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PCR primers for the estimation of contamination by seeds with normal cytoplasm in rapeseed lots bearing male‐sterility‐inducing Ogu‐INRA cytoplasm
Author(s) -
Tinchant C.,
Defrance M.C.,
Budar F.
Publication year - 1997
Publication title -
plant breeding
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.583
H-Index - 71
eISSN - 1439-0523
pISSN - 0179-9541
DOI - 10.1111/j.1439-0523.1997.tb01018.x
Subject(s) - rapeseed , cytoplasmic male sterility , biology , polymerase chain reaction , cytoplasm , nucleic acid , brassica , mitochondrial dna , dna , contamination , primer (cosmetics) , genetics , gene , microbiology and biotechnology , botany , chemistry , organic chemistry , ecology
A polymerase chain reaction (PCR)‐based test was developed to detect seeds bearing the ‘so‐called’ normal rapeseed cytoplasm in seed lots with an OGU‐INRA type cytoplasm. The test is based on the amplification of the orfB region of male fertile rapeseed mitochondrial DNA (mtDNA). The amplification reaction uses total nucleic acids of young seedlings, extracted in bulk. After the sequencing of the orfB gene region in the normal Brassica mtDNA, primers were designed for its amplification by PCR. Although the specificity of amplification for the male fertile (mf) rapeseed cytoplasm is partly impaired by the presence of tiny amounts of this fragment in the mtDNA of male sterile (ms) plants, this test proved to be applicable for the estimation of the level of contamination in seed lots in reconstituted mixes as well as in real lots.