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烟田土壤带菌量与烟草青枯病发生的相关性分析

Correlation between Ralstonia solanacearum population in soils and occurrence of tobacco bacterial wilt

  • 摘要: 为快速、准确测定烟田土壤中青枯劳尔氏菌的数量及监测田间病害的发生动态,基于实时荧光定量PCR技术,构建了一种实时监测烟田土壤青枯劳尔氏菌的方法,并运用该方法检测了临沧、文山烟田土壤中的越冬菌量及青枯病发生时的土壤带菌量。结果表明,内切葡聚糖酶基因的特异引物RSf1/RSr1在烟草青枯劳尔氏菌中的扩增效率高、特异性强,构建的标准曲线CT值与土壤带菌量具有良好的线性关系,相关系数为0.996 5,扩增效率为93.60%。田间青枯病的病情指数与土壤带菌量的CT值负相关,病害越严重,CT值越小。利用建立的qRT-PCR技术对烟田土壤越冬菌量和病害流行期间的土壤带菌量进行多次检测,结果表明越冬菌量与最终病情指数正相关,即越冬菌量越大,病害越严重。

     

    Abstract: To rapidly and accurately detect the population of Ralstonia solanacearum in tobacco-planting soils and to monitor the dynamic occurrence of tobacco bacterial wilt, a real-time fluorescent quantitative PCR (qRT-PCR) method was developed for real-timely monitoring the occurrence of R. solanacearum in tobacco-planting soils. The developed method was used to detect the overwintering population of R. solanacearum and the population of R. solanacearum when tobacco bacterial wilt occurred at Lincang and Wenshan tobacco-planting areas. The results showed that the specific primer (RSf1/RSr1) of endoglucanase gene from R. solanacearum had a high amplification efficiency and strong specificity, and there was a better linear relationship between CT value of standard curve and the population of R. solanacearum in soils with the correlation coefficient of 0.996 5 and the amplification efficiency of 93.60%. The disease index of tobacco bacterial wilt negatively correlated to CT value of R. solanacearum population in soils. The overwintering population of R. solanacearum and the population of R. solanacearum when tobacco bacterial wilt occurred were multiple detected by the developed method. This result indicated that the overwintering population of R. solanacearum positively correlated to final disease index. With the increase of overwintering population of R. solanacearum, the occurrence of tobacco bacterial wilt became more serious.

     

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