DcWRKY33 promotes petal senescence in carnation (Dianthus caryophyllus L.) by activating genes involved in the biosynthesis of ethylene and abscisic acid and accumulation of reactive oxygen species.

Carnation (Dianthus caryophyllus L.) is one of the most famous and ethylene-sensitive cut flowers worldwide, but how ethylene interacts with other plant hormones and factors to regulate petal senescence in carnation is largely unknown. Here we found that a gene encoding WRKY family transcription factor, DcWRKY33, was significantly upregulated upon ethylene treatment. Silencing and overexpression of DcWRKY33 could delay and accelerate the senescence of carnation petals, respectively. Abscisic acid (ABA) and H2 O2 treatments could also accelerate the senescence of carnation petals by inducing the expression of DcWRKY33. Further, DcWRKY33 can bind directly to the promoters of ethylene biosynthesis genes (DcACS1 and DcACO1), ABA biosynthesis genes (DcNCED2 and DcNCED5), and the reactive oxygen species (ROS) generation gene DcRBOHB to activate their expression. Lastly, relationships are existed between ethylene, ABA and ROS. This study elucidated that DcWRKY33 promotes petal senescence by activating genes involved in the biosynthesis of ethylene and ABA and accumulation of ROS in carnation, supporting the development of new strategies to prolong the vase life of cut carnation.

DNA methylation remodeled amino acids biosynthesis regulates flower senescence in carnation (Dianthus caryophyllus).

Dynamic DNA methylation regulatory networks are involved in many biological processes. However, how DNA methylation patterns change during flower senescence and their relevance with gene expression and related molecular mechanism remain largely unknown. Here, we used whole genome bisulfite sequencing to reveal a significant increase of DNA methylation in the promoter region of genes during natural and ethylene-induced flower senescence in carnation (Dianthus caryophyllus L.), which was correlated with decreased expression of DNA demethylase gene DcROS1. Silencing of DcROS1 accelerated while overexpression of DcROS1 delayed carnation flower senescence. Moreover, among the hypermethylated differentially expressed genes during flower senescence, we identified two amino acid biosynthesis genes, DcCARA and DcDHAD, with increased DNA methylation and reduced expression in DcROS1 silenced petals, and decreased DNA methylation and increased expression in DcROS1 overexpression petals, accompanied by decreased or increased amino acids content. Silencing of DcCARA and DcDHAD accelerates carnation flower senescence. We further showed that adding corresponding amino acids could largely rescue the senescence phenotype of DcROS1, DcCARA and DcDHAD silenced plants. Our study not only demonstrates an essential role of DcROS1-mediated remodeling of DNA methylation in flower senescence but also unravels a novel epigenetic regulatory mechanism underlying DNA methylation and amino acid biosynthesis during flower senescence.

Histone H3K4 methyltransferase DcATX1 promotes ethylene induced petal senescence in carnation.

Petal senescence is controlled by a complex regulatory network. Epigenetic regulation like histone modification influences chromatin state and gene expression. However, the involvement of histone methylation in regulating petal senescence remains poorly understood. Here, we found that the trimethylation of histone H3 at Lysine 4 (H3K4me3) is increased during ethylene-induced petal senescence in carnation (Dianthus caryophyllus L.). H3K4me3 levels were positively associated with the expression of transcription factor DcWRKY75, ethylene biosynthetic genes 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (DcACS1), and ACC oxidase (DcACO1), and senescence associated genes (SAGs) DcSAG12 and DcSAG29. Further, we identified that carnation ARABIDOPSIS HOMOLOG OF TRITHORAX1 (DcATX1) encodes a histone lysine methyltransferase which can methylate H3K4. Knockdown of DcATX1 delayed ethylene-induced petal senescence in carnation, which was associated with the down-regulated expression of DcWRKY75, DcACO1, and DcSAG12, whereas overexpression of DcATX1 exhibited the opposite effects. DcATX1 promoted the transcription of DcWRKY75, DcACO1, and DcSAG12 by elevating the H3K4me3 levels within their promoters. Overall, our results demonstrate that DcATX1 is a H3K4 methyltransferase that promotes the expression of DcWRKY75, DcACO1, DcSAG12 and potentially other downstream target genes by regulating H3K4me3 levels, thereby accelerating ethylene-induced petal senescence in carnation. This study further indicates that epigenetic regulation is important for plant senescence processes.

