Actinoplanes pyxinae sp. nov., a new lichen-derived rare actinobacterium exhibiting antimicrobial and anticancer activity.

A novel lichen-derived actinobacterium, designated Pm04-4T, was isolated from Pyxine cocoes (Sw.) Nyl. lichen collected from Chaiyaphum, Thailand. A polyphasic approach was used to describe the taxonomic position of the strain. The strain had morphological and chemotaxonomic properties similar to members of the genus Actinoplanes. It produced sporangia on the substrate mycelia. Meso-diaminopimelic acid, galactose, glucose and mannose were detected in the whole-cell hydrolysate of the strain. The major menaquinone was MK-9(H4). The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside. The predominant cellular fatty acids were iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. Strain Pm04-4T showed the highest 16S rRNA gene sequence similarity to Actinoplanes akusuensis TRM 8003T (99.0 %). In the phylogenomic tree, strain Pm04-4T was positioned close to A. aksuensis TRM88003T, A. maris M416T, A. polyasparticus TRM66264T, A. hotanensis TRM88002T, A. abujensis DSM 45518T, A. bogorensis NBRC 110975T, A. brasiliensis DSM 43805T, A. lichenicola LDG1-01T and A. ovalisporus LDG1-06T. The average nucleotide identity and digital DNA-DNA hybridization values between strain Pm04-4T and its closely related neighbours were below the threshold values for describing new species. Moreover, the strain could be distinguished from its closely related type strains by phenotypic properties. Based on genotypic and phenotypic evidence, it can be concluded that strain Pm04-4T is a representative of a new Actinoplanes species for which the name Actinoplanes pyxinae sp. nov. is proposed. The type strain is Pm04-4T (=TBRC 16207T=NBRC 115836T). The type strain exhibited activity against Staphylococcus aureus ATCC 25923 as well as four yeast strains, namely Candida albicans TISTR 5554, Candida glabrata TISTR 5006, Candida krusei TISTR 5351 and Candida parapsilosis TISTR 5007. It also showed cytotoxicity against Caco-2, MNT-1 and MCF-7 cancer cells.

Phytohabitans aurantiacus sp. nov., an actinomycete isolated from soil.

A novel actinomycete, designated RD004123T, was isolated from a soil sample collected in Hokkaido, Japan, and its taxonomic position was investigated by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain RD004123T fell within the cluster of the family Micromonosporaceae but did not form a reliable cluster with any member of the family. The similarity values between strain RD004123T and the type species of 29 genera in the family Micromonosporaceae were 91.7-97.7 %. Meanwhile, phylogenomic analyses indicated that strain RD004123T was closely related to members of the genus Phytohabitans. Strain RD004123T contained both meso-diaminopimelic acid and l-lysine as the diagnostic diamino acids of the peptidoglycan. The predominant isoprenoid quinones were MK-10(H8) and MK-10(H6), and the major fatty acids were anteiso-C17 :  0, iso-C16 :  0, iso-C15 :  0 and C17 :  0. The detected polar lipids were phosphatidylinositol mannosides, phosphatidylinositol, phosphatidylethanolamine and diphosphatidylglycerol. These chemotaxonomic features corresponded to those of the genus Phytohabitans. Meanwhile, the results of genome comparison analyses and phenotypic characterizations distinguished strain RD004123T from the other members of the genus Phytohabitans. Therefore, strain RD004123T should be assigned as representing a novel species of the genus Phytohabitans, for which the name Phytohabitans aurantiacus sp. nov. is proposed. The type strain is RD004123T (=NBRC 114997T=DSM 114330T).

Description of Jidongwangia harbinensis gen. nov. sp. nov.

During our previous study, strain NEAU-J3T was classified as representing a novel genus 'Wangella' within the family Micromonosporaceae. Nevertheless, it is a great pity the name cannot be validated as the proposed genus name is illegitimate (Principle 2 of the ICNP). In this study, we describe Jidongwangia as a novel genus within the family Micromonosporaceae and a polyphasic approach was used to provide evidence to support the classification. The G+C content of the genomic DNA of the type strain is 71.6 %. Digital DNA-DNA hybridization and average nucleotide identity (ANI) values could be used to differentiate NEAU-J3T from its related type strains. The phenotypic, genetic and chemotaxonomic data also indicated that NEAU-J3T occupies a branch separated from those of known genera in the family Micromonosporaceae. Therefore, NEAU-J3T represents a novel species of a novel genus in the family Micromonosporaceae, for which the name Jidongwangia harbinensis gen. nov., sp. nov. is proposed. The type strain of Jidongwangia harbinensis is NEAU-J3T (= CGMCC 4.7039T = DSM 45747T).

