Plant Cells Research Articles

Starch granules in algal cells play an inherent role to shape the popular SSC signal in flow cytometry

ObjectiveFlow cytometry (FC) is probably the most important technique for single-cell analysis. It’s precisely, rapid, and suitable for multidimensional single-cell analysis. The commonly used side scatter (SSC) intensity determined by FC is often interpreted as a measure of the internal cellular complexity of cells. In simple terms, the more structured a cell is, the higher the SSC intensity quantified by FC. Nevertheless, most of the studies that support this interpretation are based on data derived from animal or human cell lines and while it is assumed that the results can also be transferred to plant or algal cell lines, the details remain unclear. The objective of the recent work is to clarify the interpretation of the SSC signal from algal cells.ResultsAlgal lipid droplets and their starch play an inherent role to shape the popular SSC signal derived from FC. This was shown by a theoretical approach based on Lorenz–Mie theory. These results were supported by experiments with different model cultures of Chlamydomonas reinhardtii in which a high linear correlation was observed between the SSC signal and the ‘physical’ starch quantity.

Leadless pacemaker - lead parameters stability and battery longevity in the mid-term follow-up

Abstract Background A leadless pacemaker (L-PM) has been designed to avoid the need for a pacemaker pocket and transvenous lead. Long-term safety will largely depend on the need for replacements, mainly due to battery depletion (end-of-life). While the mean L-PM battery longevity predicted by the manufacturer is 12 years, real-world longevity data are still scarce. We aimed to describe L-PM parameters’ stability and battery longevity during the mid-term follow-up. Methods Single-center registry of consecutive patients undergoing L-PM implantation from May 2015 to November 2023. Procedural characteristics, lead parameters and battery longevity were collected immediately after implantation and during the most recent follow-up visit. Results Overall, 261 consecutive patients had a transcatheter L-PM implanted through a femoral vein (mean age 78±10 years; 66% male). Mean procedural duration was 44±28min and fluoroscopy time was 4,4±3,9min. The device was successfully implanted in all but one patient. The main indications for permanent pacing were high-degree atrioventricular block [N=130 (50%)], atrial fibrillation with pauses [N=78 (30%)], and sinus node disease [N=28 (11%)]. Immediately after implantation, three cases (1,2%) of pericardial effusion (one with tamponade) were reported. In one case (0,4%), a conventional pacemaker was implanted due to L-PM acute dysfunction. No other procedure-related complications, including dislodgment, were reported at discharge. After a mean follow-up of 2,8±1,7years, the pacing threshold and R-wave amplitude remained stable [0,67mV vs. 0,62mV (p=0,9); and 10,3mV vs. 16,1mV (p<0,001), respectively] and mean ventricular pacing was 54±38%. Expected battery longevity was >8 years (maximum value) in 84% of the patients. In the subgroup of patients with >5 years of follow-up (N=20; mean follow up of 6,3±1,8years), the expected battery longevity was >8 years in 85% (N=17) of the patients. In three cases, the L-PM was upgraded to cardiac resynchronization therapy or left bundle branch area pacing due to pacing-induced left ventricular dysfunction. There was one case of L-PM end-of-life 6 years after implant (pacing threshold 0,5mV; ventricular pacing 100%), in which a second L-PM was successfully implanted. Overall, 78 (31%) patients died, a mean of 2,2±1,9 years after L-PM implantation, none related to the device. Conclusion In this real-world cohort, L-PM maintained stable pacing parameters and, in the mid-term follow-up, only one battery depletion was observed while expected longevity remained above 8 years in the large majority of patients. Longer follow up will allow better understanding of both technical and clinical aspects relevant to manage end-of-life devices.Table 1Figure 1

Combined Transcriptome and Metabolome Analysis Reveals That Carbon Catabolite Repression Governs Growth and Pathogenicity in Verticillium dahliae

Carbon catabolite repression (CCR) is a common transcriptional regulatory mechanism that microorganisms use to efficiently utilize carbon nutrients, which is critical for the fitness of microorganisms and for pathogenic species to cause infection. Here, we characterized two CCR genes, VdCreA and VdCreC, in Verticillium dahliae that cause cotton Verticillium wilt disease. The VdCreA and VdCreC knockout mutants displayed slow growth with decreased conidiation and microsclerotium production and reduced virulence to cotton, suggesting that VdCreA and VdCreC are involved in growth and pathogenicity in V. dahliae. We further generated 36 highly reliable and stable ΔVdCreA and ΔVdCreC libraries to comprehensively explore the dynamic expression of genes and metabolites when grown under different carbon sources and CCR conditions. Based on the weighted gene co-expression network analysis (WGCNA) and correlation networks, VdCreA is co-expressed with a multitude of downregulated genes. These gene networks span multiple functional pathways, among which seven genes, including PYCR (pyrroline-5-carboxylate reductase), are potential target genes of VdCreA. Different carbon source conditions triggered entirely distinct gene regulatory networks, yet they exhibited similar changes in metabolic pathways. Six genes, including 6-phosphogluconolactonase and 2-ODGH (2-oxoglutarate dehydrogenase E1), may serve as hub genes in this process. Both VdCreA and VdCreC could comprehensively influence the expression of plant cell wall-degrading enzyme (PCWDE) genes, suggesting that they have a role in pathogenicity in V. dahliae. The integrated expression profiles of the genes and metabolites involved in the glycolysis/gluconeogenesis and pentose phosphate pathways showed that the two major sugar metabolism-related pathways were completely changed, and GADP (glyceraldehyde-3-phosphate) may be a pivotal factor for CCR under different carbon sources. All these results provide a more comprehensive perspective for further analyzing the role of Cre in CCR.

