为高效地获得浓香型白酒窖池中窖泥和黄水中的微生物总DNA以更好地分析其过程中微生物区系,该文设计了4种不同的总DNA提取方法,分别提取了泸州某知名浓香型白酒企业黄水和窖泥的DNA,并用紫外吸收和PCR-DGGE方法比较了不同提取方法所获得...为高效地获得浓香型白酒窖池中窖泥和黄水中的微生物总DNA以更好地分析其过程中微生物区系,该文设计了4种不同的总DNA提取方法,分别提取了泸州某知名浓香型白酒企业黄水和窖泥的DNA,并用紫外吸收和PCR-DGGE方法比较了不同提取方法所获得总DNA的纯度及其所代表的原核微生物多样性状况。结果显示,4种提取方法均能从2种样品中提取到纯度较高的DNA;以这些DNA为模板均能扩增出细菌和古菌16S r DNA的部分片段的特异性条带;其DGGE电泳图谱均呈现出较为丰富的条带多样性,但条带存在明显差异。综合评价DNA纯度、扩增效果以及DGGE谱图,对于黄水和窖泥,"酶+SDS+液氮法"提取样品中原核微生物DNA的效果最优。展开更多
The method to analyze both eukaryotic and prokaryotic microorganisms without preliminary microbial information of sample seemed to be useful not only for research and investigation of microorganisms but also for indus...The method to analyze both eukaryotic and prokaryotic microorganisms without preliminary microbial information of sample seemed to be useful not only for research and investigation of microorganisms but also for industry using microorganisms. In the present manuscript, preparation of a new DNA primers, new reference database for 18S rDNA for our newly developed method [1]- [3], and analyses of eukaryotic and prokaryotic microorganisms in fermentation products were presented. In komekouji, Aspergillus spp., was enumerated to be 46.5 × 106 MPN g<sup>-1</sup>, and Penicillium spp., was enumerated to be 1.5 × 106 MPN g<sup>-1</sup>. In dry yeast, Saccharomyces group, were enumerated to be 8600 × 106 MPN g<sup>-1</sup>. In komekouji-miso, no eukaryotic microorganism was detected, while the other Bacillus spp., was numerically dominant (21.5 × 106 MPN g<sup>-1</sup>) as prokaryotic microorganisms, followed by B. subtilis group (4.65 × 106 MPN g<sup>-1</sup>), and the other Firmicutes (3.7 × 106 MPN g<sup>-1</sup>). The komekouji-miso included lower number of Actinobacteria (0.15 × 106 MPN g<sup>-1</sup>), Burkhokderia sp. (1.5 × 106 MPN g<sup>-1</sup>), and the other α,β,γ-proteobacteria (0.12 × 106 MPN g<sup>-1</sup>). In sake-kasu, both prokaryote and eukaryote were not detected by the method. Present results indicated that using both universal primers for eukaryotic and prokaryotic microorganisms, each groups of prokaryotic and eukaryotic microorganisms were enumerated without any preliminary information nor setting up standard curve, which were required for real time PCR.展开更多
文摘为高效地获得浓香型白酒窖池中窖泥和黄水中的微生物总DNA以更好地分析其过程中微生物区系,该文设计了4种不同的总DNA提取方法,分别提取了泸州某知名浓香型白酒企业黄水和窖泥的DNA,并用紫外吸收和PCR-DGGE方法比较了不同提取方法所获得总DNA的纯度及其所代表的原核微生物多样性状况。结果显示,4种提取方法均能从2种样品中提取到纯度较高的DNA;以这些DNA为模板均能扩增出细菌和古菌16S r DNA的部分片段的特异性条带;其DGGE电泳图谱均呈现出较为丰富的条带多样性,但条带存在明显差异。综合评价DNA纯度、扩增效果以及DGGE谱图,对于黄水和窖泥,"酶+SDS+液氮法"提取样品中原核微生物DNA的效果最优。
文摘The method to analyze both eukaryotic and prokaryotic microorganisms without preliminary microbial information of sample seemed to be useful not only for research and investigation of microorganisms but also for industry using microorganisms. In the present manuscript, preparation of a new DNA primers, new reference database for 18S rDNA for our newly developed method [1]- [3], and analyses of eukaryotic and prokaryotic microorganisms in fermentation products were presented. In komekouji, Aspergillus spp., was enumerated to be 46.5 × 106 MPN g<sup>-1</sup>, and Penicillium spp., was enumerated to be 1.5 × 106 MPN g<sup>-1</sup>. In dry yeast, Saccharomyces group, were enumerated to be 8600 × 106 MPN g<sup>-1</sup>. In komekouji-miso, no eukaryotic microorganism was detected, while the other Bacillus spp., was numerically dominant (21.5 × 106 MPN g<sup>-1</sup>) as prokaryotic microorganisms, followed by B. subtilis group (4.65 × 106 MPN g<sup>-1</sup>), and the other Firmicutes (3.7 × 106 MPN g<sup>-1</sup>). The komekouji-miso included lower number of Actinobacteria (0.15 × 106 MPN g<sup>-1</sup>), Burkhokderia sp. (1.5 × 106 MPN g<sup>-1</sup>), and the other α,β,γ-proteobacteria (0.12 × 106 MPN g<sup>-1</sup>). In sake-kasu, both prokaryote and eukaryote were not detected by the method. Present results indicated that using both universal primers for eukaryotic and prokaryotic microorganisms, each groups of prokaryotic and eukaryotic microorganisms were enumerated without any preliminary information nor setting up standard curve, which were required for real time PCR.
