Molecular Characterization of Aflatoxin Biosynthesis Genes of Aspergillus flavus from Peanuts Production Area

2019 ◽  
Author(s):  
I. Lavkor
Keyword(s):  
Fragment Length Polymorphism ◽  
Intergenic Spacer ◽  
Aflatoxin Contamination ◽  
Aflatoxin Biosynthesis ◽  
Fungal Isolates ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Production Area ◽  
Polymerase Chain

In this study, molecular analysis of (100%) all fungal isolates, which were sampled from soil and air besides from infected peanut plants in the peanut planting area, were identified in â-tubulin gene by Polymerase Chain Reaction (PCR). PCR products of fungal isolates were restricted by BglII enzyme within Restriction Fragment Length Polymorphism (RFLP). The intergenic spacer (IGS) region for aflatoxin biosynthesis genes (aflJ-aflR) were determined in 254 (78.2%) A. flavus isolates using PCR-RFLP. Selected 100 isolates were detected as A. flavus by â-tubulin sequence gene fragments and comparisons of sequence showed 96–100% similarity. 254 out of 325 isolates contained aflatoxin biosynthesis genes (aflJ-aflR), whereas 213 out of 254 isolates produced aflatoxin. The results acquired in study remarked that A. flavus was the species responsible for aflatoxin contamination. Aflatoxin gene cluster in populations can be advantage for comprehension of the toxicological risk as well as the election of biocontrol isolates.

scholarly journals Characterization of a Phytoplasma Associated with Cabbage Yellows in Iran

Plant Disease ◽  
2007 ◽  
Vol 91 (5) ◽  
pp. 625-630 ◽  
Author(s):  
M. Salehi ◽  
K. Izadpanah ◽  
M. Siampour
Keyword(s):  
Nested Pcr ◽  
Fragment Length Polymorphism ◽  
Phylogenetic Analyses ◽  
Fars Province ◽  
Beet Leafhopper ◽  
Circulifer Haematoceps ◽  
Pcr Products ◽  
Disease Agent ◽  
Polymerase Chain

In 2001, a disease tentatively named Iranian cabbage yellows (ICY) was observed in cabbage fields of Zarghan (Fars Province, Iran). The major symptoms of the disease were yellowing, little leaves, plant stunting, opening of the head, and proliferation of the buds at the base of the stem into a witches'-broom. Among leafhoppers collected in cabbage fields, only Circulifer haematoceps transmitted the ICY agent. The disease agent was transmitted by the leafhopper from cabbage to cabbage, cauliflower, rape, and periwinkle, causing phytoplasma-type symptoms in these plants. Polymerase chain reaction (PCR) using phytoplasma-specific primer pair P1/P7 and nested PCR using P1/P7 and R16F2n/R16R2 primer pairs amplified products of expected size (1.8 and 1.2 kb, respectively) from symptomatic cabbage plants. Both restriction fragment length polymorphism (RFLP) of nested PCR products (1.2 kb) and phylogenetic analyses of 16S–23S rDNA spacer region sequence indicated that the ICY phytoplasma had the closest relationship to subgroup A members of the clover proliferation group, including beet leafhopper-transmitted virescence agent, ‘Candidatus Phytoplasma trifolii’, Columbia Basin potato purple top phytoplasma, and vinca virescence phytoplasma. Cabbage is reported as a new natural host to the 16SrVI group of phytoplasmas.

Characterization of genetic polymorphism in the goat β-lactoglobulin gene

2000 ◽  
Vol 67 (2) ◽  
pp. 217-224 ◽  
Author(s):  
RAMONA N. PENA ◽  
ARMAND SÁNCHEZ ◽  
JOSEP M. FOLCH
Keyword(s):  
Fragment Length Polymorphism ◽  
Gene Frequencies ◽  
Single Nucleotide ◽  
Pcr Product ◽  
Polymorphic Position ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Β Lactoglobulin ◽  
New Alleles

Two new variants have been detected and characterized for the goat β-lactoglobulin gene at the cDNA level and confirmed at the genomic level. The two polymorphisms are located on exon 7 of the gene. One of the polymorphic sites is produced by a single nucleotide substitution in position +4601, allowing a polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) genotyping procedure to be developed using SacII restriction enzyme. The other polymorphic position contains a 10 bp long insertion at position +4641 that can be detected by capillary electrophoresis of the PCR product amplified with a fluorescent primer. The association of these two polymorphisms was also investigated, resulting in the description of two new alleles. Both of these contained the point mutation at the SacII site, with or without the 10 bp insertion at position +4641. The distribution of these new polymorphisms was studied in a population of males of four different goat breeds. The gene frequencies for these variants were similar in Spanish and French breeds.