Virulence perspective genomic research unlocks the secrets of Rhizoctonia solani associated with banded sheath blight in Barnyard Millet (Echinochloa frumentacea)

IntroductionBanded sheath blight (Bsb) disease, caused by Rhizoctonia solani, is an emerging problem in barnyard millet cultivation. One of the significant goals of pathogenomic research is to identify genes responsible for pathogenicity in the fungus.MethodsA virulence profiling-based approach was employed and six R. solani isolates were collected from various ecological zones of India. The morphological parameters and virulence of all of the six R. solani isolates were investigated. The most virulent strain was designated as RAP2 and its genome has been sequenced, assembled, and annotated.ResultsThe RAP2 genome is 43.63 megabases in size and comprises 10.95% repetitive DNA, within which 46% are retroelements, 8% are DNA transposons, and 46% are unidentified DNA. The Gene Ontology (GO) annotation of RAP2 proteins revealed that “phosphorylation”, “membrane”, and “ATP binding” have the highest gene enrichment in the “biological process”, “cellular component” and “molecular function” domains, respectively. The genome comprises a majority of secretory proteins in the pectin lyase fold/virulence factor superfamily, which break down plant cell wall polymers to extract saccharides. The RAP2 genome is comparable to R. solani, which infects maize and rice, but it diverges further from soybean in terms of nucleotide-level genetic similarity. Orthologous clustering of RAP2 protein sequences with R. solani infecting maize, rice, and soybean yields 5606 proteins shared across all genomes. GO analysis of 25 proteins specific to the RAP2 genome found enrichment in the ethylene response, which can cause spore germination and infection in host plants.DiscussionInterestingly, a 28-bp deletion in the RAP2 strain’s cutinase domain was discovered in the cutinase protein, which might be important in the infection process, perhaps rendering the enzyme inactive or allowing the pathogen to infect barnyard millet while avoiding host defense. This study sheds light on the genetic makeup of R. solani, allowing researchers to discover critical genes related with pathogenicity as well as potential targets for fungicide development.

Potassium extrusion by plant cells: evolution from an emergency valve to a driver of long-distance transport.

The ability to accumulate nutrients is a hallmark for living creatures and plants evolved highly effective nutrient transport systems, especially for the uptake of potassium (K+). However, plants also developed mechanisms that enable the rapid extrusion of K+ in combination with anions. The combined release of K+ and anions is probably an ancient extrusion system, as it is found in the Characeae that are closely related to land plants. We postulate that the ion extrusion mechanisms have developed as an emergency valve, which enabled plant cells to rapidly reduce their turgor, and prevent them from bursting. Later in evolution, seed plants adapted this system for various responses, such as the closure of stomata, long-distance stress waves, dropping of leaves by pulvini, and loading of xylem vessels. We discuss the molecular nature of the transport proteins that are involved in ion extrusion-based functions of plants and describe the functions that they obtained during evolution.

The Potential Role of PeMAP65-18 in Secondary Cell Wall Formation in Moso Bamboo

Microtubule-associated proteins (MAPs) play a pivotal role in the assembly and stabilization of microtubules, which are essential for plant cell growth, development, and morphogenesis. A class of plant-specific MAPs, MAP65, plays largely unexplored roles in moso bamboo (Phyllostachys edulis). This study identified 19 PeMAP65 genes in moso bamboo, systematically examining their phylogenetic relationships, conserved motifs, gene structures, collinearity, and cis-acting elements. Analysis of gene expression indicated that PeMAP65s exhibit tissue-specific expression patterns. Functional differentiation was investigated among the members of different PeMAP65 subfamilies according to their expression patterns in different development stages of bamboo shoots. The expression of PeMAP65-18 was positively correlated with the expression of genes involved in secondary cell wall (SCW) biosynthesis. Y1H and Dual-LUC assays demonstrated that the transcription of PeMAP65-18 was upregulated by PeMYB46, a key transcription factor of SCW biosynthesis. The result of subcellular localization showed that PeMAP65-18 was located in cortical microtubules. We speculate that PeMAP65-18 may play a crucial role in the SCW deposition of moso bamboo. This comprehensive analysis of the MAP65 family offers novel insights into the roles of PeMAP65s in moso bamboo, particularly in relation to the formation of SCWs.

