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利用真菌通用引物ITS1和ITS4扩增红掌根腐病菌转录间隔区并进行序列测定,通过序列比较,设计了1对红掌根腐病菌的特异引物SF1/SR2,对30个红掌根腐病病原菌、8种其它真菌和2种细菌基因组DNA进行PCR扩增。结果表明,只有红掌根腐病菌获得572bp的特异带。使用引物SF1/SR2对华丽腐霉进行PCR扩增,其检测灵敏度在DNA水平上可达1pg。运用设计的引物从红掌根腐病菌基因组DNA以及人工接种和自然发病的红掌植株中扩增到572bp的特异片段,实现了对红掌根腐病菌的快速可靠的检测。
The common fungal primer ITS1 and ITS4 were used to amplify the transcribed spacer of Rhizoctonia solani and sequenced. A pair of primers SF1 / SR2 was designed and used to detect Rhizoctonia solani, Pathogenic bacteria, eight other fungi and two bacterial genomic DNA for PCR amplification. The results showed that only 572bp specific bands were obtained from Rhizoctonia solani. PCR amplification of Pythium rapae was carried out using primer SF1 / SR2 with detection sensitivity of 1 pg at the DNA level. A 572bp fragment was amplified from genomic DNA of Rhizoctonia solani and anthracnose from artificial inoculation and naturally occurring plants by using the designed primers. The rapid and reliable detection of Rhizoctonia solani was achieved.