scholarly journals Rhizoctonia spp. Recovered from Strawberry Roots in Central Coastal California

2000 ◽  
Vol 90 (4) ◽  
pp. 345-353 ◽  
Author(s):  
Frank N. Martin
Keyword(s):  
Polymerase Chain Reaction ◽  
Fragment Length Polymorphism ◽  
Coastal Region ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Molecular Techniques ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Collection Sites ◽  
Restriction Fragment Length

Rhizoctonia spp. were commonly recovered from the roots of strawberry plants growing in nonfumigated soil in the central coastal region of California. With the exception of one multinucleate isolate of R. solani (frequency of recovery of 0.8%), all other isolates were binucleate and were in anastomosis groups (AG) A, G, or I. AGs-A and -I were recovered from all five collection sites, whereas AG-G was recovered from only two sites. AG-A was the most commonly isolated AG, followed by AGs-I and -G. Similar levels of virulence were observed among the different AGs, but differences in virulence were observed among isolates in the same AG. Evaluating anastomosis grouping by pairing isolates recovered from strawberry with known tester isolates did not always yield a positive anastomosis reaction, even though both isolates anastomosed with other members of the same AG. Subsequent investigations with multiple isolates in the same AG from the same collection location confirmed that there was a lack of anastomosis or weak anastomosis reactions for some combinations of pairings, highlighting the need for to use multiple tester isolates or molecular techniques for AG determination. Restriction fragment length polymorphism (RFLP) analysis of a polymerase chain reaction-amplified region of the rDNA was effective for differentiating AGs. Sixteen RFLP groups were observed after cluster analysis with data for the size of the amplified products and fragment sizes after digestion with four restriction enzymes. Although each AG had isolates in multiple RFLP groups, any one individual RFLP group contained isolates of only a single AG. There was no consistent correlation between RFLP group and location of isolate collection.

Genetic analysis of nuclear DNA restriction fragment patterns

Genome ◽  
1991 ◽  
Vol 34 (5) ◽  
pp. 693-703 ◽  
Author(s):  
Elizabeth M. Gillet
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Genetic Analysis ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Restriction fragment length polymorphism (RFLP) analysis in the broad sense is the analysis of differences in restriction fragment pattern produced by defined target segments within or between cell compartments, cell types, etc., in a single individual or in different individuals. Thus both molecular hybridization and DNA amplification by two-primer extension using the polymerase chain reaction can define target segments for RFLP analysis. The two techniques are outlined with special consideration of characteristics important for genetic analysis. The mode of inheritance of restriction fragment patterns as a prerequisite for their use as genetic markers in inheritance studies is explained, leading to criticism of common usage. The importance of internal restriction sites for the determination of allelic variation is stressed. It is shown that, if target segments are under the control of a single nuclear diploid restriction fragment locus, then complete reconstruction of all parental target segments requires controlled crosses between individuals of like restriction fragment pattern.Key words: genetic analysis, inheritance, restriction fragment length polymorphism, controlled cross, polymerase chain reaction.

scholarly journals Identitas dan Keragaman Genetik Begomovirus yang Berasosiasi dengan Penyakit Keriting pada Tomat Berdasarkan Teknik Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP)

2016 ◽  
Vol 4 (1) ◽  
pp. 9 ◽  
Author(s):  
Tri Joko Santoso ◽  
Sri H. Hidayat ◽  
M. Herman ◽  
H. Aswidinnoor ◽  
Sudarsono Sudarsono
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Restriction Enzymes ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Leaf Curl Virus ◽  
Restriction Fragment Length

