Sequence characterized amplified region markers tightly linked to the mating factors ofLentinula edodes

Genome ◽  
2004 ◽  
Vol 47 (1) ◽  
pp. 156-162 ◽  
Author(s):  
Ayako Tanaka ◽  
Kazuhiro Miyazaki ◽  
Haruki Murakami ◽  
Susumu Shiraishi
Keyword(s):  
Mating Type ◽  
Lentinula Edodes ◽  
Mating Types ◽  
Randomly Amplified Polymorphic Dna ◽  
Chain Reaction ◽  
Identification Procedure ◽  
Polymerase Chain Reaction Condition ◽  
Sequence Characterized Amplified Region ◽  
Polymerase Chain ◽  
Mating Factor

Detecting the mating types in shiitake, Lentinula edodes (Berk.) Pegler, is important for making progress in the breeding of this mushroom and determining the compatibility of the pair to cross. Shiitake is a tetrapolar fungus with two unlinking mating factors, A factor and B factor. We screened molecular markers linked to the mating factors using the randomly amplified polymorphic DNA (RAPD) method to develop the mating type identification procedure. Using 147 oligonucleotide primers, a total of 6 linkage markers for the shiitake mating factors, 4 markers for the A factor and 2 markers for the B factor, were discovered with a logarithm of the odds threshold of 3.0 for linkage. Two RAPDs that perfectly segregated with each mating factor among 72 basidiospore strains were detected. Both of these RAPDs were cloned and sequenced to convert them to the sequence characterized amplified region (SCAR) markers. Four primers, two sets of primers, were designed according to the internal sequences of two RAPDs tightly linking to the A factor or B factor. Consequently, we determined the polymerase chain reaction condition for multiplex analyses of these SCAR markers.Key words: Lentinula edodes, SCAR, diagnostic, mating type.

scholarly journals A PCR-based Assay for Distinguishing between A1 and A2 Mating Types of Phytophthora capsici

2017 ◽  
Vol 142 (4) ◽  
pp. 260-264
Author(s):  
Ping Li ◽  
Dong Liu ◽  
Min Guo ◽  
Yuemin Pan ◽  
Fangxin Chen ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Mating Type ◽  
Phytophthora Capsici ◽  
Mating Types ◽  
Sequence Repeat ◽  
Chain Reaction ◽  
Plant Parasite ◽  
Dna Fragment ◽  
Polymerase Chain ◽  
Microsatellite Diversity

Sexual reproduction in the plant parasite Phytophthora capsici Leonian requires the interaction of two distinct mating types, A1 and A2. Co-occurrence of these mating types can enhance the genetic diversity of P. capsici and alter its virulence or resistance characteristics. Using an intersimple sequence repeat (ISSR) screen of microsatellite diversity, we identified, cloned, and sequenced a novel 1121-base pair (bp) fragment specific to the A1 mating type of P. capsici. Primers Pcap-1 and Pcap-2 were designed from this DNA fragment to specifically detect the A1 mating type. Polymerase chain reaction (PCR) using these primers amplified an expected 997-bp fragment from known A1 mating types, but yielded a 508-bp fragment from known A2 mating types. This PCR-based assay could be adapted to accurately and rapidly detect the co-occurrence of A1 and A2 P. capsici mating types from field material.

scholarly journals Validation of Quantitative and Digital Polymerase Chain Reaction Assays Targeting the Mating Types of Phyllosticta citricarpa, the Causal Agent of Citrus Black Spot

2021 ◽  
Author(s):  
Hector Urbina ◽  
Taylor Smith ◽  
Callie Jones ◽  
Xiaoan Sun ◽  
John McVay ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Mating Type ◽  
Citrus Fruit ◽  
Black Spot ◽  
Dispersal Ability ◽  
Mating Types ◽  
Chain Reaction ◽  
Citrus Black Spot ◽  
Limited Dispersal ◽  
Polymerase Chain

Citrus black spot (CBS) is a disease caused by the ascomycetous fungus Phyllosticta (formerly Guignardia) citricarpa (Botryosphaeriales, Pezizomycotina) currently present in citrus groves in five counties in southwest Florida. Within Florida, P. citricarpa shows limited reproduction via asexual sporulation due to the presence of only one (MAT1-2-1) of the two required mating types for sexual reproduction. Here we present two novel polymerase chain reaction (PCR) assays standardized in quantitative (qPCR) and digital (dPCR) platforms to distinguish both mating types (MAT1-1-1 and MAT1-2-1) of P. citricarpa, to monitor for the potential introduction of the MAT1-1-1 mating type into Florida and a novel protocol for DNA extraction from asymptomatic leaves. During citrus harvesting season 2018−2019, fruit lesions, as well as asymptomatic leaves adjacent to symptomatic fruit and asymptomatic trees in CBS-infected groves were surveyed for P. citricarpa presence and mating types. Results support the presence of only MAT1‑2-1 mating type in Florida, after surveying more than 1,145 citrus fruit lesions. We also confirmed the limited dispersal ability of the asexual state of P. citricarpa in Florida in ten groves using the enhanced capabilities of the dPCR platform in the detection of P. citricarpa directly from asymptomatic leaves with low pathogen inoculum.

