与番茄颈腐根腐病紧密连锁的SCAR标记开发

Development of SCAR Markers Closely Linked with Fusarium Crown and Root Rot in Tomato

利用与番茄颈腐根腐病抗性基因Frl紧密连锁的CAPS标记C2-25,开发设计了新的SCAR标记,用于快速检测Frl抗性材料。使用C2-25引物,在25份已知表型的番茄材料中进行PCR,产物单克隆测序后发现,19份纯合抗病材料存在1条完全相同的抗病序列;4份纯合感病材料存在1条完全相同的感病序列;2份杂合抗病材料均存在前述的抗病序列和感病序列。根据抗病、感病序列SNP差异设计引物,最终筛选得到可扩增出370 bp抗病特异片段和520 bp感病特异片段的番茄颈腐根腐病连锁标记R-6F/2R、S-3F/4R。在500株F2材料中进行验证,检测到122株纯合抗病单株、257株杂合抗病单株和121株纯合感病单株。F2材料人工接种鉴定结果显示,473株单株抗性与分子鉴定结果一致,分子标记辅助鉴定的准确率达到94.6%。开发的SCAR标记准确度高、成本低、操作简便,可用于Frl基因的分子标记辅助育种。

Taking advantage of the CAPS marker C2-25 closely linked to the resistant gene Frl to tomato Fusarium crown and root rot,this paper developed and designed new SCAR markers to quickly detect the dominant resistant gene Frl.Using C2-25 as the primer,this experiment conducted PCR among the 25 known phenotypic tomato material through monoclonal sequencing,of which 19 homozygous resistant material all containing one same resistant sequence,4 homozygous susceptible material all containing one same susceptible sequence,both 2 heterozyous resistant material containing the above mentioned resistant and susceptible sequences.According to SNP differences between the resistant and susceptible sequences,we designed primers and finally found out primer combination R-6F/2R,S-3F/4R linked to Frl gene and produced 370 bp resistant fragment and 520 bp susceptible fragment.Testing the primer combination among 500 F2 plants,the genotype of 473 plants were consistent with their phenotype,meaning the accuracy rate was 94.6%.This SCAR marker was characterized by high accuracy,low cost and simple operation.Therefor,it can be used in gene Frl marker assisted breeding.