Genotyping Icelandic Isolates of Rhizobia Based on rDNA-RFLP
Simple PCR amplification using universal rhizobial oligonucleotide primers for intergenic spacer (IGS) regions of 16S-23S ribosomal genes (rDNA) was able to discriminate species, biovars and strains of rhizobial isolates. Six isolates of Rhizobium and Bradyrhizobium species from Iceland were analyse...
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ftciteseerx:oai:CiteSeerX.psu:10.1.1.564.9976 2023-05-15T16:51:38+02:00 Genotyping Icelandic Isolates of Rhizobia Based on rDNA-RFLP Ægir Thór Thórsson The Pennsylvania State University CiteSeerX Archives application/pdf http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.564.9976 http://www.landbunadur.is/landbunadur/wgsamvef.nsf/8bbba2777ac88c4000256a89000a2ddb/c58acb353b31e93700256de100536842/$FILE/gr-bu13-aegir.pdf en eng http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.564.9976 http://www.landbunadur.is/landbunadur/wgsamvef.nsf/8bbba2777ac88c4000256a89000a2ddb/c58acb353b31e93700256de100536842/$FILE/gr-bu13-aegir.pdf Metadata may be used without restrictions as long as the oai identifier remains attached to it. http://www.landbunadur.is/landbunadur/wgsamvef.nsf/8bbba2777ac88c4000256a89000a2ddb/c58acb353b31e93700256de100536842/$FILE/gr-bu13-aegir.pdf Key words rhizobia ribosomal DNA PCR amplification RFLP. YFIRLIT Greining íslenskra ísólata af Rhizobium-bakteríum með rDNA-RFLP aðferð text ftciteseerx 2016-01-08T12:13:25Z Simple PCR amplification using universal rhizobial oligonucleotide primers for intergenic spacer (IGS) regions of 16S-23S ribosomal genes (rDNA) was able to discriminate species, biovars and strains of rhizobial isolates. Six isolates of Rhizobium and Bradyrhizobium species from Iceland were analysed in comparison with four isolates from Scandinavia and Siberia. The rhizobia in pure cultures were isolated from nodules of leguminous species in the genera Trifolium, Lathyrus, Lupinus, Astragalus and Thermopsis. The amplified DNA fragments were between 400 and 1400-bp long and all the isolates could be dif-ferentiated except one pair of samples. The discrimination of isolates corresponded reasonably well to the rhizobial classification and was independent of their geographic origin. Further analysis was per-formed by fractionation of the PCR amplified products with restriction endonucleases. Twelve restric-tion enzymes were used, one at a time. Six of these (i.e. EcoRI, Ha III, MboI, TaqI, PstI and PvuII) were able to identify all isolates except the pair that could not be differentiated by the PCR amplification. This pair was likely to be of the same genotype. The use of single enzyme digestion provided a simple means to verify rhizobial identification, whereas combinations of two or more restriction enzymes, used simul-taneously, maximised resolution of the restriction fragment length polymorphism (RFLP) profiles. The genetic distance analysis based on rDNA-RFLP profiles showed two clusters: one of these included essentially R. leguminosarum bv. trifolii and the other, which was a more diverse group, included other species. The rDNA-RFLP used in this study identified rhizobial species and strains, and detected mis-identification of some isolates. Text Iceland Siberia Unknown |
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Open Polar |
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Unknown |
op_collection_id |
ftciteseerx |
language |
English |
topic |
Key words rhizobia ribosomal DNA PCR amplification RFLP. YFIRLIT Greining íslenskra ísólata af Rhizobium-bakteríum með rDNA-RFLP aðferð |
spellingShingle |
Key words rhizobia ribosomal DNA PCR amplification RFLP. YFIRLIT Greining íslenskra ísólata af Rhizobium-bakteríum með rDNA-RFLP aðferð Ægir Thór Thórsson Genotyping Icelandic Isolates of Rhizobia Based on rDNA-RFLP |
topic_facet |
Key words rhizobia ribosomal DNA PCR amplification RFLP. YFIRLIT Greining íslenskra ísólata af Rhizobium-bakteríum með rDNA-RFLP aðferð |
description |
Simple PCR amplification using universal rhizobial oligonucleotide primers for intergenic spacer (IGS) regions of 16S-23S ribosomal genes (rDNA) was able to discriminate species, biovars and strains of rhizobial isolates. Six isolates of Rhizobium and Bradyrhizobium species from Iceland were analysed in comparison with four isolates from Scandinavia and Siberia. The rhizobia in pure cultures were isolated from nodules of leguminous species in the genera Trifolium, Lathyrus, Lupinus, Astragalus and Thermopsis. The amplified DNA fragments were between 400 and 1400-bp long and all the isolates could be dif-ferentiated except one pair of samples. The discrimination of isolates corresponded reasonably well to the rhizobial classification and was independent of their geographic origin. Further analysis was per-formed by fractionation of the PCR amplified products with restriction endonucleases. Twelve restric-tion enzymes were used, one at a time. Six of these (i.e. EcoRI, Ha III, MboI, TaqI, PstI and PvuII) were able to identify all isolates except the pair that could not be differentiated by the PCR amplification. This pair was likely to be of the same genotype. The use of single enzyme digestion provided a simple means to verify rhizobial identification, whereas combinations of two or more restriction enzymes, used simul-taneously, maximised resolution of the restriction fragment length polymorphism (RFLP) profiles. The genetic distance analysis based on rDNA-RFLP profiles showed two clusters: one of these included essentially R. leguminosarum bv. trifolii and the other, which was a more diverse group, included other species. The rDNA-RFLP used in this study identified rhizobial species and strains, and detected mis-identification of some isolates. |
author2 |
The Pennsylvania State University CiteSeerX Archives |
format |
Text |
author |
Ægir Thór Thórsson |
author_facet |
Ægir Thór Thórsson |
author_sort |
Ægir Thór Thórsson |
title |
Genotyping Icelandic Isolates of Rhizobia Based on rDNA-RFLP |
title_short |
Genotyping Icelandic Isolates of Rhizobia Based on rDNA-RFLP |
title_full |
Genotyping Icelandic Isolates of Rhizobia Based on rDNA-RFLP |
title_fullStr |
Genotyping Icelandic Isolates of Rhizobia Based on rDNA-RFLP |
title_full_unstemmed |
Genotyping Icelandic Isolates of Rhizobia Based on rDNA-RFLP |
title_sort |
genotyping icelandic isolates of rhizobia based on rdna-rflp |
url |
http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.564.9976 http://www.landbunadur.is/landbunadur/wgsamvef.nsf/8bbba2777ac88c4000256a89000a2ddb/c58acb353b31e93700256de100536842/$FILE/gr-bu13-aegir.pdf |
genre |
Iceland Siberia |
genre_facet |
Iceland Siberia |
op_source |
http://www.landbunadur.is/landbunadur/wgsamvef.nsf/8bbba2777ac88c4000256a89000a2ddb/c58acb353b31e93700256de100536842/$FILE/gr-bu13-aegir.pdf |
op_relation |
http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.564.9976 http://www.landbunadur.is/landbunadur/wgsamvef.nsf/8bbba2777ac88c4000256a89000a2ddb/c58acb353b31e93700256de100536842/$FILE/gr-bu13-aegir.pdf |
op_rights |
Metadata may be used without restrictions as long as the oai identifier remains attached to it. |
_version_ |
1766041760399622144 |