Supattra Kitikhun

A newly discovered facultative anaerobic strain, designated as LXY-3^T, was obtained from a soil sample collected at an industrial site in Guangxi, China, known for heavy metal processing. An investigation including phenotypic, chemotaxonomic, and genomic traits was conducted. Phylogenetic analysis based on 16S rRNA showed that LXY-3^T belonged to the genus Paenibacillus. The closest phylogenetic relative of this strain was Paenibacillus anaericanus MH21^T with the similarity of 97.03%. Iso-C_15:0, antéiso-C_15:0, and C_16:1 ω7c alcohol were the major cellular fatty acids. The predominant polar lipids comprised diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), unidentified phospholipids (PL1-PL8), unidentified resistant material (RM1–RM4), and lipids (L1–L3). For genome sequencing, the genomic DNA G+C content of the strain is 51.2 mol%. Comparative genomic analysis revealed that the average nucleotide identity (ANI) values between strain LXY-3^T and its closest phylogenetic relatives within the genus Paenibacillus (represented by type strains) were consistently below the 95% species demarcation threshold. Nitrogen fixation gene cluster (nifB, nifE, nifK, nifN, nifV, nifX, nifD, and nifH) was conserved in the strain. Correspondingly, digital DNA–DNA hybridization (dDDH) values remained below the 70% cutoff for species delineation. These genomic metrics provide compelling evidence that strain LXY-3^T represents a novel species within the genus Paenibacillus. The type strain LXY-3^T (=CGMCC 1.64949^T = JCM 37600^T) is proposed.

Culture-independent and culture-dependent approaches were employed to investigate the taxonomic diversity and biosynthetic gene cluster potential of Actinomycetota in the Mekong River. Through 16S rRNA gene metabarcoding, 21,103 OTUs were revealed to represent 190 genera and at least 595 species of Actinomycetota, including putatively novel taxa. Conventional and in situ cultivation (IC) methods provided 75 Actinomycetota isolates representing 72 species from 21 genera. Of these, 45 species in 4 genera were new to the Thailand Bioresource Research Center (TBRC), a collection of 20,079 Actinomycetota strains from 660 species. Applying both culture-independent and culture-dependent approaches to the same sample revealed greater diversity among the Actinomycetota in the Mekong River than one approach alone.