Diversity of the desert truffle Terfezia boudieri Chatin. in southern Tunisia

This study reports the genetic diversity of Terfezia boudieri collected from southern Tunisia. The study was carried out using 135 truffle fruiting bodies harvested from seven different locations. Twenty-eight Terfezia claveryi fruiting bodies were also sampled from one of the seven locations. A PCR-based technique was used to amplify the internal transcribed spacer (ITS) region of the rDNA, including the ITS1–5.8S–ITS2. The PCR products were digested with the four restriction enzymes RsaI, HhaI, AluI, and HinfI. Based on the HinfI patterns, T. boudieri specimens were separated into two different haplotypes (I and II). Nucleotide sequences of some representative amplicons were also obtained. Based on the phylogenetic results, three T. boudieri genotypes could be differentiated. One sequence, SKtb1, retrieved from PCR–RFLP of haplotype I, was obtained from a low pH soil in association with Helianthemum kahiricum . Based on the results presented in the current study, this isolate may represent a novel taxa within the Terfezia genus.

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Genetic variation among pathogens causing "Helminthosporium" diseases of rice, maize and wheat

Fitopatologia Brasileira ◽

10.1590/s0100-41582002000600015 ◽

2002 ◽

Vol 27 (6) ◽

pp. 639-643 ◽

Cited By ~ 9

Author(s):

RITA C. B. WEIKERT-OLIVEIRA ◽

M. APARECIDA DE RESENDE ◽

HENRIQUE M. VALÉRIO ◽

RACHEL B. CALIGIORNE ◽

EDILSON PAIVA

Keyword(s):

Triticum Aestivum ◽

Genetic Variation ◽

Climatic Factors ◽

Rapd Analysis ◽

Its Region ◽

Restriction Enzymes ◽

Fungal Species ◽

High Similarity ◽

Pcr Rflp ◽

Upgma Phenogram

Twenty isolates of four fungal species, agents of "Helminthosporium" diseases in cereals, were collected from different regions: nine Bipolarisoryzae isolated from rice (Oryza sativa), seven B.sorokiniana from wheat (Triticum aestivum), two B. maydis, and two Exserohilumturcicum from maize (Zea mays). The strains were compared by PCR-RFLP and RAPD analysis. Size polymorphism among the isolates in the ITS region comprising the 5.8 S rDNA indicated genetic differences among the isolates, while a UPGMA phenogram constructed after the digestion of this region with restriction enzymes showed inter- and intra-specific polymorphism. The RAPD profiles indicated an expressive level of polymorphism among different species, compared with a low level of polymorphism among isolates of the same species. A UPGMA phenogram grouped the isolates according to the species and their host plant. RAPD profiles did not reveal polymorphism that directly correlated climatic factors with geographic source of the isolates of B. sorokiniana, and B. oryzae. Teleomorphic species revealed high similarity with their correspondent anamorphs.

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Identification of restriction enzyme in the FSHR gene of indonesian local cattle

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/888/1/012024 ◽

2021 ◽

Vol 888 (1) ◽

pp. 012024

Author(s):

P W Prihandini ◽

A Primasari ◽

M Luthfi ◽

D Pamungkas ◽

A P Z N L Sari ◽

...

Keyword(s):

Restriction Enzyme ◽

Restriction Site ◽

Follicle Stimulating Hormone ◽

Restriction Enzymes ◽

Future Studies ◽

Fshr Gene ◽

Pcr Rflp ◽

Mapping Analysis ◽

Pcr Products ◽

Enzyme Mapping

Abstract The restriction enzyme is important for genotyping using the PCR-RFLP technique. Therefore, this study aims to identify the restriction enzyme mapping in the partial sequence of the follicle-stimulating hormone receptor (FSHR) gene in Indonesian local cattle. A total of 29 samples sized 306 bp, were aligned with Genbank sequence acc no. NC_032660, resulting three polymorphic sites, namely g.193G>C, g.227T>C, and g.275A>C. Furthermore, the restriction mapping analysis using the NEBcutter program V2.0 showed that no enzyme recognized the SNP g.275A>C, while the SNP g.193G>C and g.227T>C were identified by the AluI and MscI enzymes, respectively. The AluI enzyme cuts at two positions (193 bp and 243 bp) in the G allele sample producing three fragments namely 50 bp, 63 bp, and 193 bp, meanwhile, in the C allele, the AluI cuts only in position 243 bp, hence, the fragment products are 63 bp and 243 bp. In contrast, the MscI enzyme was only recognized in the T allele, producing fragments sized 77 bp and 229 bp but failed to identify the restriction site along with the PCR products in the C allele. Based on the results, the SNPs (g.193G>C and g.227T>C) and restriction enzymes (AluI and MscI) are applicable for genotyping local Indonesian cattle using the PCR-RFLP technique in future studies.

