《植物生理学报》 2011, 47(1): 102-106

番茄耐低温相关基因的SRAP 标记筛选

郭彩杰¹, 侯丽霞^2,*, 崔娜^1,*, 韩明利¹

1 沈阳农业大学生物科学技术学院, 沈阳110161; 2 山东省农科院蔬菜研究所, 山东省设施蔬菜生物学重点实验室, 国家蔬菜改良中心山东分中心, 济南250100

通信作者：侯丽霞；E-mail: houlx2006@126.com, syaua@163.com；Tel: 0531-83179184;024-88487163

摘　要：

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 273 bp, 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。

关键词：番茄; 耐低温; SRAP; 分子标记

收稿：2010-09-13   修定：2010-11-03

资助：国家“863” 计划项目(2006AA100108-3-1)和辽宁省教育厅科学技术研究项目(2008623)。

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

GUO Cai-Jie¹, HOU Li-Xia^2,*, CUI Na^1,*, HAN Ming-Li¹

¹Biological Science and Technology College, Shenyang Agricultural University, Shenyang 110161, China; ²Institute of Vegetables,Shandong Academy of Agricultural Science, Key Laboratory for Biology of Greenhouse Vegetable of Shandong Province, National Center for Vegetable Improvement (Shandong), Jinan 250100, China

Corresponding author: HOU Li-Xia; E-mail: houlx2006@126.com, syaua@163.com; Tel: 0531-83179184;024-88487163

Abstract:

SRAP (sequence-related amplified polymorphism) was used to analyze the genome DNA of coldresistanced 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.

Key words: tomato; cold-resistanced; SRAP; molecular marker