Identification of a mutant fatty acid elongase allele from zero-percent erucic acid Sinapis alba |
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Authors: | W J Drost D J Somers G Rakow J P Raney |
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Institution: | (1) Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon, Saskatchewan, S7N 0X2, Canada e-mail: rakowg@em.agr.ca Tel.: 1-306-956-7235, CA;(2) Agriculture and Agri-Food Canada, Cereal Research Centre, 195 Dafoe Road, Winnipeg, Manitoba, R3T 2M9, Canada, CA |
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Abstract: | The fae1 gene codes for KCS (β-keto-acyl-CoA synthase), the candidate enzyme for elongation of oleic acid to eicosenoic acid and erucic
acid (C22:1) in various oilseed species. Degenerate primers for the fae1 gene were used to amplify and clone fae1 gene homologs in high and zero C22:1 Sinapis alba. Under stringent PCR conditions, a polymorphism was revealed between the two genotypes and was mapped as a fae1 marker in an F2 population derived from a cross between high and zero C22:1 S. alba. The fae1 marker co-segregated with C22:1 content and the C22:1 phenotypic locus. In addition, a set of 11 RAPD markers for C22:1 in
S. alba was identified. Cloning and sequencing of the fae1 alleles in high and zero C22:1 S. alba revealed two amino-acid substitutions specific to zero C22:1 S. alba. The underlying nucleotide substitution for one of the amino-acid substitutions and an adjacent silent nucleotide substitution
were used to design primers for allele-specific amplicons for both the wild-type and zero C22:1 alleles. The two diagnostic
PCR tests are reliable selection tools to identify S. alba carrying one or both of the wild-type and mutant C22:1 alleles of the KCS gene.
Received: 27 July 2000 / Accepted: 1 February 2001 |
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Keywords: | RAPD fae1 Erucic acid Sinapis alba |
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