Development of a PCR test for the detection of Curtobacterium flaccumfaciens pv. flaccumfaciens

被引：22

作者：

Guimaraes P.M. ^{[1
]}

Palmano S. ^{[1
]}

Smith J.J. ^{[1
]}

Grossi de Sá M.F. ^{[1
]}

Saddler G.S. ^{[1
]}

机构：

[1] Scottish Agricultural Science Agency, Edinburgh EH12 8NJ, 82 Craigs Road, East Craigs

来源：

Antonie van Leeuwenhoek | 2001年 / 80卷 / 1期

关键词：

Curtobacterium flaccumfaciens pv. flaccumfaciens; PCR detection;

D O I：

10.1023/A:1012077425747

中图分类号：

学科分类号：

摘要：

A chromosomal DNA library of the bacterial pathogen of bean, Curtobacterium flaccumfaciens pv. flaccumfaciens NCPPB 559 was constructed in the plasmid pGEM-7Zf(+). Several clones were identified that hybridised to all Curtobacterium flaccumfaciens pathovars including: C. f. betae, C. f. flaccumfaciens, C. f. oortii, C. f. poinsettiae and, in addition, to some strains of Clavibacter michiganensis subsp. insidiosus and Clavibacter michiganensis subsp. One of these clones (pPMP-26), after subsequent digestion with restriction endonucleases EcoRI/SacI, yielded a fragment of approximately 0.2 Kb (pPMP-26D) that hybridised specifically to C. f. flaccumfaciens and not to any of the other plant pathogenic members of the order Actinomycetales or any of the other prokaryotic bean pathogens tested. This fragment was subcloned and sequenced, analysis of the resultant 198 bp sequence showed that no significant homology existed with any other sequence currently deposited in public databases. Further analysis of these data facilitated the design of PCR primers which were subsequently tested against a wide range of plant pathogenic actinomycetes and other prokaryotic bean pathogens. Results show that these primers are highly specific for all strains of C. f. flaccumfaciens with no cross-reaction to strains from any other bacterial taxa tested.

引用

页码：1 / 10

页数：9

共 45 条

[1]

Altenburger P., Kampfer P., Akimov V.N., Lubitz W., Busse H.J., Polyamine distribution in actinomycetes with group B peptidoglycan and species of the genera Brevibacterium, Corynebacterium and Tsukamurella. Int. J. Syst. Bacteriol., 47, pp. 270-277, (1997)

[2]

Altschul S.F., Gish W., Miller W., Myers E.W., Lipman D.J., Basic local alignment search tool, J. Molec. Biol., 215, pp. 403-410, (1990)

[3]

Ball S.F.L., Reeves J.C., Application of rapid techniques to seed health testing - Prospects and potential, Techniques for the Rapid Detection of Plant Pathogens, pp. 193-207, (1992)

[4]

Bradbury J.F., Guide to Plant Pathogenic Bacteria., (1986)

[5]

Calzolari A., Tomenasi M., Mazzucchi V., Comparison of immunofluorescence staining and indirect isolation for the detection of Corynebacterium flaccumfaciens in bean seeds, Bulletin OEPP/EPPO, 17, pp. 157-163, (1987)

[6]

Carlson R.R., Vidaver A.K., Taxonomy of Corynebacterium plant pathogens, including a new pathogen of wheat, based on polyacrylamide gel electrophoresis of cellular proteins, Int. J. Syst. Bacteriol., 32, pp. 315-326, (1982)

[7]

Collins M.D., Goodfellow M., Minnikin D.E., Fatty acid, isoprenoid quinone and polar lipid composition in the classification of Curtobacterium and related taxa, J. Gen. Microbiol., 118, pp. 29-37, (1980)

[8]

Collins M.D., Jones D., Lipids in the classification of coryneform bacteria containing peptidoglycan based on 2,4-diaminobutyric acid, J. Appl. Bacteriol., 48, pp. 459-470, (1980)

[9]

Collins M.D., Jones D., Reclassification of Corynebacterium flaccumfaciens, Corynebacterium betae, Corynebacterium oortii and Corynebacterium poinsettiae in the genus Curtobacterium as Curtobacterium flaccumfaciens comb. nov, Journal of General Microbiology, 129, pp. 3545-3548, (1983)

[10]

Coyne D.P., Schuster M.L., Bacterial diseases of legumes: Breeding and resistance, Advances in Legume Science, pp. 225-233, (1979)

← 1 2 3 4 5 →