RFLP analysis of rRNA intergenic spacer regions of 23 isolates of the entomopathogenPaecilomyces farinosus

1997 ◽  
Vol 75 (12) ◽  
pp. 2038-2044 ◽  
Author(s):  
J. S. K. Chew ◽  
D. B. Strongman ◽  
R. M. MacKay
Keyword(s):  
Ribosomal Rna ◽  
Population Biology ◽  
Genetic Relationships ◽  
Intergenic Spacer ◽  
Rflp Analysis ◽  
Restriction Enzyme Digestion ◽  
Paecilomyces Fumosoroseus ◽  
Chain Reaction ◽  
Ecological Requirements ◽  
Polymerase Chain

Genetic relationships between 23 eastern Canadian isolates of the entomopathogen Paecilomyces farinosus (Holm ex S.F. Gray) Brown & Smith were investigated by comparison of DNA fragments produced by restriction enzyme digestion of polymerase chain reaction amplified ribosomal RNA intergenic spacer regions. The variation observed was limited to 40% or less of these regions. All P. farinosus isolates were very dissimilar to isolates of the entomopathogens Beauveria bassiana (Bals.) Vuill. and Paecilomyces fumosoroseus (Wize) Brown & Smith. Seventeen P. farinosus isolates from six different hosts and diverse habitats yielded identical or nearly identical results. Two groups, each with three isolates from two different hosts, were distinct from the main group of isolates. Each of the three P. farinosus groups included some isolates that produced synnemata and some that did not, indicating multiple evolutionary losses of the ability to produce this sporulation structure. We conclude that eastern Canadian P. farinosus, while genetically and phenotypically variable, is not composed largely of strains with strict ecological requirements. Key words: entomopathogenic fungus, population biology, RFLP, ribosomal RNA intergenic spacer.

scholarly journals Differentiation of Moraxella Bovoculi sp. nov. from other Coccoid Moraxellae by the Use of Polymerase Chain Reaction and Restriction Endonuclease Analysis of Amplified DNA

2007 ◽  
Vol 19 (5) ◽  
pp. 532-534 ◽  
Author(s):  
John A. Angelos ◽  
Louise M. Ball
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequence ◽  
Restriction Analysis ◽  
Intergenic Spacer ◽  
Restriction Enzyme Digestion ◽  
Chain Reaction ◽  
Gram Negative ◽  
Intergenic Spacer Region ◽  
Polymerase Chain ◽  
Moraxella Bovoculi

Moraxella oris was historically the only coccoid Moraxella identified in cultures of ocular fluid from cattle with infectious bovine keratoconjunctivitis (IBK) and could be morphologically and biochemically differentiated from Moraxella bovis. Moraxella bovoculi sp. nov. is a recently characterized Moraxella isolated from ulcerated eyes of calves with IBK in northern California in 2002. Like Moraxella ovis, M. bovoculi sp. nov. is a gram-negative coccus/diplococcus. All 18 original isolates of M. bovoculi sp. nov. possessed phenylalanine deaminase (PADase) activity and could therefore be differentiated from M. ovis and M. bovis. During the characterization of 44 additional isolates of hemolytic gram-negative cocci that were cultured from ulcerated eyes of IBK-affected calves, 2 PADase-negative isolates were identified that could not be differentiated biochemically from M. ovis; however, the DNA sequence of the 16S-23S intergenic spacer region (ISR) of the isolates matched the 16S-23S ISR DNA sequence of M. bovoculi sp. nov. To facilitate the identification of PADase-negative moraxellae, a polymerase chain reaction (PCR) coupled with restriction enzyme digestion analysis of amplified DNA was developed. Amplification of the 16S-23S ISR followed by AfaI digestion of amplified DNA could differentiate M. bovoculi sp. nov. from M. ovis and other moraxellae. The DNA sequence analysis of the amplified 16S-23S ISR from the 42 PADase-positive isolates of hemolytic gram-negative cocci indicated that all were M. bovoculi sp. nov. and all possessed an AfaI site. A PCR coupled with restriction analysis of amplified DNA can aid in identifying M. bovoculi sp. nov.

