[Effects of effective microorganisms on growth promotion and the rhizosphere eukaryotic community structure of pepper in Xinjiang, China]. / å¤åˆèŒå‰‚å¯¹æ–°ç–†è¾£æ¤’çš„ä¿ƒç”Ÿæ•ˆæžœå’Œæ ¹é™ çœŸæ ¸ç”Ÿç‰©ç¾¤è½ç»“æž„çš„å½±å“.

Effective microorganisms (EM) might alleviate deterioration of soil environmental quality and yield decline of pepper (Capsicum annuum) caused by continuous replanting and imbalanced fertilizer application in Xinjiang. We investigated the effects of applying EM microbial agent on the growth of pepper plants, yield, soil nutrient content, soil enzyme activity, and rhizosphere eukaryotic community. The results showed that the application of EM microbial agent increased plant height, stem diameter, leaf length, leaf width and root length by 22.6%, 35.3%, 33.3%, 29.7% and 15.1%, respectively. It also increased fruit width, individual fruit weight, and yield by 5.3%, 42.9%, and 74.7%, respectively. After the application of EM microbial agent, the levels of soil available nitrogen increased by 10.2% and 5.8% during the flowering and maturity stages, respectively. Similarly, available phosphorus increased by 10.4% and 13.4%, respectively. The soil sucrase activity was increased by 40.7%, 14.6%, and 9.3% during the seedling, flowering, and maturity stages, respectively. Urease activity was also increased by 7.9%, 10.2%, and 11.5%, respectively. Furthermore, the application of EM microbial agent increased soil peroxidase activity by 16.8% and 44.6% at flowering and maturity stages, respectively. The application of microbial agent significantly altered the ß-diversity of the rhizosphere eukaryotic community in pepper plants. Specifically, microbial agent increased the relative abundances of populations belonging to Enchytraeus and Sminthurides genera, which could contribute to soil improvement and nutrient cycling. Compared to the CK, the relative abundance of pathogenic microorganisms including Olpidium and Aplanochytrium genera decreased by 98.0% and 89.3%, and the relative abundance of the Verticillium decreased to 0. These results demonstrated that EM microbial agent could increase soil nutrient content, enhance soil enzyme activity, and reduce soil pathogenic fungi in the pepper cultivation areas of Xinjiang, thus achieving beneficial effects on pepper growth and fruit yield.

First Report of 'Candidatus Phytoplasma ziziphi'-Related Strains Infecting Peony (Paeonia suffruticosa Andr.) Showing Leaf Yellows Symptoms in China.

