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Paenibacillus macquariensis subsp. defensor subsp. nov., isolated from boreal soil

¹ Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), Sapporo 062-8517, Japan
² Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan
³ School of Engineering, Hokkaido Tokai University, Sapporo 005-8601, Japan
⁴ Graduate School of Agriculture, Hokkaido University, Sapporo 060-0890, Japan
⁵ Main Botanical Garden named after N.V. Tsitsin, Russian Academy of Sciences, Moscow 127276, Russia

Correspondence
Isao Yumoto
i.yumoto{at}aist.go.jp

Two Gram-variable, aerobic, motile, rod-shaped, endospore-forming bacterial strains, M4-2^T and M4-1, were isolated from soil samples collected from Oblast Magadan, Russian Far East, as micro-organisms antagonistic to the psychrophilic phytopathogenic fungus Typhula ishikariensis. Strains M4-2^T and M4-1 were identified as members of the genus Paenibacillus by phenotypic and phylogenetic analyses based on 16S rRNA gene sequences. The strains contained anteiso-C_{15 : 0} as the major fatty acid (63.0–64.7 %) and MK-7 as the major isoprenoid quinone. The DNA G+C contents were 42.8 and 41.7 mol%, respectively. 16S rRNA gene sequence analysis showed that strains M4-2^T and M4-1 exhibited high similarities with Paenibacillus macquariensis DSM 2^T (99.5 and 99.7 %, respectively) and Paenibacillus antarcticus LMG 22078^T (99.4 and 99.5 %, respectively). There were no clear differences in the phenotypic characteristics and chemotaxonomic and phylogenetic data between the novel isolates and P. macquariensis DSM 2^T. DNA–DNA hybridization experiments between strain M4-2^T and P. macquariensis DSM 2^T and P. antarcticus LMG 22078^T revealed reassociation values of 56 and 49 %, respectively. Multilocus sequence analysis confirmed the differences between the new isolates and reference strains that were observed with the DNA–DNA hybridization studies. On the basis of the results described above, it is proposed that the isolates represent a novel subspecies of P. macquariensis, Paenibacillus macquariensis subsp. defensor subsp. nov. The type strain is M4-2^T (=JCM 14954^T=NCIMB 14397^T).

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains M4-2^T and M4-1 are AB360546 and AB360547, and of strains M4-1 and M4-2^T, P. macquariensis DSM 2^T and P. antarcticus LMG 22078^T, for the rpoB sequences are AB468054–AB468057, for the 16S rRNA gene and 16S–23S ITS sequences are AB468952–AB468955, and for the gyrB sequences are AB469015–AB469018, respectively.

A table of DNA–DNA reassociation values and figures showing the macromorphology of Typhula ishikariensis co-cultured with Paenibacillus macquariensis subsp. defensor subsp. nov. and an extended phylogenetic tree of the genus Paenibacillus based on 16S rRNA gene sequences are available as supplementary material with the online version of this paper.