摘要
根据黑麦（ＳｅｃａｌｅｃｅｒｅａｌｅＬ．）与小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）ｒＲＮＡ基因间隔区序列差异，按Ｋｏｅｂｎｅｒ设计的引物序列，合成了黑麦特异引物ＮＯＲＲ１。运用该引物对不同植物材料进行ＰＣＲ扩增。观察表明，含有黑麦１Ｒ染色体的植物材料均扩增出黑麦的特异带，但含有其他黑麦染色体的小麦种质、普通小麦品种及其近缘物种长穗偃麦草（Ａｇｒｏｐｙｒｏｎｅｌｏｎｇａｔｕｍ（Ｈｏｓｔ）Ｂｅａｕｖ．）、簇毛麦（ＨａｙｎａｌｄｉａｖｉｌｏｓａＳｈｕｒ．）及大麦（ＨｏｒｄｅｕｍｖｕｌｇａｒｅＬ．）皆无任何扩增产物。荧光原位杂交进一步显示，ＮＯＲＲ１引物的结合位点仅位于１Ｒ染色体上核仁组织区。因此可以认为，ＮＯＲＲ１引物是在小麦遗传背景中鉴别黑麦１Ｒ染色体可靠的分子标记。
Based on the differences of rRNA intergenic sequences between wheat ( Triticum aestivum L.) and rye ( Secale cereale L.), rye specific primer set NOR R1 was synthesized according to Koebner' design. PCR analyses were carried out on different DNA substrates of common wheat and its relatives such as Agropyron elongataum (Host) Beauv., Haynaldia villosa Shur. and Hordeum vulgare L. The results confirmed that NOR R1 primer set is specific to rye. It was found that PCR using DNAs from wheat materials containing 1R chromosome resulted in the specific amplification products of rye, whereas no amplification product was detected in PCR when using DNAs with other rye chromosomes. FISH (Fluorescent in situ Hybridization) further revealed that the binding sites for the primer set NOR R1 were only on nucleolar organizing region of chromosome 1R. These results indicated that the primer set NOR R1 provides a useful means for molecular tagging of rye chromosomes 1R in wheat genetic background.
基金
中国科学院"九.五"重大项目
关键词
核仁组织区
PCR
分子标记
黑麦
小麦
Nucleolar organizing region, PCR, Molecular marker, Rye, Wheat
