Francki M. G., Berzonsky W. A., Ohm H. W., Anderson J. M.
Department of Agriculture, Locked BMG 4, Bently Delivery Centre WA 6983, Australia.
Theor Appl Genet. 2002 Feb;104(2-3):184-191. doi: 10.1007/s00122-001-0781-4.
Cereal centromeres consist of a complex organization of repetitive DNA sequences. Several repetitive DNA sequences are common amongst members of the Triticeae family, and others are unique to particular species. The organization of these repetitive elements and the abundance of other types of DNA sequences in cereal centromeres are largely unknown. In this study, we have used wheat-rye translocation lines to physically map 1BL.1RS centromeric breakpoints and molecular probes to obtain further information on the nature of other types of centromeric DNA sequences. Our results, using the rye-specific centromeric sequence, pAWRC.1, indicate that 1BL.1RS contains a small portion of the centromere from 1R of rye. Further studies used molecular markers to identify centromeric segments on wheat group-1 chromosomes. Selected RFLP markers, clustered around the centromere of wheat homoeologous group-1S chromosomes, were chosen as probes during Southern hybridization. One marker, PSR161, identified a small 1BS segment in all 1BL.1RS lines. This segment maps proximal to pAWRC.1 in 1BL.1RS and on the centromere of 1B. Sequence analysis of PSR161 showed high homology to HSP70 genes and Northern hybridization showed that this gene is constitutively expressed in leaf tissue and induced by heat shock and light stimuli. The significance of this work with respect to centromere organization and the possible significance of this HSP70 gene homologue are discussed.
谷类着丝粒由复杂的重复DNA序列组织构成。几种重复DNA序列在小麦族成员中很常见,而其他一些则是特定物种所特有的。这些重复元件的组织方式以及谷类着丝粒中其他类型DNA序列的丰度在很大程度上尚不清楚。在本研究中,我们利用小麦-黑麦易位系对1BL.1RS着丝粒断点进行了物理定位,并使用分子探针获取有关其他类型着丝粒DNA序列性质的更多信息。我们使用黑麦特异性着丝粒序列pAWRC.1的结果表明,1BL.1RS包含来自黑麦1R着丝粒的一小部分。进一步的研究使用分子标记来鉴定小麦第1组染色体上的着丝粒区段。在Southern杂交过程中,选择了围绕小麦同源第1组染色体着丝粒聚集的选定RFLP标记作为探针。一个标记PSR161在所有1BL.1RS系中鉴定出一个小的1BS区段。该区段在1BL.1RS中位于pAWRC.1的近端,在1B的着丝粒上。PSR161的序列分析显示与HSP70基因具有高度同源性,Northern杂交表明该基因在叶组织中组成性表达,并受热激和光刺激诱导。讨论了这项工作对着丝粒组织的意义以及该HSP70基因同源物可能的意义。