Oat-maize chromosome addition lines: A new system for mapping the maize genome
Oat-maize chromosome addition lines: A new system for mapping the maize genome
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DOI：
10.1073/pnas.94.8.3524
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发表时间：
1997-04-15
影响因子：
11.1
通讯作者：
Phillips, RL
中科院分区：
文献类型：
--
作者：
Ananiev, EV;RieraLizarazu, O;Phillips, RL
通过对燕麦（Avena sativa L., 2n = 6x = 42）与玉米（Zen mays L., 2n = 20）杂交的胚胎修复，在完整的燕麦基因组中添加了单个玉米染色体的新植株。利用具有21对燕麦染色体和1对玉米第9染色体的燕麦-玉米二体附加系构建了一个cosmid文库。使用高度重复的玉米特异性序列的多探针（标记片段的混合物用作探针）选择性地分离出含有玉米基因组DNA的cosmid克隆。将单个玉米cosmid克隆或其亚克隆片段与玉米和燕麦基因组DNA进行杂交，发现大多数高、中、低拷贝数DNA序列是玉米特有的。这种DNA标记允许在燕麦基因组背景下鉴定玉米基因组DNA。没有发现嵌合宇宙克隆体；显然，在这些新植物或克隆过程中，玉米附加染色体与燕麦基因组之间没有发生显著的遗传物质交换。从玉米基因组库中随机选择的无性系中，约95%的无性系可被多探针检测到。在燕麦背景下选择性检测玉米序列的能力使我们能够考虑将燕麦作为克隆特定玉米染色体或玉米染色体片段的宿主。通过辐照将玉米染色体片段渗入到燕麦基因组中，可以建立每个玉米染色体重叠片段的文库，用于开发玉米基因组的物理图谱。
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 (Zen 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.