SCAR molecular markers of the B biotype and two non-B populations of the whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae)

2006 ◽  
Vol 3 (3) ◽  
pp. 189-194 ◽  
Author(s):  
Zang Lian-Sheng ◽  
Jiang Tong ◽  
Xu Jing ◽  
Liu Shu-Sheng ◽  
Zhang You-Jun
Keyword(s):  
Bemisia Tabaci ◽  
Random Amplified Polymorphic Dna ◽  
Trialeurodes Vaporariorum ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Sequence Characterized Amplified Region ◽  
Scar Primer ◽  
Polymerase Chain ◽  
Scar Primers ◽  
B Biotype

AbstractRandom amplified polymorphic DNA (RAPD) analyses were performed with random primer H16 for the B biotype and two non-B populations of Bemisia tabaci collected from Zhejiang (China). The specific sequence fragments containing 446, 390 and 1317 nucleotides were amplified for the B biotype, ZHJ-1, ZHJ-2 populations, respectively. The three specific fragments were cloned and sequenced, and three pairs of SCAR primers were designed according to the sequences determined. With improvement of the conditions of the polymerase chain reaction (PCR), the specific fragments of B biotype, ZHJ-1 and ZHJ-2 populations, namely 439, 366 and 1238 nucleotides, respectively, were amplified with the sequence characterized amplified region (SCAR) primer of the corresponding population, while specific fragments of the other populations of B. tabaci or Trialeurodes vaporariorum could not be amplified.

scholarly journals Identification of the B, Q, and native Brazilian biotypes of the Bemisia tabaci species complex using Scar markers

2016 ◽  
Vol 51 (5) ◽  
pp. 555-562 ◽  
Author(s):  
Paulo Roberto Queiroz ◽  
Erica Soares Martins ◽  
Nazaré Klautau ◽  
Luzia Lima ◽  
Lilian Praça ◽  
...  
Keyword(s):  
Bemisia Tabaci ◽  
Species Complex ◽  
Scar Marker ◽  
Random Amplified Polymorphic Dna ◽  
Scar Markers ◽  
Sequence Characterized Amplified Region ◽  
Field Samples ◽  
Scar Primer ◽  
Q Biotype ◽  
B Biotype

Abstract: The objective of this work was to develop sequence-characterized amplified region (Scar) markers to identify the B, Q, and native Brazilian biotypes of the sweet potato whitefly [Bemisia tabaci (Hemiptera: Aleyrodidae)]. Random amplified polymorphic DNA (RAPD) amplification products, exclusive to the B and Brazilian biotypes, were selected after the analysis of 12,000 samples, in order to design a specific Scar primer set. The BT-B1 and BT-B3 Scar markers, used to detect the B biotype, produced PCR fragments of 850 and 582 bp, respectively. The BT-BR1 Scar marker, used to identify the Brazilian biotype, produced a PCR fragment of 700 bp. The Scar markers were tested against the Q biotype, and a flowchart was proposed to indicate the decision steps to use these primers, in order to correctly discriminate the biotypes. This procedure allowed to identify the biotypes that occur in field samples, such as the B biotype. The used set of primers allowed to discriminate the B, Q, and native Brazilian biotypes of B. tabaci. These primers can be successfully used to identify the B biotype of B. tabaci from field samples, showing only one specific biotype present in all cultures.

Identification of Meloidogyne spp. from North East Libya and comparison of their inter- and intra-specific genetic variation using RAPDs

Nematology ◽  
2005 ◽  
Vol 7 (4) ◽  
pp. 599-609 ◽  
Author(s):  
Mark Phillips ◽  
Mohamed Adam ◽  
Vivian Blok
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Egg Mass ◽  
Root Knot Nematode ◽  
Chain Reaction ◽  
Specific Sequence ◽  
North East ◽  
Geographic Origins ◽  
Identification Of Species ◽  
Meloidogyne Spp ◽  
Polymerase Chain

AbstractFour techniques, i.e., perineal patterns, isozymes, specific sequence characterised amplified region-polymerase chain reaction (SCAR-PCR) and random amplified polymorphic DNA (RAPD), were compared for the identification of species of root-knot nematode (RKN) from Libya. The RAPD technique proved superior for species diagnosis. A population from Massa that could not, because of atypical patterns, be identified using either perineal patterns or isozyme phenotypes, showed amplification patterns consistent with those of M. incognita. A new esterase phenotype is described for this population. Single egg mass lines from two other locations, Elhamma and Durnah, were identified as M. javanica, whereas those from Aun Mare were identified as M. incognita. RAPDs were used to examine the relationships between the Libyan isolates and isolates of Meloidogyne spp. from elsewhere. This technique revealed differences in the Libyan isolates of M. incognita that distinguished them from isolates from other geographic origins. The work demonstrates the potential to utilise the RAPD technique in an integrated programme to control RKN.

scholarly journals Characterization of Xanthomonas axonopodis pv. phaseoli isolates

