Goring D R, Banks P, Beversdorf W D, Rothstein S J
Department of Molecular Biology and Genetics, University of Guelph, Ontario, Canada.
Mol Gen Genet. 1992 Aug;234(2):185-92. doi: 10.1007/BF00283838.
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.
通过将自交不亲和的白菜型油菜植株的S位点渗入到Westar品种中，培育出了一个自交不亲和的油菜品质甘蓝型油菜品系（W1）。使用从S位点糖蛋白（SLG）等位基因的保守区域衍生的引物进行聚合酶链反应（PCR），获得了活性SLG基因（910）的中心区域。随后使用PCR- cDNA末端快速扩增（RACE）程序克隆了该910基因cDNA的其余部分。序列分析表明，910 cDNA与I类自交不亲和系相关的SLG等位基因具有高度的序列相似性。发现原始自交亲和的cv. Westar（甘蓝型油菜）中不存在910基因，并且在自交不亲和的W1和自交亲和的Westar杂交产生的混合群体中，910基因与自交不亲和性发生分离。RNA印迹分析表明，随着花蕾接近开花，柱头中存在高水平的910 mRNA。因此，已鉴定出通过回交从白菜型油菜转移到cv. Westar品系的W1的SLG等位基因。
