Department of Microbial Ecology, Centre for Ecological and Evolutionary Studies, University of Groningen, RUG, Groningen, The Netherlands.
PLoS One. 2013 Jul 3;8(7):e67948. doi: 10.1371/journal.pone.0067948. Print 2013.
In this study, we assessed the actively metabolizing bacteria in the rhizosphere of potato using two potato cultivars, i.e. the genetically-modified (GM) cultivar Modena (having tubers with altered starch content) and the near-isogenic non-GM cultivar Karnico. To achieve our aims, we pulse-labelled plants at EC90 stage with (13)C-CO2 and analysed their rhizosphere microbial communities 24 h, 5 and 12 days following the pulse. In the analyses, phospholipid fatty acid/stable isotope probing (PLFA-SIP) as well as RNA-SIP followed by reverse transcription and PCR-DGGE and clone library analysis, were used to determine the bacterial groups that actively respond to the root-released (13)C labelled carbonaceous compounds.
METHODOLOGY/PRINCIPAL FINDINGS: The PLFA-SIP data revealed major roles of bacteria in the uptake of root-released (13)C carbon, which grossly increased with time. Gram-negative bacteria, including members of the genera Pseudomonas and Burkholderia, were strong accumulators of the (13)C-labeled compounds at the two cultivars, whereas Gram-positive bacteria were lesser responders. PCR-DGGE analysis of cDNA produced from the two cultivar types showed that these had selected different bacterial, alpha- and betaproteobacterial communities at all time points. Moreover, an effect of time was observed, indicating dynamism in the structure of the active bacterial communities. PCR-DGGE as well as clone library analyses revealed that the main bacterial responders at cultivar Karnico were taxonomically affiliated with the genus Pseudomonas, next to Gluconacetobacter and Paracoccus. Cultivar Modena mainly attracted Burkholderia, next to Moraxella-like (Moraxellaceae family) and Sphingomonas types.
CONCLUSIONS/SIGNIFICANCE: Based on the use of Pseudomonas and Burkholderia as proxies for differentially-selected bacterial genera, we conclude that the selective forces exerted by potato cultivar Modena on the active bacterial populations differed from those exerted by cultivar Karnico.
在这项研究中,我们使用两个马铃薯品种,即具有改变淀粉含量的块茎的转基因(GM)品种 Modena 和近等基因非 GM 品种 Karnico,评估了马铃薯根际中活跃代谢的细菌。为了实现我们的目标,我们在 EC90 阶段用(13)C-CO2 脉冲标记植物,并在脉冲后 24 小时、5 天和 12 天分析它们的根际微生物群落。在分析中,我们使用磷脂脂肪酸/稳定同位素探测(PLFA-SIP)以及 RNA-SIP 随后进行反转录和 PCR-DGGE 和克隆文库分析,以确定对根释放的(13)C 标记碳化合物做出积极响应的细菌群体。
方法/主要发现:PLFA-SIP 数据显示细菌在吸收根释放的(13)C 碳方面起着重要作用,这种作用随着时间的推移而大大增加。革兰氏阴性菌,包括假单胞菌属和伯克霍尔德氏菌属的成员,在两个品种中都是(13)C 标记化合物的强烈积累者,而革兰氏阳性菌则反应较弱。从两个品种类型的 cDNA 产生的 PCR-DGGE 分析表明,这些品种在所有时间点都选择了不同的细菌、α-和β变形杆菌群落。此外,还观察到时间的影响,表明活跃细菌群落的结构具有动态性。PCR-DGGE 以及克隆文库分析表明,在 Karnico 品种上的主要细菌反应体在分类上与假单胞菌属有关,其次是 Gluconacetobacter 和 Paracoccus。Modena 品种主要吸引 Burkholderia,其次是 Moraxella-like(Moraxellaceae 科)和 Sphingomonas 类型。
结论/意义:基于使用假单胞菌属和伯克霍尔德氏菌属作为差异选择细菌属的代表,我们得出结论,马铃薯品种 Modena 对活跃细菌种群施加的选择力与品种 Karnico 施加的选择力不同。