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Tracing of Two Pseudomonas Strains in the Rootand Rhizoplane of Maize, as Related to Their Plant Growth-Promoting Effectin Contrasting Soils

机译:玉米根际和根际平面中两个假单胞菌菌株的追踪及其在不同土壤中对植物生长的促进作用

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摘要

TaqMan-based quantitative PCR (qPCR) assays were developed to study the persistence of two well-characterized strains of plant growth-promoting rhizobacteria (PGPR), Pseudomonas fluorescens Pf153 and Pseudomonas sp. DSMZ 13134, in the root and rhizoplane of inoculated maize plants. This was performed in pot experiments with three contrasting field soils (Buus, Le Caron and DOK-M). Potential cross-reactivity of the qPCR assays was assessed with indigenous Pseudomonas and related bacterial species, which had been isolated from the rhizoplane of maize roots grown in the three soils and then characterized by Matrix-Assisted Laser Desorption Ionization (MALDI) Time-of-Flight (TOF) mass spectrometry (MS). Sensitivity of the qPCR expressed as detection limit of bacterial cells spiked into a rhizoplane matrix was 1.4 × 102 CFU and 1.3 × 104 CFU per gram root fresh weight for strain Pf153 and DSMZ 13134, respectively. Four weeks after planting and inoculation, both strains could readily be detected in root and rhizoplane, whereas only Pf153 could be detected after 8 weeks. The colonization rate of maize roots by strain Pf153 was significantly influenced by the soil type, with a higher colonization rate in the well fertile and organic soil of Buus. Inoculation with strain DSMZ 13134, which colonized roots and rhizoplane to the same degree, independently of the soil type, increased yield of maize, in terms of biomass accumulation, only in the acidic soil of Le Caron, whereas inoculation with strain Pf153 reduced yield in the soil Buus, despite of its high colonization rate and persistence. These results indicate that the colonization rate and persistence of inoculated Pseudomonas strains can be quantitatively assessed by the TaqMan-based qPCR technique, but that it cannot be taken for granted that inoculation with a well-colonizing and persistent Pseudomonas strain has a positive effect on yield of maize.
机译:开发了基于TaqMan的定量PCR(qPCR)分析方法,以研究两个生长良好的植物生长根瘤菌(PGPR)菌株(荧光假单胞菌Pf153和假单胞菌sp。)的持久性。 DSMZ 13134,在接种的玉米植物的根和根际中。这是在盆栽实验中使用三种对比田间土壤(Buus,Le Caron和DOK-M)进行的。 qPCR分析的潜在交叉反应性是用本地假单胞菌和相关细菌物种评估的,这些物种已从生长在三种土壤中的玉米根的根际平面中分离出来,然后通过基质辅助激光解吸电离(MALDI)进行了表征。飞行(TOF)质谱(MS)。对于菌株Pf153和DSMZ 13134,每克根鲜重,qPCR的灵敏度表示为加标到根际平面基质中的细菌细胞的检出限,分别为1.4×102 CFU和1.3×104 CFU。播种和接种后四个星期,根系和根际上都容易检测到这两个菌株,而8周后只能检测到Pf153。 Pf153菌株对玉米根的定殖率受到土壤类型的显着影响,在Buus肥沃的有机土壤中定殖率较高。 DSMZ 13134菌株在根部和根际平面上均能定殖,而与土壤类型无关,仅在勒卡隆酸性土壤中,就生物量积累而言,增加了玉米的产量,而在土壤生物量方面,Pf153菌株却降低了玉米的产量。尽管土壤Buus具有很高的定殖率和持久性,但仍能保持稳定。这些结果表明,可以通过基于TaqMan的qPCR技术定量评估接种的假单胞菌菌株的定殖率和持久性,但是不能认为将具有良好定殖性和持久性的假单胞菌菌株接种对产量产生积极影响玉米

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