Jung C, Koch R, Fischer F, Brandes A, Wricke G, Herrmann R G
Botanisches Institut, Ludwig-Maximilians-Universität München, FRG.
Mol Gen Genet. 1992 Mar;232(2):271-8. doi: 10.1007/BF00280006.
Genes conferring resistance to the beet cyst nematode (Heterodera schachtii Schm.) have been transferred to sugar beet (Beta vulgaris L.) from three wild species of the Procumbentes section using monosomic addition and translocation lines, because no meiotic recombination occurs between chromosomes of cultured and wild species. In the course of a project to isolate the nematode resistance genes by strategies of reverse genetics, probes were cloned from DNA of a fragmented B. procumbens chromosome carrying a resistance gene, which had been isolated by pulsed-field gel electrophoresis. One probe (pRK643) hybridized with a short dispersed repetitive DNA element, which was found only in wild beets, and thus may be used as a molecular marker for nematode resistance to progeneis of monosomic addition lines segregating resistant and susceptible individuals. Additional probes for the resistance gene region were obtained with a polymerase chain reaction (PCR)-based strategy using repetitive primers to amplify DNA located between repetitive elements. One of these probes established the existence of at least six different chromosomes from wild beet species, each conferring resistance independently of the others. A strict correlation between the length of the wild beet chromatin introduced in fragment addition and translocation lines and the repeat copy number has been used physically to map the region conferring resistance to a chromosome segment of 0.5-3 Mb.
利用单体附加系和易位系,已将来自Procumbentes组三个野生种的抗甜菜孢囊线虫(Heterodera schachtii Schm.)基因转移到甜菜(Beta vulgaris L.)中,因为栽培种和野生种的染色体之间不发生减数分裂重组。在通过反向遗传学策略分离线虫抗性基因的项目过程中,从通过脉冲场凝胶电泳分离出的携带抗性基因的裂叶B. procumbens染色体的DNA中克隆了探针。一个探针(pRK643)与一个短分散重复DNA元件杂交,该元件仅在野生甜菜中发现,因此可作为线虫抗性的分子标记,用于分离抗性和感病个体的单体附加系后代。利用基于聚合酶链反应(PCR)的策略,使用重复引物扩增重复元件之间的DNA,获得了抗性基因区域的其他探针。其中一个探针证实存在至少六条来自野生甜菜物种的不同染色体,每条染色体独立赋予抗性。在片段附加系和易位系中引入的野生甜菜染色质长度与重复拷贝数之间的严格相关性已被用于将抗性区域物理定位到0.5 - 3 Mb的染色体片段。