
Plant Host ◽  
First Report ◽  
Coat Protein Region ◽  
Pcr Product ◽  
First Time

In July 2003, noticeable deformations of leaves were observed on a local variety of Capsicum chinense, also called ‘Aji dulce’, from a pepper plantation located in Venezuela, (Monagas State). ‘Aji dulce’ is a basic ingredient of the Venezuelan gastronomy with an estimated cultivated area of 2,000 ha. The seeds of this local pepper are obtained by the growers who reproduce and multiply their own seeds every year. Seeds of affected plants were sent to our laboratory, and a group of approximately 100 seeds was sown in a controlled greenhouse that belongs to the Polytechnic University of Valencia, Spain. Three months later, obvious curling and bubbling developed on the leaves of the plants. Extracts of symptomatic plants tested negative for Tomato mosaic virus (ToMV), Tobacco mosaic virus (TMV), Pepper mild mottle virus (PMMV), and Tobacco etch virus (TEV) by double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) with policlonal antibodies specific to each virus (Loewe Biochemica GMBH, Sauerlach, Germany; Phyto-Diagnostics, INRA, France). Total RNA was isolated from 0.5 g of original seed sent from Venezuela and from 25 samples of leaves of plants grown in the greenhouse with an RNeasy Plant Mini Kit (Qiagen Sciences, Germantown, Maryland). The RNA isolated was used in reverse transcription-polymerase chain reaction (RT-PCR) with specific primers for Tobacco mild green mosaic virus (TMGMV) (1) predicted to amplify a 530 bp of the coat protein region. From all samples, a RT-PCR product of the expected size was obtained and then sequenced. BLAST analysis of one sequence (GenBank Accession No. DQ460731) showed high levels of identity with TMGMV isolates, with more than 99% nucleotide identity with the DSMZ PV-112 isolate (GenBank Accession No. AJ429096). The symptomatology observed on pepper plants, the TMGMV RT-PCR assay, and the consensus of sequenced regions with TMGMV lead us to conclude that TMGMV was the causal agent of the diseased C. chinense plants. Although TMGMV has a wide plant host range occurring worldwide (1), to our knowledge, this is not only the first time TMGMV has been detected in Venezuela, but also the first report of TMGMV in C. chinense in Venezuela and the first reliable probe of the TMGMV seed transmission. Reference: (1) J. Cohen et al. Ann. Appl. Biol. 138:153, 2001.

scholarly journals Efficiency of Randomly Amplified Polymorphic DNA to Sequence Characterized Amplified Region Marker Conversion and their Comparative Polymerase Chain Reaction Sensitivity in Cucumber

1999 ◽  
Vol 124 (2) ◽  
pp. 128-135 ◽  
Author(s):  
Thomas Horejsi ◽  
Jodie M. Box ◽  
Jack E. Staub
Keyword(s):  
Rapd Markers ◽  
Scar Marker ◽  
Scar Markers ◽  
Sequence Characterized Amplified Region ◽  
Pcr Product ◽  
Rapd Pcr ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Template Dna ◽  
Pcr Conditions

The conversion of randomly amplified polymorphic DNA (RAPD) markers to sequence characterized amplified region (SCAR) markers, and the effects of differing polymerase chain reaction (PCR) conditions were studied in cucumber (Cucumis sativus L.). Attempts were made to clone and sequence 75 RAPD PCR products to produce SCAR primers (16 to 22 nucleotides) designed to amplify original RAPD PCR products. The influence of template DNA source, purity, and concentration, MgCl2 concentration, Taq polymerase source, and type of thermocycler upon RAPD and SCAR marker performance was evaluated. Conversion of RAPD to SCAR markers was not universally successful, and SCAR primers reacted differently to varying PCR conditions. Only 48 (64%) of 75 RAPD markers were successfully converted to SCAR markers and 11 (15%) of these reproduced the polymorphism observed with the original RAPD PCR product. Moreover, some SCAR primer pairs produced multiple polymorphic PCR products. The band intensity of SCAR markers were brighter (P = 0.05) than their corresponding RAPD markers with only one exception. The SCAR markers examined were less influenced (P = 0.05) by MgCl2 concentration than their corresponding RAPD markers. However, some SCAR markers were more sensitive to reaction impurities than their RAPD counterparts and SCAR markers tended to be less readily visualized (decrease in frequency of visible PCR product) with low concentrations (1 and 2 mm) of template DNA than their corresponding RAPD markers. Neither the source of Taq nor the type of thermocycler used affected the performance of SCAR and RAPD markers. These data suggest that although SCAR markers may demonstrate enhanced performance over the RAPD markers from which they are derived, careful consideration must be given to both the costs and potential benefits of SCAR marker development in cucumber.

