Various Phyllosphere and Soil Bacterial Communities of Natural Grasses and the Impact Factors in a Copper Tailings Dam.

Copper mining caused severe damage to the ecological environment of mining areas. The combination of microbe and plant remediation has an application potential in improving the absorption and transformation efficiency of heavy metals. The phyllosphere is the largest biointerface on the planet, and bacteria are the dominant microbial inhabitants of the phyllosphere, believed to be critical to plant growth and health. This study investigated the phyllospheric and soil bacteria communities using high-throughput sequencing, and endophyte infection statuses of four natural grasses by toluidine blue heparin assay. Results showed variation in phyllospheric bacterial community structure. Gammaproteobacteria were the most abundant bacterial population. Bacilli were found in the phyllosphere of Bothriochloa ischaemum and Imperata cylindrica, while Clostridia were only found in Calamagrostis epigejos. Alphaproteobacteria were the dominant bacteria in soil. In addition, bacterial communities were influenced by endophytic infection statuses. Oxalobacteraceae was associated with soil carbon and sulfur. Enterobacteriaceae had negative correlation with the ratio of soil carbon and nitrogen, and had positive correlation with Cd content. These results offer useful insights into phyllospheric bacterial community variance in four different natural grasses in a copper tailings dam.

Distribution pattern and influencing factors of bacterial communities in different soil depths of Caragana jubata shurb in Luya Mountain, China.

Soil microorganisms are important components of terrestrial ecosystems, affecting soil formation and fertility, plant growth and stress tolerance, nutrient turnover and carbon storage. In this study, we collected soil samples (humus layer, 0-10 cm, 10-20 cm, 20-40 cm, and 40-80 cm) from Caragana jubata shrubland in Shanxi subalpine to explore the composition, diversity, and assembly of soil bacterial communities at different depths across the soil profile. The results showed that Actinomycota (19%-28%), Chloromycota (10%-36%) and Acidobacteria (15%-24%), and Proteobacteria (9%-25%) were the dominant bacterial phyla. α-diversity of soil bacterial community significantly decreased with the increases of soil depth. Soil bacterial ß-diversity varied across different soil depths. Soil pH, water content, and enzyme activity were the main ecological factors affecting the distribution of soil bacterial communities. Soil bacterial communities had more complex interactions in humus layer and 0-10 cm layer. On the whole, soil bacterial communities were dominated by coexistence in C. jubata shrubland, and the soil bacterial community assembly was driven by random process.

[Diversity Patterns and Influencing Factors of Epibiotic in Vallisneria natans and Planktonic Bacteria Communities].

Planktonic and epiphytic bacterial communities play an important role in wetland nitrogen pollutant removal and water purification, yet their community dynamics are far from understood compared with those of the wetland soil bacterial community. Taking the planktonic bacterial community in the Yuguqiao constructed wetland and the epiphytic bacterial community on the leaf surface of the common submerged plant Vallisneria natans as the research objects, the composition, structure, and functional diversity of planktonic and epiphytic bacterial communities were analyzed using high-throughput sequencing. The results showed that the compositions of the planktonic and epiphytic bacterial communities were significantly different, with more heterotrophic and denitrifying bacteria present in the epiphytic bacterial community than in the planktonic bacterial community. The α diversity of the planktonic bacterial community was significantly different among the three sampling sites but not in the epiphytic bacterial community. In general, the OTU index and Shannon index of the epiphytic bacterial community were significantly higher than those of the planktonic bacterial community, and they had obvious spatial heterogeneity. RDA analysis showed that DO, IC, TP, NH+4, and TOC had important effects on the structural changes of both planktonic and epiphytic bacterial communities but had a greater impact on planktonic bacterial communities. Co-occurrence network analysis showed that the epiphytic bacterial community had more niche differentiation, a more stable network, and stronger resistance to external disturbance. The results of FAPROTAX functional prediction analysis showed that the nitrogen cycling, especially denitrification of the epiphytic bacterial community, was significantly greater than that of the planktonic bacterial community. The results of this study revealed the driving mechanism for maintaining the diversity of planktonic and epiphytic bacterial communities, which can provide a scientific basis for excavating and utilizing planktonic and epiphytic bacterial community resources in the construction of constructed wetlands to improve the efficiency of water purification.

[Distribution Pattern of Bacterial Community in Soil Profile of Larix principis-rupprechtii Forest in Luya Mountain].

