巢式PCR检测琯溪蜜柚黑斑病病原菌
Chinese Journal of Tropical Crops(2013)
Abstract
利用特异性引物从琯溪蜜柚黑斑病菌基因组DNA中扩增出一条分子量为487 bp的特异性条带,准确地与疮痂病、炭疽病和黄斑病菌等蜜柚常发性真菌病害区分开。采用巢式PCR对该引物的检测灵敏度进行测定,结果显示,对于黑斑病菌基因组DNA,巢式PCR的检测灵敏度为100 fg/μL,灵敏度比常规PCR至少提高100倍;对于发病组织,巢式PCR可从病斑组织含量为100μg的DNA样品中检测到黑斑病菌。采用巢式PCR检测技术,可从蜜柚的黑斑病显症组织和未显症组织特异性地检测到病原菌,且检出率分别为96.67%和90%。
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Key words
Nested-PCR,Detection,ITS,Citrus black spot,Guanxi honey pomelo
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