Descriptions of Micromonospora grosourdyae nom. nov., Micromonospora sonchi comb. nov. and Micromonospora thawaii sp. nov. to resolve problems with the taxonomy and nomenclature of strains named Micromonospora endophytica.

The name Micromonospora endophytica has been used for three different organisms. The first organism with this name is the species represented by strain DCWR9-8-2T, a species published in 2015 but whose name was never validated. In 2019 the type species of the genus Jishengella was reclassified into the genus Micromonospora, while maintaining its original epithet, thus establishing the second group of organisms known as M. endophytica, but the first for which the name was validated. Additionally, in 2018 the reclassification of the genus Verrucosispora into the genus Micromonospora was proposed, but a new epithet has not been specified for the species named Verrucosispora endophytica, which remains an orphaned species. Therefore, it is necessary to propose new names that can unequivocally identify these taxa. We have analysed the taxonomic position of the strains, comparing them with the species with valid published names of the genus Micromonospora. We here propose Micromonospora thawaii sp. nov. for the species represented by strain DCWR9-8-2T, and Micromonospora grosourdyae nom. nov. and Micromonospora sonchi comb. nov. for the two orphaned species of Verrucosispora, V. endophytica and Verrucosispora sonchi, respectively. Genomic analysis also showed that M. trujilloniae is a later heterotypic synonym of M. andamanensis.

Allorhizocola rhizosphaerae gen. nov., sp. nov., a new member of Micromonosporaceae isolated from rhizosphere soil of the plant Calligonum mongolicum.

The taxonomic position of an actinobacterium, designated CPCC 204380T, which was isolated from a rhizosphere soil sample of the plant Calligonum mongolicum collected from Xinjiang Province, China, was established using a polyphasic approach. Vegetative hyphae developed well and globose bodies formed from aged hyphae. Spore chains that differentiated from the vegetative hyphae contained non-motile rod-shaped spores. The peptidoglycan contained meso-diaminopimelic acid and 3-hydroxydiaminopimelic acid as the diagnostic amino acids. The acyl type of the peptidoglycan was glycolyl. Glucose, mannose, ribose and xylose were detected in whole-cell hydrolysates. The predominant menaquinone was MK-10(H8), followed by MK-10(H6) and MK-10(H4). The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major fatty acids were iso-C15 : 0, iso-C16 : 0 and C17 : 1ω9c. The genomic G+C content was 64.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CPCC 204380T should be placed in the family Micromonosporaceae, in which it formed a distinct lineage next to the genera Rhizocola, Catellatospora, Catelliglobosispora, Hamadaea and Allocatelliglobosispora. It shared the highest 16S rRNA gene sequence similarities with Rhizocola hellebori K12-0602T (96.1 %), Catellatospora chokoriensis 2-25/1T (95.9 %), Catelliglobosispora koreensis DSM 44566T (95.9 %), Hamadaea tsunoensis DSM 44101T (95.3 %) and Allocatelliglobosispora scoriae Sco-B14 T (94.2 %), and less than 94.0 % sequence similarity with other validly described species. The combination of phylogenetic analysis and phenotypic characteristics supported the proposal of strain CPCC 204380T as representing a novel species of a new genus in the family Micromonosporaceae, for which the name Allorhizocola rhizosphaerae gen. nov., sp. nov. is proposed. CPCC 204380T (=DSM 102292T=KCTC 39746 T) is the type strain of the type species.

Micromonospora azadirachtae sp. nov., isolated from roots of Azadirachta indica A. Juss. var. siamensis Valeton.

