Molecular identification of a bacterium associated with gall formation in the marine red alga Prionitis lanceolata
Molecular identification of a bacterium associated with gall formation in the marine red alga Prionitis lanceolata
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DOI：
10.1111/j.0022-3646.1996.00286.x
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发表时间：
1996-04-01
影响因子：
2.9
通讯作者：
Goff, LJ
中科院分区：
文献类型：
--
作者：
Ashen, JB;Goff, LJ
采用聚合酶链反应（PCR）技术从海洋红藻褐刺藻（proonitis lanceolata Harvey (Gigartinales)）的胃中扩增真菌性小亚基（16S）核糖体DNA （rDNA）基因。这些肿瘤由肥大的藻细胞组成，其中含有大量无法培养的细胞间细菌。从表面灭菌的胆组织塞中克隆pcr扩增的16S rRNA，测序，并通过与现有的小亚基rRNA序列比对（伊利诺伊大学核糖体数据库项目）进行分析。鉴定了可变区域，并用于构建荧光标记的物种特异性寡脱氧核苷酸探针，用于对胆囊共生体进行全细胞原位杂交。探针949 （PLANC.949）在成熟胆组织制剂中定位了杉木假单胞菌共生体。在相同的杂交反应中，该探针不与作为对照的密切相关细菌的rDNA杂交。原位杂交显示同一细菌的存在与杉木瘿形成从加利福尼亚中部的三个地方。距离和简约分析表明，该生物是变形菌门（α分支；红杆菌群）的一员，与反硝化玫瑰杆菌最密切相关。
The polymerase chain reaction (PCR) was used to amplify eubacterial small-subunit (16S) ribosomal DNA (rDNA) genes from galls of the marine red alga Prionitis lanceolata Harvey (Gigartinales). These tumors consist of hypertrophied algal cells containing large numbers of intercellular bacteria that remain uncultivable. PCR-amplified 16S rDNAs from surface-sterilized gall tissue plugs were cloned, sequenced, and analyzed by alignment to available small-subunit rRNA sequences (University of Illinois Ribosomal Database Project). Variable regions were identified and used to construct a fluorescently labeled, species-specific oligodeoxynucleotide probe for whole cell in situ hybridization to the gall symbiont. Probe 949 (PLANC.949) localized the P. lanceolata bacterial symbiont in preparations from mature gall tissue. This probe did not hybridize to the rDNA of closely related bacteria included as controls in the same hybridization reactions. In situ hybridization revealed the presence of the same bacterium in association with P. lanceolata gall formation from three central California localities. Distance and parsimony analyses suggest that this organism is a member of the Proteobacteria (alpha subdivision; Rhodobacter group) and is most closely related to Roseobacter denitrificans.