Microbial communities are the key component to maintaining the structure and function of forest soil ecosystems. The vertical distribution of bacterial communities on the soil profile has an important impact on forest soil carbon pools and soil nutrient cycling. Using Illumina MiSeq high-throughput sequencing technology, we analyzed the characteristics of bacterial communities in the humus layer and 0-80 cm soil layer of Larix principis-rupprechtii in Luya Mountain, China, to explore the driving mechanisms affecting the structure of bacterial communities in soil profiles. The results showed that the α diversity of bacterial communities decreased significantly with increasing soil depth, and community structure differed significantly across soil profiles. The relative abundance of Actinobacteria and Proteobacteria decreased with increased soil depth, whereas the relative abundance of Acidobacteria and Chloroflexi increased with the increase in soil depth. The results of RDA analysis showed that soil NH+4, TC, TS, WCS, pH, NO-3, and TP were important factors determining the bacterial community structure of the soil profile, among which soil pH had the most significant effect. Molecular ecological network analysis showed that the complexity of bacterial communities in the litter layer and subsurface soil (10-20 cm) was relatively high, whereas the complexity of bacterial communities in deep soil (40-80 cm) was relatively low. Proteobacteria, Acidobacteria, Chloroflexi, and Actinobacteria played important roles in the structure and stability of soil bacterial communities in Larch. The species function prediction of Tax4Fun showed a gradual decline in microbial metabolic capacity along the soil profile. In conclusion, soil bacterial community structure showed a certain distribution pattern along the vertical profile of soil, the community complexity gradually decreased, and the unique bacterial groups of deep soil and surface soil were significantly different.

Distribution patterns and driving mechanism of soil protozoan community at the different depths of Larix principis-chinensis forest in the Luya Mountain, China.

To reveal the assembly mechanisms of soil protozoan community in subalpine forest ecosystems, we analyzed the composition and diversity of protozoan communities and their drivers at the six strata (the litter profile, humus profile, 0-10 cm, 10-20 cm, 20-40 cm and 40-80 cm) of soil profiles in subalpine Larix principis-rupprechtii forest in Luya Mountain using Illumina Miseq high-throughput sequencing technology. The results showed that protozoa in the soil profiles belonged to 335 genera, 206 families, 114 orders, 57 classes, 21 phyla, and 8 kingdoms. There were five dominant phyla (relative abundance >1%) and 10 dominant families (relative abundance >5%). The α diversity decreased significantly with increasing soil depth. Results of PCoA analysis showed that the spatial composition and structure of protozoan community differed significantly across soil depths. The results of RDA analysis showed that soil pH and soil water content were important factors driving protozoan community structure across soil profile. Null model analysis suggested that the heterogeneous selection dominated the processes of protozoan community assemblage. Molecular ecological network analysis revealed that the complexity of soil proto-zoan communities decreased continuously with increasing depth. These results elucidate the assembly mechanism of soil microbial community in subalpine forest ecosystem.

[Taxonomic and Functional Diversity of Soil Microbial Communities in Subalpine Meadow with Different Degradation Degrees in Mount Wutai].

Although soil microbes play a key role in grassland ecosystem functioning, the response of their diversity to grassland degradation has not been fully investigated. Here, we used shotgun metagenomic sequencing to analyze the characteristics and influencing factors of soil microbial taxonomic and functional diversity at four different degradation stages[i.e., non-degraded (ND), lightly degraded (LD), moderately degraded (MD), and heavily degraded (HD)]of subalpine meadow in the Mount Wutai. The results showed that there were significant differences in the relative abundances of Actinobacteria, Bacteroidetes, Nitrospirae, and Parcubacteria among the four subalpine grasslands with different degradation degrees (P<0.05).Compared with that in ND, the degraded meadows increased the proportion of genes related to carbon metabolism, biosynthesis of amino acids, pyruvate metabolism, citric acid cycle, propanoate metabolism, butanoate metabolism, and fatty acid metabolism (P<0.05), indicating that the degradation of subalpine grassland changed the metabolic potential of energy metabolism and the nutrient cycle of the soil microbial community. Grassland degradation changed soil microbial taxonomic and functional α diversity, especially in MD and HD.Grassland degradation resulted in significant changes in the taxonomic and functional compositions of the microbial communities. The total nitrogen, pH, and soil organic carbon significantly affected the taxonomic and functional compositions of the microbial communities.The ß diversity of the plant community was significantly correlated with the taxonomic and functional ß diversity of the microbial community (P<0.05), indicating strong coupling. The results of this study revealed the changes and driving mechanisms of subsurface microbial taxonomic and functional diversity during grassland degradation, which can provide a theoretical basis for subalpine meadow protection and ecological restoration.