Peony (Paeonia suffruticosa Andr.), belonging to family Paeoniaceae, is an important medicinal and ornamental plant. During August of each year from 2016 to 2023, peony plants at Heze city were found to exhibit leaf yellows symptoms. The incidence rate of the symptomatic plant was recorded from 10% to 30% in four peony gardens with about 200 acres. Total DNA was extracted from 0.10 g fresh plant leaf tissues from 24 symptomatic and 8 asymptomatic samples using rapid plant genomic DNA isolation kit (Aidlab Biotechnology, Beijing, China). The extracted DNA was amplified by nested polymerase chain reaction using universal primers R16mF2/R16mR1 followed by R16F2/R16R2 (Lee et al., 1993; Gundersen and Lee, 1996) specific for the 16S rRNA gene and new designed tuf gene specific primers JWB-tuforfF1 (5'-ATGGCTGAAATATTTTCAAGAG-3') and JWB-tuforfR1 (5'-TTATTCTATGATTTTAATAACAG-3') followed by JWB-tuforfF2 (5'-ATGTAAACGTAGGAACTATTGG-3') and JWB-tuforfR2 (5'- TCCGATAGTTCTTCCACCTTCAC-3'). Amplicons of about 1.25 kb and 1.02 kb (16S rRNA gene and tuf gene, respectively) were obtained in 8 symptomatic samples from four peony gardens. However, no amplification was obtained in any of the asymptomatic samples. The representative amplicons of 16S rRNA and tuf genes of three positive samples (Heze-9, -18 and -27) were cloned into a zero background pLB-simple vector (Tiangen Biotechnology, Beijing, China) and sequenced by Taihe Biotechnology, Beijing, China. Sequences obtained in the study were deposited in NCBI GenBank with accession numbers PP504882, PP504883 and PP504884 for the 16S rRNA gene as well as PP530237, PP530238 and PP530239 for the tuf gene. The phytoplasma strain under the study was described as peony yellows (PeY) phytoplasma, PeY-Heze strain. Alignment analysis by DNAMAN software showed that three 16S rRNA gene sequences obtained in the study shared 99.36% to 99.60% sequence identity and three tuf gene sequences obtained in the study were identical. BLAST analysis of the 16S rRNA gene sequences of the PeY-Heze phytoplasma strains showed 99.60%-99.84% sequence identity with 'Candidatus Phytoplasma ziziphi' (GenBank accession: CP025121). And tuf sequences of the strains showed 100% similarity with 'Ca. P. ziziphi' (CP025121). Interestingly, the virtual RFLP patterns derived from three 16Sr RNA gene sequences obtained in the study by iPhyClassifier (Zhao et al., 2009) were different from the reference patterns of all previously established 16Sr groups/subgroups. The most similar are the reference pattern of the 16Sr group VII, subgroup E (AY741531), with a similarity coefficient of 0.72, which is less than 0.85. These phytoplasma strains may represent a new 16Sr group. Phylogenetic analysis based on 16S rRNA genes using MEGA 7.0 by neighbor-joining (NJ) method with 1000 bootstrap value indicated that PeY-Heze strains clustered into one clade with the phytoplasma strains of 'Ca. P. ziziphi' with 68% bootstrap value. Although there are several reports available on 'Ca. P. solani' infecting peony in Shandong Province, China (Gao et al., 2013). To our knowledge, this is the first report of 'Ca. P. ziziphi'-related strains infecting peony in China. The findings in this study will be beneficial to the detection, quarantine, and prevention of peony yellows phytoplasmas in China.

3,4-Dichlorophenylacetic acid acts as an auxin analog and induces beneficial effects in various crops.

Auxins and their analogs are widely used to promote root growth, flower and fruit development, and yield in crops. The action characteristics and application scope of various auxins are different. To overcome the limitations of existing auxins, expand the scope of applications, and reduce side effects, it is necessary to screen new auxin analogs. Here, we identified 3,4-dichlorophenylacetic acid (Dcaa) as having auxin-like activity and acting through the auxin signaling pathway in plants. At the physiological level, Dcaa promotes the elongation of oat coleoptile segments, the generation of adventitious roots, and the growth of crop roots. At the molecular level, Dcaa induces the expression of auxin-responsive genes and acts through auxin receptors. Molecular docking results showed that Dcaa can bind to auxin receptors, among which TIR1 has the highest binding activity. Application of Dcaa at the root tip of the DR5:GUS auxin-responsive reporter induces GUS expression in the root hair zone, which requires the PIN2 auxin efflux carrier. Dcaa also inhibits the endocytosis of PIN proteins like other auxins. These results provide a basis for the application of Dcaa in agricultural practices.

A prognostic model based on DNA methylation-related gene expression for predicting overall survival in hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) continues to increase in morbidity and mortality among all types of cancer. DNA methylation, an important epigenetic modification, is associated with cancer occurrence and progression. The objective of this study was to establish a model based on DNA methylation risk scores for identifying new potential therapeutic targets in HCC and preventing cancer progression. Methods: Transcriptomic, clinical, and DNA methylation data on 374 tumor tissues and 50 adjacent normal tissues were downloaded from The Cancer Genome Atlas-Liver Hepatocellular Carcinoma database. The gene expression profiles of the GSE54236 liver cancer dataset, which contains data on 161 liver tissue samples, were obtained from the Gene Expression Omnibus database. We analyzed the relationship between DNA methylation and gene expression levels after identifying the differentially methylated and expressed genes. Then, we developed and validated a risk score model based on the DNA methylation-driven genes. A tissue array consisting of 30 human hepatocellular carcinoma samples and adjacent normal tissues was used to assess the protein and mRNA expression levels of the marker genes by immunohistochemistry and qRT-PCR, respectively. Results: Three methylation-related differential genes were identified in our study: GLS, MEX3B, and GNA14. The results revealed that their DNA methylation levels were negatively correlated with local gene expression regulation. The gene methylation levels correlated strongly with the prognosis of patients with liver cancer. This was confirmed by qRT-PCR and immunohistochemical verification of the expression of these genes or proteins in tumors and adjacent tissues. These results revealed the relationship between the level of relevant gene methylation and the prognosis of patients with liver cancer as well as the underlying cellular and biological mechanisms. This allows our gene signature to provide more accurate and appropriate predictions for clinical applications. Conclusion: Through bioinformatics analysis and experimental validation, we obtained three DNA methylation marker: GLS, MEX3B, and GNA14. This helps to predict the prognosis and may be a potential therapeutic target for HCC patients.

