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The Copy Number of the spoVA2mob Operon Determines Pressure Resistance of Bacillus Endospores

机译：spoVA2mob操纵子的拷贝数决定了芽孢杆菌内生孢子的耐压性

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The spoVA2mob operon confers heat resistance to Bacillus spp., and the resistance correlates to the copy number of the operon. Bacillus endospores also exhibit a strong variation in resistance to pressure, but the underlying mechanisms of endospore resistance to pressure are not fully understood. We determined the effects of multiple spoVA2mob operons on high-pressure resistance in Bacillus endospores. The copy numbers of the spoVA2mob operon in 17 strains of Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus cereus, Bacillus velezensis, and Bacillus pumilus were determined via droplet digital PCR (ddPCR) and genome sequencing. These strains contained between 0 and 3 copies of the spoVA2mob operon; the quantification of the gene copy number by ddPCR was as accurate as whole-genome sequencing. We further tested the pressure resistance of 17 Bacillus endospores at 600?MPa and 80°C. Strains with one or no spoVA2mob operon had significantly lower pressure resistance than strains with two or three copies of the operons (P spoVA2mob operons in Bacillus contributed to higher pressure resistance of endospores. The copy number of the spoVA2mob operon was not related to the dipicolinic acid (DPA) content of endospores. Overall, the copy number of the spoVA2mob operon contributes to pressure resistance of Bacillus endospores. This improves our understanding of the pressure resistance mechanisms in Bacillus spp. and may inform the development of high-pressure sterilization in food processing.IMPORTANCEBacillus spp. are considered pressure-resistant microorganisms, but the resistance mechanisms remain unknown. The spoVA2mob operon is a mobile genetic element, and it can transfer to pathogenic or spoilage organisms by horizontal gene transfer. Results in this study indicate that multiple copies of the spoVA2mob operon mediate high-pressure resistance of Bacillus endospores, and it might contribute to the identification of the source of pressure-resistant pathogens and spoilage organisms that may contaminate the food supply. The droplet digital PCR (ddPCR) system is well suited for analysis in some human diseases due to its high efficiency and capability to provide high precision; however, no relevant studies in food microbiology have been reported so far. This study demonstrates a novel application of ddPCR in food microbiology.
机译：spoVA2mob操纵子赋予芽孢杆菌耐热性，并且该耐热性与操纵子的拷贝数相关。芽孢杆菌内生孢子对压力的抵抗力也表现出很大的变化，但是内生孢子对压力的抵抗的潜在机制尚不完全清楚。我们确定了多个spoVA2mob操纵子对芽孢杆菌内生芽孢杆菌高压耐性的影响。通过液滴数字PCR（ddPCR）和基因组测序确定了17种枯草芽孢杆菌，解淀粉芽孢杆菌，蜡状芽孢杆菌，velezensis芽孢杆菌和pumilus芽孢杆菌中spoVA2mob操纵子的拷贝数。这些菌株含有0至3个spoVA2mob操纵子拷贝。 ddPCR对基因拷贝数的定量与全基因组测序一样准确。我们进一步测试了17种芽孢杆菌内生孢子在600？MPa和80°C的耐压性。带有一个或没有spoVA2mob操纵子的菌株的耐压性比带有两个或三个拷贝的操纵子的菌株（芽孢杆菌中的P spoVA2mob操纵子有助于内生孢子的耐压性更高。spoVA2mob操纵子的拷贝数与吡啶二酸无关）总体而言，spoVA2mob操纵子的拷贝数有助于芽孢杆菌内生芽孢杆菌的耐压性，从而增进了我们对芽孢杆菌属芽孢杆菌耐压机理的理解，并可能有助于食品加工中高压灭菌的发展。重要提示芽孢杆菌属被认为是耐压微生物，但其耐药机制尚不清楚，spoVA2mob操纵子是一种可移动的遗传元件，可以通过水平基因转移而转移到致病或腐败的生物中。 spoVA2mob操纵子介导芽孢杆菌内生芽孢杆菌的高压抗性，它可能有助于识别可能污染食品供应的耐压病原体和腐败生物的来源。液滴数字PCR（ddPCR）系统具有高效率和提供高精度的功能，非常适合某些人类疾病的分析。但是，到目前为止，尚无有关食品微生物学的相关报道。这项研究证明了ddPCR在食品微生物学中的新应用。

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