Microbacterium tenebrionis sp. nov. and Microbacterium allomyrinae sp. nov., isolated from larvae of Tenebrio molitor L. and Allomyrina dichotoma, respectively

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Abstract

Two Gram-positive bacterial strains, designated as YMB-B2 ^T and BWT-G7 ^T, were isolated from larvae of Tenebrio molitor L. and Allomyrina dichotoma, respectively, and their taxonomic positions examined by a polyphasic approach. Both of the isolates contained ornithine as the cell-wall diamino acid. The acyl type of murein was N-glycolyl. The predominant menaquinones were MK-11 and MK-12. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unidentified glycolipid. Both of the isolates contained C _{15 : 0} anteiso and C _{17 : 0} anteiso as the major fatty acids. Strain YMB-B2 ^T also had C _{16 : 0} iso as an additional major fatty acid. The 16S rRNA gene phylogeny showed that the novel strains formed two distinct sublines within the genus Microbacterium . Strain YMB-B2 ^T was most closely related to the type strains of Microbacterium aerolatum (99.1 % sequence similarity) and Microbacterium ginsengiterrae (99.0 %) , whereas strain BWT-G7 ^T formed a tight cluster with the type strain of Microbacterium thalassium (98.9 %). The phylogenomic analysis based on 92 core genes supported their relationships in 16S rRNA gene phylogeny. Overall genomic relatedness indices warranted that the isolates represent two new species of the genus Microbacterium . Based on the results obtained here, Microbacterium tenebrionis sp. nov. (type strain YMB-B2 ^T=KCTC 49593 ^T=CCM 9151 ^T) and Microbacterium allomyrinae sp. nov. (type strain BWT-G7 ^T=KACC 22262 ^T=NBRC 115127 ^T) are proposed.

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The neighbor-joining method: a new method for reconstructing phylogenetic trees.

N Saitou, M Nei (1987)

A new method called the neighbor-joining method is proposed for reconstructing phylogenetic trees from evolutionary distance data. The principle of this method is to find pairs of operational taxonomic units (OTUs [= neighbors]) that minimize the total branch length at each stage of clustering of OTUs starting with a starlike tree. The branch lengths as well as the topology of a parsimonious tree can quickly be obtained by using this method. Using computer simulation, we studied the efficiency of this method in obtaining the correct unrooted tree in comparison with that of five other tree-making methods: the unweighted pair group method of analysis, Farris's method, Sattath and Tversky's method, Li's method, and Tateno et al.'s modified Farris method. The new, neighbor-joining method and Sattath and Tversky's method are shown to be generally better than the other methods.

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A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences.

Motoo Kimura (1980)

Some simple formulae were obtained which enable us to estimate evolutionary distances in terms of the number of nucleotide substitutions (and, also, the evolutionary rates when the divergence times are known). In comparing a pair of nucleotide sequences, we distinguish two types of differences; if homologous sites are occupied by different nucleotide bases but both are purines or both pyrimidines, the difference is called type I (or "transition" type), while, if one of the two is a purine and the other is a pyrimidine, the difference is called type II (or "transversion" type). Letting P and Q be respectively the fractions of nucleotide sites showing type I and type II differences between two sequences compared, then the evolutionary distance per site is K = -(1/2) ln [(1-2P-Q) square root of 1-2Q]. The evolutionary rate per year is then given by k = K/(2T), where T is the time since the divergence of the two sequences. If only the third codon positions are compared, the synonymous component of the evolutionary base substitutions per site is estimated by K'S = -(1/2) ln (1-2P-Q). Also, formulae for standard errors were obtained. Some examples were worked out using reported globin sequences to show that synonymous substitutions occur at much higher rates than amino acid-altering substitutions in evolution.

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CONFIDENCE LIMITS ON PHYLOGENIES: AN APPROACH USING THE BOOTSTRAP.

Joseph Felsenstein (1985)

The recently-developed statistical method known as the "bootstrap" can be used to place confidence intervals on phylogenies. It involves resampling points from one's own data, with replacement, to create a series of bootstrap samples of the same size as the original data. Each of these is analyzed, and the variation among the resulting estimates taken to indicate the size of the error involved in making estimates from the original data. In the case of phylogenies, it is argued that the proper method of resampling is to keep all of the original species while sampling characters with replacement, under the assumption that the characters have been independently drawn by the systematist and have evolved independently. Majority-rule consensus trees can be used to construct a phylogeny showing all of the inferred monophyletic groups that occurred in a majority of the bootstrap samples. If a group shows up 95% of the time or more, the evidence for it is taken to be statistically significant. Existing computer programs can be used to analyze different bootstrap samples by using weights on the characters, the weight of a character being how many times it was drawn in bootstrap sampling. When all characters are perfectly compatible, as envisioned by Hennig, bootstrap sampling becomes unnecessary; the bootstrap method would show significant evidence for a group if it is defined by three or more characters.