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利用基因组探测微阵列对泡菜发酵过程中的乳酸菌进行宏转录组分析。

Metatranscriptome analysis of lactic acid bacteria during kimchi fermentation with genome-probing microarrays.

作者信息

Nam Young-Do, Chang Ho-Won, Kim Kyoung-Ho, Roh Seong Woon, Bae Jin-Woo

机构信息

University of Science & Technology, KRIBB, Daejeon, Republic of Korea.

出版信息

Int J Food Microbiol. 2009 Mar 31;130(2):140-6. doi: 10.1016/j.ijfoodmicro.2009.01.007. Epub 2009 Jan 20.

DOI:10.1016/j.ijfoodmicro.2009.01.007
PMID:19215993
Abstract

We constructed genome probing microarrays (GPM) that are specific to 39 lactic acid bacteria (LAB) in an effort to monitor microbial diversity and biological activity during the fermentation of kimchi, a traditional Korean vegetable product known to contain various health-promoting and immunity-boosting factors. Metagenomes and metatranscriptomes extracted from periodically sampled kimchi soup were labeled, hybridized and comparatively analyzed using GPMs. Each metatranscriptome was prepared by subtracting 16S rRNA and 23S rRNA from the total RNA, and selectively synthesizing mRNA-specific cDNAs from the rRNA-subtracted samples. Metagenomic analysis revealed 23 LAB related to kimchi fermentation [defined as bacteria with more than a 1% average relative composition (ARC)]. Metatranscriptome analysis revealed that, with the exception of two microorganisms, all LAB probed in the microarray contributed to kimchi fermentation. Moreover, the relative compositions of the major LAB remained unchanged (there was less than a 1.5% difference between the maximum and minimum values) in our metagenome analysis, while our metatranscriptome analysis revealed significant differences in the relative compositions of major LAB during fermentation (relative compositions changed by 2.4% to 9.5%). These data indicate that microorganisms that are less abundant in the flora (those with less than a 5% ARC in the metagenomic analysis) also participated in kimchi fermentation with relatively high activities.

摘要

我们构建了针对39种乳酸菌(LAB)的基因组探测微阵列(GPM),以监测泡菜发酵过程中的微生物多样性和生物活性。泡菜是一种传统的韩国蔬菜制品,已知含有各种促进健康和增强免疫力的因子。从定期采样的泡菜汤中提取的宏基因组和宏转录组进行标记、杂交,并使用GPM进行比较分析。每个宏转录组是通过从总RNA中减去16S rRNA和23S rRNA,并从减去rRNA的样品中选择性合成mRNA特异性cDNA来制备的。宏基因组分析揭示了23种与泡菜发酵相关的乳酸菌(定义为平均相对组成超过1%的细菌)。宏转录组分析表明,除了两种微生物外,微阵列中检测的所有乳酸菌都对泡菜发酵有贡献。此外,在我们的宏基因组分析中,主要乳酸菌的相对组成保持不变(最大值和最小值之间的差异小于1.5%),而我们的宏转录组分析显示发酵过程中主要乳酸菌的相对组成存在显著差异(相对组成变化了2.4%至9.5%)。这些数据表明,菌群中丰度较低的微生物(在宏基因组分析中平均相对组成低于5%的微生物)也以相对较高的活性参与了泡菜发酵。

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