Improving the regeneration rate of deep lethal mutant protoplasts by fusion to promote efficient L-lysine fermentation.

BACKGROUND: L-lysine is widely used for feed and special diet products. The transformation of fermentation strains plays a decisive role in the development of these industries. Based on the mutation breeding theory and metabolic engineering methods, this study aimed to improve the regeneration rate of high-lethality protoplasts by combining multiple mutagenesis and homologous cell fusion techniques to efficiently concentrate multiple dominant mutations and optimize the L-lysine production strain Escherichia coli QDW. RESULTS: In order to obtain the best protoplasts, the optimal enzymolysis time was selected as 4 h. The optimal lysozyme concentration was estimated at 0.8 mg/mL, because the protoplast formation rate and regeneration rate reached 90% and 30%, respectively, and their product reached the maximum. In this study, it was necessary that UV mutagenesis be excessive to obtain an expanded mutation library. For high lethality protoplasts, under the premise of minimal influence on its recovery, the optimal time for UV mutagenesis of protoplasts was 7 min, and the optimal time for thermal inactivation of protoplasts at 85 ℃ was 30 min. After homologous fusion, four fusion strains of E. coli were obtained, and their stability was analyzed by flow cytometry. The L-lysine yield of QDW-UH3 increased by 7.2% compared with that of QDW in a fermentation experiment, which promoted the expression of key enzymes in L-lysine synthesis, indicating that the combination of ultraviolet mutagenic breeding and protoplast fusion technology improved the acid-production level of the fusion strain. CONCLUSION: This method provides a novel approach for the targeted construction of microbial cell factories.

An insertion of transposon in DcNAP inverted its function in the ethylene pathway to delay petal senescence in carnation (Dianthus caryophyllus L.).

Petal senescence is the final stage of flower development. Transcriptional regulation plays key roles in this process. However, whether and how post-transcriptional regulation involved is still largely unknown. Here, we identified an ethylene-induced NAC family transcription factor DcNAP in carnation (Dianthus caryophyllus L.). One allele, DcNAP-dTdic1, has an insertion of a dTdic1 transposon in its second exon. The dTdic1 transposon disrupts the structure of DcNAP and causes alternative splicing, which transcribes multiple domain-deleted variants (DcNAP2 and others). Conversely, the wild type allele DcNAP transcribes DcNAP1 encoding an intact NAC domain. Silencing DcNAP1 delays and overexpressing DcNAP1 accelerates petal senescence in carnation, while silencing and overexpressing DcNAP2 have the opposite effects, respectively. Further, DcNAP2 could interact with DcNAP1 and interfere the binding and activation activity of DcNAP1 to the promoters of its downstream target ethylene biosynthesis genes DcACS1 and DcACO1. Lastly, ethylene signalling core transcriptional factor DcEIL3-1 can activate the expression of DcNAP1 and DcNAP2 in the same way by binding their promoters. In summary, we discovered a novel mechanism by which DcNAP regulates carnation petal senescence at the post-transcriptional level. It may also provide a useful strategy to manipulate the NAC domains of NAC transcription factors for crop genetic improvement.

Recent insights into function, structure and modification of cytochrome P450 153 a family.

Cytochrome P450 153 A (CYP153A) is a versatile enzyme that can catalyze a wide range of oxidation reactions on various substrates. This review provides a comprehensive overview of the current state of knowledge on CYP153A, including its classification, structure, function, and potential applications in biotechnology and pharmaceuticals. The CYP153A family encompasses many enzymes with different functions on a variety of substrates. We also discuss the structural features that are responsible for the different substrate specificities. Additionally, the enzyme has been engineered to increase its catalytic activity and modifications have been made to enhance its properties further. Despite its potential, challenges and limitations associated with studying and exploiting CYP153A remain, such as low expression levels and substrate inhibition. Nonetheless, ongoing research is exploring new ways to harness the enzyme's capabilities, particularly in synthetic biology, biocatalysis, and drug discovery, making it an exciting target for future research.

