Myxococcus xanthus viability depends on groEL supplied by either of two genes, but the paralogs have different functions during heat shock, predation, and development.

Myxococcus xanthus DK1622 contains two paralogous groEL gene loci that possess both different sequences and different organizations within the genome. Deletion of either one of these two genes alone does not affect cell viability. However, deletion of both groEL genes results in cell death unless a complemented groEL1 or groEL2 gene is present. The groEL1 gene was determined to be essential for cell survival under heat shock conditions; a strain with mutant groEL2 caused cells to be more sensitive than the wild-type strain to higher temperatures. Mutants with a single deletion of either groEL1 (MXAN_4895) or groEL2 (MXAN_4467) had a growth curve similar to that of the wild-type strain DK1622 in medium containing hydrolyzed proteins as the substrate. However, when cells were cultured on medium containing either Escherichia coli cells or casein as the substrate, deletion of groEL2, but not groEL1, led to a deficiency in cell predation and macromolecular feeding. Furthermore, groEL1 was found to play an indispensable role in the development and sporulation of cells, but deletion of groEL2 had no visible effects. Our results suggest that, although alternatively required for cell viability, the products of the two groEL genes have divergent functions in the multicellular social life cycle of M. xanthus DK1622.

Dissecting cell diversity and connectivity in skeletal muscle for myogenesis.

Characterized by their slow adhering property, skeletal muscle myogenic progenitor cells (MPCs) have been widely utilized in skeletal muscle tissue engineering for muscle regeneration, but with limited efficacy. Skeletal muscle regeneration is regulated by various cell types, including a large number of rapidly adhering cells (RACs) where their functions and mechanisms are still unclear. In this study, we explored the function of RACs by co-culturing them with MPCs in a biomimetic skeletal muscle organoid system. Results showed that RACs promoted the myogenic potential of MPCs in the organoid. Single-cell RNA-Seq was also performed, classifying RACs into 7 cell subtypes, including one newly described cell subtype: teno-muscular cells (TMCs). Connectivity map of RACs and MPCs subpopulations revealed potential growth factors (VEGFA and HBEGF) and extracellular matrix (ECM) proteins involvement in the promotion of myogenesis of MPCs during muscle organoid formation. Finally, trans-well experiments and small molecular inhibitors blocking experiments confirmed the role of RACs in the promotion of myogenic differentiation of MPCs. The RACs reported here revealed complex cell diversity and connectivity with MPCs in the biomimetic skeletal muscle organoid system, which not only offers an attractive alternative for disease modeling and in vitro drug screening but also provides clues for in vivo muscle regeneration.

Cloning and characterization of an rRNA methyltransferase from Sorangium cellulosum.

A locus (kmr) responsible for aminoglycosides-resistance of Sorangium cellulosum was cloned and characterized in Myxococcus xanthus. The gene kmr encodes a putative rRNA methyltransferase. Expression of the complete ORF endowed the Myxococcus transformants with the resistance to aminoglycosidic antibiotics of kanamycin, apramycin, gentamycin, neomycin, and tobramycin at an extraordinary high-level (MIC, higher than 500 microg/ml). However, the gene did not function in Escherichia coli cells. In Sorangium genome, the gene kmr was followed by a putative integrase gene, and was highly homologous in different Sorangium strains. The Sorangium rRNA methyltransferase sequence was in low similarity to the reported 16S rRNA methyltransferases, and their resistance spectrums were also different. The results indicate that the rRNA methyltransferase (Kmr) in Sorangium strains is a new member of the rRNA methyltransferases family.

Myxobacterial community is a predominant and highly diverse bacterial group in soil niches.

Although many molecular ecological surveys have been conducted, there is little concerning the details of specific bacterial groups, resulting in an incomplete understanding of the microorganismal composition and community structures in the environment. Myxobacteria are micropredators that are metabolically active in the soil microbial food web and have typically been considered minority components of soil bacterial communities. In this study, we surveyed the percentage of myxobacteria in a single soil sample via pyrosequencing on combined universal libraries of the V3-V4 and V6-V8 hypervariable regions of the 16S rRNA gene. Surprisingly, myxobacteria accounted for 4.10% of the bacterial community and 7.5% of the total operational taxonomic units at the 3% similarity level in the soil, containing almost all of the cultivated myxobacterial families or genera. To testify the appearance of myxobacteria in soil niches, we retrieved myxobacteria-related 16S rRNA gene sequences of 103 high-throughput sequencing data sets obtained from public databases. The results indicated that myxobacteria-related sequences were among the predominant groups in these data sets accounting for 0.4-4.5% of bacterial communities. The abundance of myxobacterial communities were correlated with site temperature, carbon-to-nitrogen ratio and pH values. Based on these results, we discussed the survival strategies of myxobacterial community in soil.