A Gram-stain positive actinomycete, strain AZ1-19T, isolated from roots of Azadirachta indica A. Juss. var. siamensis Valeton, collected from Chachoengsao province, Thailand, was characterised taxonomically by using a polyphasic approach. Strain AZ1-19T was found to have characteristics consistent with those of members of the genus Micromonospora. The cell wall peptidoglycan of the strain was found to contain meso-diaminopimelic acid. The predominant phospholipids were identified as diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The characteristic whole-cell sugars were identified as glucose, xylose, galactose and mannose. The major menaquinones were found to be MK-9(H6), MK-10(H6) and MK-10(H8), and the major cellular fatty acids were identified as iso-C16:0, iso-C15:0, anteiso-C15:0 and anteiso-C17:0. Comparative analysis of 16S rRNA gene sequences revealed that strain AZ1-19T is closely related to Micromonospora costi CS1-12T (98.75% similarity), Micromonospora avicenniae 268506T (98.75%), Micromonospora haikouensis 232617T (98.68%) and Micromonospora siamensis TT2-4T (98.61%), whilst the corresponding phylogenetic analysis based on partial gyrase subunit B (gyrB) gene sequences indicated that strain AZ1-19T forms a clade with M. avicenniae 268506T with a high bootstrap value. The DNA G + C content was determined to be 69.8 mol%. Moreover, a combination of DNA-DNA relatedness values and some phenotypic and chemotaxonomic properties indicated that the strain could be distinguished from closely related species. Therefore, it is considered that strain AZ1-19T represents a novel Micromonospora species for which the name Micromonospora azadirachtae sp. nov. is proposed. The type strain is AZ1-19T (= KCTC 39941T = NBRC 112784T = JCM 32148T = TISTR 2559T).

Micromonospora globbae sp. nov., an endophytic actinomycete isolated from roots of Globba winitii C. H. Wright.

A novel endophytic actinomycete, strain WPS1-2T, isolated from a root of Globba winitii C. H. Wright, was characterized taxonomically by using a polyphasic approach. Strain WPS1-2T exhibited identical characteristics to the members of the genus Micromonospora. Single spores were observed directly on substrate mycelia. The cell-wall peptidoglycan of the strain contained meso-diaminopimelic acid and 3-OH-meso-diaminopimelic acid. Whole-cell hydrolysates contained glucose, ribose, arabinose and xylose. The predominant menaquinones were MK-10(H8) and MK-10(H10). The major cellular fatty acids consisted of iso-C15 : 0, iso-C16 : 0 and anteiso-C15 : 0. According to the 16S rRNA gene sequence of the strain, WPS1-2T showed highest similarity to Micromonospora costi CS1-12T (99.02 %). Phylogenetic analysis of the gyrase subunit B (gyrB) gene indicated that the strain was related to M. costi CS1-12T. The DNA G+C content was 73.7 mol%. The strain could be distinguished from closely related type strains by using a combination of morphological, chemotaxonomic, physiological and biochemical data together with DNA-DNA relatedness values. Based on these observations, strain WPS1-2T is considered to represent a novel species of the genus Micromonospora, for which the name Micromonospora globbae sp. nov. is proposed. The type strain is WPS1-2T (=KCTC 39787T=NBRC 112325T=TISTR 2405T).

Micromonospora soli sp. nov., isolated from rice rhizosphere soil.

An actinomycete strain SL3-70(T) was isolated from a rice field and characterised using a polyphasic approach. The morphological and chemotaxonomical characteristics of strain SL3-70(T) indicate that it belongs to the genus Micromonospora. The phylogenetic analysis of the nearly complete 16S rRNA gene sequence revealed that strain SL3-70(T) is a member of the genus Micromonospora, and is closely related to Micromonospora echinaurantica DSM 43904(T) (99.1 % 16S rRNA gene sequence similarity) and Micromonospora kangleipakensis MBRL 34(T) (98.8 %). DNA-DNA relatedness between strain SL3-70(T) and its relatives ranged from 21.2 % ± 0.6 to 38.7 % ± 0.4. The results obtained from our study indicate that strain SL3-70(T) represents a novel species of the genus Micromonospora, for which the name Micromonospora soli sp. nov. is proposed. The type strain is SL3-70(T) (=BCC 67268(T); =NBRC 110009(T)).

Phytomonospora cypria sp. nov., isolated from soil.

A Gram-stain positive actinobacterial strain, designated KT1403(T), was isolated from a soil sample, collected from Karpaz, Magusa, Northern Cyprus, and characterised using a polyphasic approach. Morphological characteristics and chemotaxonomic data indicated that the strain belongs to the genus Phytomonospora. The cell wall of the novel strain contained meso-diaminopimelic acid and galactose, glucose and mannose as the major sugars in whole cell hydrolysates. The polar lipids in the cell membrane were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, an unidentified aminophospholipid and three unidentified glycolipids. The predominant menaquinones were MK-10(H6) and MK-10(H4). The major fatty acids were found to be iso C15:0 , anteiso C15:0 and anteiso C17:0. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain KT1403(T) belongs to the genus Phytomonospora with a sequence similarity of 99.73 % with Phytomonospora endophytica, the type species of the genus. DNA-DNA hybridization further differentiated strain KT1403(T) from its near phylogenetic neighbour, P. endophytica DSM 45386(T) (29.0 ± 2.2 % DNA relatedness). Therefore, it is proposed that strain KT1403(T) represents a novel species of the genus Phytomonospora, for which the name Phytomonospora cypria sp. nov. is proposed. The type strain is KT1403(T) (=KCTC 29479(T) = DSM 46767(T)).