Substituent Dependence on Series of Cationic Gyroidal MOFs in utc-c Topology with High CO2 Affinity and Ultrahigh Anionic Dye Adsorption Capacity.

A substituent decorating strategy for modification of the functional cavity is of great importance in the design of metal-organic frameworks (MOFs). Herein, three new isostructural cationic MOFs, [Cu3(Xpip)2]·NO3·nH2O (Xpip stands for X-substituted phenylimidazophenanthroline, where X = adm (SCNU-2), f (SCNU-3), and none for SCNU-4), have been successfully synthesized and shown gyroidal utc-c topology and large pore sizes which can be adjusted by different substituents (-N(CH3)2, -F, and -H). Interestingly, the differences of the substituents (sizes and proton donor/acceptor) show essential effects on the adsorption abilities of carbon dioxide and dyes, where SCNU-4 exhibits the highest CO2 affinity and the biggest adsorption capacity for anionic dyes Fluorescein Sodium, and SCNU-3 adsorbs the largest amount (1503.6 mg/g) of Acid Fuchsin to date for the reported porous materials. The detailed studies in adsorption kinetics, adsorption isotherms, and theoretical calculation of the binding energies between the structures and dye molecules confirm that the electric properties of the frameworks (cationic) and substituents directed to the pore surface are two important factors dramatically affecting the selective dye adsorption.

Site-Specific N-Glycan Characterization of Grass Carp Serum IgM.

Immunoglobulin M (IgM) is the major antibody in teleost fish and plays an important role in humoral adaptive immunity. The N-linked carbohydrates presenting on IgM have been well documented in higher vertebrates, but little is known regarding site-specific N-glycan characteristics in teleost IgM. In order to characterize these site-specific N-glycans, we conducted the first study of the N-glycans of each glycosylation site of the grass carp serum IgM. Among the four glycosylation sites, the Asn-262, Asn-303, and Asn-426 residues were efficiently glycosylated, while Asn-565 at the C-terminal tailpiece was incompletely occupied. A striking decrease in the level of occupancy at the Asn-565 glycosite was observed in dimeric IgM compared to that in monomeric IgM, and no glycan occupancy of Asn-565 was observed in tetrameric IgM. Glycopeptide analysis with liquid chromatography-electrospray ionization tandem mass spectrometry revealed mainly complex-type glycans with substantial heterogeneity, with neutral; monosialyl-, disialyl- and trisialylated; and fucosyl-and non-fucosyl-oligosaccharides conjugated to grass carp serum IgM. Glycan variation at a single site was greatest at the Asn-262 glycosite. Unlike IgMs in other species, only traces of complex-type and no high-mannose glycans were found at the Asn-565 glycosite. Matrix-assisted laser desorption ionization analysis of released glycans confirmed the overwhelming majority of carbohydrates were of the complex-type. These results indicate that grass carp serum IgM exhibits unique N-glycan features and highly processed oligosaccharides attached to individual glycosites.

Effect of sodium ascorbate on degree of conversion and bond strength of RealSeal SE to sodium hypochlorite treated root dentin.