[Effects of aboveground and belowground competition between grass and tree on elm seedlings growth in Horqin Sandy Land].

Elm sparse woodland steppe plays an important role in vegetation restoration and landscape protection in Horqin Sandy Land. In this paper, a two-factor and two-level field experiment was conducted to explore the effects of aboveground and belowground competition between grass and tree on the growth of elm seedlings in the Sandy Land. Five aspects were considered, i.e., seedling biomass, belowground biomass/aboveground biomass, stem height, ratio of root to stem, and leaf number. For the one-year-old elm seedlings, their biomass showed a trend of no competition > aboveground competition > full competition > belowground competition, belowground biomass / aboveground biomass showed a trend of belowground competition > full competition > no competition > aboveground competition, stem height showed a trend of aboveground competition > no competition > full competition > belowground competition, root/stem ratio showed a trend of belowground competition > full competition > no competition > aboveground competition, and leaf number showed a trend of aboveground competition > no competition > belowground competition > full competition. Belowground competition had significant effects on the growth of one-year-old elm seedlings, while aboveground competition did not have. Neither belowground competition nor aboveground competition had significant effects on the growth of two-year-old elm seedlings. It was suggested that in Horqin Sandy Land, grass affected the growth of elm seedlings mainly via below-ground competition, but the belowground competition didn' t affect the resource allocation of elm seedlings. With the age increase of elm seedlings, the effects of grass competition on the growth of elm seedlings became weaker.

Phylogeographic separation of marine and soil myxobacteria at high levels of classification.

Microorganisms are globally dispersed and are able to proliferate in any habitat that supports their lifestyles, which, however, has not yet been explored in any specific microbial taxon. The social myxobacteria are considered typical soil bacteria because they have been identified in various terrestrial samples, a few in coastal areas, but none in other oceanic environments. To explore the prevalence of marine myxobacteria and to investigate their phylogenetic relationships with their terrestrial counterparts, we established myxobacteria-enriched libraries of 16S rRNA gene sequences from four deep-sea sediments collected at depths from 853 to 4675 m and a hydrothermal vent at a depth of 204 m. In all, 68 different myxobacteria-related sequences were identified from randomly sequenced clones of the libraries of different samples. These myxobacterial sequences were diverse but phylogenetically similar at different locations and depths. However, they were separated from terrestrial myxobacteria at high levels of classification. This discovery indicates that the marine myxobacteria are phylogeographically separated from their terrestrial relatives, likely because of geographic separation and environment selection.

Taxonomic analysis of Sorangium strains based on HSP60 and 16S rRNA gene sequences and morphology.

The taxonomy of myxobacteria is based mainly on their morphological characteristics. The genus Sorangium belongs to the myxobacterial suborder Sorangiineae. Strains in the genus were classified either as one species, Sorangium cellulosum, by ignoring divergent morphological characteristics, or into several species; however, the latter classification is based on some dubious morphological characteristics and is inconsistent with the phylogeny constructed from 16S rRNA gene sequences. In this study, two HSP60 (groEL1 and groEL2) genes were amplified and sequenced from 22 Sorangium strains. The groEL1 and groEL2 gene sequences were highly conserved in Sorangium strains, suggesting that these two paralogous genes both play important roles in the life cycle. The phylogeny constructed by the groEL genes was rather consistent with the morphological characteristics of sporangioles. Including information from the phylogenetic analysis and morphological characteristics, it is suggested that the genus Sorangium includes two species.

Fruiting and non-fruiting myxobacteria: a phylogenetic perspective of cultured and uncultured members of this group.