[Responses of Soil Fungal Communities to Subalpine Meadow Degradation in Mount Wutai].

Grassland degradation has become a worldwide ecological problem. Although soil microorganisms, as the main participants in the process of grassland degradation, play a key role in maintaining ecosystem function and improving soil productivity, little is known about the changes in microbial communities caused by grassland degradation and their relationship with soil properties and plant communities. In this study, we used Illumina MiSeq sequencing to analyze the soil fungal communities of subalpine meadow soil at four different degradation stages[i.e., non-degraded (ND), lightly degraded (LD), moderately degraded (MD), and heavily degraded (HD)] on Mount Wutai. The results showed that Ascomycota, Basidiomycota, and Zygomycota were the dominant phyla of soil fungi in the subalpine meadow, regardless of degradation stage. LEfSe showed that the subalpine meadows with different degradation degrees were enriched with different biomarkers. Compared with ND, MD and HD were enriched with more pathogenic fungi. Moreover, HD apparently decreased the richness and Shannon indexes of soil fungal communities compared with those of ND. Non-metric multidimensional scaling (NMDS) and similarity analysis (ANOSIM) indicated that the compositions and structures of fungal communities were significantly different among meadows with different degradation degrees (P<0.05). Redundancy analysis (RDA) showed that soil water content, total nitrogen, plant richness, and ammonium nitrogen were significantly correlated with the compositions and structures of fungal communities (P<0.05). There were significant correlations between α diversity and ß diversity between plant and fungal communities (P<0.05), indicating strong coupling. The results of our study provide a theoretical basis for further research on the changes in soil fungal communities and their driving mechanism in different degradation stages of subalpine meadows.

[Altered Splicing in Stable Cell Strains Expressing Mini-hF9 Gene with Nonsense Mutation].

OBJECTIVE: To investigate the molecular mechanism in stable cell strains expressing Mini-hF9 gene with nonsense mutation. METHODS: Mini-hF9 gene and its nonsense mutants were transfected into HeLa cells independently, and stable cell strains were obtained after G418 resistance screening and monoclonal transformation. The altered splicing and protein expression of mRNA in Mini-hF9 gene in stable cell strains were detected by using RT-PCR and Western blot. RESULTS: The wild type and nonsense mutated human coagulation factor IX stable cell strains were constructed successfully, which were named HeLa-F9-WT, HeLa-F9-M1 and HeLa-F9-M2. Only normal splicing Norm was detected in the wild-type cell strain HeLa-F9-WT; Norm and Alt-S1 splicing were detected in HeLa-F9-M1; while Norm, Alt-S1 and Alt-S2 splicing were detected in HeLa-F9-M2. CONCLUSION: The nonsense associated altered splicing (NAS) pathway, which generated alternately spliced transcripts, might be triggered in coagulation factor IX gene with nonsense mutation.

GTPase activity analysis of eRF3 in Euplotes octocarinatus.

In eukaryotes, eRF3 participates translation termination and belongs to the superfamily of GTPase. In this work, dissociation constants for E. octocarinatus eRF3 binding to nucleosides in presence and absence of eRF1a were determined using fluorescence spectra methods. Furthermore, the GTP hydrolyzing assay of Eo-eRF3 was carried out by HPLC methods and the kinetic parameter for GTP hydrolysis by eRF3 was determined. The results showed eRF1a could promote GTP binding to eRF3 and hydrolyzing GTP activity of eRF3. The observation is consistent with the data from human. Whereas E. octocarinatus eRF3 alone can bind GTP in contrast to no GTP binding observed in the absence of eRF1 in human eRF3. The affinity for Eo-eRF3 binding nucleotides is different from that in human. Structure model and amino acids sequence alignment of potential G domains indicated these different may be due to Valine 317 and Glutamate 452 displacing conserved Glycine and Lysine, which were involved in GTP binding.

[Effects of Heavy Metal Contents on Phyllosphere and Rhizosphere Fungal Communities for Bothriochloa ischaemum in Copper Tailings Area].