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Detection of Theileria annulata by the PCR-RFLP in ticks (Acari, Ixodidae) collected from cattle in West and North-West Iran

Acta Parasitologica ◽

10.2478/s11686-011-0001-6 ◽

2011 ◽

Vol 56 (1) ◽

Cited By ~ 11

Author(s):

Mosa Tavassoli ◽

Mohammad Tabatabaei ◽

Bijan Nejad ◽

Mehran Tabatabaei ◽

Amin Najafabadi ◽

...

Keyword(s):

Restriction Enzymes ◽

Theileria Annulata ◽

Gene Encoding ◽

Hyalomma Anatolicum Anatolicum ◽

North West ◽

Blood Smears ◽

Pcr Rflp ◽

Single Pattern ◽

Pcr Products ◽

Bovine Theileriosis

AbstractThe presence of potential vectors, ticks, and susceptible hosts of bovine malignant theileriosis in all parts of Iran pose a real threat to food animal industry. The present study was conducted to determine the infection rate of ticks collected from naturally occurring bovine theileriosis in West and North-West Iran. Two hundred and thirty seven cattle suspected of suffering from theileriosis were investigated for the presence of Theileria annulata in the blood smears and any tick species on their body. In this study, 402 ticks were obtained from 99 cattle. The examination of 402 ticks by polymerase chain reaction (PCR) using primers derived from the gene encoding heat shock protein70 (Hsp70) revealed that 39.9% of Hyalomma anatolicum anatolicum, 3.5% of H. asiaticum asiaticum, and 18.2% H. anatolicum excavatum, were infected with T. annulata. The results suggest that H. a. anatolicum may play a major role in transmission of T. annulata infection in Iran. Finally, digestion of the PCR products of T. annulata with two different restriction enzymes produced only a single pattern.

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Determination of Types of Enterocytozoon bieneusiStrains Isolated from Patients with Intestinal Microsporidiosis

Journal of Clinical Microbiology ◽

10.1128/jcm.36.7.1882-1885.1998 ◽

1998 ◽

Vol 36 (7) ◽

pp. 1882-1885 ◽

Cited By ~ 40

Author(s):

Olivier Liguory ◽

Felicia David ◽

Claudine Sarfati ◽

Francis Derouin ◽

Jean-Michel Molina

Keyword(s):

Rflp Analysis ◽

Its Region ◽

Restriction Enzymes ◽

Epidemiological Studies ◽

Enterocytozoon Bieneusi ◽

Type I ◽

Pcr Rflp ◽

Intestinal Microsporidiosis ◽

Rflp Typing

To determine the types of Enterocytozoon bieneusistrains associated with intestinal microsporidiosis, we developed a rapid and efficient approach for typing parasites obtained from stool specimens by PCR-restriction fragment length polymorphism (PCR-RFLP). Typing was based on DNA polymorphism of the ribosomal DNA internal transcribed spacer (ITS) region of E. bieneusi. RFLPs generated with two restriction enzymes (NlaIII andFnu4HI) in PCR-amplified ITS products were used to classify strains into different lineages. This approach was successfully used to differentiate 78 strains that had been obtained from the stools of 65 patients with intestinal microsporidiosis. Among the 78 strains, we found four genetically unrelated lineages, showing the genetic diversity of E. bieneusi. Type I strains of E. bieneusi were found in a majority of the samples, accounting for 51 (78%) of the 65 microsporidiosis cases. In contrast, type II, III, and IV strains were found in only 8 (12%), 3 (5%), and 3 (5%) cases, respectively. All strains of E. bieneusi we have tested so far fall into one of four different lineages, and this study shows that human intestinal microsporidiosis is most often associated with type I strains. PCR-RFLP analysis of the ITS region of E. bieneusishould be useful for epidemiological studies.

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Molecular Profile of Trichophyton mentagrophytes and Microsporum canis based on PCR-RFLP of Internal Transcribed Spacer

Jurnal Ilmu Ternak dan Veteriner ◽

10.14334/jitv.v26i1.2546 ◽

2021 ◽

Vol 26 (1) ◽

pp. 10

Author(s):

Dwi Endrawati ◽

Eni Kusumaningtyas

Keyword(s):

