氧化胁迫诱导苍白杆菌JP1形成VBNC状态及其特征

石油降解菌在各种有害环境因素作用下会进入活的非可培养(viable but non-culturable, VBNC)状态，从而影响其生长及石油降解率。为了研究有害环境因素对石油降解菌生长及石油降解率的影响，采用分光光度法、荧光染色-激光共聚焦显微镜观测H₂O₂胁迫下苍白杆菌(Ochrobactrum sp.)JP1细胞的生长及VBNC状态形成情况。结果表明，不同浓度H₂O₂对其生长有一定抑制作用，当培养液中H₂O₂浓度为75.0 mmol/L时，可有效抑制苍白杆菌JP1生长，处理12 h后苍白杆菌JP1进入VBNC状态。VBNC状态的苍白杆菌JP1细胞缩小变成球体，周质间隙增大；在适宜条件下，VBNC状态苍白杆菌JP1能够复苏为可培养状态，添加丙酮酸钠能够促进VBNC状态细菌细胞的复苏。复苏后的苍白杆菌RJP1具有良好的环境适应性和石油降解能力，为石油污染生物修复的菌种筛选及应用提供了新的策略。

VBNC State Formation and Features of Ochrobactrum JP1 Induced by Oxidative Coercion

Petroleum degradable bacteria could enter the viable but non-culturable (VBNC) state under harmful environments, thus affecting their growth and petroleum degradation efficiency. In order to investigate the effect of harmful environmental factors on the growth and oil degradation rate of petroleum degrading bacteria, the growth and formation of the VBNC state of Ochrobactrum sp. JP1 under H₂O₂ coercion was observed by spectrophotometry and staining= laser confocal microscopy in this paper. The results showed that when the concentration of H₂O₂ in the culture was at 75.0 mmol/L, the growth of Ochrobactrum sp. JP1 could be effectively inhibited. The Ochrobactrum sp. JP1 cells entered the VBNC state 12 hours after treating. Ochrobactrum sp. JP1 cells in the VBNC state shrunk into spheroid and the periplasmic space increased. Ochrobactrum cells in the VBNC state of could be resuscitated and regain culturability under suitable conditions, and the addition of sodium pyruvate could promote the resuscitation of bacterial cells in VBNC state. The resuscitated Ochrobactrum sp. RJP1 cells had excellent environmental adaptability and petroleum degradation ability, which provided a new strategy for the screening and application of strains for bioremediation of oil pollution.