Use of fulvic acid-like compounds from pulp-derived black liquor for enhancing the selenium content of peanut buds.

BACKGROUND: Cleaner production involving the extraction of useful material from the black liquor by-product of straw pulp would be environmentally beneficial and would permit increased wastewater usage. RESULTS: The fulvic-acid-like components of pulp black liquor (PFA) with molecular weights below 10 kDa were isolated. The chemical and physiological characteristics of PFAs were investigated. Selenite can enhance the selenium nutrition level of crops, but excessive selenite may be toxic to plant growth. In order to explore how to increase selenite tolerance and selenium accumulation in peanut, the effects of PFA on selenium-associated properties in peanut seedlings were examined by growing seedlings with sodium selenite (0, 5, 15, and 25 mg·L- 1 Na2SeO3, 15 mg·L- 1 Na2SeO3 solution containing 60 mg-C/L PFA, and 25 mg·L- 1 Na2SeO3 containing 60 mg-C/L PFA). CONCLUSION: The results showed that with 15 mg·L- 1 Na2SeO3, PFA significantly increased both the total and hypocotyl fresh weight of the seedlings but reduced the fresh weight of the root. PFA also effectively promoted the conversion of Se from inorganic to organic compounds in the root and hypocotyl, increased the soluble total sugar and soluble protein contents of the hypocotyl, and thus improved the edible quality and food safety of the selenium-enriched peanut buds. The results suggest that PFA can be used as an innovative bio-based substance for selenium-enriched sprout vegetable production.

Block Copolymer Vesicles with Tunable Membrane Thicknesses and Compositions Prepared by Aqueous Seeded Photoinitiated Polymerization-Induced Self-Assembly at Room Temperature.

Block copolymer vesicles with diverse functionalities and intrinsic hollow structures have received considerable attention due to their broad applications in biomedical fields, including drug delivery, bioimaging, theranostics, gene therapy, etc. However, efficient preparation of block copolymer vesicles with tunable membrane thicknesses and compositions under mild conditions is still a challenge. Herein, we report an aqueous seeded photoinitiated polymerization-induced self-assembly (photo-PISA) for the precise preparation of block copolymer vesicles at room temperature. By changing the total degree of polymerization (DP) of the hydrophobic block in seeded photo-PISA, one can easily tune the membrane thickness without compromising the morphology of vesicles. Moreover, by adding different comonomers such as hydrophobic monomers, hydrophilic monomers, and cross-linkers into seeded photo-PISA, vesicles with different compositions could be prepared without compromising the morphology and colloidal stability. Polymerization kinetics show that seeded photo-PISA can skip the step of in situ self-assembly with a short homogeneous polymerization stage being observed. To demonstrate potential biological applications, enzymatic nanoreactors were constructed by loading horseradish peroxidase (HRP) inside vesicles via seeded photo-PISA. The enzymatic properties of these nanoreactors could be easily regulated by changing the membrane thickness and hydrophobicity. It is expected that this method can provide a facile platform for the precise preparation of block copolymer vesicles that may find applications in different fields.

Electrochemical Heteroatom-Heteroatom Bond Construction.

Heteroatom-heteroatom linkage, with S-S bond as a presentative motif, served a crucial role in biochemicals, pharmaceuticals, pesticides, and material sciences. Thus, preparation of the privileged scaffold has always been attracting tremendous attention from the synthetic community. However, classic protocols suffered from several drawbacks, such as toxic and unstable agents, poor functional group tolerance, multiple steps, and explosive oxidizing regents as well as the transitional metal catalysts. Electrochemical organic synthesis exhibited a promising alternative to the traditional chemical reaction due to the sustainable electricity can be employed as the traceless redox agents. Hence, toxic and explosive oxidants and/or transitional metals could be discarded under mild reaction with high efficiency. In this context, a series of electrochemical approaches for the construction of heteroatom-heteroatom bond were reviewed. Notably, most of the cases illustrated the dehydrogenative feature with the clean energy molecules hydrogen as the sole by-product.

