Monitoring of the relation between 2,4-diacetylphloroglucinol-producing Pseudomonas and Thielaviopsis basicola populations by real-time PCR in tobacco black root-rot suppressive and conducive soils
Monitoring of the relation between 2,4-diacetylphloroglucinol-producing Pseudomonas and Thielaviopsis basicola populations by real-time PCR in tobacco black root-rot suppressive and conducive soils
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DOI：
10.1016/j.soilbio.2012.09.003
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发表时间：
2013-02-01
影响因子：
9.7
通讯作者：
Muller, Daniel
中科院分区：
文献类型：
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作者：
Almario, Juliana;Moenne-Loccoz, Yvan;Muller, Daniel
瑞士土壤对烟草黑根腐病的自然抑制作用被认为主要依赖于产生抗菌剂化合物2，4-二乙酰间苯三酚的荧光假单胞菌。然而，这些phl(+)假单胞菌种群与这些土壤中的巴氏锥虫种群和病害抑制力之间的关系尚不清楚，实时荧光定量聚合酶链式反应工具被用来解决这个问题。在抑制土壤和有益土壤中都曾发现过根际显著的phl(+)假单胞菌水平，但这只能通过基于培养的方法来实现。在这里，我们开发并验证了一种针对所有phlD(+)基因的基于phlD(+)的实时聚合酶链式反应方法，不同于目前可用的菌株特异性实时聚合酶链式反应方法(检测下限约为4个对数细胞g(-1)土壤和扩增效率80%)。当在抑制或有利于黑根腐病的瑞士土壤上实施时，它澄清了这样的假设，即抑制不需要更高水平的phlD(+)假单胞菌。用实时荧光定量聚合酶链式反应(Huang and Kang，2010)对烟草根结线虫种群的平行评估表明，抑制作用不是由于病原菌不能在抑制土壤中的根际和烟草根中定植，而是phl(+)假单胞菌可能通过限制病原菌在抑制土壤中的根渗透而起作用。综上所述，本研究建立了一种检测phlD(+)假单胞菌的实时荧光定量PCR方法，可用于监测各种环境条件下的phlD(+)假单胞菌的关键功能群，从而更好地了解抑制性土壤的生态。(C)2012爱思唯尔有限公司。保留所有权利。
Natural suppressiveness of Swiss soils to Thielaviopsis basicola-mediated tobacco black root rot is thought to depend mainly on fluorescent pseudomonads producing the antimicrobial compound 2,4-diacetylphloroglucinol. However, the relation between these phl(+) Pseudomonas populations and both the T. basicola population and disease suppressiveness in these soils is unknown, and real-time PCR tools were used to address this issue. Significant rhizosphere levels of phl(+) pseudomonads had been evidenced before in suppressive as well as conducive soils, but this was done using culture-based approaches only. Here, a phlD-based real-time PCR method targeting all phlD(+) genotypes, unlike the strain-specific real-time PCR methods available so far, was developed and validated (detection limit around 4 log cells g(-1) soil and amplification efficiency >80%). When implemented on Swiss soils suppressive or conducive to black root rot, it clarified the hypothesis that suppressiveness does not require higher levels of phlD(+) pseudomonads. The parallel assessment of T. basicola population by real-time PCR (method of Huang and Kang, 2010) suggested that suppressiveness was not due to the inability of the pathogen to colonize the rhizosphere and tobacco roots in suppressive soils, but rather that phl(+) pseudomonads might act by limiting root penetration by the pathogen in suppressive soils. In conclusion, an effective real-time PCR method was achieved for phlD(+) pseudomonads and can be used to monitor this key functional group in various environmental conditions, including here to better understand the ecology of suppressive soils. (C) 2012 Elsevier Ltd. All rights reserved.