Different microbial communities in paddy soils under organic and nonorganic farming.

Organic agriculture is a farming method that provides healthy food and is friendly to the environment, and it is developing rapidly worldwide. This study compared microbial communities in organic farming (Or) paddy fields to those in nonorganic farming (Nr) paddy fields based on 16S rDNA sequencing and analysis. The predominant microorganisms in both soils were Proteobacteria, Chloroflexi, Acidobacteria, Actinobacteria, and Nitrospirota. The alpha diversity of the paddy soil microbial communities was not different between the nonorganic and organic farming systems. The beta diversity of nonmetric multidimensional scaling (NMDS) revealed that the two groups were significantly separated. Distance-based redundancy analysis (db-RDA) suggested that soil pH and electrical conductivity (EC) had a positive relationship with the microbes in organic paddy soils. There were 23 amplicon sequence variants (ASVs) that showed differential abundance. Among them, g_B1-7BS (Proteobacteria), s_Sulfuricaulis limicola (Proteobacteria), g_GAL15 (p_GAL15), c_Thermodesulfovibrionia (Nitrospirota), two of f_Anaerolineaceae (Chloroflexi), and two of g_S085 (Chloroflexi) showed that they were more abundant in organic soils, whereas g_11-24 (Acidobacteriota), g__Subgroup_7 (Acidobacteriota), and g_Bacillus (Firmicutes) showed differential abundance in nonorganic paddy soils. Functional prediction of microbial communities in paddy soils showed that functions related to carbohydrate metabolism could be the major metabolic activities. Our work indicates that organic farming differs from nonorganic farming in terms of microbial composition in paddy soils and provides specific microbes that might be helpful for understanding soil fertility.

Functional Prediction of Microbial Communities in Sediment Microbial Fuel Cells.

Sediment microbial fuel cells (MFCs) were developed in which the complex substrates present in the sediment could be oxidized by microbes for electron production. In this study, the functional prediction of microbial communities of anode-associated soils in sediment MFCs was investigated based on 16S rRNA genes. Four computational approaches, including BugBase, Functional Annotation of Prokaryotic Taxa (FAPROTAX), the Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt2), and Tax4Fun2, were applied. A total of 67, 9, 37, and 38 functional features were statistically significant. Among these functional groups, the function related to the generation of precursor metabolites and energy was the only one included in all four computational methods, and the sum total of the proportion was 93.54%. The metabolism of cofactor, carrier, and vitamin biosynthesis was included in the three methods, and the sum total of the proportion was 29.94%. The results suggested that the microbial communities usually contribute to energy metabolism, or the metabolism of cofactor, carrier, and vitamin biosynthesis might reveal the functional status in the anode of sediment MFCs.

Dynamic Changes in Soil Microbial Communities with Glucose Enrichment in Sediment Microbial Fuel Cells.

To investigate soil microbial community dynamics in sediment microbial fuel cells (MFCs), this study applied nonhydric (D) and hydric (S) soils to single-chamber and mediator-free MFCs. Glucose was also used to enrich microorganisms in the soils. The voltage outputs of both the D and S sediment MFCs increased over time but differed from each other. The initial open circuit potentials were 345 and 264 mV for the D and S MFCs. The voltage output reached a maximum of 503 and 604 mV for D and S on days 125 and 131, respectively. The maximum power densities of the D and S MFCs were 2.74 and 2.12 mW m-2, analyzed on day 50. Clustering results revealed that the two groups did not cluster after glucose supplementation and 126 days of MFC function. The change in Geobacter abundance was consistent with the voltage output, indicating that these bacteria may act as the main exoelectrogens on the anode. Spearman correlation analysis demonstrated that, in the D soils, Geobacter was positively correlated with Dialister and negatively correlated with Bradyrhizobium, Kaistobacter, Pedomicrobium, and Phascolarctobacterium; in the S soils, Geobacter was positively correlated with Shewanella and negatively correlated with Blautia. The results suggested that different soil sources in the MFCs and the addition of glucose as a nutrient produced diverse microbial communities with varying voltage output efficiencies. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00959-x.

The effect of paclobutrazol on soil bacterial composition across three consecutive flowering stages of mung bean.

Paclobutrazol, (2RS, 3RS)-1-(4-chlorophenyl)-4, 4-dimethyl-2-(1H-1,2,4-triazol-1-yl) pentan-3-ol, is a plant growth retardant that mainly inhibits gibberellins (GAs) biosynthesis. In agricultural practice, paclobutrazol is applied to arrest vegetative growth so as to increase the reproductive growth of many orchard fruit, as well as grain crops. However, due to its over-application and chemical stability, paclobutrazol accumulates in soil and inhibits the growth of subsequent crops, especially those grown for vegetative purposes. The present study focused mainly on the changes in the soil bacterial community following application of paclobutrazol. Mung bean (Vigna radiata) plants were treated with paclobutrazol and cultivated for three consecutive seasons. Soil samples were collected and analyzed by denaturing gradient gel electrophoresis (DGGE) using 16S rDNA gene fragments and clone library analyses. The results obtained through clustering and clonal sequencing analysis showed that the bacterial community was affected by paclobutrazol, and in addition, was more diverse in the third stage of mung bean plant cultivation. The results of the study showed that paclobutrazol affected bacterial composition, and the population of bacteria varied greatly across time.

