Symbiosis and Insect Diversiﬁcation: an Ancient Symbiont of Sap-Feeding Insects from the Bacterial Phylum Bacteroidetes-猫眼课题宝

DOI：

发表时间：

2005

期刊：

影响因子：

通讯作者：

中科院分区：

其他

文献类型：

作者：

J. Cryan;G. Monteith;K. Morishima;M. Whiting

一些昆虫群体有专性的、垂直传播的细菌共生体，为宿主提供饮食中有限的营养。其中一些细菌已被证明是从古代感染中遗传下来的。在这里，我们展示了包括蝉、叶蝉、树蝉、飞虱和飞虱在内的一大群相关昆虫，它们拥有一个独特的细菌共生体分支。这个新描述的共生谱系属于拟杆菌门。对16S rRNA基因的分析表明，该共生体的系统发育与目前已知的昆虫宿主的系统发育完全一致。这些结果支持这些昆虫的共同祖先在古代获得共生体，将原始感染追溯到至少2.6亿年前。从一种飞沫虫（cercopo总科）和一种神枪手虫（Cicadellinae）中可以看出，共生体的细胞非常大，呈细长状，长度通常超过30 （cid:1）米；原位杂交证实这些对应于拟杆菌门。“Candidatus Sulcia muelleri”被提议作为新的共生体的名称。将C. arizonana的晚期若虫解剖到PA缓冲液中（50 mM Tris- HCl [pH 7.6], 100 mM EDTA， 250 mM蔗糖），在盖盖下轻轻操作破坏，用SYBR-Gold染色，并在1000倍放大下使用荧光检查。同样的方法被应用于整个乳状线虫菌群。为了确定与拟杆菌门rRNA基因序列相对应的生物，进行了原位杂交。我们使用了两个探针，每个探针长20个核苷酸。其中CFB319 (5 （cid:2) -TGG TCC GTG TCT CAG TAC-3 (cid:2)）与拟杆菌门16S rRNA的319 ~ 336位完全匹配，包括我们从这些共生体中获得的所有序列；这个探针显示两个或更多的不匹配已知的细菌在其他门。另一个基因Pro319 (5 （cid:2) -TGG ACC GTG TCT CAG TTC-3 (cid:2)）在两个位点上不同，与Buchnera、Candidatus Baumannia cicadellinicola和其他一些Gammaproteobacteria的16S rRNA序列完全匹配。CFB319与6-羧基四甲基罗丹明连接（吸收/发射551/ 576nm）；Pro319连接到alexa488nm)。测定杂交细菌细胞的特异性

Several insect groups have obligate, vertically transmitted bacterial symbionts that provision hosts with nutrients that are limiting in the diet. Some of these bacteria have been shown to descend from ancient infections. Here we show that the large group of related insects including cicadas, leafhoppers, treehoppers, spittlebugs, and planthoppers host a distinct clade of bacterial symbionts. This newly described symbiont lineage belongs to the phylum Bacteroidetes . Analyses of 16S rRNA genes indicate that the symbiont phylogeny is completely congruent with the phylogeny of insect hosts as currently known. These results support the ancient acquisition of a symbiont by a shared ancestor of these insects, dating the original infection to at least 260 million years ago. As visualized in a species of spittlebug (Cercopoidea) and in a species of sharpshooter (Cicadellinae), the symbionts have extraordinarily large cells with an elongate shape, often more than 30 (cid:1) m in length; in situ hybridizations verify that these correspond to the phylum Bacteroidetes . “ Candidatus Sulcia muelleri” is proposed as the name of the new symbiont. of late-stage nymphs of C. arizonana were dissected into PA buffer (50 mM Tris- HCl [pH 7.6], 100 mM EDTA, 250 mM sucrose), disrupted by manipulating gently under a coverslip, stained with SYBR-Gold, and examined using ﬂuores- cence at a 1,000-fold magniﬁcation. The same procedure was applied to the entire bacteriome of Homalodisca lacerta . To determine the organisms corresponding to the Bacteroidetes rRNA gene sequences, in situ hybridization was performed. We used two probes, each 20 nucleotides long. One, CFB319 (5 (cid:2) -TGG TCC GTG TCT CAG TAC-3 (cid:2) ), was a perfect match to positions 319 to 336 of the 16S rRNA of Bacteroidetes species, including all of our sequences from these symbionts; this probe shows two or more mismatches to known bacteria in other phyla. The other, Pro319 (5 (cid:2) -TGG ACC GTG TCT CAG TTC-3 (cid:2) ), differed at two sites and was a perfect match to 16S rRNA sequences of Buchnera , “ Candidatus Baumannia cicadellinicola”, and some other Gammaproteobacteria . CFB319 was linked to 6-carboxytetramethyl-rhodamine (absorption/emission of 551/576 nm); Pro319 was linked to Alexa488 nm). determine the speciﬁcity hybridiza- bacteriocytes

Symbiosis and Insect Diversiﬁcation: an Ancient Symbiont of Sap-Feeding Insects from the Bacterial Phylum Bacteroidetes

文献摘要

被引文献

相似文献