Basic design of artificial membrane-less organelles using condensation-prone proteins in plant cells

Membrane-less organelles, formed by the condensation of biomolecules, play a pivotal role in eukaryotes. Artificial membrane-less organelles and condensates are effective tools for the creation of new cellular functions. However, it is poorly understood how to control the properties that affect condensate function, particularly in plants. Here, we report the construction of model artificial condensates using the condensation-prone proteins OsJAZ2 and AtFCA in a transient assay using rice (Oryza sativa) cells, and how condensate properties, such as subcellular localization, protein mobility, and size can be altered. We showed that proteins of interest can be recruited to condensates using nanobodies or chemically induced dimerization. Furthermore, by combining two types of condensation-prone proteins, we demonstrated that artificial hybrid condensates with heterogeneous material properties could be constructed. Finally, we showed that modified artificial condensates can be constructed in transgenic Arabidopsis thaliana plants. These results provide a framework for the basic design of synthetic membrane-less organelles in plants.

Molecular insights into Solanum sisymbriifolium’s resistance against Globodera pallida via RNA-seq

BackgroundThe presence of potato cyst nematodes (PCN) causes a significant risk to potato crops globally, leading to reduced yields and economic losses. While the plant Solanum sisymbriifolium is known for its resistance to PCN and can be used as a trap crop, the molecular mechanisms behind this resistance remain poorly understood. In this study, genes differentially expressed were identified in control and infected plants during the early stages of the S. sisymbriifolium - G. pallida interaction.ResultsGene expression profiles were characterized for two S. sisymbriifolium cultivars, Melody and Sis6001, uninfected and infected by G. pallida. The comparative transcriptome analysis revealed a total of 4,087 and 2,043 differentially expressed genes (DEGs) in response to nematode infection in the cultivars Melody and Sis6001, respectively. Gene ontology (GO) enrichment analysis provided insights into the response of the plant to nematode infection, indicating an activation of the plant metabolism, oxidative stress leading to defence mechanism activation, and modification of the plant cell wall. Genes associated with the jasmonic and salicylic acid pathways were also found to be differentially expressed, suggesting their involvement in the plant’s defence response. In addition, the analysis of NBS-LRR domain-containing transcripts that play an important role in hypersensitive response and programmed cell death led to the identification of ten transcripts that had no annotations from the databases, with emphasis on TRINITY_DN52667_C1_G1, found to be upregulated in both cultivars.ConclusionsThese findings represent an important step towards understanding the molecular basis underlying plant resistance to nematodes and facilitating the development of more effective control strategies against PCN.

CRISPR/Cas system-mediated base editing in crops: recent developments and future prospects.

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (CRISPR/Cas9) genome-editing system has altered plant research by allowing for targeted genome alteration, and they are emerging as powerful tools for evaluating plant gene function and improving crop yield. Even though CRISPR/Cas9 cleavage and subsequent repair are effective ways to precisely replace genes and change base pairs in plants, the dominance of the non-homologous end-joining pathway (NHEJ) and homology-directed repair's (HDR) poor effectiveness in plant cells have restricted their use. Base editing is gaining popularity as a potential alternative to HDR or NHEJ-mediated replacement, allowing for precise changes in the plant genome via programmed conversion of a single base to another without the need for a donor repair template or double-stranded breaks. In this review, we primarily present the mechanisms of base-editing system, including their distinct types such as DNA base editors (cytidine base editor and adenine base editor) and RNA base editors discovered so far. Next, we outline the current potential applications of the base-editing system for crop improvements. Finally, we discuss the limitations and potential future directions of the base-editing system in terms of improving crop quality. We hope that this review will enable the researcher to gain knowledge about base-editing tools and their potential applications in crop improvement.