Internal Transcribed Spacer ◽

Agar Plate ◽

Its Region ◽

Restriction Enzymes ◽

Trichophyton Mentagrophytes ◽

Molecular Profile ◽

Specific Marker ◽

Microsporum Canis ◽

Fungal Identification ◽

Pcr Rflp

Trichophyton mentagrophytes and Microsporum canis are dermatophytes fungi which commonly infect animal and human. Conventional and molecular methods were used for identification of the fungus. The region of internal transcribed spacer (ITS) has a high probability for fungal identification. PCR-RFLP was reported as a useful method to differentiate dermatophytes fungi. The objective of the study was to compare molecular profile of T. mentagrophytes and M. canis based on the result of ITS fragment digestion using Dde I, Hinf I and Mva I. The molds were isolated from skin scrapping of 18 animals which showed dermatophytosis lesion. The isolated molds were grown on agar plate for 14 days of incubation at 37oC and then identified based on macro and microscopic morphologies. Amplification of chitin synthase gene was used for confirmation and separation of dermatophytes from other fungi. ITS fragment was amplified and then digested using restriction enzymes Dde I, Hinf I and Mva I. The result showed that digestion products from ITS fragment of T. mentagrophytes and M. canis were different. The fragment 159 bp from Dde I, 374 bp from Hinf I and 89 bp from Mva I were present in T. mentagrophytes but absent in M. canis. Based on these results, specific RFLP profile of digestion ITS region by Dde I, Hinf I and Mva I can be used as a specific marker for species of dermatophytes fungi.

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Polymorphism of the exon 3 of leptin gene in Malpura sheep

Indian Journal of Animal Research ◽

10.18805/ijar.v0i0f.3783 ◽

2016 ◽

Author(s):

A.S. Meena ◽

A. Sahoo ◽

R. S. Bhatt ◽

A.S. Meena

Keyword(s):

Genetic Variants ◽

Genomic Dna ◽

Extraction Method ◽

Allelic Variation ◽

Restriction Enzymes ◽

Allelic Frequency ◽

Adipose Tissues ◽

Allelic Variants ◽

Pcr Rflp ◽

Pcr Products

Leptin (LEP) is primarily expressed in the adipose tissues. It regulates the feed intake, energy metabolism and body composition and plays a crucial role in regulating body weight and growth in mammals. The aim of the present study was to find out an allelic variation in leptin gene of Malpura sheep. A total of 112 Malpura sheep were selected and the genomic DNA was isolated by phenol-chloroform extraction method. PCR was carried out in order to amplify the 471 bp fragment of the exon 3 coding sequence of the leptin gene. The genotyping was done by PCR-RFLP technique. PCR products were digested with three (BcnI, SsiI and OliI) restriction enzymes. For detection of allelic variants, three non-synonymous SNPs were found in Malpura sheep. The A271G and A316C loci were found mono-morphic, while T387G locus was found polymorphic. Two genetic variants (G and T) and three genotypes (GG, GT and TT) were found in Malpura sheep. The allelic frequency of G and T allele at T387G locus was found 0.82 and 0.18, respectively.

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Characterization of Ascaris from Ecuador and Zanzibar

Journal of Helminthology ◽

10.1017/s0022149x14000431 ◽

2014 ◽

Vol 89 (4) ◽

pp. 512-515 ◽

Cited By ~ 5

Author(s):

A.M. Sparks ◽

M. Betson ◽

G. Oviedo ◽

C. Sandoval ◽

P.J. Cooper ◽

...

Keyword(s):

Fragment Length Polymorphism ◽

Rflp Analysis ◽

Its Region ◽

Restriction Enzymes ◽

Zoonotic Transmission ◽

Pcr Rflp ◽

Double Digestion ◽

Polymerase Chain ◽

Shed Light

AbstractTo shed light on the epidemiology of ascariasis in Ecuador and Zanzibar, 177 adult worms retrieved by chemo-expulsion from either people or pigs were collected, measured and subjected to polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the ribosomal internal transcribed spacer (ITS) region. Upon double digestion with RsaI and HaeIII, PCR-RFLP analysis revealed the presence of A. lumbricoides in people and A. suum in pigs in Ecuador. In contrast, while there are no pigs on Zanzibar, of the 56 worms obtained from people, one was genotyped as A. suum. No additional genetic variation was detected upon further PCR-RFLP analysis with several other restriction enzymes. Upon measurement, worm mass and length differed by location and by species, A. suum being lighter and longer. While there is no evidence to suggest zoonotic transmission in Ecuador, an enduring historical signature of previous zoonotic transmission remains on Zanzibar.