scholarly journals Molecular typing of bacteria of the genus Asaia in malaria vector Anopheles arabiensis Patton, 1905

2012 ◽  
Vol 44 (2) ◽  
pp. 7 ◽  
Author(s):  
S. Epis ◽  
M. Montagna ◽  
F. Comandatore ◽  
C. Damiani ◽  
A. Diabaté ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Internal Transcribed Spacer ◽  
Anopheles Arabiensis ◽  
Acetic Acid Bacterium ◽  
Sub Saharan Africa ◽  
Randomly Amplified Polymorphic Dna ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sub Saharan ◽  
Dna Pcr

The acetic acid bacterium <em>Asaia</em> spp. was successfully detected in <em>Anopheles arabiensis</em> Patton, 1905, one of the major vector of human malaria in Sub-Saharan Africa. A collection of 45 <em>Asaia</em> isolates in cellfree media was established from 20 individuals collected from the field in Burkina Faso. 16S rRNA universal polymerase chain reaction (PCR) and specific qPCR, for the detection of <em>Asaia</em> spp. were performed in order to reveal the presence of different bacterial taxa associated with this insect. The isolates were typed by internal transcribed spacer-PCR, BOX-PCR, and randomly amplified polymorphic DNA-PCR, proved the presence of different <em>Asaia</em> in <em>A. arabiensis</em>.

Genetic differentiation of Indian camel (Camelus dromedarius) breeds using random oligonucleotide primers

2006 ◽  
Vol 39 ◽  
pp. 77-88 ◽  
Author(s):  
S.C. Mehta ◽  
B.P. Mishra ◽  
M.S. Sahani
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetic Variability ◽  
Genetic Differentiation ◽  
Population Subdivision ◽  
Randomly Amplified Polymorphic Dna ◽  
Chain Reaction ◽  
Dna Profile ◽  
Oligonucleotide Primers ◽  
Polymerase Chain ◽  
Polymorphic Bands

SummaryThe camel population in India is facing a severe decline which demands that immediate steps are taken to ensure its conservation. Characterisation is an integral part of the conservation program. The Polymerase Chain Reaction-Randomly Amplified Polymorphic DNA profile of unrelated camels of the Bikaneri (29), Jaisalmeri (30) and Kachchhi (18) breeds were analyzed. Reproducible polymorphic bands with varying frequencies among the three breeds of camel were obtained with five oligonucleotide primers. A total of 75 bands were amplified, of which 27 (36%) were polymorphic. The probability of obtaining identical fingerprints was observed to be the lowest in primer GC-10 (5.7%) followed by OP-08 (8.7%), GT-10 (11.3%), G-2 (15.5%) and G-1 (80%). Breed informative bands were amplified. The maximum genetic variability was observed in the Bikaneri (0.80±0.05) followed by the Kachchhi (0.84±0.06) and the Jaisalmeri (0.87±0.05) breeds. The inter-breed genetic distance estimates indicated a closer relationship in the Bikaneri-Kachchhi camels, (0.075), followed by the Jaisalmeri-Kachchhi (0.106) and Bikaneri-Jaisalmeri (0.132) breeds. A similar genetic relationship was observed when the degree of population subdivision was measured between the Bikaneri-Kachchhi (0.529), Jaisalmeri-Kachchhi (0.558) and Bikaneri-Jaisalmeri (0.566) breeds.

Genetic diversity among microsporidian isolates from the silkworm, Bombyx mori, as revealed by randomly amplified polymorphic DNA (RAPD) markers

2011 ◽  
Vol 56 (4) ◽  
Author(s):  
B. Surendra Nath ◽  
W. Hassan ◽  
S. Nageswara Rao ◽  
N. Vijaya Prakash ◽  
S. Gupta ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Type Species ◽  
Rapd Markers ◽  
Randomly Amplified Polymorphic Dna ◽  
Chain Reaction ◽  
Rapd Pcr ◽  
Random Amplification ◽  
Major Cluster ◽  
Sources Of Infection ◽  
Polymerase Chain

