番茄耐低温相关基因的SRAP标记筛选

Identification of the Specific SRAP Marker Associated with Cold Resistance of Tamoto

以番茄耐低温和不耐低温基因组DNA为材料进行SRAP分析,共筛选了225对SRAP引物,其中27对引物在两池之间表现差异,经测序只有Me2Em5扩增出与番茄耐低温相关的差异性片段,大小约为273bp,该片段仅在耐低温植株中稳定扩增。经Blast分析比对,该片段与已报道的PEG和低温诱导后在沙冬青幼苗中表达基因的cDNA片段同源。根据差异片段序列设计特异引物,将M2E5-273标记成功转化为更稳定的SCAR标记。

SRAP （sequence-related amplified polymorphism） was used to analyze the genome DNA of cold- resistanced tomato and normal tomato. Total 225 pairs of SRAP primers were used. The amplification of twenty-seven primers was polymorphic in the two lines. Only the Me2Em5 primer had fragment that was highly homologous with cold-resistanced plant testing. And a 273-bp specific band M2E5-273 was detected in coldresistanced tomato but not in normal tomato. Analysis of the sequence showed that this fragment was highly homologous with Ammopiptanthus mongolicus seedling PEG and cold-induced transcript-derived fragment. After cloning and sequencing, specific primers were designed to transform the SRAP marker to more stable SCAR marker, which was named M2E5-273.