【目的】根据RAPD分析结果,对松杨栅锈菌西部和北方两大地理相关菌群RAPD扩增特异性DNA片段进行SCAR标记转换。【方法】经pGEM—T载体克隆和测序、Seqman整合后,用Primerselct分别设计了两地理菌群的简并5I物对(西部菌群简并引物对PNW0118—382-F:GAATCGGCCACAAAACTATC,PNW0118—382-R:GAATCGGCCATGACTTGAGC,北方菌群简并引物对PNW0118-400-F:GAATCGGCCACAAAACTATC,PNW0118-400-R:GAATCGGCCATGACTTGAGCTGC)。【结果】经PCR条件优化成功获得两大地理菌群的SCAR标记。西部菌群SCAR标记为382bp,北方茵群的SCAR标记为400bp。【结论】两对引物可用于松杨栅锈菌地理菌群的检测。
[ Objective ] This paper focused on achieving sequence characterized amplified region (SCARs) of geographic populations of Melampsora larici-populina, and transferring them to codominant markers. [ Method ] Based on the results of RAPD study of Melampsora larici-populina, DNA fragments of Western and Northern populations were recycled, purified, pGEM-T vector cloned and sequenced. About 400bp length DNA fragments were achieved after being integrated by software Seqman, and two pairs of primers were designed by software Primerselect (Western population primers: PNW0118-382-F: GAATCGGCCACAAAACTATC, PNW0118-382-R: GAATCGGCC ATGACTTGAGC, and Northern population primers: PNW0118-400-F: GAATCGGCCACAAAA CTATC, PNW0118-400-R: GAATCGGCCATGACTTGAGCTGC).[ Result] With these primers, SCARs of Western population (382 bp) and Northern population (400 bp) were amplified successfully by optimized conditions of polymerase chain reactions. [ Conclusion ] The two pairs of primer can be used to monitor the geographic groups of Melampsora larici-populina.