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By enrichment culturing of soil contaminated with metribuzin, a highly efficient metribuzin degrading bacterium,Bacillussp. N1, was isolated. This strain grows using metribuzin at 5.0% (v/v) as the sole nitrogen source in a liquid medium. Optimal metribuzin degradation occurred at a temperature of 30ºC and at pH 7.0. With an initial concentration of 20 mg L−1, the degradation rate was 73.5% in 120 h. If the initial concentrations were higher than 50 mg L−1, the biodegradation rates decreased as the metribuzin concentrations increased. When the concentration was 100 mg L−1, the degradation rate was only 45%. Degradation followed the pesticide degradation kinetic equation at initial concentrations between 5 mg L−1and 50 mg L−1. When the metribuzin contaminated soil was mixed with strain N1 (with the concentration of metribuzin being 20 mg L−1and the inoculation rate of 1011g−1dry soil), the degradation rate of the metribuzin was 66.4% in 30 days, while the degradation rate of metribuzin was only 19.4% in the control soil without the strain N1. These results indicate that the strain N1 can significantly increase the degradation rate of metribuzin in contaminated soil. [ABSTRACT FROM PUBLISHER]

Haematococcus pluvialis accumulates a large amount of astaxanthin under various stresses, e.g., blue light and salicylic acid (SA). However, the metabolic response of H. pluvialis to blue light and SA is still unclear. We investigate the effects of blue light and SA on the metabolic response in H. pluvialis using both transcriptomic and proteomic sequencing analyses. The largest numbers of differentially expressed proteins (DEPs; 324) and differentially expressed genes (DEGs; 13,555) were identified on day 2 and day 7 of the treatment with blue light irradiation (150 μmol photons m−2s−1), respectively. With the addition of SA (2.5 mg/L), a total of 63 DEPs and 11,638 DEGs were revealed on day 2 and day 7, respectively. We further analyzed the molecular response in five metabolic pathways related to astaxanthin synthesis, including the astaxanthin synthesis pathway, the fatty acid synthesis pathway, the heme synthesis pathway, the reactive oxygen species (ROS) clearance pathway, and the cell wall biosynthesis pathway. Results show that blue light causes a significant down-regulation of the expression of key genes involved in astaxanthin synthesis and significantly increases the expression of heme oxygenase, which shows decreased expression by the treatment with SA. Our study provides novel insights into the production of astaxanthin by H. pluvialis treated with blue light and SA. [ABSTRACT FROM AUTHOR]

Wild soybean germplasm is distributed widely in China, particularly in Heilongjiang Province. In this study, 242 wild soybean accessions from four agricultural divisions in Heilongjiang Province were evaluated based on six qualitative and eight quantitative traits. Results showed that a large amount of variation occurred in these evaluated traits. Abundant qualitative traits included the wild type (78.51%), purple flowers (90.50%), needle leaf (39.26%), black seed (83.88%), brown hilum (52.07%), and mud film (87.60). Results of multivariate analysis based on quantitative traits showed that 100-seed weight, seeds weight per plant, number of seeds per plant, number of effective pods, and number of invalid pods were significantly different among samples (p < 0.05). A total of 27 germplasms were screened. Cluster analysis identified the 242 accessions into two groups, not following the geographical distribution pattern, with rich wild soybean resources revealed in the northern site. The present study indicated that wild soybean in Heilongjiang Province should be conserved in situ. The rich genetic diversity revealed in soybeans of different sites in Heilongjiang Province suggested its significant potential utilization in genetic improvement and breakthrough for soybean breeding. This information will help to exploit and conserve wild soybean accessions in Heilongjiang Province. [ABSTRACT FROM AUTHOR]

Three species (Aconitum taipeicum, Delphinium giraldii, and Consolida ajacis) of the tribe Delphinieae (Ranunculaceae) were examined using scanning electron microscopy and histological methods. The results showed that members of Delphinieae differ from their polysymmetrical relatives by four unique features: (1) a spiral phyllotaxis of their perianth and stamens, and a series of carpels, which initiated superficially in a whorl-liked arrangement; (2) sepal 2 being the largest one among the five sepals and becoming helmet-shaped or having a spur; (3) petals 2 and 5 initiated adaxially of sepal 2 and also becoming spurred; and (4) the monosymmetry of the first flower becoming established when sepal 2 becomes the largest. Major differences among the species include the timing of development of the second series; the fusion of two petals into a single one in C. ajacis; and, during early developmental stages, the two young spurred petals giving rise to a stalk and two bulges in A. taipeicum, a single bulge in D. giraldii, or an arch blade in C. ajacis. The unequal growth of the perianth, together with the reduction and the rearrangement of the carpels, are critical in inducing the symmetrical transformation of the flowers. [ABSTRACT FROM AUTHOR]

