Bianciotto Valeria, Lumini Erica, Bonfante Paola, Vandamme Peter
Istituto per la Protezione delle Piante (Sezione di Torino) del CNR and Dipartimento di Biologia Vegetale dell'Università di Torino, Viale Mattioli 25, 10125 Torino, Italy.
Laboratorium voor Microbiologie, Faculteit Wetenschappen, Universiteit Gent, Gent, Belgium.
Int J Syst Evol Microbiol. 2003 Jan;53(Pt 1):121-124. doi: 10.1099/ijs.0.02382-0.
Arbuscular mycorrhizal fungi are obligate endosymbionts that colonize the roots of almost 80 % of land plants. The present paper describes morphological and molecular data on a bacterial endosymbiont living in the cytoplasm of dormant or germinating spores and symbiotic mycelia of the fungal species Gigaspora margarita, Scutellospora persica and Scutellospora castanea. PCR amplification of almost the entire 16S rRNA gene of the Gigaspora margarita BEG 34 endosymbiont, using universal bacterial primers, and subsequent sequence analysis demonstrated that this organism occupies a very distinct phylogenetic position within the beta-Proteobacteria, with the genera Burkholderia, Pandoraea and Ralstonia as its closest neighbours. Primers specific to the 16S rDNA of the endosymbiotic bacteria of BEG 34 allowed amplification of spore DNA from endosymbionts of Gigaspora margarita, Gigaspora decipiens, S. persica and S. castanea, but not from the Gigaspora gigantea endosymbiont (which was morphologically different) or from the cytoplasm of Gigaspora rosea (which did not contain endosymbiotic bacteria). These specific primers were successfully used as a probe for the in-situ hybridization of endobacteria in Gigaspora margarita spores. The overall rod-shaped morphology of the Gigaspora margarita, Gigaspora decipiens, S. persica and S. castanea endosymbionts was similar, and amplification and sequence analysis of the almost-complete 16S rRNA genes of several Gigaspora margarita, S. persica and S. castanea endosymbionts revealed over 98% sequence similarity. These morphological and genomic characteristics were used to assign the endosymbionts of these three species (five isolates) of arbuscular mycorrhizal fungi as 'Candidatus Glomeribacter gigasporarum' gen. nov., sp. nov.
丛枝菌根真菌是专性内生共生菌,能定殖于近80%的陆地植物根系中。本文描述了一种细菌内生共生菌的形态学和分子数据,该内生共生菌存在于巨孢球囊霉、波斯盾巨孢囊霉和栗色盾巨孢囊霉的休眠或萌发孢子以及共生菌丝体的细胞质中。使用通用细菌引物对巨孢球囊霉BEG 34内生共生菌的几乎整个16S rRNA基因进行PCR扩增,并进行后续序列分析,结果表明该生物体在β-变形杆菌中占据非常独特的系统发育位置,与伯克霍尔德氏菌属、潘多拉菌属和罗尔斯通氏菌属是最接近的邻居。针对BEG 34内共生细菌16S rDNA的特异性引物能够扩增巨孢球囊霉、欺骗性球囊霉、波斯盾巨孢囊霉和栗色盾巨孢囊霉内共生体的孢子DNA,但不能扩增巨大球囊霉内共生体(其形态不同)或玫瑰色球囊霉细胞质(不含内共生细菌)的DNA。这些特异性引物成功用作巨孢球囊霉孢子内细菌原位杂交的探针。巨孢球囊霉、欺骗性球囊霉、波斯盾巨孢囊霉和栗色盾巨孢囊霉内共生体的整体杆状形态相似,对几种巨孢球囊霉、波斯盾巨孢囊霉和栗色盾巨孢囊霉内共生体的几乎完整的16S rRNA基因进行扩增和序列分析,结果显示序列相似性超过98%。这些形态学和基因组特征被用于将这三种丛枝菌根真菌(五个分离株)的内共生体归为“新种球形球囊杆菌”新属、新种。