Peng Anping, Liu Juan, Gao Yanzheng, Chen Zeyou
Institute of Organic Contaminant Control and Soil Remediation, College of Resource and Environmental Sciences, Nanjing Agricultural University, Nanjing, People's Republic of China.
PLoS One. 2013 Dec 17;8(12):e83054. doi: 10.1371/journal.pone.0083054. eCollection 2013.
The distributions of endophytic bacteria in Alopecurus aequalis Sobol and Oxalis corniculata L. grown in soils contaminated with different levels of polycyclic aromatic hydrocarbons (PAHs) were investigated with polymerase chain reaction followed by denaturing gradient gel electrophoresis technology (PCR-DGGE) and cultivation methods. Twelve types of PAHs, at concentrations varying from 0.16 to 180 mg·kg(-1), were observed in the roots and shoots of the two plants. The total PAH concentrations in Alopecurus aequalis Sobol obtained from three different PAH-contaminated stations were 184, 197, and 304 mg·kg(-1), and the total PAH concentrations in Oxalis corniculata L. were 251, 346, and 600 mg·kg(-1), respectively. The PCR-DGGE results showed that the endophytic bacterial communities in the roots and shoots of the two plants were quite different, although most bacteria belonged to Firmicutes, Proteobacteria, Actinobacteria and Bacteroidetes. A total of 68 endophytic bacterial strains were isolated from different tissues of the two plants and classified into three phyla: Firmicutes, Proteobacteria and Bacteroidetes. In both plants, Bacillus spp. and Pseudomonas spp. were the dominant cultivable populations. With an increase in the PAH pollution level, the diversity and distribution of endophytic bacteria in the two plants changed correspondingly, and the number of cultivable endophytic bacterial strains decreased rapidly. Testing of the isolated endophytic bacteria for tolerance to each type of PAH showed that most isolates could grow well on Luria-Bertani media in the presence of different PAHs, and some isolates were able to grow rapidly on a mineral salt medium with a single PAH as the sole carbon and energy source, indicating that these strains may have the potential to degrade PAHs in plants. This research provides the first insight into the characteristics of endophytic bacterial populations under different PAH pollution levels and provides a species resource for the isolation of PAH-degrading endophytic bacteria.
采用聚合酶链反应-变性梯度凝胶电泳技术(PCR-DGGE)和培养方法,研究了生长在不同多环芳烃(PAHs)污染水平土壤中的稗草和酢浆草内生细菌的分布情况。在这两种植物的根和茎中检测到12种PAHs,浓度范围为0.16至180 mg·kg(-1)。从三个不同PAH污染站点采集的稗草中总PAH浓度分别为184、197和304 mg·kg(-1),酢浆草中的总PAH浓度分别为251、346和600 mg·kg(-1)。PCR-DGGE结果表明,尽管大多数细菌属于厚壁菌门、变形菌门、放线菌门和拟杆菌门,但这两种植物根和茎中的内生细菌群落差异很大。从这两种植物的不同组织中共分离出68株内生细菌菌株,分为三个门:厚壁菌门、变形菌门和拟杆菌门。在这两种植物中,芽孢杆菌属和假单胞菌属是主要的可培养种群。随着PAH污染水平的增加,这两种植物内生细菌的多样性和分布相应改变,可培养内生细菌菌株数量迅速减少。对分离出的内生细菌进行每种PAH耐受性测试表明,大多数分离株在含有不同PAHs的Luria-Bertani培养基上能良好生长,一些分离株在以单一PAH作为唯一碳源和能源的无机盐培养基上能快速生长,表明这些菌株可能具有降解植物中PAHs的潜力。本研究首次深入了解了不同PAH污染水平下内生细菌种群的特征,并为PAH降解内生细菌的分离提供了物种资源。