Cross-dimensional magnitude interaction is modulated by representational noise: evidence from space-time interaction.

Magnitudes along different dimensions (e.g., space and time) tend to interact with each other in perception, with some magnitude dimensions more susceptible to cross-dimensional interference than others. What causes such asymmetries in cross-dimensional magnitude interaction is being debated. The current study investigated whether the representational noise of magnitudes modulates the (a)symmetry in space-time interaction. In three experiments using different formats of length, we showed that dynamic unfilled lengths resulted in a higher representational noise than either static unfilled length or static filled length. Correspondingly, we observed that the time-on-space effect was larger for dynamic unfilled lengths than for static unfilled length or static filled length (and it did not differ between the latter two). Further correlational analyses showed that the susceptibility of a target dimension to the influence of a concurrent dimension increased as a function of participants' representational noise in the target dimension (e.g., the noisier length representations, the larger the time-on-space effect). In all, our study showed that the representational noise of space and time modulates the way the two dimensions interact. These findings suggest that cross-dimensional magnitude interactions arise as a result of memory interference, with noisier magnitudes being more prone to being nudged by concurrent magnitudes in other dimensions. Such memory interference can be seen as a result of Bayesian inference with correlated priors between magnitude dimensions.

Anti-Biofilm and Anti-Hemolysis Activities of 10-Hydroxy-2-decenoic Acid against Staphylococcus aureus.

Persistent infections caused by Staphylococcus aureus biofilms pose a major threat to global public health. 10-Hydroxy-2-decenoic acid (10-HDA), a main fatty acid in royal jelly, has been shown to possess various biological activities. The purpose of this study was to explore the effects of 10-HDA on the biofilms and virulence of S. aureus and its potential molecular mechanism. Quantitative crystal violet staining indicated that 10-HDA significantly reduced the biofilm biomass at sub-minimum inhibitory concentration (MIC) levels (1/32MIC to 1/2MIC). Scanning electron microscope (SEM) observations demonstrated that 10-HDA inhibited the secretion of extracellular polymeric substances, decreased bacterial adhesion and aggregation, and disrupted biofilm architecture. Moreover, 10-HDA could significantly decrease the biofilm viability and effectively eradicated the mature biofilms. It was also found that the hemolytic activity of S. aureus was significantly inhibited by 10-HDA. qRT-PCR analyses revealed that the expressions of global regulators sarA, agrA, and α-hemolysin gene hla were downregulated by 10-HDA. These results indicate that 10-HDA could be used as a potential natural antimicrobial agent to control the biofilm formation and virulence of S. aureus.

Research progress and the biotechnological applications of multienzyme complex.

The multienzyme complex system has become a research focus in synthetic biology due to its highly efficient overall catalytic ability and has been applied to various fields. Multienzyme complexes are formed by cascading complexes, which are multiple functionally related enzymes that continuously and efficiently catalyze the production of substrates. Compared with current mainstream microbial cell catalytic systems, in vitro multienzyme molecular machines have many advantages, such as fewer side reactions, a high product yield, a fast reaction speed, easy product separation, a tolerable toxic environment, and robust system operability, showing increasing competitiveness in the field of biomanufacturing. In this review, the research progress of multienzyme complexes in nature and multienzyme cascades in vivo or in vitro will be introduced, and the discovered enzyme cascades concerning scaffolding proteins will also be discussed. This review is expected to provide a more theoretical basis for the modification of multienzyme complexes and broaden their application in the field of synthetic biology. KEY POINTS: • The cascade reactions of some natural multienzyme complexes are reviewed. • The main approaches of constructing artificial multienzyme complexes are summarized. • The structure and application of cellulosomes are discussed and prospected.

Increasing L-lysine production in Corynebacterium glutamicum by engineering amino acid transporters.

