古井贡酒的酿造过程中,窖泥和酒醅中发现大量耐酵乳杆菌(Lactobacillus acetotolerans),对该菌快速特异性检测非常重要。该文利用L.acetotolerans菌及大量Lactobacillus类似菌种的全基因组序列与PRIMER-BLAST方法,设计了几对L.acetotolerans特异性引物,其中2对经过系列验证具有较好的特异性,可结合QPCR技术对不同质量和不同发酵阶段的窖泥进行L.acetotolerans含量的快速检测。结果表明,古井贡酒窖泥中L.acetotolerans 含量总体明显低于酒醅,老窖泥中L.acetotolerans含量总体明显低于新窖池窖泥;而酒醅中老厂区和新厂区L.acetotolerans含量平均来看都很高,但样本之间存在明显波动。所用L.acetotolerans特异性引物可对浓香型白酒发酵样本中L.acetotolerans的相对含量测定并辅助评估窖泥酒醅质量。
When brewing Gujing Tribute liquor, a large number of Lactobacillus acetotolerans were found in both pitmud and Zaopei. Therefore, it is important to rapidly detect L. acetotolerans specifically. In this paper, whole genome sequences of Lactobacillus bacteria and PRIMER-BLAST method were used to design L. acetotolerans-specific primers. Two pairs of primers were confirmed with decent specificity and combined with quantitative polymerase chain reaction (qPCR) to rapidly measure the amount of L. acetotolerans with different qualities and at different fermentation stages in pitmud and Zaopei. The results showed that the population of L. acetotolerans in Gujing Tribute pitmud was apparently lower than that of Zaopei and was apparently lower in old pitmud than that in pitmud of new plant. Moreover, L. acetotolerans in Zaopei from both new and old plants were generally high with apparent fluctuations among samples. In general, QPCR mediated by L. acetotolerans-specific primers can be used for L. acetotolerans quantification and facilitate quality assessment of pitmud or Zaopei in strong-aroma liquor production.
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