Gao Wenxiang, Chen Z Jeffrey, Yu John Z, Raska Dwaine, Kohel Russell J, Womack James E, Stelly David M
Department of Soil and Crop Sciences, Texas A&M University, College Station, Texas 77843-2474, USA.
Genetics. 2004 Jul;167(3):1317-29. doi: 10.1534/genetics.103.020479.
We report the development and characterization of a "wide-cross whole-genome radiation hybrid" (WWRH) panel from cotton (Gossypium hirsutum L.). Chromosomes were segmented by gamma-irradiation of G. hirsutum (n = 26) pollen, and segmented chromosomes were rescued after in vivo fertilization of G. barbadense egg cells (n = 26). A 5-krad gamma-ray WWRH mapping panel (N = 93) was constructed and genotyped at 102 SSR loci. SSR marker retention frequencies were higher than those for animal systems and marker retention patterns were informative. Using the program RHMAP, 52 of 102 SSR markers were mapped into 16 syntenic groups. Linkage group 9 (LG 9) SSR markers BNL0625 and BNL2805 had been colocalized by linkage analysis, but their order was resolved by differential retention among WWRH plants. Two linkage groups, LG 13 and LG 9, were combined into one syntenic group, and the chromosome 1 linkage group marker BNL4053 was reassigned to chromosome 9. Analyses of cytogenetic stocks supported synteny of LG 9 and LG 13 and localized them to the short arm of chromosome 17. They also supported reassignment of marker BNL4053 to the long arm of chromosome 9. A WWRH map of the syntenic group composed of linkage groups 9 and 13 was constructed by maximum-likelihood analysis under the general retention model. The results demonstrate not only the feasibility of WWRH panel construction and mapping, but also complementarity to traditional linkage mapping and cytogenetic methods.
我们报道了从棉花(陆地棉)构建的一个“远缘杂交全基因组辐射杂种”(WWRH)群体的开发及特征分析。通过对陆地棉(n = 26)花粉进行γ射线辐照来分割染色体,并在海岛棉卵细胞(n = 26)体内受精后拯救分割后的染色体。构建了一个5千拉德γ射线的WWRH定位群体(N = 93),并在102个SSR位点进行了基因分型。SSR标记保留频率高于动物系统,且标记保留模式具有信息性。使用程序RHMAP,102个SSR标记中的52个被定位到16个同线群中。连锁群9(LG 9)的SSR标记BNL0625和BNL2805通过连锁分析共定位,但它们的顺序通过WWRH植株间的差异保留得以解析。两个连锁群,LG 13和LG 9,被合并为一个同线群,染色体1连锁群标记BNL4053被重新定位到染色体9。细胞遗传学材料的分析支持LG 9和LG 13的同线性,并将它们定位到染色体17的短臂上。它们还支持将标记BNL4053重新定位到染色体9的长臂上。在一般保留模型下,通过最大似然分析构建了由连锁群9和13组成的同线群的WWRH图谱。结果不仅证明了构建WWRH群体和定位的可行性,还证明了其与传统连锁定位和细胞遗传学方法的互补性。