Abstract: An apple starch-branching enzyme SbeI gene (GenBank Accession No. DQ115404) has been isolated, cloned, and sequenced. The SbeI is a single copy gene in the apple genome, consisting of 14 exons and 13 introns, and covering 6075bp. As detected by RT-PCR, the apple SbeI is expressed at very low levels during early stages of fruit development; while, the highest levels of mRNA transcripts are observed at ∼44 days post-pollination. Besides fruits, the apple SbeI is also expressed in buds and flowers, and very weakly in leaves. The genomic structure of SbeI in apple is strikingly similar to those reported so far in grasses (Poaceae), with exons 4 through 13 being of identical lengths in both apple and grasses. Moreover, structure similarities in exon lengths have also been detected in SbeII genes of both grasses and eudicots. These findings prompted the investigation of the evolutionary process of the Sbe gene family in angiosperms. A total of 26 Sbe sequences, representing an array of monocots and eudicots, are investigated in this study. Phylogenetic analysis has suggested that Sbe genes have duplicated into SbeI and SbeII prior to the divergence of moncots from eudicots. The SbeII gene is further duplicated into SbeIIa and SbeIIb prior to the radiation of grasses; however, it is not yet clear whether this duplication event has occurred before or after the radiation of the eudicots. [Copyright &y& Elsevier]

The treatment of complex polluted wastewater has become an increasingly critical concern for the various types of hazardous organic compounds, including synthetic dyes and pharmaceuticals. Due to their efficient and eco-friendly advantages, the white-rot fungi (WRF) have been applied to degrade environmental pollutants. This study aimed to investigate the removal ability of WRF (i.e., Trametes versicolor WH21) in the co-contamination system composed of Azure B dye and sulfacetamide (SCT). Our study discovered that the decolorization of Azure B (300 mg/L) by strain WH21 was significantly improved (from 30.5% to 86.5%) by the addition of SCT (30 mg/L), while the degradation of SCT was also increased from 76.4% to 96.2% in the co-contamination system. Transcriptomic and biochemical analyses indicated that the ligninolytic enzyme system was activated by the enhanced enzymatic activities of MnPs and laccases, generating higher concentration of extracellular H 2 O 2 and organic acids in strain WH21 in response to SCT stress. Purified MnP and laccase of strain WH21 were revealed with remarkable degradation effect on both Azure B and SCT. These findings significantly expanded the existing knowledge on the biological treatment of organic pollutants, indicating the strong promise of WRF in the treatment of complex polluted wastewater., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Heme is an iron/porphyrin complex compound, widely used in the health care, food, and pharmaceutical industries. It is more advantageous and attractive to develop microbial cell factories to produce heme by fermentation, with lower production costs and environmentally more friendly procedures than those of the traditional extraction based on animal blood. In this study, Bacillus subtilis, a typical industrial model microorganism of food safety grade, was used for the first time as the host to synthesize heme., Results: The heme biosynthetic pathway was engineered as four modules, the endogenous C5 pathway, the heterologous C4 pathway, the uroporphyrinogen (urogen) III synthesis pathway, and the downstream synthesis pathway. Knockout of hemX encoding the negative effector of the concentration of HemA, overexpression of hemA encoding glutamyl-tRNA reductase, and knockout of rocG encoding the major glutamate dehydrogenase in the C5 pathway, resulted in an increase of 427% in heme production. Introduction of the heterologous C4 pathway showed a negligible effect on heme biosynthesis. Overexpression of hemCDB, which encoded hydroxymethylbilane synthase, urogen III synthase, and porphobilinogen synthase participating in the urogen III synthesis pathway, increased heme production by 39%. Knockouts of uroporphyrinogen methyltransferase gene nasF and both heme monooxygenase genes hmoA and hmoB in the downstream synthesis pathway increased heme production by 52%. The engineered B. subtilis produced 248.26 ± 6.97 mg/L of total heme with 221.83 ± 4.71 mg/L of extracellular heme during the fed-batch fermentation in 10 L fermenter., Conclusions: Strengthening endogenous C5 pathway, urogen III synthesis pathway and downstream synthesis pathway promoted the biosynthesis of heme in B. subtilis. The engineered B. subtilis strain has great potential as a microbial cell factory for efficient industrial heme production., (© 2023. The Author(s).)

