摘要
利用大麦 (HordeumvulgareL .)第 5染色体上RFLP探针衍生的 19个序列标志位点PCR(STS_PCR)引物对“中国春”小麦 (TriticumaestivumL .) (CS)及其ph1b突变体基因组总DNA进行PCR扩增 ,筛选出Ph1基因的一个连锁标记 ,再用“中国春”第 5部分同源群缺体_四体系和CS×ph1b突变体F2 群体证明并定位于离Ph1基因近着丝点端 5 .7cM (centiMorgan)处。然后将该标记转换成特异的序列特征扩增区 (SCAR)标记。以“阿勃”5B缺体为桥梁亲本 ,冬小麦“京 411”为受体亲本 ,“中国春”ph1b突变体为供体亲本 ,进行三轮杂交和一轮自交 ,每一轮经减数分裂分析和SCAR标记的辅助选择 ,快速地筛选出了ph1b基因型 ,并选得一个冬小麦“京 411”的ph1b中间代换系。
The genomic DNA of common wheat ( Triticum aestivum L.) “Chinese Spring” (CS) and its ph1b mutant were analyzed by using 19 sequence tagged site PCR (STS_PCR) primers, which derived from RFLP probes from barley ( Hordeum vulgare L.) chromosome 5H. One marker was identified on wheat chromosome 5BL, which is 5.7 cM (centiMorgan) proximal to Ph1 gene, using the CS homoeologous group 5 nullisomic_tetrasomic, ditelosomic 5BL line and an F 2 population from CS×ph1b mutant. This linked PCR marker was converted into a more specific sequence characterized amplified region (SCAR) marker. To obtain a new winter wheat line containing ph1b gene, the authors used a nullisomic 5B line of “Abbodanza” as a bridge parent and crossed respectively with the CS ph1b mutant (donor) and a winter wheat variety, “Jing 411” (recipient). The meiotic chromosome pairing was checked in the progeny of each cross, as well as using the marker_assistant selection of the SCAR marker identified for ph1b gene. After three inter_crossing and one selfing, a relatively stable ph1b substitution line of winter wheat with “Jing 411” background was obtained.
关键词
小麦
ph1b突变体
分子标记
ph1基因
common wheat
Chinese Spring
ph1b mutant
sequence characterized amplified region (SCAR) marker
marker_assistant selection
