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期 刊 ：植物保护学报 2007年

关键词：致病疫霉；随机扩增多态性DNA；序列特征扩增区域；甲霜灵抗性；

Keywords:Phytophthora infestans, RAPD, SCAR, metalaxyl-resistance,

摘 要 ：建立马铃薯晚疫病菌抗甲霜灵SCAR(sequenced characterized amplified region,序列特征扩增区域)标记,以马铃薯晚疫病菌对甲霜灵高抗菌株HD01-3和对甲霜灵高感菌株DK98-1为亲本,通过无性单游动孢子分离和有性杂交获得菌株HD01-3无性后代群体、菌株DK98-1无性后代群体以及F₁代分离群体,以此为试验材料,利用BSA法(bulked segregant analysis,分离群体分组分析法)构建抗感基因池对后代菌系的甲霜灵抗性进行RAPD(random amplified polymorphic DNA,随机扩增多态性DNA)分析。从178条RAPD随机引物中找到一条特异性引物S2054,其可以扩增出一个与晚疫病菌对甲霜灵抗性相关的遗传标记,将该特异条带回收、克隆、测序,发现此标记大小为457bp,根据测序结果设计特异PCR引物,用于扩增抗感基因池,成功地将特异RAPD标记转化为SCAR标记。初步建立了马铃薯晚疫病菌抗甲霜灵SCAR标记,辅助监测晚疫病菌对甲霜灵的抗性。

Abstract:A SCAR (sequenced characterized amplified region) marker for metalaxyl-resistance of Phytophthora infestans was developed. With highly metalaxyl-resistant isolate HD01-3 and highly metalaxyl-sensitive isolate DK98-1 of Phytophthora infestans as parents, strains of asexual single-zoospores of HD01-3 and DK98-1 were reproduced by single-zoospore isolation and F₁ progeny were reproduced from the sexual crosses HD01-3×DK98-1. DNA pools of metalaxyl-resistant and metalaxyl-sensitive strains were developed by BSA (bulked segregant analysis). By using RAPD (random amplified polymorphic DNA) analysis, the specific primer S2054 was screened out from 178 random 10bp primers. One genetic marker that linked with metalaxyl-resistance of Phytophthora infestans could be amplified using primer S2054. This band was collected, colned and sequenced, the length was 457bp. Primers were designed based on the sequence for amplifying DNA pools of metalaxyl-resistant and metalaxyl-sensitive strains. RAPD marker was successfully converted into SCAR marker. A SCAR marker for metalaxyl-resistance of Phytophthora infestans was preliminarily found and could monitor the metalaxyl-resistance of Phytophthora infestans.

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