A new multiplex PCR test for the determination of Vrn-B1 alleles in bread wheat (Triticum aestivum L.)

Milec Z.,Tomková L., Sumíková T., Pánková K.
MOLECULAR BREEDING 30: 317-323, 2012

Klíčová slova: Wheat, Vrn-B1, PCR marker, Multiplex PCR
Abstrakt: Winter wheat requires vernalization, a long exposure to low but non-freezing temperatures, to promote reproductive development. The vernali-zation requirement in bread wheat (Triticum aes-tivum L.) is mainly controlled by theVrn-1genes that are located on chromosomes 5A, 5B and 5D. Dominant alleles confer spring habit and are epistatic to the recessive winter alleles which means that spring varieties carry at least one dominant allele. To date, two dominant and one recessiveVrn-B1alleles have been described. Vrn-B1a(formerly designated asVrn-B1)differs from the wintervrn-B1allele by a large deletion in intron 1. Vrn-B1b has an additional small deletion and is probably derived from Vrn-B1a. The novel allele described here and designated as Vrn-B1calso has a large deletion within intron 1 but with different break-points fromVrn-B1aorb, and sequence duplication, showing that this is an independently derived spring allele. By combining an exon 1 primer with previously published PCR primers it was possible to develop a multiplex PCR that distinguished all four alleles simultaneously. The multiplex PCR was validated by testing 320 winter wheat and 137 spring wheat varieties. This demonstrated that the novel Vrn-B1c allele was present in 25 spring varieties of diverse origin, showing this allele to be widely distributed.
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