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Applied and Environmental Microbiology, July 2003, p. 3758-3766, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.3758-3766.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Dynamics of Fungal Communities in Bulk and Maize Rhizosphere Soil in the Tropics

Newton C. Marcial Gomes,1 Olajire Fagbola,1 Rodrigo Costa,2 Norma Gouvea Rumjanek,3 Arno Buchner,4 Leda Mendona-Hagler,2 and Kornelia Smalla1*

Biologische Bundesanstalt für Land- und Forstwirtschaft, Institut für Pflanzenvirologie, Mikrobiologie und biologische Sicherheit, 38104 Braunschweig,1 Lehrstuhl für Mikrobiologie, Technische Universität München, 85350 Freising, Germany,4 Instituto de Microbiologia, Cidade Universitaria-CCS Bloco I, Universidade Federal do Rio de Janeiro, 21949-590 Rio de Janeiro,2 Empresa Brasileira de Pesquisa Agropecuária Agrobiologia, Antiga Rodovia Rio-Sao Paulo, 23851-970 Seropédica, Rio de Janeiro, Brazil3

Received 26 November 2002/ Accepted 28 March 2003

The fungal population dynamics in soil and in the rhizospheres of two maize cultivars grown in tropical soils were studied by a cultivation-independent analysis of directly extracted DNA to provide baseline data. Soil and rhizosphere samples were taken from six plots 20, 40, and 90 days after planting in two consecutive years. A 1.65-kb fragment of the 18S ribosomal DNA (rDNA) amplified from the total community DNA was analyzed by denaturing gradient gel electrophoresis (DGGE) and by cloning and sequencing. A rhizosphere effect was observed for fungal populations at all stages of plant development. In addition, pronounced changes in the composition of fungal communities during plant growth development were found by DGGE. Similar types of fingerprints were observed in two consecutive growth periods. No major differences were detected in the fungal patterns of the two cultivars. Direct cloning of 18S rDNA fragments amplified from soil or rhizosphere DNA resulted in 75 clones matching 12 dominant DGGE bands. The clones were characterized by their HinfI restriction patterns, and 39 different clones representing each group of restriction patterns were sequenced. The cloning and sequencing approach provided information on the phylogeny of dominant amplifiable fungal populations and allowed us to determine a number of fungal phylotypes that contribute to each of the dominant DGGE bands. Based on the sequence similarity of the 18S rDNA fragment with existing fungal isolates in the database, it was shown that the rhizospheres of young maize plants seemed to select the Ascomycetes order Pleosporales, while different members of the Ascomycetes and basidiomycetic yeast were detected in the rhizospheres of senescent maize plants.

* Corresponding author. Mailing address: BBA, Institut PS, Messeweg 11-12, 38104 Braunschweig, Germany. Phone: 49-531-2993814. Fax: 49-531-2993013. E-mail: K.Smalla{at}bba.de.

Applied and Environmental Microbiology, July 2003, p. 3758-3766, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.3758-3766.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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