The invention discloses a preparation method and an application of a
molecular marker of a rape male
sterility restoring
gene. The preparation method comprises the following steps of A, designing degenerate primers according to PPR
gene conservative
amino acid sites, B, extracting a lamina
DNA of a rape
cytoplasm male
sterility restoring line through a cetyltrimethyl
ammonium bromide (CTAB) method and magnifying the lamina
DNA through a touchdown
polymerase chain reaction (PCR) process to obtain a specific restoring
gene candidate segment, C, carrying out
recovery, conversion, sequencing and
sequence analysis processes on PCR products, D, carrying out 5 ' RACE and 3 ' RACE processes and splicing a sequencing result to obtain a full-length cDNA sequence, and E, designing
specific primers in 5 ' UTR and 3 ' UTR zones of the full-length cDNA sequence according to the full-length cDNA sequence to obtain a
molecular marker. The invention also discloses an application of a restoring gene
molecular marker in
hybrid rape parent breeding. The preparation method has the advantages of improvements of a speed of restorer seeding and an efficiency of
verification, simple and easy method, good
operability,
rapid identification speed of restoring genes, great reduction of necessary
workload of
test cross and progeny planting observation in a restorative
verification process, and improvement of an efficiency of
hybrid seeding.