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通过 FISH 揭示的短柄草和普通小麦之间的同线性。

Synteny between Brachypodium distachyon and Hordeum vulgare as revealed by FISH.

机构信息

Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), 06466 Gatersleben, Germany.

出版信息

Chromosome Res. 2010 Nov;18(7):841-50. doi: 10.1007/s10577-010-9166-3. Epub 2010 Nov 23.

Abstract

We investigated by fluorescence in situ hybridization (FISH) the synteny between Brachypodium distachyon with a small genome (1C = 320 Mb) and barley with a large genome (1C = 5,100 Mb) at the chromosome level. Reciprocal genomic in situ hybridization (GISH) between B. distachyon and barley labeled mainly 45S ribosomal DNA loci, indicating that most high copy DNA is weakly conserved between both grasses. Of 13 BAC clones with inserts from different B. distachyon chromosomes, only two belonging to chromosome 1 yielded hybridization signals on a barley metaphase chromosome (on 7HS and 7HL, respectively), confirming synteny between both chromosomes. FISH experiments to characterize the synteny of single-copy loci were performed. Two of four Brachypodium sylvaticum BACs spanning a 223-kb interval homologous to the region of barley that harbors a gibberellic-acid-insensitive semi-dwarfing gene, sdw3, hybridized specifically to a central position of B. distachyon chromosome 1 short arm but not to the homologous region of the barley genome. Repeat-free sequences PCR amplified from four non-overlapping barley BACs linked to the core of Sdw3 region yielded signals at distinct positions in the middle of barley chromosome arm 2HS. Together, these results (1) confirmed the synteny between B. distachyon chromosome 1 and barley chromosomes 2H and 7H at the cytological level, (2) indicated mid-arm position for the Sdw3 locus genetically mapped at the centromere of barley chromosome 2H, and (3) proved that the sdw3 core interval of < 100 kb in B. distachyon corresponds to a megabase-sized syntenic region in barley.

摘要

我们通过荧光原位杂交(FISH)研究了小基因组(1C = 320 Mb)的短柄草和大基因组(1C = 5,100 Mb)大麦之间的染色体水平上的同线性。短柄草和大麦之间的相互基因组原位杂交(GISH)主要标记 45S 核糖体 DNA 基因座,表明两种禾本科植物之间大多数高拷贝 DNA 是弱保守的。在来自不同短柄草染色体的 13 个 BAC 克隆中,只有两个属于染色体 1 的克隆在大麦中期染色体上产生杂交信号(分别位于 7HS 和 7HL 上),证实了这两个染色体之间的同线性。进行了 FISH 实验以表征单拷贝基因座的同线性。在包含赤霉素不敏感半矮化基因 sdw3 的大麦区域中,有 4 个短柄草 sylvaticum BAC 跨越 223-kb 间隔,其中 2 个 BAC 特异性杂交到短柄草 1 号染色体短臂的中心位置,但不与大麦基因组的同源区域杂交。从与 Sdw3 区域核心相连的四个非重叠大麦 BAC 中扩增的无重复序列 PCR 产物在大麦染色体 2HS 臂的中间位置产生信号。这些结果(1)在细胞学水平上证实了短柄草 1 号染色体与大麦 2H 和 7H 染色体之间的同线性,(2)表明 Sdw3 基因座在大麦染色体 2H 着丝粒处遗传定位的臂中位置,(3)证明短柄草中的 sdw3 核心间隔区<100 kb 对应于大麦中一个兆碱基大小的同线性区域。

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