Ananiev E V, Riera-Lizarazu O, Rines H W, Phillips R L
Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul 55108, USA.
Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3524-9. doi: 10.1073/pnas.94.8.3524.
Novel plants with individual maize chromosomes added to a complete oat genome have been recovered via embryo rescue from oat (Avena sativa L., 2n = 6x = 42) x maize (Zea mays L., 2n = 20) crosses. An oat-maize disomic addition line possessing 21 pairs of oat chromosomes and one maize chromosome 9 pair was used to construct a cosmid library. A multiprobe (mixture of labeled fragments used as a probe) of highly repetitive maize-specific sequences was used to selectively isolate cosmid clones containing maize genomic DNA. Hybridization of individual maize cosmid clones or their subcloned fragments to maize and oat genomic DNA revealed that most high, middle, or low copy number DNA sequences are maize-specific. Such DNA markers allow the identification of maize genomic DNA in an oat genomic background. Chimeric cosmid clones were not found; apparently, significant exchanges of genetic material had not occurred between the maize-addition chromosome and the oat genome in these novel plants or in the cloning process. About 95% of clones selected at random from a maize genomic cosmid library could be detected by the multiprobe. The ability to selectively detect maize sequences in an oat background enables us to consider oat as a host for the cloning of specific maize chromosomes or maize chromosome segments. Introgressing maize chromosome segments into the oat genome via irradiation should allow the construction of a library of overlapping fragments for each maize chromosome to be used for developing a physical map of the maize genome.
通过对燕麦(Avena sativa L.,2n = 6x = 42)与玉米(Zea mays L.,2n = 20)杂交的胚胎进行挽救,已获得了添加了单个玉米染色体的完整燕麦基因组的新型植株。利用一个具有21对燕麦染色体和一对玉米9号染色体的燕麦 - 玉米二体附加系构建了一个黏粒文库。使用高度重复的玉米特异性序列的多探针(用作探针的标记片段混合物)来选择性分离含有玉米基因组DNA的黏粒克隆。将单个玉米黏粒克隆或其亚克隆片段与玉米和燕麦基因组DNA杂交,结果表明大多数高、中、低拷贝数的DNA序列是玉米特异性的。这类DNA标记可在燕麦基因组背景中鉴定玉米基因组DNA。未发现嵌合黏粒克隆;显然,在这些新型植株或克隆过程中,玉米附加染色体与燕麦基因组之间未发生显著的遗传物质交换。从玉米基因组黏粒文库中随机挑选的克隆中,约95%可被多探针检测到。在燕麦背景中选择性检测玉米序列的能力使我们能够将燕麦视为克隆特定玉米染色体或玉米染色体片段的宿主。通过辐射将玉米染色体片段渗入燕麦基因组,应该能够构建每个玉米染色体的重叠片段文库,用于绘制玉米基因组的物理图谱。