There are complex interrelationships between plant microorganisms (phyllosphere and rhizosphere) and host plants, which can promote plant growth and enhance the tolerance of host plants to stress. In this study, we selected the dominant species Bothriochloa ischaemum as the research subject in a copper tailings dam. Using high-throughput sequencing, we investigated the structures of the fungal communities and diversities in the phyllosphere and rhizosphere of B. ischaemum. This study also explored the effects of heavy metal content on fungal community characteristics. The results showed that Ascomycota and Basidiomycota were the dominant phyla in the phyllosphere and rhizosphere of B. ischaemum. The diversities and richness of the rhizosphere fungal community were higher than that of the phyllosphere fungal community. The diversities of rhizosphere and phyllosphere fungal communities was affected by different heavy metals. Phyllosphere fungal diversity was mainly affected by the content of Zn and Cu in leaves, and the content of Pb in roots was the key factor affecting the diversity of the rhizophere fungal community. Furthermore, Pleosporaceae had a very significant positive correlation with Cd in the phyllosphere, and Nectriaceae had a significant positive correlation with Zn in the rhizosphere. These fungal communities could be used as indicators of ecological recovery in areas with heavy metal pollution. The results could provide an ecological basis for the exploration and utilization of phyllosphere or rhizosphere fungi resources during ecological restoration processes. This study also provides guidance for selecting the plant-microbial symbionts during ecological restoration in areas with heavy metal pollution.

MiR-32-5p aggravates intestinal epithelial cell injury in pediatric enteritis induced by Helicobacter pylori.

BACKGROUND: Pediatric enteritis is one of the infectious diseases in the digestive system that causes a variety of digestive problems, including diarrhea, vomiting, and bellyache in children. Clinically, Helicobacter pylori (H. pylori) infection is one of the common factors to cause pediatric enteritis. It has been demonstrated that aberrant expression of microRNAs (miRNAs) is found in gastrointestinal diseases caused by H. pylori, and we discovered a significant increase of miR-32-5p in H. pylori-related pediatric enteritis. However, the exact role of miR-32-5p in it is still unknown. AIM: To investigate the role of aberrant miR-32-5p in pediatric enteritis induced by H. pylori. METHODS: MiR-32-5p expression was detected by quantitative real time-polymerase chain reaction. The biological role of miR-32-5p in H. pylori-treated intestinal epithelial cells was evaluated by Cell Counting Kit-8 assay and flow cytometry. The potential target of miR-32-5p was predicted with TargetScanHuman and verified by luciferase assay. The downstream mechanism of miR-32-5p was explored by using molecular biology methods. RESULTS: We found that miR-32-5p was overexpressed in serum of H. pylori-induced pediatric enteritis. Further investigation revealed that H. pylori infection promoted the death of intestinal epithelial cells, and increased miR-32-5p expression. Moreover, miR-32-5p mimic further facilitated apoptosis and inflammatory cytokine secretion of intestinal epithelial cells. Further exploration revealed that SMAD family member 6 (SMAD6) was the direct target of miR-32-5p, and SMAD6 overexpression partially rescued cell damage induced by H. pylori. The following experiments showed that miR-32-5p/SMAD6 participated in the apoptosis of intestinal epithelial cells induced by transforming growth factor-ß-activated kinase 1 (TAK1)-p38 activation under H. pylori infection. CONCLUSION: Our work uncovered the crucial role of aberrant expression of miR-32-5p in H. pylori-related pediatric enteritis, and suggested that the TAK1-p38 pathway is involved in it.

[Distribution Pattern and Diversity Maintenance Mechanisms of Fungal Community in Subalpine Lakes].

