Assessment of chitin decomposer diversity within an upland grassland.
The breakdown of chitin within an acidic upland grassland was studied. The aim was to provide a molecular characterisation of microorganisms involved in chitin degradation in the soil using soil microcosms and buried litter bags containing chitin. The investigation involved an examination of the effects of liming on the microbial communities within the soil and their chitinolytic activity. Microcosm experiments were designed to study the influence of lime and chitin enrichment on the grassland soil bacterial community ex situ under controlled environmental conditions. Bacterial and actinomycete counts were determined and total community DNA was extracted from the microcosms and from chitin bags buried at the experimental site. PCR based on specific 16S rRNA target sequences provided products for DGGE analysis to determine the structure of bacterial and actinomycete communities. Chitinase activity was assessed spectrophotometrically using chitin labelled with remazol brilliant violet. Both liming and chitin amendment increased bacterial and actinomycete viable counts and the chitinase activity. DGGE band patterns confirmed changes in bacterial populations under the influence of both treatments. PCR products amplified from DNA isolated from chitin bags were cloned and sequenced. Only a few matched known species but a prominent coloniser of chitin proved to be Stenotrophomonas maltophilia.
Krsek
M
M
Department of Biological Sciences, University of Warwick, Coventry, UK. kr@dna.bio.warwick.ac.uk
Wellington
E M
EM
D000192:Actinomycetales / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D002686:Chitin / Q000378:metabolism*; D002688:Chitinases / Q000378:metabolism; D015169:Colony Count, Microbial; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006863:Hydrogen-Ion Concentration; D006109:Poaceae; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012988:Soil Microbiology; D020588:Stenotrophomonas / Q000254:growth & development / Q000302:isolation & purification
D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D002686:Chitin; D002688:Chitinases