Use of Randomly Amplified Polymorphic Dna Markers as a Tool to Study Variation in Lichen-Forming Fungi

1999 ◽  
Vol 31 (3) ◽  
pp. 257-267 ◽  
Author(s):  
G. J. Murtagh ◽  
P. S. Dyer ◽  
P. C. McClure ◽  
P. D. Crittenden
Keyword(s):  
Dna Polymerase ◽  
Dna Markers ◽  
Rapd Markers ◽  
Rapd Analysis ◽  
Pcr Amplification ◽  
Randomly Amplified Polymorphic Dna ◽  
Banding Patterns ◽  
Rapd Pcr ◽  
Key Features ◽  
Axenic Cultures

AbstractA protocol is described to enable the production of reliable genetic fingerprints of lichen-forming fungi using randomly amplified polymorphic DNA (RAPD) markers. Key features of the method are the use of mycobiont DNA extracted from axenic cultures by a phenol-chloroform procedure, and PCR amplification using DyNAzyme II DNA polymerase. RAPD-PCR fingerprints of Graphis scripta, G. elegans and Phacographis dendritica were successfully generated using this protocol and individual isolates could be identified on the basis of differences in banding patterns produced. DNA extracted from whole thalli of G. scripia was also subjected to RAPD-PCR but the fingerprints produced differed from those given by axenic cultures of the mycobiont. Therefore difficulties of interpretation may arise when whole thalli are used in RAPD analysis.

scholarly journals Production of Reliable Randomly Amplified Polymorphic DNA (RAPD) Markers from DNA of Woody Plants

HortScience ◽  
1993 ◽  
Vol 28 (12) ◽  
pp. 1188-1190 ◽  
Author(s):  
Amnon Levi ◽  
Lisa J. Rowland ◽  
John S. Hartung
Keyword(s):  
Woody Plants ◽  
Rapd Markers ◽  
Woody Plant ◽  
Rapd Analysis ◽  
Pcr Amplification ◽  
Intergenic Spacer ◽  
23S Rrna ◽  
Randomly Amplified Polymorphic Dna ◽  
Polymerase Chain ◽  
Triton X

A procedure for identifying reproducible RAPD markers from woody plant DNA is presented. The procedure relies on using a PCR buffer that contains 1% Triton-X-100 and 0.1 % gelatin [previously described for successful polymerase chain reaction (PCR) amplification of 16S/23S rRNA intergenic spacer regions from eubacteria], and amplification conditions of 50 cycles: 30 sec at 94C, 70 sec at 48C, and 120 sec at 72C. The combination of this buffer and these conditions amplified consistent fragments in higher amounts, as compared to other standard PCR buffers and conditions generally used for RAPD analysis. This procedure resulted in reliable RAPD patterns for all organisms tested. Chemical name used: α-[4-(1,1,3,3,-tetramethylbutyl)phenyl]-cohydroxypoly(oxy-l,2-ethanediyl) (Triton-X-l00).

scholarly journals Characterization of Alstroemeria Species using Random Amplified Polymorphic DNA (RAPD) Analysis

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 482F-482 ◽  
Author(s):  
Deric D. Picton ◽  
Harrison G. Hughes
Keyword(s):  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Randomly Amplified Polymorphic Dna ◽  
Banding Patterns ◽  
Extraction Buffer ◽  
Rapd Pcr ◽  
High Degree ◽  
Species Hybrids ◽  
Color Variants

In this study, 11 species, hybrids, and color variants were characterized using randomly amplified polymorphic DNA (RAPD) analysis. Total genomic DNA was extracted using a 2% CTAB extraction buffer using fresh or frozen leaf material. The DNA was amplified using standard RAPD-PCR protocols utilizing 10-mer primers. All primers utilized exhibited a high degree of polymorphism in their banding patterns among the species and hybrids studied. The primers used produced ≈40 reproducible bands. It was possible to identify and uniquely distinguish all species and hybrids investigated using these bands.