The composition and diversity of fungal communities are essential to maintain the ecosystem balance of subalpine lakes. The aquatic fungal communities at different depths from the subalpine Pipahai (PPH, 0, 2, 4 m), Mayinghai (MYH, 0, 2, 4, 6 m), and Gonghai (GH, 0, 2, 4, 6, 8 m) lakes were studied. In addition to that, the distribution pattern and diversity maintenance mechanism (determination process vs. random process) of fungal communities were explored using high-throughput sequencing. The results showed that the physicochemical parameters of the water were significantly different among the three lakes. The pH, electrical conductivity (EC), ammonia nitrogen (NH4+-N), total carbon (TC), and inorganic carbon (IC) of GH were significantly higher than in the other two lakes. The fungal community was mainly composed of Ascomycota (0.82%-21.05%), Basidiomycota (1.26%-11.79%), Chytridiomycota (0.42%-4.26%), and Rozellomycota (0.11%-0.33%). Cystobasidiomycetes, Dothideomycetes, Chytridiomycetes, and Sordariomycetes were the dominant classes shared by the three lakes. The α-diversity index and the relative abundance of dominant classes were significantly different among the three lakes (P<0.05), but there were no significant differences between the various depths on each lake. The results of the ANOSIM analysis showed that the ß-diversity of the fungal communities were significantly different (r=0.99, P<0.01) among the lakes. There was also expressive differences at various depths on MYH (r=0.98, P<0.01) and GH (r=0.25, P<0.05), but no significant difference in PPH (r=0.23, P>0.05). The analysis results of redundancy and variation partitioning showed that the ß-diversity pattern of fungal communities in small region areas (among the three lakes) and local areas (different depths of MYH) were driven by environmental selection and dispersal limitation. However, the relative role of environmental selection was more significant, with water pH, dissolved oxygen (DO), TC, and EC being the main influencing factors. The results of the null model analysis showed that the interspecific interactions promoted the maintenance of the ß-diversity pattern of the fungal community in GH. In summary, the ß-diversity pattern of fungal communities in the subalpine lakes was mainly driven by a deterministic process.

[Spatio-temporal Patterns of Microbial Communities and Their Driving Mechanisms in Subalpine Lakes, Ningwu, Shanxi].

Human activities and climate change cause the degradation of subalpine lake ecosystems, which induce the shift of microbial community structure. The spatio-temporal dynamics and the diversity maintenance mechanisms of bacterial communities in Gonghai Lake in Ningwu, Shanxi, were investigated by using Q-PCR and DGGE. The results showed that the temperature), pH, dissolved oxygen, electrical conductivity salinity, and ammonium nitrogen (NH4+) contestation were significantly different among the different sampling depths during different months. Bacterial abundance was the highest in August and the lowest in November, and the abundance was higher in the middle water layer (2 m, 4 m, and 6 m depths), but relatively low in the surface layer and bottom layer (0 m and 8 m depths, respectively). The α diversity index of bacterial communities had significant differences among the different months and depths, and showed an initial decreasing trend and then an increasing trend from April to December. A PERMANOVA test showed that the spatial distribution of bacterial communities was significantly different among depths (P<0.001). The results of redundancy analysis and variation partitioning indicated that environmental selection and diffusion limitation had an effect on the maintenance of the diversity patterns of bacterial communities at the different depths of GH. However, the relative effect of environmental factors was stronger, of which the concentration of NO3-, NO2-, and NH4+ were the main influencing factors. In conclusion, the bacterial communities in GH subalpine lake showed clear spatio-temporal distribution patterns, and environmental variables had a significant effect on shaping the community diversity.

[Environmental Filters Drive the Assembly of the Soil Fungal Community in the Larix principis-rupprechtii Forests of the Guandi Mountains].

The community assembly process, which purports that the trade-off between the stochastic process and the deterministic process, is the central issue of community ecology, and is one of the most controversial ecological issues. The current research investigated the fungal community in the Larix principis-rupprechtii forests of the Guandi Mountains and elucidated the relative role of the stochastic and deterministic processes in the assembly of soil fungal community on a local scale. The correlation analysis and redundancy analysis of the physicochemical factors in soil and dominant fungal phyla, as well as the structural equation model analysis, showed that these physicochemical factors and aboveground vegetation diversity had significant effects on fungal communities. The direct effect of vegetation diversity on fungal community structure was the most (1.1858). It is inferred that the determination process (environmental selection) has a certain influence on the assembly of fungal communities. The ß-diversity of fungal community shows a distance-decay pattern; thus, it can be concluded that the stochastic process (dispersal limitation) has a certain effect on the assembly of fungal communities. Null model analysis confirmed that the deterministic process was the main driving factor for the assembly of the fungal communities in the study area, and their relative importance varied along with altitudinal gradient. The null deviations in the study area were negative, suggesting that habitat filtering was the driving factor of the assembly of fungal communities. Overall, the deterministic versus stochastic processes jointly drive the assembly of fungal communities in the study area, while the deterministic processes triumph.