The aim of this study was to investigate the effect of sodium ascorbate (Sa) on degree of conversion (DC) and bond strength (BS) of RealSeal SE to sodium hypochlorite (NaOCl) treated root dentin. Two hundreds simulated canals were prepared and irrigated with Distilled water(DW), 1.3% NaOCl (1.3% N), 5.2% NaOCl (5.2% N), MTAD, 17% EDTA (EDTA), 10% Sa, 1.3% NaOCl/MTAD (N-M), 1.3% NaOCl/Sa/MTAD(N-Sa-M), 5.2% NaOCl/EDTA(N-E), and 5.2% NaOCl/Sa/EDTA (N-Sa-E) respectively. They were subsequently bulk filled with RealSeal SE and analyzed with micro-Raman spectroscopy and universal testing machine for DC and BS respectively. One-way ANOVA and post hoc Tukey's test showed DC of 1.3% N, 5.2% N, N-M and N-E were significantly lower (p<0.01) than other six groups. BS of DW, Sa, N-M were significantly lower than 1.3% N, 5.2% N, MTAD, EDTA, N-Sa-M and N-E (p<0.01), and group N-Sa-E achieved the highest BS among all groups (p<0.01). NaOCl negatively affected DC and BS of RealSeal SE, which could be reversed with 10% Sa.

Degree of conversion of a methacrylate-based endodontic sealer: a micro-Raman spectroscopic study.

INTRODUCTION: Recently, a methacrylate-based obturation system, Resilon/RealSeal SE, has been developed to replace gutta-percha and traditional sealers. As a resin-based material, degree of conversion (DC) is one of the most important characteristics. The aim of this study was to investigate the time-dependent change of DC of RealSeal SE as well as the influence of canal moisture and root canal depth on it. METHODS: DC of RealSeal SE, either self-cured or dual-cured (n = 8 in each group), was calculated according to the Raman spectra obtained at different times after mixing. Thirty extracted teeth with single canal were instrumented and divided randomly into 2 groups in terms of different canal drying methods. In the ethanol group, excess distilled water in root canal was removed with paper points followed by 95% ethanol. In the paper points group, root canals were blot-dried with paper points until the last one appeared dry. DC of RealSeal SE was calculated from serial cross sections (2, 5, and 8 mm from the apex) obtained 1 week after obturation with Resilon/RealSeal SE. RESULTS: Significant increase in DC of RealSeal SE was observed in 1 week (P < .01), with little change afterwards (P > .05). DC of sealer in ethanol group was significantly higher than in paper points group (P < .01). However, DCs of RealSeal SE at different levels of tooth sections were not significantly different (P > .05). CONCLUSIONS: Both self-cured and dual-cured RealSeal SE achieved stable DC after 1 week. Root canal moisture was a critical factor in determining DC of RealSeal SE.

Antagonistic potential against pathogenic microorganisms and hydrogen peroxide production of indigenous lactobacilli isolated from vagina of Chinese pregnant women.

OBJECTIVE: To investigate the indigenous lactobacilli from the vagina of pregnant women and to screen the isolates with antagonistic potential against pathogenic microorganisms. METHODS: The strains were isolated from pregnant women's vagina and identified using the API50CH system. The ability of the isolates to produce hydrogen peroxide was analyzed semi-quantitatively using the TMB-HRP-MRS agar. The antagonistic effects of the isolates on pathogenic microorganisms were determined with a double layer agar plate. RESULTS: One hundred and three lactobacilli strains were isolated from 60 samples of vaginal secretion from healthy pregnant women. Among them, 78 strains could produce hydrogen peroxide, in which 68%, 80%, 80%, and 88% had antagonistic effects against Candida albicans CMCC98001, Staphylococcus aureus CMCC26003, Escherichia coli CMCC44113, and Pseudomonas aeruginosa CMCC10110, respectively. CONCLUSION: The recovery of hydrogen peroxide-producing lactobacilli decreases with the increasing pregnant age and time. The most commonly isolated species from vagina of Chinese pregnant women are Lactobacillus acidophilus and Lactobacillus crispatus. Most of L. acidophilus and L. crispatus produce a high H2O2 level.

[Improvement of two-dimensional gel electrophoresis of total proteins from rice anthers].

