Phylogenetic diversity of lactic acid bacteria associated with paddy rice silage as determined by 16S ribosomal DNA analysis
Phylogenetic diversity of lactic acid bacteria associated with paddy rice silage as determined by 16S ribosomal DNA analysis
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DOI：
10.1128/aem.69.1.444-451.2003
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发表时间：
2003-01-01
影响因子：
4.4
通讯作者：
Fujita, Y
中科院分区：
文献类型：
--
作者：
Ennahar, S;Cai, YM;Fujita, Y
对从全季水稻青贮饲料中分离到的161株低G + C含量革兰氏阳性菌进行了分类和表型及遗传分析。根据形态学和生化特征，这些乳酸菌可分为肠球菌属（Enterococcus）、乳杆菌属（Lactobacillus）、乳球菌属（Lactococcus）、明串珠菌属（Leuconostoc）、片球菌属（Pediococcus）和魏氏菌属（Weissella）10个类群。16S核糖体DNA（rDNA）的分析用于确认表型分析所示的优势群体的存在，并确定代表菌株的系统发育关系。PCR扩增16S rRNA基因并测序。来自各种LAB分离株的序列显示出与GenBank参考菌株的高度相似性（在98.7%和99.8%之间）。基于16S rDNA序列构建的系统发育树具有较高的一致性，且各节点的自举值较高。除一个物种外，遗传数据与表型鉴定一致。主要为同型发酵的LAB（66%）由植物乳杆菌（24%）、乳酸乳球菌（22%）、类肠明串珠菌（20%）、乳酸片球菌（11%）、短乳杆菌（11%）、粪肠球菌（7%）、泡菜魏氏菌（3%）和戊糖片球菌（2%）组成。本研究是第一个完全记录水稻相关LAB的研究，显示了一个非常多样化的LAB群落，其中涉及水稻青贮发酵过程的物种数量相对较高。基于16SrDNA的乳酸菌群落结构描述方法，有助于揭示水稻青贮饲料中乳酸菌的多样性，并为今后设计合适的接种剂以改善其发酵品质提供参考。
A total of 161 low-G+C-content gram-positive bacteria isolated from whole-crop paddy rice silage were classified and subjected to phenotypic and genetic analyses. Based on morphological and biochemical characters, these presumptive lactic acid bacterium (LAB) isolates were divided into 10 groups that included members of the genera Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, and Weissella. Analysis of the 16S ribosomal DNA (rDNA) was used to confirm the presence of the predominant groups indicated by phenotypic analysis and to determine the phylogenetic affiliation of representative strains. The virtually complete 16S rRNA gene was PCR amplified and sequenced. The sequences from the various LAB isolates showed high degrees of similarity to those of the GenBank reference strains (between 98.7 and 99.8%). Phylogenetic trees based on the 16S rDNA sequence displayed high consistency, with nodes supported by high bootstrap values. With the exception of one species, the genetic data was in agreement with the phenotypic identification. The prevalent LAB, predominantly homofermentative (66%), consisted of Lactobacillus plantarum (24%), Lactococcus lactis (22%), Leuconostoc pseudomesenteroides (20%), Pediococcus acidilactici (11%), Lactobacillus brevis (11%), Enterococcus faecalis (7%), Weissella kimchii (3%), and Pediococcus pentosaceus (2%). The present study, the first to fully document rice-associated LAB, showed a very diverse community of LAB with a relatively high number of species involved in the fermentation process of paddy rice silage. The comprehensive 16S rDNA-based approach to describing LAB community structure was valuable in revealing the large diversity of bacteria inhabiting paddy rice silage and enabling the future design of appropriate inoculants aimed at improving its fermentation quality.