PCR-Based Genotyping of Epidemic and PreepidemicTrichoderma Isolates Associated with Green Mold ofAgaricus bisporus

1999 ◽  
Vol 65 (6) ◽  
pp. 2674-2678 ◽  
Author(s):  
X. Chen ◽  
C. P. Romaine ◽  
Q. Tan ◽  
B. Schlagnhaufer ◽  
M. D. Ospina-Giraldo ◽  
...  
Keyword(s):  
Genetic Variation ◽  
North America ◽  
Agaricus Bisporus ◽  
Rapd Markers ◽  
Rapd Analysis ◽  
Pcr Amplification ◽  
Green Mold ◽  
Rapd Pcr ◽  
Cultivated Mushroom ◽  
Highly Virulent

ABSTRACT We used randomly amplified polymorphic DNA (RAPD)-PCR to estimate genetic variation among isolates of Trichoderma associated with green mold on the cultivated mushroom Agaricus bisporus. Of 83 isolates examined, 66 were sampled during the recent green mold epidemic, while the remaining 17 isolates were collected just prior to the epidemic and date back to the 1950s.Trichoderma harzianum biotype 4 was identified by RAPD analysis as the cause of almost 90% of the epidemic-related episodes of green mold occurring in the major commercial mushroom-growing region in North America. Biotype 4 was more closely allied to T. harzianum biotype 2, the predominant pathogenic genotype in Europe, than to the less pathogenic biotype 1 and Trichoderma atroviride (formerly T. harzianum biotype 3). No variation in the RAPD patterns was observed among the isolates within biotype 2 or 4, suggesting that the two pathogenic biotypes were populations containing single clones. Considerable genetic variation, however, was noted among isolates of biotype 1 and T. atroviride from Europe. Biotype 4 was not represented by the preepidemic isolates of Trichoderma as determined by RAPD markers and PCR amplification of an arbitrary DNA sequence unique to the genomes of biotypes 2 and 4. Our findings suggest that the onset of the green mold epidemic in North America resulted from the recent introduction of a highly virulent genotype of the pathogen into cultivated mushrooms.

scholarly journals Application of Random Amplified Polymorphic DNA Markers for Identification of Marula Genotypes

HortScience ◽  
1999 ◽  
Vol 34 (7) ◽  
pp. 1256-1258 ◽  
Author(s):  
Feiga Gutman ◽  
Avinoam Nerd ◽  
Yosef Mizrahi ◽  
Dudy Bar-Zvi ◽  
Dina Raveh
Keyword(s):  
Dna Markers ◽  
Rapd Markers ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Band Pattern ◽  
Randomly Amplified Polymorphic Dna ◽  
Distinct Band ◽  
Sclerocarya Birrea

Twenty-four genotypes of marula (Sclerocarya birrea subsp. caffra) were characterized using randomly amplified polymorphic DNA (RAPD) analysis. A distinct band pattern was obtained for each of the trees, using as few as four arbitrary 10-mer primers. Trees propagated vegetatively by grafting showed identical fingerprints. These results suggest that RAPD markers provide a useful system for documenting the identity of marula genotypes.

scholarly journals Using Randomly Amplified Polymorphic DNA (RAPD) Markers to Identify Annona Cultivars

1995 ◽  
Vol 120 (5) ◽  
pp. 726-729 ◽  
Author(s):  
C.M. Ronning ◽  
R.J. Schnell ◽  
S. Gazit
Keyword(s):  
Native American ◽  
Efficient Method ◽  
Interspecific Hybrids ◽  
Rapd Markers ◽  
Rapd Analysis ◽  
Randomly Amplified Polymorphic Dna ◽  
Banding Patterns ◽  
Polymorphic Loci ◽  
Edible Fruit ◽  
Custard Apple

The native American genus Annona contains many species that are cultivated for their edible fruit, including the custard apple (A. reticuluta L.), soursop (A. muricata L.), cherimoya (A. cherimola L.), sugar apple (A. squamosa L.), and interspecific hybrids, the atemoyas. RAPD analysis of A. cherimola. `Campa' and `Jete,' A. squamosa `Lessard,' and the atemoyas `Ubranitzki,' `Malali,' and `Kaspi' resulted in very distinctive patterns, indicating that RAPD markers, may be an efficient method of fingerprinting genotypes within and between Annona species. All 15 primers used generated repeatable, polymorphic patterns. An F1 population of `Jete' × `Lessard' was analyzed to determine the inheritance of the RAPD banding patterns. Fifty-two polymorphic loci were identified, which segregated in an expected Mendelian fashion.