[Spatiotemporal dynamics and driving forces of soil bacterial communities on the dam of Shibahe copper mine tailings in Shanxi, China.] / åå «æ²³é“œå°¾çŸ¿åº“åé¢ç»†èŒç¾¤è½æ—¶ç©ºåŠ¨æ€åŠå ¶é©±åŠ¨åŠ›.

The maintaining mechanism of community diversity is the core of community ecology. The mine tailing is a good field for studying on the underlying mechanism of community diversity, as a kind of original bare land with heavy metal pollution, where the physicochemical characteristics of soil change with the restoration periods. We examined the driving forces for bacterial community diversity based on the investigation of edaphic factors, plant community, and bacterial communities in Shibahe copper mine tailing, Shanxi. The results showed that nutrient contents in soil increased with restoration periods. The seasonal dynamics of soil nutrient in different restoration time were different. Shannon diversity and richness of bacterial community showed an increasing trend, indicating community stability was improved with restoration. Influenced by plant community, the seasonal changes of those indices differed with restoration. Results from the RDA analysis showed that the diversity and structure of bacterial communities were determined by environmental factors (edaphic, plant and heavy metals). Results from the structure equation models further confirmed that soil nutrients (TC, TN, NO3--N, NO2-N), plant community, and soil enzyme activities jointly drove bacterial community assembly on the copper mine tailings.

[Characteristics of Soil Physicochemical Properties and Enzyme Activities over Different Reclaimed Years in a Copper Tailings Dam].

Mining for metal and mineral resources lead to the rapid rise of tailings dams and caused serious damage to the ecological environment of the mining area. Soil physicochemical characteristics and enzyme activities were important indexes for ecosystem functions, and they were also important factors in evaluating soil restoration qualities. We selected nine sub-dams of the Eighteen River copper tailings in Yuanqu County, and analyzed the relationship between soil physicochemical properties and soil enzyme activities. The results showed that there were great differences in soil physicochemical properties over different reclaimed years, and as the reclaimed years passed, soil nutrient contents significantly increased. There were significant negative correlations between catalase and the ratio of soil carbon and nitrogen, and urease was positively correlated to total nitrogen and soil moisture. Phosphatase and sucrose demonstrated no significant relationships with soil physicochemical factors. Copper content gradually accumulated in soil as the restoration period of sub-dams increased. Arsenic and cadmium content increased initially and then decreased before they gradually reached a stable level. In addition, there was no significant difference in zinc content among different sub-dams. Together, these results provide the ecological basis for further studies in soil ecosystem restoration and degradation mechanisms in copper tailings.

[Driving Factors of the Dynamics of Microbial Community in a Dam of Copper Mine Tailings].

The relative importance of the deterministic versus stochastic processes underlying community dynamics has long been a central theme in community ecology, and is intensively debated in the field. Microbial communities play key roles in nutrient cycling and the flow of energy in ecosystems. The research on the structural dynamics of microbial community will provide data and theoretical support for understanding the assembly mechanisms of community, and for predicting the dynamics of microbial community under environmental stress. In this study, the Illumina MiSeq method was applied to investigate the structural dynamics of bacterial and fungal community in a dam of Shibahe mine tailings at different restoration stages (1-45 years). The results indicated that the soil physicochemical properties in the dam of mine tailings formed an ecological gradient, and the plant community showed succession along the restoration time. The diversity of plant communities was significantly correlated with soil nutrient contents but not with soil heavy metal contents. The structure of the microbial communities showed significant differences at different restoration stages of the dam land, in which Proteobacteria, Actinobacteria, Firmicutes, and Acidobacteria were the dominant bacterial phyla, and Ascomycota, Basidiomycota, and Zygomycota were the dominant fungal phyla. The assembly of the microbial community was shaped mainly by the soil nutrients and soil heavy metal contents, but plant diversity had no significant effect on the microbial community structure. It was suggested that edaphic factors drive the dynamics of microbial communities under the stress conditions of pH and heavy metals on small, local scales.

[Environmental selection and dispersal limitation drive the assemblage of bacterial community in temperate forest soils]. / 环境选择和扩散限制驱动温带森林土壤细菌群落的构建.

