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从新疆采集的向日葵黑茎病罹病植株的茎秆、叶片和花盘共分离获得20个真菌分离物。经致病性测定,证明分离物XJ011和XJ111是引起该病害的病原物。采用ITS通用引物ITS1/ITS4对XJ011和XJ111菌株的rDNA-ITS区进行PCR扩增和测序,并结合形态学特征,将该菌鉴定为麦氏茎点霉Phoma macdonaldii。同时,在rDNA-ITS的多态性丰富区域设计了一对特异性引物320FOR/320REV,建立了P.macdonaldii病菌的快速分子检测体系,能特异性检测出向日葵黑茎病菌,灵敏度可达到1fg。
Twenty fungal isolates were isolated from the stalks, leaves and faceplates of diseased plants of black sunflower collected from Xinjiang. The pathogenicity test proved that isolates XJ011 and XJ111 were the etiological agents for this disease. The rDNA-ITS region of XJ011 and XJ111 strains were amplified by PCR and sequenced by using ITS1 / ITS4 ITS primers. Phylogenetic analysis showed that the strain was identified as Phoma macdonaldii. At the same time, a pair of specific primer 320FOR / 320REV was designed in the rich region of rDNA-ITS polymorphism. A rapid molecular detection system of P.macdonaldii was established, which can detect the black stalk of sunflower with specific sensitivity of 1fg.