2008 ◽  
Vol 34 (3) ◽  
pp. 228-231 ◽  
Author(s):  
Willian Mário de Carvalho Nunes ◽  
Maria Júlia Corazza ◽  
Silvana Aparecida Crestes Dias de Souza ◽  
Siu Mui Tsai ◽  
Eiko Eurya Kuramae
Keyword(s):  
Xanthomonas Campestris ◽  
Random Amplified Polymorphic Dna ◽  
Hypersensitive Reaction ◽  
Chain Reaction ◽  
Xanthomonas Axonopodis ◽  
Paulo State ◽  
Total Dna ◽  
Polymerase Chain ◽  
Bacterial Plant Pathogen

A simple, quick and easy protocol was standardized for extraction of total DNA of the bacteria Xanthomonas axonopodis pv. phaseoli. The DNA obtained by this method had high quality and the quantity was enough for the Random Amplified Polymorphic DNA (RAPD) reactions with random primers, and Polymerase Chain Reaction (PCR) with primers of the hypersensitivity and pathogenicity gene (hrp). The DNA obtained was free of contamination by proteins or carbohydrates. The ratio 260nm/380nm of the DNA extracted ranged from 1.7 to 1.8. The hrp gene cluster is required by bacterial plant pathogen to produce symptoms on susceptible hosts and hypersensitive reaction on resistant hosts. This gene has been found in different bacteria as well as in Xanthomonas campestris pv. vesicatoria (9). The primers RST21 and RST22 (9) were used to amplify the hrp gene of nine different isolates of Xanthomonas axonopodis pv. phaseoli from Botucatu, São Paulo State, Brazil, and one isolate, "Davis". PCR amplified products were obtained in all isolates pathogenic to beans.

scholarly journals Extração de DNA a partir de sangue humano coagulado para aplicação nas técnicas de genotipagem de antígenos leucocitários humanos e de receptores semelhantes à imunoglobulina

2009 ◽  
Vol 42 (6) ◽  
pp. 651-656 ◽  
Author(s):  
Daniela Maira Cardozo ◽  
Gláucia Andréia Guelsin ◽  
Samaia Laface Clementino ◽  
Fabiano Cavalcante de Melo ◽  
Marco Antônio Braga ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Killer Cell ◽  
Human Leukocyte ◽  
Reaction Sequence ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Salting Out ◽  
Leukocyte Antigens ◽  
Polymerase Chain ◽  
Made In

O objetivo deste estudo foi padronizar uma metodologia de extração de DNA de alta qualidade a partir de amostras de sangue coagulado. Quarenta e oito amostras de sangue humano coagulado foram utilizadas para a extração de DNA pelo kit comercial EZ-DNA® (Biological Industries, Beit Haemek, Israel), pelo kit de coluna Neoscience® (One Lambda Inc., San Diego, CA) e pelo método modificado de salting out. Apenas o método de salting out foi capaz de extrair altas concentrações de DNA (média, 180ng/µL), as quais foram medidas pelo detector de fluorescência Qubit® (Invitrogen, USA). Este método permitiu a amplificação dos genes HLA (human leukocyte antigens) pela tecnologia PCR-SSO (polymerase chain reaction - specific sequence of oligonucleotides) Luminex, a qual exige DNA de boa qualidade, e de genes KIR (killer cell immunoglobulin-like receptors) pela técnica made in house PCR-SSP (polymerase chain reaction-sequence specific of primers), a qual demanda uma concentração específica de DNA (10ng/µL). Concluímos que a técnica de salting out modificada foi muito eficiente, simples e rápida para a extração de DNA de amostras de sangue humano coagulado, com o objetivo de realizar a genotipagem de genes HLA e KIR.

scholarly journals Genetic variation between susceptible and non-susceptible snails to Schistosoma infection using random amplified polymorphic DNA analysis (RAPDs)

1999 ◽  
Vol 41 (5) ◽  
pp. 291-295 ◽  
Author(s):  
Abdel-Hamid Zaki ABDEL-HAMID ◽  
Jeanne Blanco de MOLFETTA ◽  
Vania FERNANDEZ ◽  
Vanderlei RODRIGUES
Keyword(s):  
Dna Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Polyacrylamide Gel Electrophoresis ◽  
Genome Comparison ◽  
Gene Products ◽  
Chain Reaction ◽  
High Molecular Weight Dna ◽  
Polymerase Chain ◽  
Or Gene ◽  
Host Parasite

Susceptibility of snails to infection by certain trematodes and their suitability as hosts for continued development has been a bewildering problem in host-parasite relationships. The present work emphasizes our interest in snail genetics to determine what genes or gene products are specifically responsible for susceptibility of snails to infection. High molecular weight DNA was extracted from both susceptible and non-susceptible snails within the same species Biomphalaria tenagophila. RAPD was undertaken to distinguish between the two types of snails. Random primers (10 mers) were used to amplify the extracted DNA by the polymerase chain reaction (PCR) followed by polyacrylamide gel electrophoresis (PAGE) and silver staining. The results suggest that RAPD represents an efficient means of genome comparison, since many molecular markers were detected as genetic variations between susceptible and non-susceptible snails.

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