PCR primers for the estimation of contamination by seeds with normal cytoplasm in rapeseed lots bearing male-sterility-inducing Ogu-INRA cytoplasm

• Published in: Plant breeding Vol. 116; no. 4; pp. 390 - 392
• Main Authors: Tinchant, C, Defrance, M.C, Budar, F
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.09.1997; Blackwell; Wiley
• Subjects: Agronomy. Soil science and plant productions; Biological and medical sciences; Brassica napus; cytoplasmic male sterility; detection; DNA; DNA primers; estimation; Fundamental and applied biological sciences. Psychology; genes; Genetics and breeding of economic plants; genomics; Heterosis. Floral biology applications: apomixy, male sterility, incompatibility, varia; Life Sciences; male fertility; male-sterility; mitochondrial DNA; Ogu-INRA cytoplasm; orfB; orfb gene; PCR; Plant breeding: fundamental aspects and methodology; polymerase chain reaction; seed contamination; seeds; Vegetal Biology; Impurity; Primer; Mitochondrial DNA; Oil plant(vegetal); Contamination; Polymerase chain reaction; Cruciferae; Dicotyledones; Seed; Angiospermae; Brassica napus; Spermatophyta; Cytoplasmic male sterility; COLZA; CONTROLE; CYTOPLASME OGU-INRA; BIOLOGIE MOLECULAIRE
• ISSN: 0179-9541; 1439-0523
• DOI: 10.1111/j.1439-0523.1997.tb01018.x

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.