Investigation of Interspecies Interactions within Marine Micromonosporaceae Using an Improved Co-Culture Approach.

With respect to bacterial natural products, a significant outcome of the genomic era was that the biosynthetic potential in many microorganisms surpassed the number of compounds isolated under standard laboratory growth conditions, particularly among certain members in the phylum Actinobacteria. Our group, as well as others, investigated interspecies interactions, via co-culture, as a technique to coax bacteria to produce novel natural products. While co-culture provides new opportunities, challenges exist and questions surrounding these methods remain unanswered. In marine bacteria, for example, how prevalent are interspecies interactions and how commonly do interactions result in novel natural products? In an attempt to begin to answer basic questions surrounding co-culture of marine microorganisms, we have tested both antibiotic activity-based and LC/MS-based methods to evaluate Micromonosporaceae secondary metabolite production in co-culture. Overall, our investigation of 65 Micromonosporaceae led to the identification of 12 Micromonosporaceae across three genera that produced unique metabolites in co-culture. Our results suggest that interspecies interactions were prevalent between marine Micromonosporaceae and marine mycolic acid-containing bacteria. Furthermore, our approach highlights a sensitive and rapid method for investigating interspecies interactions in search of novel antibiotics, secondary metabolites, and genes.

Natronosporangium hydrolyticum gen. nov., sp. nov., a haloalkaliphilic polyhydrolytic actinobacterium from a soda solonchak soil in Central Asia.

During a cultural diversity survey on hydrolytic bacteria in saline alkaline soils, a hydrolytic actinobacterium strain ACPA39T was enriched and isolated in pure culture from a soda solonchak soil in southwestern Siberia. It forms a substrate mycelium with rod-shaped sporangia containing 1-3 exospores. The isolate is obligately alkaliphilic, growing at pH 7.5-10.3 (optimum at 8.5-9.0) and moderately halophilic, tolerating up to 3 M total Na+ in the form of sodium carbonates. It is an obligately aerobic, organoheteroterophic, saccharolytic bacterium, utilizing various sugars and alpha/beta-glucans as growth substrates. According to the 16S rRNA gene-based phylogenetic analysis, strain ACPA39T forms a distinct branch within the family Micromonosporaceae, with the sequence identities below 94.5% with type strains of other genera. This is confirmed by phylogenomic analysis based on the 120 conserved single copy protein-based markers and genomic indexes (ANI, AAI). The cell-wall of ACPA39T contained meso-DAP, glycine, glutamic acid and alanine in a equimolar ratio, characteristic of the peptidoglycan type A1γ'. The whole-cell sugars include galactose and xylose. The major menaquinone is MK-10(H4). The identified polar lipids consist of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The polar lipid fatty acids were dominated by anteiso-C17:0, iso-C16:0, iso-C17:0, 10 Me-C18:0 and C18:1ω9. Based on the distinct phylogeny, the chemotaxonomy features and unique phenotypic properties, strain ACPA39T (DSM 106523T = VKM 2772T) is classified into a new genus and species in the family Micromonosporaceae for which the name Natronosporangium hydrolitycum gen. nov., sp. nov. is proposed.

Complete genome sequence of the motile actinomycete Actinoplanes missouriensis 431(T) (= NBRC 102363(T)).

Actinoplanes missouriensis Couch 1963 is a well-characterized member of the genus Actinoplanes, which is of morphological interest because its members typically produce sporangia containing motile spores. The sporangiospores are motile by means of flagella and exhibit chemotactic properties. It is of further interest that members of Actinoplanes are prolific sources of novel antibiotics, enzymes, and other bioactive compounds. Here, we describe the features of A. missouriensis 431(T), together with the complete genome sequence and annotation. The 8,773,466 bp genome contains 8,125 protein-coding and 79 RNA genes.