scholarly journals Population Structure of Cercospora kikuchii, the Causal Agent of Cercospora Leaf Blight and Purple Seed Stain in Soybean

2008 ◽  
Vol 98 (7) ◽  
pp. 823-829 ◽  
Author(s):  
G. Cai ◽  
R. W. Schneider
Keyword(s):  
Genetic Relationships ◽  
Random Amplified Polymorphic Dna ◽  
Intergenic Spacer ◽  
Chain Reaction ◽  
Cercospora Kikuchii ◽  
Vegetative Compatibility Groups ◽  
Intergenic Spacer Region ◽  
Group I ◽  
Polymerase Chain ◽  
Purple Seed Stain

Random amplified polymorphic DNA (RAPD) and microsatellite-primed polymerase chain reaction (MP-PCR) were used to characterize 164 isolates of Cercospora kikuchii, most of which were collected from Louisiana. Plant tissue (seeds versus leaves), but not host cultivar, had a significant impact on pathogen population differentiation. Cluster analysis showed that the Louisiana population was dominated by a primary lineage (group I) with only a few Louisiana isolates belonging to the minor lineage that also included the non-Louisiana isolates (group II). A previous study showed that isolates could be differentiated according to vegetative compatibility groups (VCGs). However, RAPD and MP-PCR data demonstrated that isolates of C. kikuchii were not generally clustered according to these VCGs. Furthermore, genetic relationships within and between VCGs were examined using sequences of the intergenic spacer region of rDNA. These analyses showed that VCG is not an indicator of evolutionary lineage in this fungus. Our results suggest the likely existence of a cryptically functioning sexual stage in some portion of the C. kikuchii population.

An Assessment of Genetic Relationships between Members of the Phytophthora megasperma Complex and Phytophthora vignae using Molecular Markers

1993 ◽  
Vol 6 (4) ◽  
pp. 295 ◽  
Author(s):  
SC Whisson ◽  
BJ Howlett ◽  
ECY Liew ◽  
DJ Maclean ◽  
JM Manners ◽  
...  
Keyword(s):  
Nuclear Dna ◽  
Genetic Relationships ◽  
Random Amplified Polymorphic Dna ◽  
Intergenic Spacer ◽  
Rflp Analysis ◽  
Protein Patterns ◽  
Separate Species ◽  
Phytophthora Megasperma ◽  
Polymerase Chain ◽  
Mitochondrial Rflps

Genetic relationships between Phytophora megasperma f. sp. glycinea (Pmg) and morphologically similar taxa, P. megasperma f. sp. medicaginis (Pmm), P. megasperma f. sp. trifolii (Pmt), P. megasperma from Douglas Fir (PmDF) and asparagus (PmAS) and Phytophthora vignae, were explored by restriction fragment length polymorphism (RFLP) analysis of nuclear DNA using random genomic multi-copy, cDNA, and ribosomal DNA probes as well as random amplified polymorphic DNA (RAPDs) and RFLP analysis of ribosomal intergenic spacer regions amplified by the polymerase chain reaction (PCR). Each method detected large differences between these taxa and P. megasperma f. sp. glycinea. P. vignae was more closely related to P. megasperma f. sp. glycinea than the other taxa on the basis of the cDNA RFLPs and RFLPs of PCR amplified rDNA intergenic spacer regions. We conclude that each of the taxa examined represent separate species. This supports the most recent reclassification based on mitochondrial RFLPs and electrophoretic protein patterns of the host-specific taxa to P. sojae (Pmg), P. trifolii (Pmt) and P. medicaginis (Pmm).