<p>Begomoviruses, members of the Geminivirus,<br />are considered as emerging plant viruses. This was due to<br />the increasing incidences and severities of the diseases in a<br />number of economically important crops, including tomato.<br />Genetic diversities of the Begomovirus isolates infecting<br />tomato (Lycopersicon esculentum) of several areas in Indonesia<br />were analyzed by using Polymerase Chain Reaction-<br />Restriction Fragment Length Polymorphism (PCR-RFLP)<br />technique. A 1500 base pairs of PCR fragments amplified by<br />using degenerate primers for Begomovirus was digested<br />using four restriction enzymes, i.e., DraI, EcoRI, RsaI, and<br />PstI. The pattern of RE digested fragments of 8 Begomovirus<br />isolates and the predicted RFLP fragments of the Begomovirus<br />isolates in the GeneBank database were used to determine<br />the genetic identities and diversities among the isolates.<br />Positive results of the PCR amplifications proved that<br />diseased tomato plant samples collected from 8 locations in<br />Java and Sumatra were infected with at least one Begomovirus<br />isolate. The PCR amplification products, which were<br />digested using the four restriction enzymes indicated the<br />presence of polimorfisms among the DNA fragments of the<br />Begomovirus isolates. Identifications of the Begomovirus<br />indicated that the Brastagi, Bogor, Sragen, Ketep, and Boyolali<br />isolates were Tomato Leaf Curl Virus (ToLCV); the<br />isolates from Malang and Blitar isolates were Ageratum<br />Yellow Vein Virus (AYVV), while one isolate from Kaliurang<br />was Tomato Yellow Leaf Curl Virus (TYLCV). Results of the<br />phylogenetic analysis of the 8 Begomovirus isolates based<br />on Begomoviruses from the DNA database indicated that<br />they belonged to three different groups.</p>

Limitations on the use of polymerase chain reaction – restriction fragment length polymorphism analysis of the rDNA NTS2 region for the taxonomic classification of the speciesSaccharomyces cerevisiae

2005 ◽  
Vol 51 (9) ◽  
pp. 759-764 ◽  
Author(s):  
Andrea Pulvirenti ◽  
Lisa Solieri ◽  
Luciana De Vero ◽  
Paolo Giudici
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Polymerase Chain Reaction ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Chain Reaction ◽  
D2 Domain ◽  
Pcr Rflp ◽  
Its Regions ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Different molecular techniques were tested to determine which was the most effective in the identification of Saccharomyces cerevisiae strains. In particular, polymerase chain reaction – restriction fragment length polymorphism (PCR–RFLP) analysis of the internal transcribed spacer (ITS) regions and the nontranscribed spacer 2 (NTS2) region, sequencing of the D1/D2 domain, and electrophoretic karyotyping were applied to 123 yeast strains isolated from different sourdoughs and tentatively attributed to the species S. cerevisiae. All of the strains tested showed an identical PCR–RFLP pattern for the ITS regions, an identical nucleotide sequence of the D1/D2 domain, and the typical electrophoretic karyo type of S. cerevisiae. In contrast, 14 out of the 123 strains tested showed some polymorphism with BanI restriction analysis of the NTS2 region. Our results indicate that while the sequencing of the D1/D2 domain, the PCR–RFLP analysis of the ITS regions, and the electrophoretic karyotype can be employed successfully to identify S. cere visiae strains, PCR–RFLP analysis of the NTS2 region does not allow a consistent and accurate grouping for S. cere visiae strains. The fact that the NTS2 region of a small number of strains (8.78% of the total strains tested) is different from that of the other S. cerevisiae strains confirms that molecular methods should always be tested on a great number of strains.Key words: ribosomal DNA, Saccharomyces cerevisiae, yeast identification.

Genetic variation detectable with molecular markers among unadapted germ-plasm resources of cultivated peanut and related wild species

Genome ◽  
1991 ◽  
Vol 34 (6) ◽  
pp. 1013-1020 ◽  
Author(s):  
Tracy M. Halward ◽  
H. Thomas Stalker ◽  
Elizabeth A. Larue ◽  
Gary Kochert
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Wild Species ◽  
Germ Plasm ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Chain Reaction ◽  
Cultivated Peanut ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Peanut germ plasm consists of the cultivated allotetraploid species Arachis hypogaea L. and a large number of wild species, which are nearly all diploids. Our previous work indicated a very low level of genetic variability in American cultivars, as assayed by restriction fragment length polymorphism (RFLP) analysis. Since American cultivars might represent a narrow genetic base, we expanded our study to include unadapted germ-plasm lines from the various South American centers of origin, Africa, and China, where considerable morphological and physiological variability has been reported to exist. Wild species of section Arachis were included in the evaluations since they show a high degree of variation when assayed by RFLPs. Three methods were used to assay for RFLP variation: (i) conventional RFLP analysis using random genomic clones from peanut and cDNA clones from peanut and alfalfa (Medicago sativa); (ii) polymerase chain reaction (PCR) amplification of random primer sequences; (iii) four-cutter analysis of PCR-amplified fragments. In all cases a very low level of variability was found in cultivated peanut, while abundant variability was present among wild diploid species. The results are discussed in terms of peanut evolution and significance to peanut breeding.Key words: polymerase chain reaction, Arachis hypogaea, restriction fragment length polymorphism.

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