# Chloracidobacterium validum sp. nov., a thermophilic chlorophotoheterotrophic bacterium of the phylum Acidobacteriota from an alkaline hot spring microbial mat, represents Chloracidobacterium gen. nov., Chloracidobacteriaceae fam. nov. and Chloracidobacteriales ord. nov

## TL;DR

Scientists discovered a new thermophilic bacterium from a hot spring microbial mat and classified it as a new species, genus, family, and order.

## Contribution

The paper introduces a new species, genus, family, and order of thermophilic chlorophotoheterotrophic bacteria based on phenotypic, phylogenetic, and genomic analyses.

## Key findings

- Strain BV2-CT is a novel thermophilic chlorophotoheterotrophic bacterium requiring branched-chain amino acids for growth.
- Genomic analysis revealed two circular chromosomes and a 59.9 mol% G+C content for strain BV2-CT.
- The new genus, family, and order are proposed within the class Blastocatellia based on phylogenomic data.

## Abstract

A novel chlorophototrophic bacterium, strain BV2-CT, was isolated from a microbial mat (~40 °C) at Rupite hot springs, Bulgaria. Cells were rod-shaped, Gram-negative, with numerous fimbriae/pili, but non-motile. Optimal growth occurred under micro-oxic conditions in the light (20–50 µmol photons m−2 s−1) at 45 °C (range 35–50 °C) and pH 7.2 (range pH 5.5–9.0). Bacteriochlorophyll c, bacteriochlorophyll aP, echinenone and γ-carotene were the major pigments in cells capable of producing chlorosomes and type-1 homodimeric reaction centres. Dense liquid culture appeared greenish-brown, with cells forming small clumps and aggregates. Strain BV2-CT grows photoheterotrophically using organic compounds as carbon source (i.e. a mixture of 20 proteinogenic amino acids or peptone) and thioglycolate as reduced sulphur source. Strain BV2-CT obligately required branched-chain amino acids (l-isoleucine, l-leucine and l-valine) and l-lysine for growth. It is closely related to Chloracidobacterium thermophilum and ‘Candidatus Chloracidobacterium aggregatum’ with 16S rRNA gene sequence identities of 96.97 and 97.78%, respectively. The genome of strain BV2-CT comprises two circular chromosomes of 2,659,040 and 1,000,103 bp (total genome, 3.65 Mb) and G+C content of 59.9 mol%. Phenotypic, phylogenetic and genomic analyses demonstrated that Chloracidobacterium validum strain BV2-CT (=DSM 113832T=JCM 39534T) represents a novel species of the genus ‘Candidatus Chloracidobacterium’, which we hereby propose as Chloracidobacterium gen. nov. with Chloracidobacterium validum sp. nov. as the type species. Based on phylogenomic analyses, we also propose a new family, Chloroacidobacteriaceae fam. nov., and a new order, Chloracidobacteriales ord. nov., within the class Blastocatellia. We further propose the name ‘Ca. C. aggregatum’ for pro-valid publication with the genome sequence of strain E as nomenclatural type.