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葡萄上的植原体病害由于引起叶片黄化而被称为葡萄黄化病。由于这一病害极为严重,葡萄黄化植原体被列为我国的植物检疫对象。其中,葡萄金黄化植原体(16SrV)、维吉尼亚葡萄黄化植原体(16SrⅢ)和澳大利亚葡萄黄化植原体(16SrⅫ)是引起葡萄黄化病的主要3个株系,它们导致的病害症状相似,难以区分。本文进行了3个株系16S rRNA基因DNA序列比对,而后根据同源性相对低的序列设计了43条特异性引物、103对引物对组合,对葡萄黄化植原体3个株系各自的DNA及混合DNA进行PCR扩增,从中筛选出来特异性较强的8个引物对组合。这些引物对组合,能够同步、特异、快速地检测3种葡萄黄化植原体。
Phytoplasma disease on grapes is called yellowing disease due to yellowing of the leaves. As a result of this disease is very serious, grape phytoplankton was listed as China’s plant quarantine object. Among them, Grapevine (16SrV), Virginia (16SrIII) and Australian Grapevine (16SrⅫ) are the three main pathogens causing yellowing disease in grapevine, Symptoms are similar and indistinguishable. In this paper, 16S rRNA gene DNA sequence alignment of three strains was conducted. Then, 43 specific primers were designed according to the sequences with relatively low homology, and 103 pairs of primer pairs were used to compare the 16S rRNA gene sequences of three strains of grapevine DNA and mixed DNA for PCR amplification, screening out from the more specific eight primer combinations. These primer pairs are capable of simultaneous, specific and rapid detection of three grapevine protoplasts.