Corynebacterium glutamicum has a long and successful history in the biotechnological production of L-lysine. Besides the adjustment of metabolic pathways, intracellular and extracellular transport systems are critical for the cellular metabolism of L-lysine or its by-products. Here, three amino acid transmembrane transporters, namely, GluE, BrnE/BrnF, and LysP, which are widely present in C. glutamicum strains, were each investigated by gene knockout. In comparison with that in the wild-type strain, the yield of L-lysine increased by 9.0%, 12.3%, and 10.0% after the deletion of the gluE, brnE/brnF, and lysP genes, respectively, in C. glutamicum 23,604. Moreover, the amount of by-product amino acids decreased significantly when the gluE and brnE/brnF genes were deleted. It was also demonstrated that there was no effect on the growth of the strain when the gluE or lysP gene was deleted, whereas the biomass of C. glutamicum WL1702 (ΔbrnE/ΔbrnF) in the fermentation medium was significantly reduced in comparison with that of the wild type. These results also provide useful information for enhancing the production of L-lysine or other amino acids by C. glutamicum.

Improving enzyme activity of glucosamine-6-phosphate synthase by semi-rational design strategy and computer analysis.

OBJECTIVE: To improve enzyme activity of Glucosamine-6-phosphate synthase (Glms) of Bacillus subtilis by site saturation mutagenesis at Leu593, Ala594, Lys595, Ser596 and Val597 based on computer-aided semi-rational design. RESULTS: The results indicated that L593S had the greatest effect on the activity of BsGlms and the enzyme activity increased from 5 to 48 U/mL. The mutation of L593S increased the yield of glucosamine by 1.6 times that of the original strain. The binding energy of the mutant with substrate was reduced from - 743.864 to - 768.246 kcal/mol. Molecular dynamics simulation results showed that Ser593 enhanced the flexibility of the protein, which ultimately led to increased enzyme activity. CONCLUSION: We successfully improved BsGlms activity through computer simulation and site saturation mutagenesis. This combination of methodologies may fit into an efficient workflow for improving Glms and other proteins activity.

Chinese character handwriting: A large-scale behavioral study and a database.

We report on a psycholinguistic database of Chinese character handwriting based on a large-scale study that involved 203 participants, each handwriting 200 characters randomly sampled from a cohort of 1,600 characters. Apart from collecting writing latencies, durations, and accuracy, we also compiled 14 lexical variables for each character. Regressions showed that frequency, age of acquisition, and the word context (in which a character appears) are all-around and influential predictors of orthographic access (as reflected in writing latency), motor execution of handwriting (as reflected in writing duration), and accuracy. In addition, phonological factors (phonogram status, spelling regularity, and homophone density) impacted orthographic access but not handwriting execution. Semantic factors (imageability and concreteness) only affected accuracy. These results suggest, among other things, that phonology is consulted in orthographic access while handwriting. As the first of its kind, this database can be used as a source of secondary data analyses and a tool for stimulus construction in handwriting research.

Perceptual Representations in L1, L2 and L3 Comprehension: Delayed Sentence-Picture Verification.

We used delayed sentence-picture verification tasks to investigate multilingual perceptual representations. In experiment 1, participants listened to sentences with implied shapes. After a 10-min interval, they judged whether pictures had been mentioned in the preceding sentences or not. Results in experiment 1 showed significant match effect in L1, but not in high proficient L2 or low proficient L3. In experiment 2, Participants listened to one language block, then immediately judged one picture block, totally three language-picture blocks. Results in experiment 2 were parallel to results in experiment 1. Our study supports the view of distributed conception: L2 and L3 are associated with less perceptual symbols than L1, indicating great impact of acquisition styles on perceptual representations. Our results show little impact of language proficiency levels on perceptual representations in delayed tasks.

Dissociating the neural substrates for inhibition and shifting in domain-general cognitive control.