Assessment of the genetic diversity ofFrankiamicrosymbionts ofElaeagnus angustifoliaL. plants growing in a Tunisian date-palm oasis by analysis of PCR amplifiednifD-Kintergenic spacer

2007 ◽  
Vol 53 (3) ◽  
pp. 440-445 ◽  
Author(s):  
Maher Gtari ◽  
Daniele Daffonchio ◽  
Abdellatif Boudabous
Keyword(s):  
Climatic Condition ◽  
Date Palm ◽  
Fragment Length Polymorphism ◽  
Intergenic Spacer ◽  
Climatic Conditions ◽  
Elaeagnus Angustifolia ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Diversity of Frankia microsymbionts of non-native Elaeagnus angustifolia L. plants spontaneously growing in a Tunisian desertic retreat area, the date-palm oasis of Tozeur, was investigated by polymerase chain reaction – restriction fragment length polymorphism (PCR–RFLP) and PCR-sequencing techniques targeting the nifD-K intergenic spacer. Three PCR–RFLP haplotypes (I, II, and III) were detected among collected nodules. Haplotype I was detected at all five sampling sites and dominated the other haplotypes present at these sites. This haplotype was also exhibited by strain BMG5.10, which was isolated by a plant-capturing assay in 1998 from soil collected in the same locality, qualifying it to be the most competitive haplotype in the edapho-climatic condition of the studied desertic date-palm oasis. nifD-K sequences of the three haplotypes formed a closely related phylogenetic subgroup. These results suggest that Frankia variability is constrained by severe edapho-climatic conditions of retreated desert in Tunisian area.

scholarly journals Survey of mutations in prolificacy genes in Santa Ines and Morada Nova sheep

2017 ◽  
Vol 69 (4) ◽  
pp. 1047-1053
Author(s):  
G.M.L. Holanda ◽  
J.C. Oliveira ◽  
D.M.F. Silva ◽  
S.S.N. Rocha ◽  
V. Pandolfi ◽  
...  
Keyword(s):  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
Nucleotide Sequencing ◽  
Chain Reaction ◽  
Specific Primers ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Santa Inês ◽  
Restriction Fragment Length

ABSTRACT Polymorphisms in the BMP-15 gene related to Galway (FecXG) and Inverdale (FecXI) and in the BMPR-1B gene known as Booroola (FecB) mutations were investigated using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) method, on sheep from the breeds Santa Inês (n= 574) and Morada Nova (n=282). DNA was extracted and amplified through PCR with specific primers that introduced a restriction site in association with the mutation. The PCR products were submitted to endonucleases. The experiment found no FecXG and FecXI mutations. Six samples of animals with multiple offspring/birth history presented polymorphism for FecB similar to control samples, but this pattern was not confirmed by nucleotide sequencing. Although the absence of these mutations in the studied breeds, other factors related to prolificacy should be investigated to explain the inherent prolificity mechanisms.

Application of riboprinting for the identification of isolates of cutaneousLeishmaniaspp.

Parasitology ◽  
2003 ◽  
Vol 127 (3) ◽  
pp. 201-205 ◽  
Author(s):  
L. F. NIMRI ◽  
H. D. F. H. SCHALLIG
Keyword(s):  
Fragment Length Polymorphism ◽  
Restriction Pattern ◽  
Chain Reaction ◽  
Restriction Patterns ◽  
Single Genome ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Selection Of ◽  
Restriction Fragment Length

Riboprinting is one of several molecular methods that can generate comparative data independently of the complexity of the organism's morphology. Restriction fragment length polymorphism (RFLP) profiles derived from digestion of polymerase chain reaction (PCR) products of the ribosomal 18S fromLeishmaniaspp. yields a typical ‘riboprint’ profile that can vary intraspecifically. A selection of 76 stocks ofL. majorandL. tropica, isolated from patients with cutaneous leishmaniasis, was analysed by riboprinting to assess divergence within and between species.L. majorandL. tropicacould be easily differentiated from each other. Analysis of PCR–RFLP profiles indicated that stocks ofLeishmaniaspp. could be broadly partitioned into 2 species corresponding toL. majorandL. tropica. To test if ribosomal 18S sequences were homogeneous within each species, several isolates of each of theLeishmaniaspp. were digested. Interpretation of the riboprint profiles of the 18S independently amplified by PCR, there would appear to be one restriction pattern present within eachLeishmaniaspp. Homogeneity within copies of the ribosomal 18S within a single genome has, therefore, been demonstrated. The species designation established by riboprinting results were in agreement with the zymodeme analysis of the same isolates. The restriction patterns produced were simple, reproducible and easy to interpret.

scholarly journals Characterization of Ascaris from Ecuador and Zanzibar

2014 ◽  
Vol 89 (4) ◽  
pp. 512-515 ◽  
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.

