Construction and use of a computerized DNA fingerprint database for lactic acid bacteria from silage.
Efficient selection of new silage inoculant strains from a collection of over 10,000 isolates of lactic acid bacteria (LAB) requires excellent strain discrimination. Toward that end, we constructed a GelCompar II database of DNA fingerprint patterns of ethidium bromide-stained EcoRI fragments of total LAB DNA separated by conventional agarose gel electrophoresis. We found that the total DNA patterns were strain-specific; 56/60 American Type Culture Collection strains of 33 species of LAB could be distinguished. Enterococcus faecium strains ATCC19434 and ATCC35667 had identical total DNA patterns and RiboPrints. Lactobacillus rhamnosus strains ATCC7469 and ATCC27773 also had identical total DNA patterns, but different RiboPrints. EcoRI RiboPrint patterns could distinguish only about 9/23 Lactobacillus plantarum strains and about 6/10 Lactobacillus buchneri strains, whereas all 33 strains could be distinguished by EcoRI total DNA patterns. Despite gel-to-gel variation, new DNA patterns can be readily grouped with existing patterns using GelCompar II. The database contains large homogenous clusters of L. plantarum, E. faecium, L. buchneri, Lactobacillus brevis and Pediococcus species that can be used for tentative taxonomic assignment. We routinely use the DNA fingerprint database to identify and characterize new strains, eliminate duplicate isolates and for quality control of inoculant product strains. The GelCompar II database has been in continuous use for 7 years and contains more than 3600 patterns representing approximately 700 unique patterns from over 300 gels and is the largest computerized DNA fingerprint database for LAB yet reported.
Chan
Russell K
RK
Forage Additives Research, Crop Genetics Research and Development, Pioneer Hi-Bred International, Inc., a DuPont Business, DuPont Agriculture and Nutrition, P.O. Box 1004, Johnston, IA 50131, USA. Russell.Chan@pioneer.com
Wortman
Cora R
CR
Smiley
Brenda K
BK
Hendrick
Carol A
CA
D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D030561:Databases, Nucleic Acid; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D007091:Image Processing, Computer-Assisted; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics; D010802:Phylogeny; D021521:Ribotyping; D012820:Silage / Q000382:microbiology*
D004269:DNA, Bacterial; D015246:Deoxyribonuclease EcoRI