Inhibition and shifting are two key components of domain-general cognitive control. Numerous studies have investigated the neural substrates of both components, but it is still unclear whether the relevant brain regions are specifically involved in one specific component or commonly engaged in both components. Here, we addressed this question by using functional magnetic resonance imaging and a modified saccade paradigm that was effective to disentangle inhibition and shifting in one experiment. The results showed that both the middle frontal gyrus and left parietal lobe were involved in both components but the middle frontal gyrus was more active for the inhibition while the inferior parietal lobe was more active for the shifting processing. The outcome suggests that, although both regions are engaged in inhibition and shifting, each plays a dominant role in one component. These findings provide a further insight into the neural dissociation in inhibition and shifting, as well as a better explanation on the framework of unity and diversity from a neuropsychological viewpoint.

The Roles of Relative Linguistic Proficiency and Modality Switching in Language Switch Cost: Evidence from Chinese Visual Unimodal and Bimodal Bilinguals.

The current study investigated the mechanism of language switching in unbalanced visual unimodal bilinguals as well as balanced and unbalanced bimodal bilinguals during a picture naming task. All three groups exhibited significant switch costs across two languages, with symmetrical switch cost in balanced bimodal bilinguals and asymmetrical switch cost in unbalanced unimodal bilinguals and bimodal bilinguals. Moreover, the relative proficiency of the two languages but not their absolute proficiency had an effect on language switch cost. For the bimodal bilinguals the language switch cost also arose from modality switching. These findings suggest that the language switch cost might originate from multiple sources from both outside (e.g., modality switching) and inside (e.g., the relative proficiency of the two languages) the linguistic lexicon.

Fully Conjugated Two-Dimensional sp2 -Carbon Covalent Organic Frameworks as Artificial Photosystem†I with High Efficiency.

The synthesis of fully conjugated sp2 -carbon covalent organic frameworks (COF) is extremely challenging given the difficulty of the formation of very stable carbon-carbon double bonds (-C=C-). Here, we report the successful preparation of a 2D COF (TP-COF) based on triazine as central planar units bridged by sp2 -carbon linkers through the -C=C- condensation reaction. High-resolution-transmission electron microscopy (HRTEM) clearly confirmed the tessellated hexagonal pore structure with a pore center-to-center distance of 2 nm. Powder X-ray diffraction (PXRD) together with structural simulations revealed an AA stacking mode of the obtained layered structure. TP-COF turned out to be an excellent semiconductor material with a LUMO energy of -3.23 eV and a band gap of 2.36 eV. Excitingly, this novel sp2 -carbon conjugated TP-COF exhibited unprecedented coenzyme regeneration efficiency and can significantly boost the coenzyme-assisted synthesis of l-glutamate to a record-breaking 97 % yield within 12 minutes.

Cross-dimensional magnitude interactions arise from memory interference.

Magnitudes from different dimensions (e.g., space and time) interact with each other in perception, but how these interactions occur remains unclear. In four experiments, we investigated whether cross-dimensional magnitude interactions arise from memory interference. In Experiment 1, participants perceived a constant-length line consisting of two line segments of complementary lengths and presented for a variable stimulus duration; then they received a cue about which of the two segment lengths to later reproduce. Participants were to first reproduce the stimulus duration and then the cued length. Reproduced durations increased as a function of the cued length if the cue was given before duration was retrieved from memory for reproduction (i.e. before duration reproduction; Experiment 1) but not if it was given after the duration memory had been retrieved from memory (i.e. after the start of duration reproduction; Experiment 2). These findings demonstrate that space-time interaction arises as a result of memory interference when length and duration information co-exist in working memory. Experiment 3 further demonstrated spatial interference on duration memories from memories of filled lengths (i.e. solid line segments) but not from noisier memories of unfilled lengths (demarcated empty spatial intervals), thus highlighting the role of memory noise in space-time interaction. Finally, Experiment 4 showed that time also exerted memory interference on space when space was presented as (relatively noisy) unfilled lengths. Taken together, these findings suggest that cross-dimensional magnitude interactions arise as a result of memory interference and the extent and direction of the interaction depend on the relative memory noises of the target and interfering dimensions. We propose a Bayesian model whereby the estimation of a magnitude is based on the integration of the noisily encoded percept of the target magnitude and the prior knowledge that magnitudes co-vary across dimensions (e.g., space and time). We discuss implications for cross-dimensional magnitude interactions in general.