Census of the bacterial community of the gypsy moth larval midgut by using culturing and culture-independent methods
Census of the bacterial community of the gypsy moth larval midgut by using culturing and culture-independent methods
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DOI：
10.1128/aem.70.1.293-300.2004
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发表时间：
2004-01-01
影响因子：
4.4
通讯作者：
Handelsman, J
中科院分区：
文献类型：
--
作者：
Broderick, NA;Raffa, KF;Handelsman, J
人们对与鳞翅目相关的细菌知之甚少，鳞翅目是一大类主要为植食性昆虫，包括飞蛾和蝴蝶。我们通过使用传统培养和独立于培养的方法清点了一种多食性食叶动物吉普赛蛾（Lymantria dispar L.）的幼虫中肠细菌，其肠道呈高碱性。我们还研究了饮食对微生物组成的影响。对个体三龄幼虫的分析表明，以相同饮食喂养的昆虫之间的微生物组成高度相似。 DNA序列分析表明，大多数PCR扩增的16S rRNA基因属于γ-变形菌门和低G+C革兰氏阳性菌，并且培养的成员代表了已鉴定的系统发育型的一半以上。较少检测到的分类群包括α-变形杆菌属、放线杆菌属和噬细胞菌属/柔韧杆菌属/拟杆菌属的成员。 15 个培养生物中的 7 个以及通过 PCR 扩增鉴定的 9 个序列中的 8 个的 16S rRNA 基因序列与之前报道的细菌序列不同。以无菌人工饲料、白杨、落叶松、白橡树或柳树为食的幼虫的中肠微生物组成有很大差异。培养分离株的 16S rRNA 分析表明肠球菌属物种和独立于培养物的分析表明肠杆菌属 sp。两者都存在于所有幼虫中，无论其摄食基质如何；这两种系统发育型的序列在昆虫个体之间的差异小于 1%。这些结果首次全面描述了鳞翅目中肠的微生物多样性，并证明饮食中的植物种类影响肠道细菌群落的组成。
Little is known about bacteria associated with Lepidoptera, the large group of mostly phytophagous insects comprising the moths and butterflies. We inventoried the larval midgut bacteria of a polyphagous foliivore, the gypsy moth (Lymantria dispar L.), whose gut is highly alkaline, by using traditional culturing and culture-independent methods. We also examined the effects of diet on microbial composition. Analysis of individual third-instar larvae revealed a high degree of similarity of microbial composition among insects fed on the same diet. DNA sequence analysis indicated that most of the PCR-amplified 16S rRNA genes belong to the gamma-Proteobacteria and low G+C gram-positive divisions and that the cultured members represented more than half of the phylotypes identified. Less frequently detected taxa included members of the alpha-Proteobacterium, Actinobacterium, and Cytophaga/Flexibacter/Bacteroides divisions. The 16S rRNA gene sequences from 7 of the 15 cultured organisms and 8 of the 9 sequences identified by PCR amplification diverged from previously reported bacterial sequences. The microbial composition of midguts differed substantially among larvae feeding on a sterilized artificial diet, aspen, larch, white oak, or willow. 16S rRNA analysis of cultured isolates indicated that an Enterococcus species and culture-independent analysis indicated that an Enterobacter sp. were both present in all larvae, regardless of the feeding substrate; the sequences of these two phylotypes varied less than 1% among individual insects. These results provide the first comprehensive description of the microbial diversity of a lepidopteran midgut and demonstrate that the plant species in the diet influences the composition of the gut bacterial community.