scholarly journals Genetic relationship among indigenous sheep population of Bangladesh

2019 ◽  
Vol 48 (1) ◽  
pp. 17-22
Author(s):  
GK Deb ◽  
MP Choudhury ◽  
MA Kabir ◽  
MYA Khan ◽  
M Ershaduzzaman ◽  
...  
Keyword(s):  
Genetic Distance ◽  
Microsatellite Markers ◽  
River Basin ◽  
Genetic Relationships ◽  
Genetic Distances ◽  
Genetic Group ◽  
Sheep Population ◽  
Blood Samples ◽  
Indigenous Sheep ◽  
Pcr Product

The study was conducted to investigate the genetic relationships among indigenous sheep population of Bangladesh (Barind, Jamuna river basin, Coastal and Garole sheep) using microsatellite markers. A total of 96 blood samples were collected from adult sheep of Barind (24), Jamuna River Basin (24), Coastal (24), Garole(10) and available Chotanagpuri (10) sheep. Chotanagpuri sheep was used as an outgroup population. DNA was extracted from blood samples using QIAGEN DNA Mini extraction kit and was quantified using a nanodrop. FAO recommended 13 labeled microsatellite markers were used for polymerase chain reaction (PCR). PCR product was confirmed by 2% agarose gel electrophoresis and visualized by staining with ethidium bromide.The exact allele sizes in each primer were determined by GeneMaker V1.85 demo. Microsatellite tool kit and Dispan software package were used for calculation of allele frequency, number of alleles per locus, observed and expected heterozygosity and genetic distances (DA). The Dispan software was used to calculate inter-individual genetic distances. These distance values were used to construct an UPGMA tree. Results showed that average number of polymorphic alleles per locus varied from4 in HUJ616 to 12 in MAF70. Observed heterozygosity was also varied from 0.54±0.04 in Coastal to 0.63±0.03 in Barind sheep population. Genetic distance between Jamuna river basin and Barind was lowest (0.01) and between Garole and Costal was highest (0.17). Garoleand Chotonagpuri sheep has higher genetic distance from other three sheep populations. Phylogenetic dendogram showed that sheep of Jamuna river basin and barind were belonged to same genetic group. Whereas, coastal, garole and Nagpur sheeps were shown higher genetic distances from Jamuna river basin and coastal sheep. Considering findings of this study it may be concluded that the Barind and Jamuna river basin sheep belongs to a similar genetic group while, Garole and Coastal sheep are belonging to two distinct genetic groups. Bang. J. Anim. Sci. 2019. 48 (1): 17-22

Identification and authentication of commercial mi-iuy croaker (Miichthys miiuy) products by two PCR-based methods

2020 ◽  
Author(s):  
tae sun kang
Keyword(s):  
Economic Value ◽  
Cost Effective ◽  
Cross Reactivity ◽  
Duplex Pcr ◽  
Consensus Sequences ◽  
Pcr Product ◽  
Otolithes Ruber ◽  
Pcr Method ◽  
First Time ◽  
Miichthys Miiuy

Mi-iuy croaker (Miichthys miiuy) is one of the most important ingredients of Korean cuisine and thus has the highest economic value. However, the similar morphological traits among croaker fish belonging to family Sciaenidae are often exploited for seafood fraud. In this study, M. miiuy-specific primer set was designed and further improved by the development of a rapid and cost-effective duplex PCR method. The specificity of M. miiuy-specific duplex PCR was tested using 22 seafood species, and no cross-reactivity was observed. The sensitivity of the PCR assay was found to be 0.1 ng/µL. For the first time, labeling compliance of 43 commercial m-iuy croaker products was verified using both full DNA barcoding and M. miiuy-specific duplex PCR methods. For species identification, BOLDSYSTEMS and GenBank database were screened with the consensus sequences of each PCR product as a query. This identification result was further confirmed using the M. miiuy-specific duplex PCR method. The findings of this study revealed that principal species substituted were law croaker (Pseudotolithus senegallus, n=4), bigeye croaker (Micropogonias megalops, n=3), whitemouth croaker (Micropogonias furnieri, n=1), and tigertoothed croaker (Otolithes ruber, n=1). A significant percentage (21%) of mislabeling was present in commercial mi-iuy products sold on the South Korea market.

