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Combined Molecular and Biochemical Approach Identifies Aspergillus japonicus and Aspergillus aculeatus as Two Species

Combined Molecular and Biochemical Approach Identifies Aspergillus japonicus and Aspergillus... Combined Molecular and Biochemical Approach Identifies Aspergillus japonicus and Aspergillus aculeatus as Two Species Lucie Pařenicová 1 , Pernille Skouboe 2 , Jens Frisvad 3 , Robert A. Samson 4 , Lone Rossen 2 , Marjon ten Hoor-Suykerbuyk 1 , and Jaap Visser 1 , * Section of Molecular Genetics of Industrial Microorganisms, Wageningen University, NL-6703 HA Wageningen, 1 and Centraalbureau voor Schimmelcultures, NL-3740 AG Baarn, 4 The Netherlands; and Division of Applied Molecular Biology, Biotechnological Institute, DK-2970 Hørsholm, 2 and Department of Biotechnology, Technical University of Denmark, DK-2800 Lyngby, 3 Denmark ABSTRACT We examined nine Aspergillus japonicus isolates and 10 Aspergillus aculeatus isolates by using molecular and biochemical markers, including DNA sequences of the ITS1-5.8S rRNA gene-ITS2 region, restriction fragment length polymorphisms (RFLP), and secondary-metabolite profiles. The DNA sequence of the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene could not be used to distinguish between A. japonicus and A. aculeatus but did show that these two taxa are more closely related to each other than to other species of black aspergilli. Aspergillus niger pyruvate kinase ( pki A) and pectin lyase A ( pel A) and Agaricus bisporus 28S rRNA genes, which were used as probes in the RFLP analysis, revealed clear polymorphism between these two taxa. The A. niger pki A and pel A probes placed six strains in an A. japonicus group and 12 isolates in an A. aculeatus group, which exhibited intraspecific variation when they were probed with the pel A gene. The secondary-metabolite profiles supported division of the isolates into the two species and differed from those of other black aspergilli. The strains classified as A. japonicus produced indole alkaloids and a polar metabolite, while the A. aculeatus isolates produced neoxaline, okaramins, paraherquamidelike compounds, and secalonic acid. A. aculeatus CBS 114.80 showed specific RFLP patterns for all loci examined. The secondary-metabolite profile of strain CBS 114.80 also differed from those of A. japonicus and A. aculeatus . Therefore, this strain probably represents a third taxon. This study provides unambiguous criteria for establishing the taxonomic positions of isolates of black aspergilli, which are important in relation to industrial use and legal protection of these organisms. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied and Environmental Microbiology American Society For Microbiology

Combined Molecular and Biochemical Approach Identifies Aspergillus japonicus and Aspergillus aculeatus as Two Species

Combined Molecular and Biochemical Approach Identifies Aspergillus japonicus and Aspergillus aculeatus as Two Species

