Stahl RJ, Arnoldo M, Glavin TL, Goring DR, Rothstein SJ
Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Plant Cell. 1998 Feb;10(2):209-18. doi: 10.1105/tpc.10.2.209.
The self-incompatible (SI) Brassica napus line W1, which carries the 910 S allele, was transformed with an inactive copy of the 910 S locus receptor kinase (SRK) gene. Two transformed lines were analyzed based on their heritable ability to set self-seed. The first line was virtually completely self-compatible (SC), and reciprocal pollinations with the original W1 line demonstrated that only the stigma side of the SI phenotype was altered. An analysis of the expression of endogenous SRK-910 demonstrated that the mechanism of transgene action is via gene suppression. Furthermore, the expression of the S locus glycoprotein gene present in the 910 allele (SLG-910), SLG-A10, which is derived from a nonfunctional S allele, and an S locus-related gene were also suppressed. When the transgene was crossed into another SI line carrying the A14 S allele, it was also capable of suppressing the expression of the endogenous genes and of making this line SC. The second transgenic line studied was only partly SC. In this case as well, only the stigma phenotype was affected, although no gene suppression was detected for endogenous SRK-910 or SLG-910. In this line, the expression of the transgene most likely was causing the change in phenotype, and no effect was observed when this transgene was crossed into the other SI line. Therefore, this work reinforces the hypothesis that the SRK gene is required, but only for the stigma side of the SI phenotype, and that a single transgene can alter the SI phenotype of more than one S allele.
携带910 S等位基因的自交不亲和(SI)甘蓝型油菜品系W1用910 S位点受体激酶(SRK)基因的无活性拷贝进行了转化。基于它们产生自交种子的遗传能力分析了两个转化株系。第一个株系几乎完全自交亲和(SC),与原始W1品系的正反交表明只有SI表型的柱头一侧发生了改变。对内源SRK - 910表达的分析表明转基因作用机制是通过基因抑制。此外,910等位基因中存在的S位点糖蛋白基因(SLG - 910)、源自无功能S等位基因的SLG - A10以及一个S位点相关基因的表达也受到了抑制。当将该转基因导入另一个携带A14 S等位基因的SI品系时,它也能够抑制内源基因的表达并使该品系变为SC。研究的第二个转基因株系只是部分自交亲和。在这种情况下,同样只有柱头表型受到影响,尽管未检测到内源SRK - 910或SLG - 910的基因抑制。在这个株系中,转基因的表达很可能导致了表型变化,当将该转基因导入另一个SI品系时未观察到影响。因此,这项工作强化了以下假设:SRK基因是必需的,但仅对于SI表型的柱头一侧是必需的,并且单个转基因可以改变不止一个S等位基因的SI表型。