Purpose: Increasing evidences have revealed the anti-cancer effect of disulfiram. Current disulfiram-based cancer therapies still have limitations, such as poor tumor-targeting ability and insufficient studies on anti-tumor mechanisms., Methods: In the present study, tumor-targeting liposomes were prepared as drug carriers to increase retention of disulfiram in tumor cells. Then, anti-tumor efficacy of liposomes and the underlying mechanisms were investigated in in vitro, in vivo, and transcriptomic level., Results: The results showed that disulfiram enhanced sensitivity of human hepatocellular carcinoma cells to doxorubicin by 15-27-fold, and increased reactive oxygen species (ROS) production as well as caspase-dependent apoptosis. Inhibition of tumor migration and invasion by doxorubicin were further enhanced by disulfiram. In vivo study showed that disulfiram additive doxorubicin liposomes had better performance in suppressing tumor growth than single doxorubicin liposomes. Gene expression profiling found that cellular components destruction, cell stress, check point regulation, and immunoregulation were the main anti-tumor mechanisms of disulfiram. More importantly, disulfiram possessed a great potential to be a protein ubiquitination and murine double minute 4 (MDM4) targeting compound., Conclusions: Due to its low price and good safety, it is worth to repurposing disulfiram as a chemotherapeutic drug. Furthermore, MDM4 may act as a biomarker for observation the clinical effect of disulfiram-based treatment., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Degradation efficiency of Atrazine by Klebsiella variicola FH-1 is improved by the addition of Zn 2+ . Both the chromosome and plasmid genomes of strain FH-1 were sequenced and annotated to identify genes involved in the degradation of Atrazine. Four open reading frames (ORFs) 1040, 2582, 3597, and 4043 encoding Zn 2+ -dependent hydrolases were knocked out to verify their predicted functions in the degradation of Atrazine. In the presence of Zn 2+ , the biodegradation efficiency of Atrazine by knockout mutant ∆ORF 3597 was 13.7% lower than that of wild type (WT) of strain FH-1 but still 9.4% higher than that of WT without Zn 2+ . These results indicated that ORF 3597 played a synergistic role but may not be the sole factor involved in the degradation of Atrazine. The enzymatic activities of pydC encoded by ORF 3597 were further characterized in the degradation of Atrazine. Results of fluorescence staining and flow cytometry analyses showed that the survival of bacterial cells and cell membrane permeability were increased in the presence of Zn 2+ at different concentrations. Our study provided a scientific foundation for further investigation of the biological mechanisms of improving the degradation of Atrazine by strain FH-1 with the presence of Zn 2+ ., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The vacuolar H + -ATPase (V-ATPase) plays many important roles in cell growth and in response to stresses in plants. The V-ATPase subunit H (VHA-H) is required to form a stable and active V-ATPase. Genome-wide analyses of VHA-H genes in crops contribute significantly to a systematic understanding of their functions. A total of 22 VHA-H genes were identified from 11 plants representing major crops including cotton, rice, millet, sorghum, rapeseed, maize, wheat, soybean, barley, potato, and beet. All of these VHA-H genes shared exon-intron structures similar to those of Arabidopsis thaliana . The C-terminal domain of VHA-H was shorter and more conserved than the N-terminal domain. The VHA-H gene was effectively used as a genetic marker to infer the phylogenetic relationships among plants, which were congruent with currently accepted taxonomic groupings. The VHA-H genes from six species of crops ( Gossypium raimondii , Brassica napus , Glycine max , Solanum tuberosum , Triticum aestivum , and Zea mays ) showed high gene structural diversity. This resulted from the gains and losses of introns. Seven VHA-H genes in six species of crops ( Gossypium raimondii , Hordeum vulgare , Solanum tuberosum , Setaria italica , Triticum aestivum , and Zea mays ) contained multiple transcript isoforms arising from alternative splicing. The study of cis-acting elements of gene promoters and RNA-seq gene expression patterns confirms the role of VHA-H genes as eco-enzymes. The gene structural diversity and proteomic diversity of VHA-H genes in our crop sampling facilitate understanding of their functional diversity, including stress responses and traits important for crop improvement.

Objective To explore the optimal method for in vitro cultivation of SD fetal rat cardiomyocytes. Methods Cardiomyocytes of SD fetal rats from pregnant rats on gestational day 19 were obtained by digesting cardiac tissues with 0.8 g/L trypsin and 0.4 g/L collagenase II. With differential centrifugation and different animal serums (bovine fetal serum or horse serum), we tried to look for the optimal culture conditions. The morphology and beat frequency of cardiomyocytes were observed under an inverted phase-contrast microscope. In addition, the expressions of alpha-sarcomeric actinin (α-SA) and cardiac troponin I (cTnI) in the cultured cardiomyocytes were detected by immunofluorescence staining to identify cardiomyocytes and the purity of these cells after 24, 48, 72 and 96 hours of cultivation. Results After 24 hours of cultivation, we had seen a majority of adherent cells and a few of suspension cells, and also seen some pulsating cardiomyocytes. The positive rates of α-SA and cTnI in the cardiomyocytes of the horse serum-cultured group were obviously higher than those in the bovine fetal serum-cultured group after 48, 72 and 96 hours of cultivation. Conclusion Co-digestion with trypsin and collagenaseII in combination with horse serum cultivation are easier to cultivate fetal rat cardiomyocytes.