Genetic diversity analysis of diazotrophs in the rice rhizosphere

2007 ◽  
Vol 4 (3) ◽  
pp. 253-258 ◽  
Author(s):  
Chen Bin ◽  
Zheng Si-Ping ◽  
Zhou Li-Juan ◽  
Lin Zhi-Min ◽  
Song Ya-Na ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Fragment Length Polymorphism ◽  
Rhizospheric Soil ◽  
Chain Reaction ◽  
Rice Roots ◽  
Rice Rhizosphere ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Diazotrophic Community

SUMMARYThe genetic diversity of dinitrogen-fixing bacteria associated with rice (Oryza sativa) was assessed by a polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) approach on thenifHgene amplified directly from DNA extracted from washed rice roots and rhizospheric soil. Restriction digestion with the enzymesMnlI andHaeIII was performed to characterize 54 clonednifHPCR products. RFLP profiles were clustered and analysed with the UPGMA program. Eight pairs of similar RFLP patterns (similarity>50%) and two pairs of homologous RFLP patterns (100% identity) were found from the washed roots and the rhizospheric soil, respectively. Three specific diazotrophic patterns were found from rhizospheric soil and rice roots. The analyses have revealed the presence of differentnifHtypes, which appear to be significant components of the diazotrophic community in paddy fields, indicating that some of the diazotrophs may colonize the inside and the surface of the rice roots.

scholarly journals Polymorphism of alpha s1-casein gene in a dairy goat herd in the southeastern region of Brazil

2009 ◽  
Vol 38 (6) ◽  
pp. 1026-1032 ◽  
Author(s):  
Maria Amélia Menck Soares ◽  
Marcelo Teixeira Rodrigues ◽  
Giuliana Patrícia Mognol ◽  
Lucinéia de Fátima Chasko Ribeiro ◽  
José Luis da Conceição Silva ◽  
...  
Keyword(s):  
Fragment Length Polymorphism ◽  
Complete Characterization ◽  
Dairy Goat ◽  
Dairy Goats ◽  
Casein Gene ◽  
Pcr Rflp ◽  
Exon 9 ◽  
Polymerase Chain ◽  
Southeastern Region

Three different regions of the alpha s1-casein gene (CSN1S1) were investigated to determine the frequencies of major alleles for null, low, intermediate and high milk protein expression in a herd of dairy goats raised in the southeastern region of Brazil. Genomic DNA samples were obtained from leukocytes of 145 dairy goats and regions of interest in the gene were amplified through Polymerase Chain Reaction (PCR), then evaluated in both agarose (O and E allele) and polyacrylamide gels (F allele). For better characterization of the F allele, a PCR-RFLP (PCR-Restriction Fragment Length Polymorphism) study was performed employing the endonuclease XmnI. The allelic frequencies in the herd of 62 Saanen goats studied were: CSN1S1E = 0.35; CSN1S1F = 0.30; CSN1S1O1 = 0.02; CSN1S1A+B+C = 0.30, other alleles = 0.03. In another group of 83 Alpine animals, the frequencies were: CSN1S1E = 0.48; CSN1S1F = 0.28; CSN1S1O1 = 0.01; CSN1S1A+B+C = 0.20, other alleles = 0.03. In the region of exon 9 and intron downstream, where mutations that characterize the F allele occur, it was verified that different intragenic haplotypes may exist, involving the deletion of the 23rd nucleotide in the ninth exon in addition to the insertion of 11bp on intron. These haplotypes may be used to make direct association with other alleles. Although rare, a higher number of combinations were found in this work by evaluating in conjunction the region of the insertion of 3bp in the referred intron, which may allow a higher number of associations. A complete characterization of these combinations will allow elaborating simplified protocols to identify animals concerning the alleles of CSN1S1 gene in goats.

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