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土壤中烟草根黑腐病菌的real-time PCR分子检测

Dafeng XU, Lei YE, Lina ZHANG,Yongjing HAN,Benguo ZHOU,Genjia TAN

Acta Tabacaria Sinica（2019）

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Abstract
根据烟草根黑腐菌与其它近缘真菌在ITS序列上的差异,设计PCR检测引物TbF/TbR,并通过对各菌株基因组DNA扩增验证其特异性;以接种10倍浓度梯度的根黑腐菌孢子的土壤DNA为模板,建立土壤烟草根黑腐菌real-time PCR检测的标准曲线,得出每个反应体系中分生孢子数目的对数(x)和对应的Ct值(y)之间呈线性关系:y=-1.5373x+24.955,R2=0.9909 (P＜0.01).通过对5块烟田土壤烟草根黑腐菌检测和烟田烟草根黑腐病的调查,证实上述方法可以准确检测土壤中烟草根黑腐菌孢子数量.
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