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Titolo:
TRANSFER-RNA INTERGENIC SPACERS REVEAL POLYMORPHISMS DIAGNOSTIC FOR XANTHOMONAS ALBILINEANS
Autore:
HONEYCUTT RJ; SOBRAL BWS; MCCLELLAND M;
Indirizzi:
CTR APPLICAT MOLEC BIOL INT AGR AMER,11099 N TORREY PINES RD,STE 295 LA JOLLA CA 92037 SIDNEY KIMMEL CANC CTR LA JOLLA CA 92037
Titolo Testata:
Microbiology
, volume: 141, anno: 1995,
parte:, 12
pagine: 3229 - 3239
SICI:
1350-0872(1995)141:<3229:TISRPD>2.0.ZU;2-0
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE CHAIN-REACTION; DEOXYRIBONUCLEIC-ACID RELATEDNESS; LENGTH POLYMORPHISMS; ARBITRARY PRIMERS; PCR; DNA; STRAINS; PATHOVARS; GENOMES;
Keywords:
XANTHOMONAS ALBILINEANS; TRANSFER-RNA; POLYMERASE CHAIN REACTION; PHYTOPATHOGEN; LEAF SCALD DISEASE; SUGARCANE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
29
Recensione:
Indirizzi per estratti:
Citazione:
R.J. Honeycutt et al., "TRANSFER-RNA INTERGENIC SPACERS REVEAL POLYMORPHISMS DIAGNOSTIC FOR XANTHOMONAS ALBILINEANS", Microbiology, 141, 1995, pp. 3229-3239

Abstract

A PCR-based detection system was developed for Xanthomonas albilineans, a pathogen of sugarcane, and other related xanthomonads, using the conserved sequence of two adjacent tRNA genes and the variable length and sequence of the spacer region between them. An appropriate region was identified as follows: tRNA genes with the same anticodon from a wide variety of bacteria were aligned and the most frequent base at each position was chosen to derive primers that would anneal to the gene in either orientation. Pairs of such primers were screened against various Xanthomonas species and members of related genera using PCR at low to moderate annealing stringency. A subset of these pairs of tRNA consensus primers gave one or more PCR products which generally displayed interspecific length variability. The primer pair 5'-3' tRNA(ala) and 3'-5' tRNA(ile) Showed interspecific length polymorphisms between X.albilineans and all other Xanthomonas species examined. These PCR products were cloned and sequenced from four isolates of X. albilineans and four isolates from different pathovars of X. campestris, and the spacer length variation confirmed. Specific tRNA gene primers were derived from the tRNA gene sequences. These primers yielded a PCR product of a characteristic length within most Xanthomonas species and pathovars tested, When a primer that projected from tRNA(ala) into the 3' end of the variable intergenic spacer was used with a tRNA(ile)-specific primer, PCR was a very sensitive diagnostic test for X. albilineans-infected sugarcane and gave no product or only a faint product with otherspecies of bacteria. The specificity of this PCR-based detection system was further enhanced by a nested PCR reaction that took advantage of the fact that the tRNA(ala)-tRNA(ile) region was found to be embedded in a 16S rRNA-23S rRNA gene spacer. By amplifying the region betweenthe 16S rRNA gene and tRNA(ile) or between the tRNA(ala) and the 23S rRNA gene, the subsequent nested PCR product was shown to be X. albilineans-specific.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/09/20 alle ore 04:31:43