Seasonal changes in bacterial communities associated with healthy and diseased Porites coral in southern Taiwan.

We compared the bacterial communities associated with healthy scleractinian coral Porites sp. with those associated with coral infected with pink spot syndrome harvested during summer and winter from waters off the coast of southern Taiwan. Members of the bacterial community associated with the coral were characterized by means of denaturing gradient gel electrophoresis (DGGE) of a short region of the 16S rRNA gene and clone library analysis. Of 5 different areas of the 16S rRNA gene, we demonstrated that the V3 hypervariable region is most suited to represent the coral-associated bacterial community. The DNA sequences of 26 distinct bands extracted from DGGE gels and 269 sequences of the 16S rRNA gene from clone libraries were determined. We found that the communities present in diseased coral were more heterogeneous than the bacterial communities of uninfected coral. In addition, bacterial communities associated with coral harvested in the summer were more diverse than those associated with coral collected in winter, regardless of the health status of the coral. Our study suggested that the compositions of coral-associated bacteria communities are complex, and the population of bacteria varies greatly between seasons and in coral of differing health status.

Cultivation of the culinary-medicinal Lung Oyster mushroom, Pleurotus pulmonarius (Fr.) Quél. (Agaricomycetideae) on grass plants in Taiwan.

Cultivation of the culinary-medicinal Lung Oyster mushroom, Pleurotus pulmonarius, on the stalks of three grass plants, i.e., Panicum repens, Pennisetum purpureum, and Zea mays were investigated. The effects of various combinatorial substrates on mushroom mycelial growth and yield calculated as biological efficiency (BE) were determined. Among 9 experimental substrates, the most suitable substrate for mycelial growth was 45ZMS:45S, followed by 45PRS:45S; their mycelial growth rates were obviously quicker than that of the control substrate. The BEs of all the experimental substrates respectively containing P. repens stalk, P. purpureum stalk and Z. mays stalk were higher than that of the control (39.55%) during the 2.5 months of cultivation period. The best substrate in terms of BE was 60ZMS:30S (58.33%), followed by 45PRS:45S (57.16%), 45ZMS:45S (49.86%), and 30ZMS:60S (47.20%). Based on the BE of the tested substrates, Z mays stalk appeared to be the best alternative material for the production of P. pulmonarius.

Analysis of Codon Usage Patterns in Toxic Dinoflagellate Alexandrium tamarense through Expressed Sequence Tag Data.

We have analyzed synonymous codon usage in the genome of A. tamarense CCMP 1598 for protein-coding sequences from 10865 expressed sequence tags (ESTs). We reconstructed a total of 4284 unigenes, including 74 ribosomal protein and 40 plastid-related genes, from ESTs using FrameDP, an open reading frame (ORF) prediction program. Correspondence analysis of A. tamarense genes based on codon usage showed that the GC content at the third base of synonymous codons (GC3s) was strongly correlated with the first axis (r = 0.93 with P < .001). On the other hand, the second axis discriminated between presumed highly and low expressed genes, with expression levels being confirmed by the analysis of EST frequencies (r = -0.89 with P < .001). Our results suggest that mutational bias is the major factor in shaping codon usage in A. tamarense genome, but other factors, namely, translational selection, hydropathy, and aromaticity, also appear to influence the selection of codon usage in this species.

Bacterial diversity in paclobutrazol applied agricultural soils.

The aim of this study was to investigate the bacterial communities on paclobutrazol [(2RS, 3RS)-1-(4-Chlorophenyl)-4, 4-dimethyl-2-(1H-1,2,4-triazol-1-yl) pentan-3-ol]-applied agricultural soils by denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rDNA gene fragments. Three different agricultural soil samples were collected from paclobutrazol applied mango and waxapple orchards, peanut fields and untreated rice fields as a control for DGGE analysis. The DGGE pattern of PCR- generated 16S rDNA gene fragments indicated that the bacterial populations from four paclobutrazol-applied soils of peanut fields were closely related to each other and two paclobutrazol-applied soils of mango and waxapple orchards harbored closely related bacterial communities. But, paclobutrazol-free agricultural soils comprised relatively a different bacterial group. However, the bacterial populations of mango and waxapple orchard are completely different from the bacterial communities of peanut field. Further purification and sequence analysis of 40 DGGE bands followed by phylogenetic tree assay showed similar results that soil bacteria from paclobutrazol applied mango and waxapple orchard are phylogenetically related. Based on the phylogenetic analysis, the clone M-4 was clad 100 % (bootstrap value) with Mycobacterium sp. The Mycobacterium sp. has been proved to degrade the phenolic compounds such as phenol, 4-chlorphenol, 2,4-dichlorophenol and paclobutrazol molecule containing chlorobenzene ring.

Circulating apoptotic factors in patients with acute cerebral infarction.

