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Pyruvate-amended modified SMSA medium: improved sensitivity for detection of Ralstonia solanacearum

机译:丙酮酸修饰的改良SMSA培养基:提高检测青枯雷尔氏菌的灵敏度

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摘要

The viable but nonculturable (VBNC) state is induced in the bacterial wilt pathogen Ralstonia solanacearum under prolonged environmental stress. These VBNC cells lose their ability to grow on standard media such as CPG agar, but some of the cells can recover this ability on media supplemented with sodium pyruvate (SP), that degrades hydrogen peroxide. Recently, we suggested that some of the cells in the low-temperature-induced SP-recoverable VBNC state regained their ability to grow on CPG agar after exposure to moderate temperature. These revived cells also retained their virulence on tomato. Although R. solanacearum is detectable on semiselective media, VBNC cells are not detectable on any known semiselective media for the pathogen. To create a suitable medium to detect VBNC cells, we therefore added various compounds that can either degrade hydrogen peroxide or serve an antioxidant function in a semiselective medium, modified SMSA. SP at 5 g/l most improved the sensitivity of R. solanacearum detection. Furthermore, counts on modified SMSA plates for R. solanacearum that had been added to field soil also increased after the addition of 5 g/l SP. SP thus improved the medium's sensitivity for the detection of R. solanacearum by rescuing a portion of the VBNC cells.
机译:在长时间的环境压力下,在细菌枯萎病原体青枯雷尔氏菌中诱导了存活但不可培养的状态。这些VBNC细胞失去了在标准培养基(例如CPG琼脂)上生长的能力,但是一些细胞可以在补充有丙酮酸钠(SP)的培养基上恢复这种能力,丙酮酸钠可降解过氧化氢。最近,我们建议低温暴露于SP可恢复的VBNC状态的某些细胞在暴露于中等温度后可恢复其在CPG琼脂上的生长能力。这些复活的细胞还保留了对番茄的毒力。尽管青枯菌可在半选择培养基上检测到,但在病原体的任何已知半选择培养基上都无法检测到VBNC细胞。为了创建合适的培养基来检测VBNC细胞,我们在半选择性培养基(改良的SMSA)中添加了可以降解过氧化氢或具有抗氧化功能的各种化合物。 5 g / l的SP可以最大程度地提高青枯菌检测的灵敏度。此外,在添加5 g / l SP后,已添加到田间土壤中的青枯菌的改良SMSA平板上的计数也增加了。因此,SP抢救了一部分VBNC细胞,从而提高了培养基对青枯菌的检测灵敏度。

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