Steck N, Messmer M, Schaffner W, Bueter K B
Planta Med. 2001 Jun;67(4):384-5. doi: 10.1055/s-2001-14320.
The aim of this project was to establish RAPD markers to determine the percentage of sexual off-spring of the facultative apomictic plant species H. perforatum. We did reciprocal crosses between four different accessions (A x B, B x A, C x D, D x C) by mechanical emasculation and hand pollination. Genomic DNA of the parents and the off-spring was isolated and PCR conditions were optimized in order to obtain reproducible bands with RAPD markers. Of the 260 screened RAPD primers 127 revealed polymorphism between the parental lines of A and B, whereas 53 revealed no amplification products. Each progeny was tested for the presence of paternal bands with three primers. We found no sexual off-spring among the 22 progenies of A x B, the nine progenies of B x A and the ten progenies of D x C. However, we detected six sexual off-spring among the 45 progenies of C x D. We have proved that RAPD markers can be used to distinguish between sexual and apomictic off-spring in H. perforatum and that sexual off-spring can be obtained from intraspecific crosses. The percentage of sexual progeny might depend on the genotype of the parental lines.
本项目的目的是建立随机扩增多态性DNA（RAPD）标记，以确定 facultative apomictic 植物物种贯叶连翘有性后代的比例。我们通过机械去雄和人工授粉，对四个不同的种质（A×B、B×A、C×D、D×C）进行了正反交。提取了亲本和后代的基因组DNA，并优化了PCR条件，以便用RAPD标记获得可重复的条带。在筛选的260个RAPD引物中，127个在A和B的亲本系之间显示出多态性，而53个没有扩增产物。用三个引物对每个后代进行父本条带检测。我们在A×B的22个后代、B×A的9个后代和D×C的10个后代中未发现有性后代。然而，在C×D的45个后代中检测到6个有性后代。我们已经证明，RAPD标记可用于区分贯叶连翘的有性和无融合生殖后代，并且有性后代可从种内杂交中获得。有性后代的比例可能取决于亲本系的基因型。
