Open AccessObservation of high seasonal variation in community structure of denitrifying bacteria in arable soil receiving artificial fertilizer and cattle manure by determining T‐RFLP of nir gene fragments
Author(s) -
Wolsing Martin,
Priemé Anders
Publication year - 2004
Publication title -
fems microbiology ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.377
H-Index - 155
eISSN - 1574-6941
pISSN - 0168-6496
DOI - 10.1016/j.femsec.2004.02.002
Subject(s) - denitrifying bacteria , biology , restriction fragment length polymorphism , manure , terminal restriction fragment length polymorphism , bacteria , nitrite reductase , veterinary medicine , fertilizer , soil microbiology , agronomy , denitrification , ecology , nitrate , nitrite , gene , genotype , genetics , nitrogen , medicine , physics , quantum mechanics
Temporal and spatial variation of communities of soil denitrifying bacteria at sites receiving mineral fertilizer (60 and 120 kgNha −1 year −1 ) and cattle manure (75 and 150 kgNha −1 year −1 ) were explored using terminal restriction fragment length polymorphism (T‐RFLP) analyses of PCR amplified nitrite reductase ( nirK and nirS ) gene fragments. The analyses were done three times during the year: in March, July and October. nirK gene fragments could be amplified in all three months, whereas nirS gene fragments could be amplified only in March. Analysis of similarities in T‐RFLP patterns revealed a significant seasonal shift in the community structure of nirK ‐containing bacteria. Also, sites treated with mineral fertilizer or cattle manure showed different communities of nirK ‐containing denitrifying bacteria, since the T‐RFLP patterns of soils treated with these fertilizers were significantly different. Also, these sites significantly differed from the control plot (no fertilizer treatment), whereas the patterns for low and high N‐additions were barely separable from each other. Sequencing and phylogenetic analysis of 54 nirK clones revealed that the major part of the nirK ‐containing bacteria investigated belonged to a yet uncultivated cluster of denitrifying bacteria.