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Titolo:
A one step PCR-based method for the detection of economically important soft rot Erwinia species on micropropagated potato plants
Autore:
Toth, IK; Hyman, LJ; Wood, JR;
Indirizzi:
Scottish Crop Res Inst, Dept Fungal & Bacterial Plant Pathol, Dundee DD2 5DA, Scotland Scottish Crop Res Inst Dundee Scotland DD2 5DA Dundee DD2 5DA, Scotland Scottish Agr Sci Agcy, Edinburgh, Midlothian, Scotland Scottish Agr Sci Agcy Edinburgh Midlothian Scotland Midlothian, Scotland
Titolo Testata:
JOURNAL OF APPLIED MICROBIOLOGY
fascicolo: 1, volume: 87, anno: 1999,
pagine: 158 - 166
SICI:
1364-5072(199907)87:1<158:AOSPMF>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
CAROTOVORA SUBSP ATROSEPTICA; POLYMERASE CHAIN-REACTION; SENSITIVE DETECTION; TUBERS; DIFFERENTIATION; CONTAMINATION; CHRYSANTHEMI; SEQUENCES; BLACKLEG; MEDIA;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
27
Recensione:
Indirizzi per estratti:
Indirizzo: Toth, IK Scottish Crop Res Inst, Dept Fungal & Bacterial Plant Pathol, Dundee DD2 5DA, Scotland Scottish Crop Res Inst Dundee Scotland DD2 5DA D2 5DA, Scotland
Citazione:
I.K. Toth et al., "A one step PCR-based method for the detection of economically important soft rot Erwinia species on micropropagated potato plants", J APPL MICR, 87(1), 1999, pp. 158-166

Abstract

A simple, rapid and sensitive PCR-based method was developed for the detection of all fire subspecies Of Erwinia carotovora, including subsp. carotovora and subsp. atroseptica, and all pathovars/biovars of Erwinia chrysanthemi, on plant tissue culture material. Primers SR3F and SR1cR, based on a conserved region of the 16S rRNA gene, amplified a DNA fragment of 119 bp from all 65 such strains tested. Detection limits of the method in vitro were 2.0 x 10(2)-3.4 x 10(3) cfu ml(-1) (equivalent to 1-17 cfu per PCR) and, following extraction of genomic DNA from plant extract, detection limits were2.3 x 10(2)-1.9 x 10(4) cfu per microplant sample (equivalent: to 5 cfu - 3.8 x 10(2) cfu per PCR). To improve the sensitivity of the method in planta, to obviate the need fur complex and laborious DNA extractions, and to remove inhibitory substances present in the plant extract, an enrichment stepwas included prior to PCR. Following enrichment, the sensitivity of detection was < 10 cfu per microplant sample. This method provides the first sensitive means of detecting latent: infection caused by several economically important soft rot erwinias simultaneously on potato tissue culture material.

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Documento generato il 24/09/20 alle ore 05:27:26