Rapid control of purity for the cytoplasm of male-sterile seed stocks by means of a dot hybridization assay.
To produce hybrids, one member of the parental line is genetically made male-sterile. This male-sterile trait is encoded by mitochondria so that it is maternally inherited. Consequently, the progeny of a male-sterile plant is fully sterile. Nevertheless, during the handling of cytoplasmic male-sterile seed stocks, some mixture with seeds of the maintainer lines can occur. Up to the present time, the only way to check the homogeneity of the cytoplasmic male-sterile seed stock was to grow the plants until flowering time. We have developed a method which can be used immediately after the harvest, allowing us to check samples from both sunflower and sugar beet. We used the mitochondrial plasmid, present only in the maintainer lines, as a probe for the total nucleic acids prepared from the cytoplasmic male-sterile seed stocks which might be contaminated. The signals compared to those of samples artificially contaminated allow us to measure as few as one male-fertile seed in 1000 seeds in a rapid and accurate manner.
Santoni
S
S
INRA, Station de Génétique et d'Amélioration des Plantes, Dijon, France.
Faivre-Rampant
P
P
Moreau
E
E
Bervillé
A
A
D015152:Blotting, Northern; D015139:Blotting, Southern; D015342:DNA Probes; D004272:DNA, Mitochondrial / Q000235:genetics*; D003720:Densitometry; D004587:Electrophoresis, Agar Gel; D006368:Helianthus / Q000166:cytology / Q000235:genetics* / Q000502:physiology; D009693:Nucleic Acid Hybridization; D010945:Plants, Edible / Q000166:cytology / Q000235:genetics* / Q000502:physiology; D010957:Plasmids; D012639:Seeds / Q000166:cytology / Q000235:genetics* / Q000502:physiology
D015342:DNA Probes; D004272:DNA, Mitochondrial