Ecotypic variation of Gremmeniella abietina in northern Europe: disease patterns reflected by DNA variation

1995 ◽  
Vol 73 (10) ◽  
pp. 1531-1539 ◽  
Author(s):  
M. Hellgren ◽  
N. Högberg
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetic Variation ◽  
Ribosomal Dna ◽  
Intergenic Spacer ◽  
Principal Component ◽  
Nuclear Ribosomal Dna ◽  
Restriction Enzyme Digestion ◽  
Chain Reaction ◽  
Northern Fennoscandia ◽  
Polymerase Chain

Genetic variation in Gremmeniella abietina isolated from Pinus sylvestris, Pinus contorta, and Picea abies in southern and northern Fennoscandia was studied with arbitrary primed polymerase chain reaction. Fennoscandian G. abietina isolates were clearly separated into two ecotypically distinct groups based on their amplified banding patterns. Analysis of variance based on amplified fragments, AMOVA, and principal component analysis confirmed the separation of the isolates into the two groups. One group contained isolates associated with a disease syndrome affecting young trees covered by deep snow during winter in northern Fennoscandia. The second group of isolates was found on trees between 15 and 40 years old, scattered throughout the crowns. It occurs throughout Fennoscandia but is most frequent in the southern parts. No size polymorphism was found in fragments resulting after restriction enzyme digestion of internal transcribed spacer and intergenic spacer regions of nuclear ribosomal DNA. An estimate of gene flow between populations calculated based on amplified band frequencies, FST, indicated that there was restricted genetic exchange between populations of the two groups of isolates. Key words: Gremmeniella abietina, arbitrary primed polymerase chain reaction, genetic variation, ecotypes, ribosomal DNA.

scholarly journals Clinical Evaluation of a 16S Ribosomal RNA Polymerase Chain Reaction Test for the Diagnosis of Lymph Node Tuberculosis

2006 ◽  
Vol 43 (7) ◽  
pp. 855-859 ◽  
Author(s):  
Fernando Osores ◽  
Oscar Nolasco ◽  
Kristien Verdonck ◽  
Jorge Arevalo ◽  
Juan Carlos Ferrufino ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Lymph Node ◽  
Rna Polymerase ◽  
Ribosomal Rna ◽  
Polymerase Chain Reaction Test ◽  
Chain Reaction ◽  
Reaction Test ◽  
Polymerase Chain ◽  
Lymph Node Tuberculosis ◽  
Chain Reaction Test

scholarly journals Detection of Periprosthetic Infections With Use of Ribosomal RNA-Based Polymerase Chain Reaction

2010 ◽  
Vol 92 (3) ◽  
pp. 654-663 ◽  
Author(s):  
Patrick F Bergin ◽  
Jason D Doppelt ◽  
William G Hamilton ◽  
Gudrun E Mirick ◽  
Angela E Jones ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Ribosomal Rna ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Periprosthetic Infections

scholarly journals Molecular Characterization and Classification of Phytoplasmas Associated with Canola Yellows and a New Phytoplasma Strain Associated with Dandelions

Plant Disease ◽  
2001 ◽  
Vol 85 (1) ◽  
pp. 76-79 ◽  
Author(s):  
Keri Wang ◽  
Chuji Hiruki
Keyword(s):  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Universal Primers ◽  
Dna Fragments ◽  
Chain Reaction ◽  
Aster Yellows ◽  
Distinct Subgroup ◽  
16S Ribosomal Dna ◽  
Polymerase Chain

DNA isolated from symptomatic canola (Brassica napus, Brassica rapa) and dandelion (Taraxacum officinale) was used to amplify 16S ribosomal DNA fragments by polymerase chain reaction using two pairs of universal primers P1/P6 and R16F2n/R2. Restriction fragment length polymorphism (RFLP) analysis of the amplified DNA fragments using endonucleases AluI, HhaI, HpaII, MseI, RsaI, and Sau 3AI revealed two distinct types of phytoplasmas in canola with similar symptoms. One had the same RFLP profiles as the phytoplasmas in subgroup 16SrI-A, whereas the other one had RFLP profiles similar to those of phytoplasmas in subgroup 16SrI-B. Phytoplasmas were detected in symptomatic dandelion plants that were collected from canola and alfalfa fields where severe alfalfa witches'-broom occurred. Comparative studies indicated that two different phytoplasmas were associated with the dandelion plants. One was identified as a member of subgroup 16SrI-A, whereas another one was classified as a member of a distinct subgroup in the aster yellows group on the basis of the unique RFLP patterns.

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