Community structure of actively growing bacterial populations in plant pathogen suppressive soil.
The bacterial community in soil was screened by using various molecular approaches for bacterial populations that were activated upon addition of different supplements. Plasmodiophora brassicae spores, chitin, sodium acetate, and cabbage plants were added to activate specific bacterial populations as an aid in screening for novel antagonists to plant pathogens. DNA from growing bacteria was specifically extracted from the soil by bromodeoxyuridine immunocapture. The captured DNA was fingerprinted by terminal restriction fragment length polymorphism (T-RFLP). The composition of the dominant bacterial community was also analyzed directly by T-RFLP and by denaturing gradient gel electrophoresis (DGGE). After chitin addition to the soil, some bacterial populations increased dramatically and became dominant both in the total and in the actively growing community. Some of the emerging bands on DGGE gels from chitin-amended soil were sequenced and found to be similar to known chitin-degrading genera such as Oerskovia, Kitasatospora, and Streptomyces species. Some of these sequences could be matched to specific terminal restriction fragments on the T-RFLP output. After addition of Plasmodiophora spores, an increase in specific Pseudomonads could be observed with Pseudomonas-specific primers for DGGE. These results demonstrate the utility of microbiomics, or a combination of molecular approaches, for investigating the composition of complex microbial communities in soil.
Hjort
Karin
K
Department of Microbiology, Swedish University of Agricultural Sciences, Box 7025, SE-750 07, Uppsala, Sweden.
Lembke
Antje
A
Speksnijder
Arjen
A
Smalla
Kornelia
K
Jansson
Janet K
JK
D001419:Bacteria / Q000235:genetics / Q000254:growth & development*; D001483:Base Sequence; D044822:Biodiversity; D001937:Brassica; D001973:Bromodeoxyuridine / Q000032:analysis / Q000378:metabolism; D002686:Chitin / Q000378:metabolism / Q000494:pharmacology; D016000:Cluster Analysis; D017753:Ecosystem; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005658:Fungi / Q000187:drug effects / Q000472:pathogenicity / Q000502:physiology*; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D018517:Plant Roots / Q000382:microbiology; D012150:Polymorphism, Restriction Fragment Length; D011549:Pseudomonas / Q000187:drug effects / Q000254:growth & development; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D019346:Sodium Acetate / Q000494:pharmacology; D012988:Soil Microbiology; D013172:Spores, Fungal
D012336:RNA, Ribosomal, 16S; D002686:Chitin; D019346:Sodium Acetate; D001973:Bromodeoxyuridine