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Titolo:
A DIRECTED SEARCH FOR DNA-SEQUENCES TIGHTLY LINKED TO CEREAL CYST-NEMATODE RESISTANCE GENES IN TRITICUM-TAUSCHII
Autore:
EASTWOOD RF; LAGUDAH ES; APPELS R;
Indirizzi:
VICTORIAN INST DRYLAND AGR,DEPT AGR,PB 260 HORSHAM VIC 3400 AUSTRALIA VICTORIAN INST DRYLAND AGR,DEPT AGR HORSHAM VIC 3400 AUSTRALIA CSIRO,DIV PLANT IND CANBERRA ACT 2601 AUSTRALIA
Titolo Testata:
Genome
fascicolo: 2, volume: 37, anno: 1994,
pagine: 311 - 319
SICI:
0831-2796(1994)37:2<311:ADSFDT>2.0.ZU;2-Q
Fonte:
ISI
Lingua:
ENG
Soggetto:
FRAGMENT-LENGTH-POLYMORPHISM; POLYMERASE CHAIN-REACTION; NEAR-ISOGENIC LINES; HETERODERA-AVENAE; ENZYMATIC AMPLIFICATION; ARBITRARY PRIMERS; ADDITION LINES; MARKERS; WHEAT; IDENTIFICATION;
Keywords:
GENE TARGETING; LOW COPY SEQUENCES; RAPD; RFLP; BULK SEGREGANTS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
52
Recensione:
Indirizzi per estratti:
Citazione:
R.F. Eastwood et al., "A DIRECTED SEARCH FOR DNA-SEQUENCES TIGHTLY LINKED TO CEREAL CYST-NEMATODE RESISTANCE GENES IN TRITICUM-TAUSCHII", Genome, 37(2), 1994, pp. 311-319

Abstract

An improved system for identifying DNA sequences linked to a targetedregion was developed by fractionating DNA sequences prior to polymerase chain reaction (PCR) analysis. In an attempt to identify DNA markers linked to a strong CCN resistance gene, Ccn-D1, in Triticum tauschii, DNA samples from individuals homozygous for resistance and susceptibility at the Ccn-DI locus in a segregating progeny were bulked separately to produce ''near isogenic'' DNA pools. The polymerase chain reaction was employed to generate several DNA amplification products from each of the bulked DNA segregants using 240 random (RAPD) and 4 semirandom (consensus sequences of intron-splice junctions) primers: A DNA polymorphic fragment was apparent between the resistant and susceptible bulks using one of the semirandom primers. Hydroxylapatite chromatography of reannealed DNA (to C(0)t values > 100) was used to enrich low copy DNA sequences in the bulk DNA segregants (resistant and susceptible DNA pools). PCR analysis on the low copy enriched DNA pool increasedthe level of polymorphism detected between bulked segregants. One of the RAPD fragments present in only the resistant low copy DNA pool wascloned and mapped to the distal region of the long arm of chromosome 2D. By using the cloned RAPD fragment, csE20-2, to assay an RFLP locusin three independent F-2 progenies, complete cosegregation was obtained with the Ccn-DI locus. Joint segregation analysis from a genome-wide mapping of RFLP markers and a second CCN resistance in 2: tauschii, Ccn-D2, showed this locus to be loosely linked to the proximal region of chromosome 2.

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Documento generato il 29/11/20 alle ore 18:15:56