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首页> 美国政府科技报告 >Monitoring Rates and Heterogeneity of High-Pressure Germination of Bacillus Spores by Phase-Contrast Microscopy of Individual Spores
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Monitoring Rates and Heterogeneity of High-Pressure Germination of Bacillus Spores by Phase-Contrast Microscopy of Individual Spores

机译：通过个体孢子的相衬显微镜监测芽孢杆菌孢子高压萌发率和异质性

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Germination of Bacillus spores with a high pressure (HP) of150 MPa is via activation of spores germinant receptors (GRs). The HP germination of multiple individual Bacillus subtilis spores in a diamond anvil cell (DAC) was monitored with phasecontrast microscopy. Major conclusions were that (i)>95% of wild-type spores germinated in 40 min in a DAC at approximately 150 MPa and 37 C but individual spores germination kinetics were heterogeneous; (ii) individual spores HP germination kinetic parameters were similar to those of nutrient- triggered germination with a variable lag time (T sub lag) prior to a period of the rapid release (delta T(sub release)) of the spores dipicolinic acid in a1:1 chelate with Ca2+ (CaDPA); (iii) spore germination at 50 MPa had longer average T(sub lag) values than that at approximately 150 MPa, but the deltaT(sub release) values at the two pressures were identical and HPs of < 10 MPa did not induce germination; (iv) B, subtillis spores that lacked the cortex-lytic enzyme CwlJ and that were germinated with an HP of 150 MPs exhibited average Delta T release values 15-fold longer than those for wild-type spores, but the two types of spores exhibited similar average T lag values; and (v) the germination of wild-type spores given a>30-s 140-MPa HP pulse followed by a constant pressure of 1 MPa was the same as that of spores exposed to a constant pressure of 140 MPa that was continued for > 35 min; (vi) however, after short 150-MPa HP pulses and incubation at 0.1 MPa (ambient pressure), spore germination stopped 5 to 10 min after the HP was released. These results suggest that an HP of 150 MPa for <30 s is sufficient to fully activate spores GRs, which remain activated at 1 MPa but can deactivate at ambient pressure.

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