Abstract：
OBJECTIVE: The increasingly widespread use of beneficial microbial inocula in agriculture gives rise to two primary needs: i) the assessment of the environmental risk, i.e. their impact on local soil microbiome and soil properties; ii) being able to track them and monitor their persistence and fate to both optimize their formulation and application method. In previous years, PCR-based methods have detected bacterial or fungal bioinoculant at the species or strain level. However, the selective detection, quantification, and monitoring of target microbial species in a complex ecosystem such as soil require that the tests possess high specificity and sensitivity.
RESULTS: The work proposes a quantitative real-time PCR detection method using TaqMan chemistry, showing high specificity and sensitivity for the Paenibacillus polymyxa K16 strain. The primer and probe sets were designed using the polymyxin gene cluster targeting pmxC and pmxE sequences. Validation tests showed that these assays allowed a discriminant and specific detection of P. polymyxa K16 in soil.
CONCLUSIONS: The TaqMan-assay developed could thus ensure the necessary level of discrimination required by commercial and regulatory purposes to detect and monitor the bioinoculant in soil.
RESULTS: The work proposes a quantitative real-time PCR detection method using TaqMan chemistry, showing high specificity and sensitivity for the Paenibacillus polymyxa K16 strain. The primer and probe sets were designed using the polymyxin gene cluster targeting pmxC and pmxE sequences. Validation tests showed that these assays allowed a discriminant and specific detection of P. polymyxa K16 in soil.
CONCLUSIONS: The TaqMan-assay developed could thus ensure the necessary level of discrimination required by commercial and regulatory purposes to detect and monitor the bioinoculant in soil.
摘要：
目的：在农业中越来越广泛地使用有益的微生物接种物引起了两个主要需求：i）环境风险评估，即它们对当地土壤微生物组和土壤性质的影响;ii)能够跟踪它们并监测它们的持久性和命运,以优化它们的配方和施用方法。在前几年,基于PCR的方法已经在物种或菌株水平上检测到细菌或真菌生物接种剂。然而,选择性检测，量化，和监测目标微生物物种在复杂的生态系统，如土壤要求测试具有高的特异性和灵敏度。
结果：工作提出了一种使用TaqMan化学的定量实时PCR检测方法，对多粘类芽孢杆菌K16菌株具有较高的特异性和敏感性。使用靶向pmxC和pmxE序列的多粘菌素基因簇设计引物和探针组。验证测试表明，这些测定允许对土壤中的多粘菌K16进行判别和特异性检测。
结论：因此，开发的TaqMan测定法可以确保商业和监管目的所需的必要歧视水平，以检测和监测土壤中的生物接种剂。
结果：工作提出了一种使用TaqMan化学的定量实时PCR检测方法，对多粘类芽孢杆菌K16菌株具有较高的特异性和敏感性。使用靶向pmxC和pmxE序列的多粘菌素基因簇设计引物和探针组。验证测试表明，这些测定允许对土壤中的多粘菌K16进行判别和特异性检测。
结论：因此，开发的TaqMan测定法可以确保商业和监管目的所需的必要歧视水平，以检测和监测土壤中的生物接种剂。
