松杨栅锈菌两菌群RAPD特异序列的标记转换

Specific RAPD-DNA Markers Transfer of Two Populations of Melampsora larici-populina

【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引物对(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,北方菌群的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.