The diversity of myxobacteria present in campus garden soil was surveyed by both cultivation-based and cultivation-independent methods. Detailed phylogenetic analysis of cultured and uncultured myxobacteria 16S rRNA gene sequences revealed that many undescribed relatives of the myxobacteria exist in nature. Molecular systematic analyses also revealed that myxobacterial genera described to date on the basis of the morphology of multi-cellular fruiting bodies were mostly monophyletic. However, these known taxa comprised only in a small part of the sequences recovered directly from soil in a cultivation-independent approach, indicating that the group is much more diverse than previously thought. We propose that the myxobacteria exist in two forms: the fruiting and the non-fruiting types. Most of the uncultured myxobacteria may represent taxa which rarely form fruiting bodies, or may lack some or all of the developmental genes needed for fruiting body formation. In order to identify non-fruiting myxobacteria, new morphology-independent cultivation and isolation techniques need to be developed.

Adaptation of salt-tolerant Myxococcus strains and their motility systems to the ocean conditions.

More and more studies have indicated that myxobacteria are able to live in seawater conditions, which, however, can decrease the fruiting body formation ability and also the adventurous (A) and social (S) motility systems of the myxobacteria. To learn the adaptation mechanism of the salt-tolerant myxobacteria to marine conditions, we analyzed 10 salt-tolerant Myxococcus strains of their fruiting body formation and motility. The isolates were from marine samples and possessed different levels of salt tolerance. They had the dual motility system and formed fruiting bodies in the presence of suitable seawater concentrations. Some high salt-tolerant strains even lost their fruiting abilities in the absence of seawater. In response to the presence of seawater, the S-motility was found to be increased in the high salt-tolerants but decreased in the low salt-tolerants. The A-motility, on the other hand, was observed in all the salt-tolerant Myxococcus strains, but increased or decreased in response to the presence of seawater. Perceived shifts of fruiting body formation abilities and motilities discovered in the salt-tolerant Myxococcus strains suggested an ecological adaptation of myxobacterial social behaviors to the marine environments.

[Amelioration effect of sand-fixing Hedysarum fruticosum plantations on soil nutrient contents and biological activities].

With adjacent semi-moving dune as the control, this paper studied the effects of 5-, 10- and 22-year old Hedysarum fruticosum plantations on the nutrient status, microbial biomass, and enzyme activities at the soil depths 0-10, 10-20 and 20-30 cm. The results showed that with the establishment of H. fruticosum plantation on moving dune, soil C, N, P and K contents and biological activities increased obviously with the increasing age of the plantation, and the increment was much higher at 0-10 cm than at 10-20 and 20-30 cm. At 0-30 cm, soil C/N increased from 7.3 to 8.5, and microbial biomass C, N and P as well as the activities of urease, protease, saccharase, phosphomonoesterase, dehydrogenase, polyphenol oxidase and nitrate reductase all increased. Among the test enzyme activities, saccharase activity had the most significant increase, with its value at 0-10 cm being 49.7-284.5 times of the control. There were significant positive correlations between soil microbial biomass C, N and P and organic C, total N and total P, respectively, and between soil microbial biomass and enzyme activities.

Exploring the diversity of myxobacteria in a soil niche by myxobacteria-specific primers and probes.

The diversity of myxobacteria in a soil niche was explored using culture-dependent and -independent methods. Conventional cultivation for bacteriolytic myxobacteria produced six types of myxobacteria, which were identified as two Myxococcus spp., two Corallococcus spp., a Cystobacter sp. and a Nannocysts sp. Hybridization analysis of the soil bacterial 16S rRNA gene library with myxobacteria-specific probes revealed that myxobacteria accounted for less than 1% in the bacterial community. A Cystobacterineae 16S rRNA genes-rich library was further established from the soil DNA by polymerase chain reaction amplification with a Cystobacterineae-specific primer combined with a universal bacterial primer. Screening of the special library using Cystobacterineae- and Sorangineae-specific probes produced approximately 45% and 3% positive signals respectively. Sixty-four positive clones were randomly selected for sequencing. Except three repeats, the sequences were diverse ranging from 0.3% to 21.3%, and homologous with the known myxobacteria at 77.6-99.8%, including 57 in Cystobacterineae, one close to Nannocystis and three much more distant from the known myxobacteria. The sequences in the Cystobacterineae can further be divided into at least 12 groups, of which most were unreported. The results suggest that myxobacteria in nature are much more diverse than were ever known, even in one soil niche.