降香黄檀黑痣病菌的快速分子检测

<正>黄檀黑痣菌(Phyllachora dalbergiicola)能引起降香黄檀黑痣病,该病菌在世界分布十分广泛,且寄主植物种类繁多,除了降香黄檀以外,还可侵染大果紫檀、檀香紫檀等多种紫檀属植物。该菌所致病害会在寄主表面形成黑色凸起的盾片,严重影响寄主光合作用,致使其提前落叶~(1])。通过对黄檀黑痣菌进行早期检测,可以及时采取防治措施,控制病害的进一步发展。而黄檀黑痣菌是活体营养真菌,不能离体培养,使用常规的病害诊断技术

Rapid Molecular diagnosis of Phyllachora dalbergiicola in Dalbergia odorifera

Tar spot of Dalbergia odorifera, caused by Phyllachora dalbergiicola, is a common disease, and seriously affected the rate of photosynthesis. Here we developed a species-specific Nested-PCR approach for rapid and accurate detection of P. dalbergiicola, based on the differences in internal transcribed spacer (ITS) sequences of P. dalbergiicola and another P. spp., an endophytic fungus, from which a pair of species-specific primers P1/P2(P1:5'-CGAGGTCAGAATCAAACG-3', P2:5'-TGAAGAACGCAGCGAAAT-3'), was designed. P1/P2 amplified only a unique 273 bp band from the genomic DNA of P. dalbergiicola. A Nested-PCR procedure using ITS4/ITS5 as the first-round primers, followed by P1/P2 primers, increased detection sensitivity 10 000 fold to 100 ag. Using the Fast DNA-kit to extract DNA from the diseased plant tissues, the detection of the pathogen by Nested-PCR assay could be completed within 1 day. The results suggested that the PCR-based methods here could simplify both plant disease diagnosis and pathogen detection