This paper reported an improvement in 2-D gel electrophoresis of the proteome in Honglian cytoplasmic male sterile rice. An IPGphor unit with immobile pH gradient strips was used as the first dimension and SDS-PAGE as the second. The total anther proteins were extracted using TCA/acetone and then were washed 5-6 times with acetone till the proteins were white and clean, and then tributylphosphine and DTT were added into the rehydration buffer to improve the solubility of the proteins. The 2-D gel was stained by both methods of coomassie blue G-250 and silver. Extraction of proteins, pH of the strips and rehydration of the strips were optimized and compared. Higher repeatability and better separating protein pattern could be gained by this technique.

[Preliminary proteomics analysis of the total proteins of HL Type cytoplasmic male sterility rice anther].

The proteins of HL type cytoplasmic male sterility rice anther of YTA (CMS) and YTB (maintenance line) were separated by two-dimensional electrophoresis with immobilized ph (3-10 non-linear) gradients as the first dimension and SDS-PAGE as the second. The silver-stained proteins spots were analyzed using Image Master 2D software, there were about 1800 detectable spots on each 2D-gel, and about 85 spots were differential expressed. With direct MALDI-TOF mass spectrometry analysis and protein database searching, 9 protein spots out of 16 were identified. Among those proteins, there were Putative nucleic acid binding protein, glucose-1-phosphate adenylyltransferase (ADP-glucose pyrophosphorylase, AGPase) (EC: 2.7.7.27) large chain, UDP-glucuronic acid decarboxylase, putative calcium-binding protein annexin, putative acetyl-CoA synthetase and putative lipoamide dehydrogenase etc. They were closely associated with metabolism, protein biosynthesis, transcription, signal transduction and so on, all of which are cell activities that are essential to pollen development. Some of the identified proteins, i.e. AGPase, putative lipoamide dehydrogenase and putative acetyl-CoA synthetase were deeply discussed on the relationship to CMS. AGPase catalyzes a very important step in the biosynthesis of alpha 1,4-glucans (glycogen or starch) in bacteria and plants: synthesis of the activated glucosyl donor, ADP-glucose, from glucose-1-phosphate and ATP. The lack of the AGPase in male sterile line might directly result in the reduction of starch, and the synthesis of starch was the most important processes during the development of pollen. In present research, the descent or reduction of putative lipoamide dehydrogenase and putative acetyl-CoA synthetase seemed involved in pollen sterility in rice. The degeneration and formation of various tissues during pollen development may impose high demands for energy and key biosynthetic intermediates. Under such conditions, the TCA cycle needs to operate fully, because the TCA cycle is an important source for many intermediates required for biosynthetic pathways, in addition to performing an oxidative, energy-producing role. Thus, it seemed reasonable to infer that the decrease of putative lipoamide dehydrogenase and putative acetyl-CoA synthetase in anther might prevent the conversion of pyruvate into acetyl-CoA, and as a result, the TCA cycle could no longer operate at a sufficient rate to meet all requirements in anther cells, leading to pollen sterility. This study gave new insights into the mechanism of CMS in rice and demonstrated the power of the proteomic approach in plant biology studies.

[Study on the editing sites of the coxII gene transcripts of rice mitochondria].

The term RNA editing is generally used to describe those molecular processes in which the information content is altered in an RNA molecule. This process is not limited to mRNA since alterations of non-informational RNA have also been found. RNA editing exists extensively in the higher plant mitochondria, and is the necessary step for forming functional proteins. In this paper, the research materials are the gametopthyte male sterility line (A), maintainer line (B) and F1 hybrid (F1) of HL-type CMS. 15 editing sites are found in the transcripts of coxII by comparing cDNAs and DNAs sequences. A,B and F1 have same Editing sites. When editing occurs at the first or second position of codons, the encoded amino acid is likely to be altered. As a result, the conservation of the predicted protein is improved as compared with other organisms.

[A RAPD method acceptable for the analysis of mitochondrial gene expression].

Several techniques are available in detecting variations in gene expression between different samples, such as SSH, RACE, etc. However, they can not be applied to analyze mitochondrial gene expression due to the specific characteristics of mitochondrial RNA. So some modifications were made to the conventional techniques. Here we reported a demonstration of this modified technique, taking rice mitochondria as materials. In this technique, using random hexamers to prime the RT, the resultant cDNA likely included coding regions because it was not locked to the poly(A) tail of the messenger RNA.