02390naa a2200277 a 450000100080000000500110000800800410001910000170006024500720007726000090014952016490015865000220180765000090182965000140183865000130185265000180186565000090188370000180189270000220191070000170193270000200194970000180196970000170198770000200200477300880202416209451997-12-01       1996    bl uuuu     u00u1 u    #d1 aBORNEMAN, J.  aMolecular microbial diversity of an agricultural soil in Wisconsin.  c1996  aA culture-independent survey of the soil microbial diversity in a clover-grass pasture in souther wisconsin was conducted by sequence analysis of a universal clone library of genes coding for small-subunit rRNA(rDNA). A rapid and efficient method for extraction of DNA from sils which resulted in highly purified DNA wit minimal shearing was developed. universal small-subunit-rRNA primers were used to amplify DNA extracted from the pasture soil. the PCR products were  cloned into pGEM-T,and either hypervariable or conseverd regions were sequenced. the relationships of 124 sequences to those of cultured orgnisms of known pylogeny were determined. of the 124 clones sequenced,98.4% were from the domain bacteria. two of the rDNA sequences were derived from eukaryotic organelles. two of the 124 sequences were of nuclear origin, one being fungal and the other a plant sequences. no sequences of the domain ARCHEA were found. within the domain bacteria,three kingdoms were highly representend:the PROTEOBACTERIA (16.1%),the CYTOPHAGA-FLEXIBACTER-BACTEROIDES GROUP(21.8%), and the low-g+c-contet gram-positive group(31.8%). some kingdoms,such as the thermotogales,the dreen nonsulfer groupthe fusobacteria,and the spirochaetes,were absent.a large number of the sequences(39.4%) were distributed among several clades that are not among the major taxa describ by olsen et al.(G.J.olsen,C.R.Woese,and Overbeek,J.Bacteriol.,176:1-6,1994).From the alignments of the sequencedata,distance matrices were calculated to display the enormous microbial diversity foud in this soil in two wals,as phylogenetic trees and as multidimencional-scaling plots.  amicrobial ecology  asoil  aWisconsin  aEcologia  aMicrorganismo  aSolo1 aSKROCH, P. W.1 aO'SULLIVAN, K. M.1 aPALUS, J. A.1 aRUMJANEK, N. G.1 aJANSEN, J. L.1 aNIENHUIS, J.1 aTRIPLETT, E. W.  tApplied and Environmental Microbiology, Wisconsingv. 62, n. 6, p. 1935-1943, 1996.