Applied and Environmental Microbiology , Volume 67 (2): 521 – Feb 1, 2001

Abstract

Combined Molecular and Biochemical Approach Identifies Aspergillus japonicus and Aspergillus aculeatus as Two Species Lucie Pařenicová 1 , Pernille Skouboe 2 , Jens Frisvad 3 , Robert A. Samson 4 , Lone Rossen 2 , Marjon ten Hoor-Suykerbuyk 1 , and Jaap Visser 1 , * Section of Molecular Genetics of Industrial Microorganisms, Wageningen University, NL-6703 HA Wageningen, 1 and Centraalbureau voor Schimmelcultures, NL-3740 AG Baarn, 4 The Netherlands; and Division of Applied Molecular Biology, Biotechnological Institute, DK-2970 Hørsholm, 2 and Department of Biotechnology, Technical University of Denmark, DK-2800 Lyngby, 3 Denmark ABSTRACT We examined nine Aspergillus japonicus isolates and 10 Aspergillus aculeatus isolates by using molecular and biochemical markers, including DNA sequences of the ITS1-5.8S rRNA gene-ITS2 region, restriction fragment length polymorphisms (RFLP), and secondary-metabolite profiles. The DNA sequence of the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene could not be used to distinguish between A. japonicus and A. aculeatus but did show that these two taxa are more closely related to each other than to other species of black aspergilli. Aspergillus niger pyruvate kinase ( pki A) and pectin lyase A ( pel A) and Agaricus bisporus 28S rRNA genes, which were used as probes in the RFLP analysis, revealed clear polymorphism between these two taxa. The A. niger pki A and pel A probes placed six strains in an A. japonicus group and 12 isolates in an A. aculeatus group, which exhibited intraspecific variation when they were probed with the pel A gene. The secondary-metabolite profiles supported division of the isolates into the two species and differed from those of other black aspergilli. The strains classified as A. japonicus produced indole alkaloids and a polar metabolite, while the A. aculeatus isolates produced neoxaline, okaramins, paraherquamidelike compounds, and secalonic acid. A. aculeatus CBS 114.80 showed specific RFLP patterns for all loci examined. The secondary-metabolite profile of strain CBS 114.80 also differed from those of A. japonicus and A. aculeatus . Therefore, this strain probably represents a third taxon. This study provides unambiguous criteria for establishing the taxonomic positions of isolates of black aspergilli, which are important in relation to industrial use and legal protection of these organisms.

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Publisher
American Society For Microbiology
Copyright
Copyright © 2001 by the American society for Microbiology.
ISSN
0099-2240
eISSN
1098-5336
DOI
10.1128/AEM.67.2.521-527.2001
pmid
11157212
Publisher site
See Article on Publisher Site

Abstract

Combined Molecular and Biochemical Approach Identifies Aspergillus japonicus and Aspergillus aculeatus as Two Species Lucie Pařenicová 1 , Pernille Skouboe 2 , Jens Frisvad 3 , Robert A. Samson 4 , Lone Rossen 2 , Marjon ten Hoor-Suykerbuyk 1 , and Jaap Visser 1 , * Section of Molecular Genetics of Industrial Microorganisms, Wageningen University, NL-6703 HA Wageningen, 1 and Centraalbureau voor Schimmelcultures, NL-3740 AG Baarn, 4 The Netherlands; and Division of Applied Molecular Biology, Biotechnological Institute, DK-2970 Hørsholm, 2 and Department of Biotechnology, Technical University of Denmark, DK-2800 Lyngby, 3 Denmark ABSTRACT We examined nine Aspergillus japonicus isolates and 10 Aspergillus aculeatus isolates by using molecular and biochemical markers, including DNA sequences of the ITS1-5.8S rRNA gene-ITS2 region, restriction fragment length polymorphisms (RFLP), and secondary-metabolite profiles. The DNA sequence of the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene could not be used to distinguish between A. japonicus and A. aculeatus but did show that these two taxa are more closely related to each other than to other species of black aspergilli. Aspergillus niger pyruvate kinase ( pki A) and pectin lyase A ( pel A) and Agaricus bisporus 28S rRNA genes, which were used as probes in the RFLP analysis, revealed clear polymorphism between these two taxa. The A. niger pki A and pel A probes placed six strains in an A. japonicus group and 12 isolates in an A. aculeatus group, which exhibited intraspecific variation when they were probed with the pel A gene. The secondary-metabolite profiles supported division of the isolates into the two species and differed from those of other black aspergilli. The strains classified as A. japonicus produced indole alkaloids and a polar metabolite, while the A. aculeatus isolates produced neoxaline, okaramins, paraherquamidelike compounds, and secalonic acid. A. aculeatus CBS 114.80 showed specific RFLP patterns for all loci examined. The secondary-metabolite profile of strain CBS 114.80 also differed from those of A. japonicus and A. aculeatus . Therefore, this strain probably represents a third taxon. This study provides unambiguous criteria for establishing the taxonomic positions of isolates of black aspergilli, which are important in relation to industrial use and legal protection of these organisms.

Journal

Applied and Environmental MicrobiologyAmerican Society For Microbiology

Published: Feb 1, 2001

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