AbstractRandom amplification of polymorphic DNA polymerase chain reaction (RAPD-PCR) was carried out to assess the genetic diversity of five new microsporidian isolates viz., NIWB-11bp, NIWB-12n, NIWB-13md, NIWB-14b and NIWB-15mb identified from the silkworms. A type species, NIK-1s_mys was used as control for comparison. Differences in the spore shape, length and width were observed. Of the 30 decamer random primers tested, 22 primers gave repeatable RAPD profiles and yielded a total of 143 fragments, of which 78 were polymorphic (55%). The resulting data was used to derive genetic similarity values for constructing a dendrogram. The neighbour joining method based on Dice coefficients indicate a major cluster comprising NIK-1s_mys, NIWB-11bp and NIWB-12n, whereas NIWB-13md, NIWB-14b and NIWB-15mb appear to be different from each other as well from the major cluster mentioned above which includes the type species (NIK-1s_mys). Based on the reproducibility of RAPD profiles, we are able to identify these microsporidians as different isolates. The RAPD technique may be useful in detecting sources of infection of this economically important domestic insect.

scholarly journals A Pseudomonas syringae Diversity Survey Reveals a Differentiated Phylotype of the Pathovar syringae Associated with the Mango Host and Mangotoxin Production

2013 ◽  
Vol 103 (11) ◽  
pp. 1115-1129 ◽  
Author(s):  
José A. Gutiérrez-Barranquero ◽  
Víctor J. Carrión ◽  
Jesús Murillo ◽  
Eva Arrebola ◽  
Dawn L. Arnold ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Randomly Amplified Polymorphic Dna ◽  
Chain Reaction ◽  
Catabolic Diversity ◽  
Rapd Pcr ◽  
Geographical Regions ◽  
Extensive Collection ◽  
Polymerase Chain ◽  
Apical Necrosis ◽  
Dna Pcr

Pseudomonas syringae pv. syringae, the causal agent of bacterial apical necrosis (BAN) in mango crops, has been isolated in different mango-producing areas worldwide. An extensive collection of 87 P. syringae pv. syringae strains isolated from mango trees affected by BAN from different countries, but mainly from Southern Spain, were initially examined by repetitive sequence-based polymerase chain reaction (rep-PCR) to analyze the genetic diversity with an epidemiological aim. rep-PCR was powerful in assessing intrapathovar distribution and also allowing clustering of the P. syringae pv. syringae strains isolated from mango, depending on the isolation area. A clear pattern of clustering was observed for all the P. syringae pv. syringae strains isolated from mango distinct from strains from other hosts, including strains for the same geographical regions as the mango isolates. For this reason, a representative group of 51 P. syringae pv. syringae strains isolated from mango and other hosts, as well as some P. syringae strains from other pathovars, were further characterized to determine their possible genetic, phenotypic, and phylogenetic relationships. Similar to the rep-PCR results, the randomly amplified polymorphic DNA PCR (RAPD-PCR) and catabolic diversity analysis using the Biolog GN2 profile grouped 90% of the mango isolates together in a unique cluster. Interestingly, the majority of P. syringae pv. syringae strains isolated from mango produced mangotoxin. The analysis of the phylogenetic distribution using the multilocus sequence typing analysis strongly supports the existence of a differentiated phylotype of the pathovar syringae mainly associated with the mango host and characterized by the mangotoxin production.

scholarly journals Mating Type Locus-Specific Polymerase Chain Reaction Markers for Differentiation of Pyrenophora teres f. teres and P. teres f. maculata, the Causal Agents of Barley Net Blotch

2010 ◽  
Vol 100 (12) ◽  
pp. 1298-1306 ◽  
Author(s):  
Shunwen Lu ◽  
Gregory J. Platz ◽  
Michael C. Edwards ◽  
Timothy L. Friesen
Keyword(s):  
Polymerase Chain Reaction ◽  
Mating Type ◽  
Pathogen Detection ◽  
Epidemiological Studies ◽  
Pyrenophora Teres ◽  
Nucleotide Polymorphisms ◽  
Net Blotch ◽  
Chain Reaction ◽  
Type Locus ◽  
Polymerase Chain

Fourteen single nucleotide polymorphisms (SNPs) were identified at the mating type (MAT) loci of Pyrenophora teres f. teres (Ptt), which causes net form (NF) net blotch, and P. teres f. maculata (Ptm), which causes spot form (SF) net blotch of barley. MAT-specific SNP primers were developed for polymerase chain reaction (PCR) and the two forms were differentiated by distinct PCR products: PttMAT1-1 (1,143 bp) and PttMAT1-2 (1,421 bp) for NF MAT1-1 and MAT1-2 isolates; PtmMAT1-1 (194 bp) and PtmMAT1-2 (939 bp) for SF MAT1-1 and MAT1-2 isolates, respectively. Specificity was validated using 37 NF and 17 SF isolates collected from different geographic regions. Both MAT1-1 and MAT1-2 SNP primers retained respective specificity when used in duplex PCR. No cross-reactions were observed with DNA from P. graminea, P. tritici-repentis, or other ascomycetes, or barley. Single or mixed infections of the two different forms were also differentiated. This study provides the first evidence that the limited SNPs at the MAT locus are sufficient for distinguishing closely related heterothallic ascomycetes at subspecies levels, thus allowing pathogenicity and mating type characteristics of the fungus to be determined simultaneously. Methods presented will facilitate pathogen detection, disease management, and epidemiological studies.