Environmental selection and dispersal limitation are two basic processes underlying community assembly. The relative importance of those two processes differs across scales, community identities, and community types. The processes responsible for structuring microbial communities in soil of temperate subalpine forest are poorly understood. Here, we investigated the relationship between soil bacterial community structure and environmental factors, and quantified the relative role of edaphic factors, vegetation, and spatial variables in shaping the structure of six soil bacterial communities (LpMC1, LpMC2, PwMC, PmMC, PtMC, and BMC) in five forest types including Larix principis-rupprechtii, Picea wilsonii, Picea meyeri, Pinus tabulaeformis, and Betula platyphylla in Pangquangou Nature Reserve by using PCR-DGGE technology. Our results showed that the structure and biodiversity of bacterial communities were significantly different among six communities. The biodiversity of bacterial community were higher in LpMC2 and PtMC, lowest in PmMC, and highest in LpMC1. Soil environmental factors, such as pH, soil water content, total carbon, total nitrogen, soil organic matter, available phosphorous, and soil enzymes, were significantly correlated with biodiversity and structure of soil bacterial community. The beta diversity of bacterial communities were significantly correlated with geographic distance, indicating the influence of dispersal limitation on the structure of bacterial community. The order of driving force on the structure of bacterial community was edaphic factors (0.27), spatial factor (0.19) and vegetation (0.15) in six samples. Using regional soil microbes from 10 samples around reserve as source community, results from the microcosm experiments showed that the edaphic factors were the predominant driving factors (0.35) on structure of artificial dispersal bacterial community, while the high diversity of source microbial community affected the structure of microcosm soil. In summary, at local scale, environmental selection predominantly determined the structural and biodiversity of soil bacterial communities in temperate subalpine forest, while dispersal limitation played a significant role. Such a result indicated that deterministic processes and stochastic processes played important roles in shaping the structure of soil bacterial community at local scale, with the former having the leading role. The composition of dispersal soil bacteria community was source-dependent but also modulated by local environmental selection.

Therapeutic potential of chlorotoxin-like neurotoxin from the Chinese scorpion for human gliomas.

Chlorotoxin, one of the key toxins in scorpion Leiurus quinquestriatus venom, has been shown to bind specifically to glioma cell surface as a specific chloride channel blocker. In this study, a purified, recombinant chlorotoxin-like peptide from the scorpion Buthus martensii Karsch (named rBmK CTa) was characterized by in vivo and in vitro studies. The results from cell proliferation assay with human glioma (SHG-44) cells showed that rBmK CTa inhibits the growth of glioma cells in a dose-dependent manner, with an IC(50) value of approximately 0.28microM. Under the same conditions, the IC(50) value for normal astrocytes increased to 8microM. This clearly indicated that rBmK CTa had specific toxicity against glioma cells but not astrocytes. Results from whole-cell patch-clamp recording showed that chloride current in SHG-44 was inhibited by rBmK CTa in a voltage-dependent manner and percent inhibitions for the blocking action of rBmK CTa (0.07 and 0.14microM) on I(Cl) was 17.64+/-3.06% and 55.86+/-2.83%, respectively. Histological analysis of rBmK CTa treated mice showed that brain, leg muscle and cardiac muscle were the target organs of this toxin. These results suggest that rBmK CTa may have potential therapeutic application in clinical treatment of human glioma. It represents an approach for developing a novel therapeutic agent.

[Cloning and characterization of a novel trinucleotide repeat-containing gene GARP from Euplotes octocarinatus].

The expansion of trinucleotide repeats in genome is related to the phthogenesis of several neurodegenerative diseases. A GARP (glutamic acid-rich protein) gene was isolated from the macronuclear plasmid mini library of Euplotes octocarinatus. A micronuclear version of the GARP gene was amplified by polymerase chain reaction. The macronuclear DNA molecule carrying the GARP gene is 460 bp long and shows the characteristics of macronuclear chromosomes of hypotrichous ciliates. One of the three cysteines is encoded by the opal codon TGA(88-90). The predicted open reading frame encodes a 112-amino acid polypeptide, with a predicted molecular mass of 13 kDa and an isoelectric point of 3.82. Micronuclear version of the GARP gene contains two internal eliminated sequences (IES), IES1 and IES2. IES1 is 41 bp long and is flanked by 5'-GA-3' direct repeats. IES2 is 41 bp long and flanked by 5'-TA-3' direct repeats. Transcriptional activity of GARP gene was confirmed by reverse transcription polymerase chain reaction (RT-PCR).