Gros O, Darrasse A, Durand P, Frenkiel L, Mouëza M
Département de Biologie, Université des Antilles et de la Guyane, Pointe-à-Pitre, French West Indies.
Appl Environ Microbiol. 1996 Jul;62(7):2324-30. doi: 10.1128/aem.62.7.2324-2330.1996.
Codakia orbicularis is a large tropical member of the bivalve mollusk family Lucinidae which inhabits shallow-water sea-grass beds (Thalassia testudinum environment) and harbors sulfur-oxidixing endosymbiotic bacteria within bacteriocytes of its gill filaments. When a C. orbicularis-specific 16S rDNA (DNA encoding rRNA) primer is used with a bacterium-specific 16S rDNA reverse primer in amplifications by PCR, the primer set was unsuccessful in amplifying symbiont DNA targets from ovaries, eggs, veligers, and metamorphosed juveniles (600 microns to 1 mm in shell length) cultivated in sterile sand, whereas successful amplifications were obtained from gill tissue of adult specimens and from metamorphosed juveniles (600 microns to 1 mm in shell length) cultivated in unsterilized sea-grass bed sand. To ascertain the presence of the symbiont target in juveniles, restriction fragment length polymorphism analysis, Southern blotting, and transmission electron microscopy were used. Specific hybridizations and observation of endosymbiotic bacteria in the gills of numerous juveniles cultivated in unsterilized sea-grass bed sand showed that the sulfur-oxidizing endosymbionts of C. orbicularis are environmentally transmitted to the new generation after larval metamorphosis.
圆拟枣贝是双壳贝类动物科露齿蛤科的一种大型热带贝类,栖息于浅水海草床(泰来藻环境),并在其鳃丝的细菌细胞内寄生着硫氧化内共生细菌。当在聚合酶链式反应(PCR)扩增中使用圆拟枣贝特异性16S核糖体DNA(编码核糖体RNA的DNA)引物与细菌特异性16S核糖体DNA反向引物时,该引物组未能从在无菌沙子中培养的卵巢、卵、面盘幼虫和变态后的幼体(壳长600微米至1毫米)中扩增出共生体DNA靶点,而从成年标本的鳃组织以及在未灭菌的海草床沙子中培养的变态后幼体(壳长600微米至1毫米)中成功扩增出了靶点。为了确定幼体中共生体靶点的存在,采用了限制性片段长度多态性分析、Southern印迹法和透射电子显微镜技术。对在未灭菌的海草床沙子中培养的众多幼体鳃内的内共生细菌进行的特异性杂交和观察表明,圆拟枣贝的硫氧化内共生体在幼体变态后通过环境传递给新一代。