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首页> 外文期刊>Plant and Soil >Culture-independent molecular approaches reveal a mostly unknown high diversity of active nitrogen-fixing bacteria associated with Pennisetum purpureum-a bioenergy crop
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Culture-independent molecular approaches reveal a mostly unknown high diversity of active nitrogen-fixing bacteria associated with Pennisetum purpureum-a bioenergy crop

机译：与文化无关的分子方法揭示了与紫草—生物能源作物有关的活性固氮细菌的高度未知的多样性

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Previous studies have shown that elephant grass is colonized by nitrogen-fixing bacterial species; however, these results were based on culture-dependent methods, an approach that introduces bias due to an incomplete assessment of the microbial community. In this study, we used culture-independent methods to survey the diversity of endophytes and plant-associated bacterial communities in five elephant grass genotypes used in bioenergy production. The plants of five genotypes of elephant grass were harvested from the experimental area of Embrapa Agrobiologia and divided into stem and root tissues. Total DNA and RNA were extracted from plant tissues and the bacterial communities were analyzed by DGGE and clone library of the 16S rRNA and nifH genes at both the cDNA and DNA levels. Overall, the patterns based on DNA- and RNA-derived DGGE-profiles differed, especially within tissue samples. DNA-based DGGE indicated that both total bacterial and diazotrophic communities associated with roots (rhizoplane + endophytes) differed clearly from those obtained from stems (endophytes). These results were confirmed by the phylogenetic analyses of RNA-derived sequences of 16S rRNA (total bacteria; 586 sequences), but not for nifH (186). In fact, rarefaction analyses showed a higher diversity of diazotrophic organisms associated with stems than roots. Based on 16S rRNA sequences, the clone libraries were dominated by sequences affiliated to members of Leptotrix (12.8 %) followed by Burkholderia (9 %) and Bradyrhizobium (6.5 %), while most of the nifH clones were closely related to the genus Bradyrhizobium (26 %). Our results revealed an unexpectedly large diversity of metabolically active bacteria, providing new insights into the bacterial species predominantly found in association with elephant grass. Furthermore, these results can be very useful for the development of new strategies for selection of potential bacteria that effectively contribute to biological nitrogen fixation and enhance the sustainable production of elephant grass as bioenergy crop.
机译：先前的研究表明，象草被固氮细菌物种所定殖。然而，这些结果是基于依赖于培养物的方法，该方法由于对微生物群落的不完全评估而引入了偏见。在这项研究中，我们使用了与文化无关的方法来调查生物能源生产中使用的五种大象草基因型中内生植物和与植物相关的细菌群落的多样性。从Embrapa农业生物试验区收获了五种基因型的象草植物，并将其分为茎和根组织。从植物组织中提取总DNA和RNA，并通过DGGE分析细菌群落，并在cDNA和DNA水平上分析16S rRNA和nifH基因的克隆文库。总体而言，基于DNA和RNA的DGGE谱的模式有所不同，尤其是在组织样品中。基于DNA的DGGE表明，与根部相关的总细菌和重氮营养群落（根际平面+内生菌）与从茎（内生菌）获得的群落明显不同。这些结果已通过对16S rRNA的RNA序列（总细菌; 586个序列）进行系统发育分析而得到证实，但对nifH则没有（186）。实际上，稀疏分析显示与茎相关的重氮营养生物比根部具有更高的多样性。根据16S rRNA序列，克隆文库以隶属于钩端螺旋体（12.8％），伯克霍尔德氏菌（9％）和缓生根瘤菌（6.5％）的序列为主，而大多数nifH克隆与缓生根瘤菌属（Bradyrhizobium（ 26％）。我们的研究结果揭示了代谢活性细菌的出乎意料的多样性，从而为主要与象草相关的细菌种类提供了新见解。此外，这些结果对于开发新的策略以选择潜在细菌非常有用，这些细菌有效地有助于生物固氮并增强象草作为生物能源作物的可持续生产。

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