scholarly journals ITS-2 sequences-based identification of Trichogramma species in South America

2015 ◽  
Vol 75 (4) ◽  
pp. 974-982 ◽  
Author(s):  
R. P. Almeida ◽  
R. Stouthamer
Keyword(s):  
South America ◽  
Species Identification ◽  
Internal Transcribed Spacer ◽  
Restriction Analysis ◽  
Molecular Technique ◽  
Reliable Identification ◽  
Trichogramma Species ◽  
Dichotomous Key ◽  
Pcr Product ◽  
First Time

Abstract ITS2 (Internal transcribed spacer 2) sequences have been used in systematic studies and proved to be useful in providing a reliable identification of Trichogramma species. DNAr sequences ranged in size from 379 to 632 bp. In eleven T. pretiosum lines Wolbachia-induced parthenogenesis was found for the first time. These thelytokous lines were collected in Peru (9), Colombia (1) and USA (1). A dichotomous key for species identification was built based on the size of the ITS2 PCR product and restriction analysis using three endonucleases (EcoRI, MseI and MaeI). This molecular technique was successfully used to distinguish among seventeen native/introduced Trichogramma species collected in South America.

scholarly journals Characterization of Nuclear Microsatellite Markers for the Narrow Endemic Syringa josikaea Jacq. fil. ex Rchb.

2013 ◽  
Vol 41 (1) ◽  
pp. 301 ◽  
Author(s):  
Bertalan LENDVAY ◽  
Andrzej PEDRYC ◽  
Mária Höhn
Keyword(s):  
Microsatellite Markers ◽  
Ornamental Plants ◽  
Enriched Library ◽  
Microsatellite Primers ◽  
Genetic Lineages ◽  
Nuclear Microsatellite ◽  
Rare Endemic ◽  
Two Populations ◽  
Pcr Conditions

The species of the genus Syringa L. are among the most popular ornamental plants worldwide. One particular species, Syringa josikaea, a rare endemic of the Carpathian Mountains, is of great conservation interest. Although microsatellite markers may be useful for studying the genetic variability of varieties and populations, no microsatellites have previously been characterized for any species of the Syringa genus. Our aim was therefore to test the applicability of microsatellite primers developed for neighboring genera (Olea and Ligustrum) and to complement these with markers isolated and characterized for S. josikaea. Twelve primer pairs of Olea and Ligustrum were tested by optimizing PCR conditions and checking the variability in 40 samples of two populations of S. josikaea. Two of them proved to be easy to PCR amplify and variable at the same time. To develop new primers we constructed a microsatellite enriched library and sequenced 48 clones. 18 sequences contained microsatellite motifs, and three of the designed primer pairs presented high allele variability. The five primer pairs characterized for S. josikaea proved to be highly informative and sufficient to distinguish between individuals. These microsatellite primers are valuable tools to study genetic variation of native populations, genetic lineages of hybrids and cultivars of S. josikaea.

scholarly journals Channels of Product Formation and Excited Molecules Relaxation at Multiphoton Dissociation of Chlorodifluoromethane

1993 ◽  
Vol 13 (1) ◽  
pp. 29-41 ◽  
Author(s):  
V. A. Dimand ◽  
K. K. Maltzev ◽  
A. A. Nadeikin ◽  
A. I. Nikitin ◽  
A. M. Velichko ◽  
...  
Keyword(s):  
Product Formation ◽  
Vibrationally Excited ◽  
Fast Relaxation ◽  
Multiphoton Dissociation ◽  
Excited Molecules ◽  
First Time

C2F4, molecules have been for the first time directly proved to be generated in the reaction of vibrationally-excited molecules CF2HCl. The contribution of this reaction to CF2HCl multiphoton dissociation (MPD) products formation was shown to become predominant at the initial CF2HCl pressures about ten Torr. Also fast relaxation of the highly excited molecules CF2HCl on the product molecules C2F4 was discovered.

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