降香黄檀黑痣病菌的快速分子检测  

Rapid Molecular diagnosis of Phyllachora dalbergiicola in Dalbergia odorifera

作　　者：刘倩丽 周国英[1] 李河[1] 董文统[1] 刘成锋[1] 刘君昂[1] 

机构地区：[1]中南林业科技大学经济林培育与保护教育部重点实验室,长沙410004

出　　处：《植物病理学报》2016年第1期135-139,共5页Acta Phytopathologica Sinica

基　　金：国家林业公益性行业科研专项(201304402)

摘　　要：Tar spot of Dalbergia odorifera, caused by Phyllachora dalbergiicola, is a common disease, and se-riously affected the rate of photosynthesis. Here we developed a species-specific Nested-PCR approach for rapidand accurate detection of P. dalbergiicola, based on the differences in internal transcribed spacer （ITS） se-quences of P. dalbergiicola and another P. spp., an endophytic fungus, from which a pair of species-specificprimers 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 proce-dure using ITS4/ITS5 as the first-round primers, followed by P1/P2 primers, increased detection sensitivity10 000 fold to 100 ag. Using the Fast DNA-kit to extract DNA from the diseased plant tissues, the detection ofthe pathogen by Nested-PCR assay could be completed within I day. The results suggested that the PCR-basedmethods here could simplify both plant disease diagnosis and pathogen detection.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 Pl/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 sensitivity10 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.

关 键 词：DALBERGIA odorifera Phyllachora dalbergiicola MOLECULAR detection CONVENTIONAL PCR NESTED-PCR 

分 类 号：S[农业科学]