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Rapid and reliable identification of rice genomes by RFLP analysis of PCR-amplified Adh genes

Genome ◽

10.1139/g01-086 ◽

2001 ◽

Vol 44 (6) ◽

pp. 1136-1142 ◽

Cited By ~ 6

Author(s):

Song Ge ◽

Tao Sang ◽

Bao-rong Lu ◽

De-yuan Hong

Keyword(s):

Rflp Analysis ◽

Restriction Enzymes ◽

Pcr Primers ◽

Specific Pcr ◽

Adh Genes ◽

Pcr Rflp ◽

Restriction Sites ◽

Pcr Products ◽

Adh Gene ◽

Genus Oryza

The rice genus (Oryza L.) consists of 24 species with 10 recognized genome types. With the realization of many useful genes in species of wild rice, continuous efforts have been made to understand their genomic composition and relationships. However, the identification of rice genomes has often been difficult owing to complex morphological variation and formation of allotetraploids. Here we propose a rapid and reliable method for identifying rice genomes based on the restriction sites of PCR-amplified Adh genes. The experimental procedure was as follows: (i) amplify a portion of Adh1 and Adh2 genes with the locus-specific PCR primers; (ii) digest PCR products with restriction enzymes that distinguish different genomes; and (iii) run the digested products on 1.4% agarose gel, and photograph. Using various combinations of restriction digestion of the two Adh genes, all of the rice genomes can be identified.Key words: Adh gene, genome, identification, Oryza L., PCRRFLP.

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Identification and differentiation among chicken’s, duck’s, quail’s, rabbit’s and turkey's meat using PCR-RFLP technique

Biotechnology in Animal Husbandry ◽

10.2298/bah1501101a ◽

2015 ◽

Vol 31 (1) ◽

pp. 101-108 ◽

Cited By ~ 1

Author(s):

S.M. Abdel-Rahman ◽

A.M. Elmaghraby ◽

A.S. Haggag

Keyword(s):

Mitochondrial Dna ◽

Cytochrome B ◽

Fragment Length ◽

Restriction Analysis ◽

Restriction Enzymes ◽

Pcr Product ◽

Pcr Rflp ◽

Pcr Products ◽

Species Specific ◽

Rflp Assay

PCR-RFLP technique was developed for identification and differentiation among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat. DNA from small amount of muscles (0.05 g) was extracted and a region of mitochondrial DNA (cytochrome-b gene) in chicken, duck, quail, rabbit and turkey was amplified by PCR. Fragment length of the PCR product was 371 bp in chicken, 374 bp in duck and rabbit and 377 bp in both quail and turkey. Six nucleotides different makes it difficult to differentiate among these five species-specific meat. For differentiation, three different restriction enzymes (DdeI, MspI and TaqI) were used to digest the PCR products. Restriction analysis showed difference among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat. Where, DdeI yielded two fragments (291 and 83 bp) only in rabbit?s meat. MspI yielded three fragments (221, 85 and 65 bp) in chicken?s meat and two fragments (290 and 87 bp) in both quail?s and turkey's meat. TaqI yielded three fragments (146, 134 and 94 bp) in duck?s meat and two fragments (226 and 151 bp) in quail?s meat. The use of Cytb- PCR-RFLP assay allowed a direct and fast authentication and differentiation among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat.

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Genetic Homogenity of Commerson’s Anchovy (Stolephorus commersonnii) in Segara Anakan Cilacap Central Java Inferred from PCR-RFLP Markers

Biogenesis Jurnal Ilmiah Biologi ◽

10.24252/bio.v7i1.6352 ◽

2019 ◽

Vol 7 (1) ◽

pp. 14

Author(s):

Agus Nuryanto ◽

Rani Eva Dewi ◽

Hendro Pramono

Keyword(s):

Genetic Diversity ◽

Restriction Enzymes ◽

Coi Gene ◽

Survey Method ◽

Rflp Markers ◽

Small Pelagic Fish ◽

Central Java ◽

Low Genetic Diversity ◽

Pcr Rflp ◽

Pcr Products

Commerson’s anchovy (Stolephorus commersonnii) is a small pelagic fish that live in a group and its existence is very abundant in Segara Anakan Cilacap. This anchovy is widely consumed by communities live around Segara Anakan. This leads to a high exploitation rate. Exploited populations generally have low genetic diversity. This study aims to evaluate genetic diversity of commerson’s anchovy population in Segara Anakan Cilacap inferred from PCR-RFLP of the cytochrome c oxidase 1 (CO1) gene. This study was conducted from January to April 2018 and used survey method by applying random sampling. As many as 30 samples of anchovy were taken. Genomic mtDNA was isolated using modified Chelex method. Partial sequences of the COI gene were amplified using a pair forward commercially available primer. The lengths of 650 base pair of the PCR products were digested with four restriction enzymes. The HindIII enzyme produces PCR-RFLP fragment with the size of 416 bp and 234 bp lengths, VspI produces 435 bp and 214 bp, CO1-TaqI produces 556 bp and 94 bp and RsaI produces 319 bp, 183 bp, and 148 bp fragments, respectively. The PCR-RFLP fragments were obtained from all samples but they produced uniform band pattern for all 30 anchovy individuals. These results indicated that the anchovy population in Segara Anakan Cilacap has monomorphic allele for all PCR-RFLP markers. Hence, it can be concluded that genetic homogenity was observed on anchovy population in Segara Anakan Cilacap as inferred from PCR-RFLP COI gene.

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