OBJECTIVE: Studies on animal models have shown that apoptosis plays a role in acute ischemic stroke, but the clinical significance of apoptosis in human stroke has not been well established. DESIGN AND METHODS: Acute ischemic stroke patients who had been examined by magnetic resonance imaging (MRI) were recruited, and their blood samples were collected after informed consent obtained. Sixteen samples were taken from 7 patients and 4 control subjects. DNA was extracted from the plasma and analyzed. The plasma level of tumor necrosis factor-alpha (TNF-alpha) was analyzed by western blot, and relative caspase 3/7 activity was assayed. RESULTS: Soluble and membrane-associated TNF-alpha were found to be higher in the plasma from ischemic stroke patients than in that from the controls. Further, this result was more significant in patients with grey matter lesions than white matter lesions. CONCLUSIONS: Apoptotic factors are detected in the plasma of patients with acute ischemic stroke and might be related to infarct area.

Characterization of the 5'-flanking regions of the sea anemone ADP ribosylation factor 1 and actin genes.

The 5'-flanking regions of the sea anemone, Aiptasia pulchella (ap) ARF1 gene showed the absence of a TATA box. The transcriptional start site determined by 5' rapid amplification of cDNA ends (5'RACE) is located 75 base pairs upstream of the translational start site. Transfection experiments in HeLa and COS-7 cells demonstrate that all the elements required to achieve significant basal transcription activity are located between position -208 and -88 relative to the transcriptional start site. There are three consensus initiator (Inr) elements for TATA-less promoter around the transcriptional start site of the apARF1 gene (+29, -158, and -226) that are likely to play roles in the regulation. For the apactin gene, the 5'-flanking region contains a TATA box located 30 base pairs upstream of the transcriptional start site. The transient transfection of apactin/luciferase deletion constructs revealed that the TATA box indeed is necessary for full expression.

Comparative gene expression in the symbiotic and aposymbiotic Aiptasia pulchella by expressed sequence tag analysis.

Intracellular symbiotic relationships are prevalent between cnidarians, such as corals and sea anemones, and the photosynthetic dinoflagellate symbionts. However, there is little understanding about how the genes express when the symbiotic relationship is set up. To characterize genes involved in this association, the endosymbiosis between sea anemone, Aiptasia pulchella, and dinoflagellate zooxanthellae, Symbiodinium spp., was employed as a model. Two complementary DNA (cDNA) libraries were constructed from RNA isolated from symbiotic and aposymbiotic A. pulchella. Using single-pass sequencing of cDNA clones, a total of 870 expressed sequence tags (ESTs) clones were generated from the two libraries: 474 from symbiotic animal and 396 from aposymbiotic animal. The initial ESTs consisted of 143 clusters and 231 singletons. A BLASTX search revealed that 147 unique genes had similarities with protein sequences available from databases; 120 of these clones were categorized according to their putative function. However, many ESTs could not assign functionally. The putative roles of some of the identified genes relative to endosymbiosis were discussed. This is the first report of the use of EST analysis to examine the gene expression in symbiotic and aposymbiotic states of the cnidarians. The systematic analysis of EST from this study provides a useful database for future investigations of the molecular mechanisms involved in algal-cnidarian symbiosis.

Cloning and characterization of the first cnidarian ADP-ribosylation factor, and its involvement in the Aiptasia-Symbiodinum endosymbiosis.

Marine cnidarian-microalgal endosymbiosis is a form of intracellular association that contributes greatly to the high primary productivity of reefs; however, little is known about its molecular mechanisms. Since the ADP-ribosylation factor (ARF) family proteins are key regulators of host intracellular vesicle transport systems, which are critical to many endosymbiotic interactions, we set out to clone and characterize ARF proteins in the symbiotic sea anemone Aiptasia pulchella. Experiments indicated that at least 3 ARF protein classes (class I, class II and class III) were present and expressed as a single messenger RNA species in Aiptasia, with highest mRNA expression levels for apARF1, medium for apARF5, and lowest for apARF6. Quantitative analysis revealed a great reduction at both the RNA and the protein levels in apARF1, but not apARF5 and apARF6, in the symbiotic animals. The apARF1 protein was highly homologous in sequence to other known ARF1 proteins and displayed a Golgi-like localization pattern. Overall, our study identified apARF1 as a potential negative regulator of Aiptaisia-microalgal endosymbiosis.

EST-based identification of genes expressed in the hypothalamus of adult tilapia, Oreochromis mossambicus.

The hypothalamus is involved in many physiological functions in teleosts. To accelerate the molecular analysis of hypothalamic functions, a list of transcripts expressed in the hypothalamus of adult tilapia, Oreochromis mossambicus, was compiled using the expressed sequence tag (EST) strategy. Of 161 clones, 130 clones were unique genes and 31 clones were found to be redundant. Of 130 unique genes, 32.3% (42/130 ESTs) were identified as known genes and 67.7% (88/130 ESTs) as unknown genes. The functional categorization of the known genes was analyzed. Bioinformatic analysis revealed that 62 of 88 unknown genes (62/130 ESTs, 47.7%) showed a significant homology to neither nucleotide nor translated peptide sequences in the public database. These genes might be particularly expressed in the tilapia hypothalamus.