scholarly journals Restriction fragment length polymorphisms in isolates of Aspergillus fumigatus probed with part of the intergenic spacer region from the ribosomal RNA gene complex of Aspergillus nidulans

1990 ◽  
Vol 136 (10) ◽  
pp. 1991-1994 ◽  
Author(s):  
C. L. Spreadbury ◽  
B. W. Bainbridge ◽  
J. Cohen
1999 ◽  
Vol 77 (9) ◽  
pp. 1261-1269 ◽  
Author(s):  
Erica Lumini ◽  
Marco Bosco

During the last few years, some Frankia culture collections that maintained a large number of unidentified and uncharacterized Frankia strains were closed because of funding shortages. To reduce the costs of maintenance, we evaluated the biodiversity of half of the Frankia strains from our collection, by polymerase chain reaction - restriction fragment length polymorphisms (PCR-RFLPs) of nifD-nifK intergenic spacer and 16S-23S rDNA intergenic spacer regions. In this way we were able to reduce the number of strains without reducing the biodiversity of the whole collection. In general the nifD-nifK target proved to be more polymorphic than the rrn target. From 51 isolates of Elaeagnus frankiae, PCR-RFLP results allowed us to detect 13 identical strains, and to predict that the genomic species P8 of Akimov and Dobritsa (1992) very likely agrees with genomic species 5 of Fernandez et al. (1989). Moreover, we revealed genomic groups not yet described, as well as intraspecific variability. For Alnus frankiae, the polymorphisms shown by both the nif and the rrn PCR-RFLPs revealed three host plant species-specific subgroups inside Frankia alni. An expandable data base was created to serve as reference for future biodiversity evaluations on both culture collections and unisolated Frankia populations. It will be accessible by Internet at the International Frankia Website (http://www.unifi.it/unifi/distam/frankia/international.html).Key words: Frankia, PCR-RFLP, nifD-nifK intergenic spacer, rrn 16S-23S intergenic spacer, biodiversity, culture collections.


Parasitology ◽  
1995 ◽  
Vol 111 (1) ◽  
pp. 87-94 ◽  
Author(s):  
C. O. Cunningham ◽  
D. M. McGillivray ◽  
K. MacKenzie ◽  
W. T. Melvin

SUMMARYThe small subunit ribosomal RNA (srRNA) gene was amplified from Gyrodactylus salaris using the polymerase chain reaction (PCR), cloned, and the complete gene sequence of 1966 bp determined. The V4 region of the srRNA gene was identified and amplified from single specimens of G. salaris, G. derjavini and G. truttae. Comparison of the V4 sequences from these three species revealed sequence differences from which restriction fragment length polymorphisms (RFLPs) were predicted and an oligonucleotide probe (GsV4) specific to G. salaris designed. Digestion of the amplified V4 region of the srRNA gene with Hae III and either Alw I, BstY I, Dde I or Mbo I provided a means of discriminating between G. salaris, G. derjavini and G. truttae. The GsV4 probe was used to detect the srRNA gene from G. salaris in Southern and dot blots of the amplified V4 region. The nucleotide sequences reported in this paper have been submitted to the EMBL Data Library under accession numbers Z26942 (G. salaris), Z35128 (G. derjavini) and Z35129 (G. truttae).


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