Coram Tristan E, Settles Matthew L, Wang Meinan, Chen Xianming
Wheat Genetics, Quality, Physiology and Disease Research Unit, U.S. Department of Agriculture, Agricultural Research Service, Pullman, WA 99163, USA.
Theor Appl Genet. 2008 Aug;117(3):401-11. doi: 10.1007/s00122-008-0784-5. Epub 2008 May 10.
DNA polymorphisms are valuable for several applications including genotyping, molecular mapping and marker-assisted selection. The 55 K Affymetrix Wheat GeneChip was used to survey expression level polymorphisms (ELPs) and single-feature polymorphisms (SFPs) between two near-isogenic wheat genotypes (BC(7):F(4)) that differ for the Yr5 stripe rust resistance locus, with the objective of developing genetic markers linked to Yr5. Ninety-one probe sets showing ELPs and 118 SFP-containing probe sets were identified between isolines, of which just nine ELP probe sets also contained SFPs. The proportion of the transcriptome estimated to be variable between isolines from this analysis was 0.30% for the ELPs and 0.39% for the SFPs, which was highly similar to the theoretical genome difference between isolines of ~0.39%. Using wheat-rice synteny, both ELPs and SFPs mainly clustered on long arms of rice chromosomes four and seven, which are syntenous to wheat chromosomes 2L (Yr5 locus) and 2S, respectively. The strong physical correlation between the two types of polymorphism indicated that the ELPs may be regulated by cis-acting DNA polymorphisms. Twenty SFPs homologous to rice 4L were used to develop additional genetic markers for Yr5. Physical mapping of the probe sets containing SFPs to wheat chromosomes identified nine on the target chromosome 2BL, thus wheat-rice synteny greatly enhanced the selection of SFPs that were located on the desired wheat chromosome. Of these nine, four were converted into polymorphic cleaved amplified polymorphic sequence (CAPS) markers between Yr5 and yr5 isolines, and one was mapped within 5.3 cM of the Yr5 locus. This study represents the first array-based polymorphism survey in near-isogenic genotypes, and the results are applied to an agriculturally important trait.
DNA多态性在包括基因分型、分子图谱构建和标记辅助选择等多种应用中具有重要价值。使用55K Affymetrix小麦基因芯片来检测两个在抗条锈病基因Yr5位点存在差异的近等基因小麦基因型(BC(7):F(4))之间的表达水平多态性(ELP)和单特征多态性(SFP),目的是开发与Yr5连锁的遗传标记。在近等基因系之间鉴定出91个显示ELP的探针组和118个包含SFP的探针组,其中只有9个ELP探针组也包含SFP。通过该分析估计,近等基因系之间转录组的可变比例对于ELP为0.30%,对于SFP为0.39%,这与近等基因系之间约0.39%的理论基因组差异高度相似。利用小麦-水稻共线性关系,ELP和SFP主要聚集在水稻染色体4号和7号的长臂上,它们分别与小麦染色体2L(Yr5位点)和2S同源。两种多态性之间的强物理相关性表明ELP可能受顺式作用DNA多态性调控。使用20个与水稻4L同源的SFP开发了更多Yr5的遗传标记。将包含SFP的探针组物理定位到小麦染色体上,在目标染色体2BL上鉴定出9个,因此小麦-水稻共线性关系极大地增强了对位于所需小麦染色体上的SFP的选择。在这9个中,4个被转化为Yr5和yr5近等基因系之间的多态性酶切扩增多态性序列(CAPS)标记,其中1个被定位在Yr5位点5.3 cM范围内。本研究代表了首次在近等基因基因型中基于芯片的多态性检测,其结果应用于一个重要的农业性状。