Department of Forest Mycology and Pathology, Uppsala BioCenter, SLU, Uppsala, Sweden.
J Appl Microbiol. 2009 Sep;107(3):736-45. doi: 10.1111/j.1365-2672.2009.04265.x. Epub 2009 Mar 9.
To detect and quantify the plant drought tolerance enhancing bacterium Paenibacillus polymyxa in a collection of 160 Hordeum spontaneum rhizosphere samples at the 'Evolution Canyon' ('EC'), Israel.
PCR primers and a FAM-TAMRA probe (6-carboxyfluorescein, 6-carboxy-tetramethyl-rhodamine) targeting 16S rRNA genes were designed and used to detect and quantify the target strain. Two commercial kits, Bio101 Fast Spin and Mo Bio Ultra Clean Soil DNA, were tested for DNA isolation from the rhizosphere and surrounding soil. Population densities of P. polymyxa were studied in the rhizosphere of wild barley and surrounding soil from the contrasting climatic slopes at the 'EC' using the real-time PCR and culture based methods.
Paenibacillus polymyxa is one of the best established species in wild barley rhizosphere at the 'EC' slopes. With the real-time PCR assay we are able to detect 1 pg of DNA per PCR corresponding to 100 cells per ml. The results at the 'EC' correlate well to bacterial estimations by culture based methods.
Significantly higher P. polymyxa cell number was detected in the rhizosphere of arid 'African' microclimate indicating possible role of adaptive co-evolution with plants.
在以色列“进化峡谷”（‘EC’）的 160 个野生大麦根际样本中，检测和定量具有植物耐旱增强作用的芽孢杆菌 Paenibacillus polymyxa。
设计了针对 16S rRNA 基因的 PCR 引物和 FAM-TAMRA 探针（6-羧基荧光素，6-羧基四甲基罗丹明），用于检测和定量目标菌株。测试了两种商业试剂盒，Bio101 Fast Spin 和 Mo Bio Ultra Clean Soil DNA，用于从根际和周围土壤中分离 DNA。使用实时 PCR 和基于培养的方法，在‘EC’的不同气候斜坡的野生大麦根际及其周围土壤中，研究了 P. polymyxa 的种群密度。
芽孢杆菌 Paenibacillus polymyxa 是‘EC’斜坡野生大麦根际中最成熟的物种之一。通过实时 PCR 检测，我们能够检测到每个 PCR 对应 1 pg DNA，相当于每毫升 100 个细胞。‘EC’的结果与基于培养的方法的细菌估计值很好地相关。
在干旱的“非洲”微气候的根际中，检测到更高数量的 P. polymyxa 细胞，这表明与植物的适应性协同进化可能具有重要作用。