scholarly journals Phylogenetic Analysis of Culex tritaeniorhynchus and Culex vishnui Vector of Japanese Encephalitis Virus

2019 ◽  
Vol 7 (3) ◽  
Author(s):  
Raden Roro Upiek Ngesti Wibawaning Astuti ◽  
Raden Wisnu Nurcahyo ◽  
R.C. Hidayat Soesilohadi ◽  
Suwarno Hadisusanto ◽  
Budi Mulyaningsih
Keyword(s):  
Polymerase Chain Reaction ◽  
Japanese Encephalitis ◽  
Genetic Similarity ◽  
Pcr Amplification ◽  
Randomly Amplified Polymorphic Dna ◽  
Chain Reaction ◽  
Human Bite ◽  
Culex Tritaeniorhynchus ◽  
Polymerase Chain ◽  
Dna Pcr

Culex tritaeniorhynchus and Culex vishnui are medically essential mosquitoes that transmit the Japanese encephalitis (JE) virus. There is less information about the recording data and research due to genetic character differences among them. The objective of this study was to examine the genetic variation of Cx. tritaeniorhynchus and Cx. vishnui in 3 sites of Central Java using polymerase chain reaction randomly amplified polymorphic DNA (PCR-RAPD). The study was done in January to November 2017 in Pekalongan city, Pekalongan regency, and Semarang regency. Adult female mosquitoes collected by human bite method. DNA of ten Cx. tritaeniorhynchus samples and fifteen samples of Cx. vishnui purified using DNA extraction kit. Furthermore, PCR amplification was conducted with 5 RAPD primers (OPA 11, 12, 15, 16, and 20) and would run into 2% gel electrophoresis for 45 minutes. Cluster analysis was using MVSPTM software (version 3.1). The results showed 213 genetic characters of Cx. vishnui, while 142 characters shown by Cx. tritaeniorhynchus. The dendrograms showed three distinct groups of Cx. vishnui from 2 sites of Pekalongan and one site of Semarang, while Cx. tritaeniorhynchus showed two distinct groups, which were 1 group from Pekalongan and 1 group from Semarang. Low genetic similarity (<10%) shown Cx. vishnui from Pekalongan city and Pekalongan district, and there was no genetic similarity in Cx. tritaeniorhynchus from Pekalongan and Semarang. It concluded that the polymorphism of Cx. tritaeniorhynchus and Cx. vishnui reached 100%. ANALISIS FILOGENETIK CULEX TRITAENIORHYNCHUS DAN CULEX VISHNUI VEKTOR VIRUS JAPANESE ENCEPHALITISNyamuk Culex tritaeniorhynchus dan Culex vishnui memiliki peran penting di bidang medis terutama dalam penularan virus Japanese  encephalitis (JE). Sampai saat ini data dan riset tentang karakter genetik vektor JE masih sangat terbatas. Penelitian ini bertujuan menjelaskan variasi genetik Cx. tritaeniorhynchus dan Cx. vishnui di 3 lokasi di Jawa Tengah berdasar polymerase chain reaction randomly amplified polymorphic DNA (PCR-RAPD). Studi ini dilakukan dari bulan Januari sampai November 2017 di Kota Pekalongan, Kabupaten Pekalongan, dan Kabupaten Semarang. Metode human bite digunakan untuk koleksi nyamuk. Ekstraksi DNA nyamuk dilakukan pada 10 ekor Cx. tritaeniorhynchus dan 15 ekor Cx. vishnui menggunakan kit ekstraksi DNA. Selanjutnya, diamplifikasi dengan 5 macam primer RAPD (OPA 11, 12, 15, 16, dan 20), serta dielektroforesis pada 2% agar selama 45 menit. Analisis klaster dilakukan menggunakan program MVSPTM (versi 3.1). Ditemukan 213 dan 142 karakter genetik masing-masing pada Cx. vishnui dan Cx. tritaeniorhynchus. Analisis dendogram menunjukkan 3 grup yang berbeda untuk Cx. vishnui, sedangkan untuk Cx. tritaeniorhynchus terdapat 2 grup yang berbeda, yaitu 1 grup dari Pekalongan dan 1 grup dari Semarang. Similaritas genetik yang rendah (<10%) ditunjukkan Cx. vishnui dari Kota Pekalongan dan Kabupaten Pekalongan, bahkan tidak ada persamaan genetik pada Cx. tritaeniorhynchus dari Pekalongan dengan Semarang. Disimpulkan bahwa polimorfisme Cx. tritaeniorhynchus dan Cx. vishnui mencapai 100%.

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