Use of the polymerase chain reaction to isolate an S-locus glycoprotein cDNA introgressed from Brassica campestris into B. napus ssp. oleifera.
A self-incompatible canola-quality Brassica napus ssp. oleifera line (W1) was generated by introgressing the S-locus from a self-incompatible B. campestris plant into the Westar cultivar. Using the polymerase chain reaction (PCR) with primers derived from conserved regions in S-locus glycoprotein (SLG) alleles, the central region of the active SLG gene (910) was obtained. The remaining portions of the cDNA for this 910 gene were subsequently cloned using the PCR-rapid amplification of cDNA ends (RACE) procedure. Sequence analysis revealed that the 910 cDNA show a high degree of sequence similarity to SLG alleles associated with Class I self-incompatible lines. The 910 gene was found to be absent in the original self-compatible cv. Westar (B. napus) and segregated with self-incompatibility in a mixed population generated from a cross between self-incompatible W1 and self-compatible Westar. RNA blot analysis indicated that high levels of 910 mRNAs were present in the stigma as buds approached anthesis. Thus, the SLG allele of W1 transferred from B. campestris via backcrosses to a line of cv. Westar has been identified.
Goring
D R
DR
Department of Molecular Biology and Genetics, University of Guelph, Ontario, Canada.
Banks
P
P
Beversdorf
W D
WD
Rothstein
S J
SJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D001937:Brassica / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
D006023:Glycoproteins; D010940:Plant Proteins; C064156:self-incompatibility locus specific glycoprotein, Brassica; D004247:DNA