ANALYSIS OF GENETIC DIVERSITY OF 26 CASSAVA VARIETIES (Manihot esculenta CRANTZ) WITH DIFFERENT RESPONSES TO WATERLOGGING CONDITIONS BY USING RAPD MARKERS

2021 ◽  
Vol 66 (3) ◽  
pp. 170-179
Author(s):  
Sengsoulichan Dethvongsa ◽  
Vu Nguyen Anh ◽  
Van Tran Khanh
Keyword(s):  
Genetic Diversity ◽  
Phylogenetic Tree ◽  
Manihot Esculenta ◽  
Rapd Markers ◽  
Flood Tolerance ◽  
Randomly Amplified Polymorphic Dna ◽  
Manihot Esculenta Crantz ◽  
Rapd Pcr ◽  
Cassava Production ◽  
Cassava Varieties

RAPD (Randomly Amplified Polymorphic DNA) is an indicator for high and stable polymorphism, widely used in the study of the diversity of cassava. In this paper, the results of using 20 polymorphic primers OPK combined with the establishment of the phylogenetic tree to analyze the genetic diversity of 26 cassava varieties with different responses to waterlogging conditions by using the RAPD-PCR technique were presented. The purpose of this experiment was to show the genetic relevance of the studied cassava varieties. The results showed that the flood tolerance of cassava was not related to the polymorphism and branching characteristics of the stem. This information may be use as a basis for selecting flood-tolerant cassava varieties for cassava production, as well as the basis for selecting genetically different parents for breeding.

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 Seasonal Dynamics and Metagenomic Characterization of Estuarine Viriobenthos Assemblages by Randomly Amplified Polymorphic DNA PCR

2009 ◽  
Vol 75 (8) ◽  
pp. 2259-2265 ◽  
Author(s):  
Rebekah R. Helton ◽  
K. Eric Wommack
Keyword(s):  
Chesapeake Bay ◽  
Oligonucleotide Primer ◽  
Viral Diversity ◽  
Randomly Amplified Polymorphic Dna ◽  
Aquatic Sediments ◽  
Banding Patterns ◽  
Rapd Pcr ◽  
Viral Sequences ◽  
Dna Pcr

ABSTRACT Direct enumeration and genetic analyses indicate that aquatic sediments harbor abundant and diverse viral communities. Thus far, synecological analysis of estuarine sediment viral diversity over an annual cycle has not been reported. This oversight is due in large part to a lack of molecular genetic approaches for assessing viral diversity within a large collection of environmental samples. Here, randomly amplified polymorphic DNA PCR (RAPD-PCR) was used to examine viral genotypic diversity within Chesapeake Bay sediments. Using a single 10-mer oligonucleotide primer for all samples, RAPD-PCR analysis of sediment viral assemblages yielded unique banding patterns across spatial and temporal scales, with the occurrence of specific bands varying among the sample set. Cluster analysis of RAPD-PCR amplicon banding patterns indicated that sediment viral assemblages changed with season and to a lesser extent with geographic location. Sequence analysis of RAPD-PCR amplicons revealed that 76% of sediment viral sequences were not homologous to any sequence in the GenBank nonredundant protein database. Of the GenBank sequence homologs, the majority belonged to viruses within the Podoviridae (24%) and Myoviridae (22%) viral families, which agrees with the previously observed frequencies of these morphological families in Chesapeake Bay sediments. Furthermore, the majority of the sediment viral sequences homologous to GenBank nonredundant protein sequences were phages or prophages (57%). Hence, RAPD-PCR proved to be a reliable and useful approach for characterization of viral assemblages and the genetic diversity of viruses within aquatic sediments.

scholarly journals Use of Random Amplified Polymorphic DNA Markers for Cultivar Identification in Mint

HortScience ◽  
2001 ◽  
Vol 36 (4) ◽  
pp. 761-764 ◽  
Author(s):  
A.L. Fenwick ◽  
S.M. Ward
Keyword(s):  
United States ◽  
Genetic Diversity ◽  
Dna Markers ◽  
Geographical Origin ◽  
Cultivar Identification ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
The United States ◽  
Randomly Amplified Polymorphic Dna ◽  
Commercial Oil

Seventeen mint accessions representing the three species grown for commercial oil production in the United States were characterized using randomly amplified polymorphic DNA (RAPD) analysis. The RAPD profiles readily identified the different Mentha species; calculation of genetic distance, based on the number of shared bands, indicated that M. spicata L. is more closely related to M. × gracilis than to M. × piperita. The RAPD profiles also distinguished among eight peppermint accessions of different geographical origin. However, only limited polymorphism was observed among the most widely grown peppermint and Scotch spearmint cultivars. These results indicate a potential lack of genetic diversity in mint cultivars grown for oil in the United States.

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