A CC-Type Glutaredoxins GRX480 Functions in Cadmium Tolerance by Maintaining Redox Homeostasis in Arabidopsis

Cadmium (Cd) toxicity causes oxidative stress damage in plant cells. Glutaredoxins (GRXs), a type of small oxidoreductase, play a crucial role in modulating thiol redox states. However, whether GRXs act in Cd stress remains to be identified. Here, we reveal that Arabidopsis GRX480, a member of the CC-type family, enhances plant Cd stress tolerance. The GRX480 mutants exhibit enhanced sensitivity to Cd stress, manifested by shortened root, reduced biomass, lower chlorophyll and proline levels, and decreased photosynthetic efficiency compared with the wild type. The Cd concentration in GRX480 mutants is higher than the wild type, resulting from the inhibition of Cd efflux and transport genes transcription. Lower levels of GSH were detected in Cd-treated GRX480 mutants than in the wild type, indicating that GRX480 regulates plant Cd tolerance by influencing the balance between GSH and GSSG. Furthermore, the hyperaccumulation of reactive oxygen species (ROS) is associated with decreased expression of H2O2 scavenging genes in Cd-treated GRX480 mutants. Additionally, more toxic reactive carbonyl species (RCS), produced during oxidative stress, accumulate in Cd-treated GRX480 mutants than in wild type. Overall, our study establishes a critical role of GRX480 in response to Cd stress, highlighting its multifaceted contributions to detoxification and the maintenance of redox homeostasis.

The secreted protein FonCHRD is essential for vegetative growth, asexual reproduction, and pathogenicity in watermelon Fusarium wilt fungus

Fungal pathogens often secrete numerous effectors to interfere with and/or suppress plant immunity to promote their infection. Watermelon Fusarium wilt, caused by the soil-borne fungus Fusarium oxysporum f. sp. niveum (Fon), is one of the devastating diseases that severely affect the watermelon industry. Here, we report the function of a candidate effector protein, FonCHRD, in Fon. FonCHRD harbors a chordin (CHRD) domain of unknown function and has a signal peptide with secretion activity. FonCHRD shows a relatively high expression level in Fon marcoconidia and is inducible by watermelon root tissues. Phenotypic analysis of the targeted deletion mutant revealed that FonCHRD plays roles in vegetative growth, asexual reproduction, and conidial morphology of Fon, while it is not involved in spore germination as well as cell wall, oxidative and salt stress responses. Deletion of FonCHRD impaired the ability to colonize and spread within host plants, significantly reducing its virulence on watermelon. FonCHRD is distributed across multiple compartments of plant cells but can target to the apoplast space in plants. FonCHRD inhibits the INF1- and Bcl2-associated X protein-triggered cell death and defense gene expression in transiently expressed Nicotiana benthamiana leaves. These findings suggest that FonCHRD is essential for Fon pathogenicity by modulating invasive growth and spreading abilities as well as by suppressing plant immune responses.

Integration of fungal transcriptomics and metabolomics provides insights into the early interaction between the ORM fungus Tulasnella sp. and the orchid Serapias vomeracea seeds

In nature, germination of orchid seeds and early plant development rely on a symbiotic association with orchid mycorrhizal (ORM) fungi. These fungi provide the host with the necessary nutrients and facilitate the transition from embryos to protocorms. Despite recent advances in omics technologies, our understanding of this symbiosis remains limited, particularly during the initial stages of the interaction. To address this gap, we employed transcriptomics and metabolomics to investigate the early responses occurring in the mycorrhizal fungus Tulasnella sp. isolate SV6 when co-cultivated with orchid seeds of Serapias vomeracea. The integration of data from gene expression and metabolite profiling revealed the activation of some fungal signalling pathways before the establishment of the symbiosis. Prior to seed contact, an indole-related metabolite was produced by the fungus, and significant changes in the fungal lipid profile occurred throughout the symbiotic process. Additionally, the expression of plant cell wall-degrading enzymes (PCWDEs) was observed during the pre-symbiotic stage, as the fungus approached the seeds, along with changes in amino acid metabolism. Thus, the dual-omics approach employed in this study yielded novel insights into the symbiotic relationship between orchids and ORM fungi and suggest that the ORM fungus responds to the presence of the orchid seeds prior to contact.

Pengembangan Multimedia Pembelajaran Berbasis Web Pada Mata Pelajaran IPA Kelas VIII SMP

The lack of utilization of learning media is one of the factors in students' low understanding of the material discussed and students feel bored during the learning process. Science subjects are one of the subjects that have a high level of understanding, one of the materials is animal cells and plant cells, therefore multimedia learning can help visualize this concept so that it is easier for students to understand. This research aims to find out how the process of developing multimedia learning and find out the results of validity tests and practicality tests. The development used is Research & Development (R&D). with a development model, namely a 4D model consisting of define, design, develop and disseminate. The results of the data analysis showed that the media validity with validator 1 got an average result of 4.93 in the "very valid" category and the result with validator 2 got an average of 4.80 in the "very valid" category. The acquisition of material validity by material validators obtained an average result of 4.80 with the category of "very valid". This learning multimedia is practical based on a practicality test with an average of 4.52. Based on the results of the practicality test, it was concluded that multimedia learning in science subjects in grade VIII junior high school can be used in the teaching and learning process.

Highest Occurring Vascular Plants from Ladakh Provide Wood Anatomical Evidence for a Thermal Limitation of Cell Wall Lignification.

As an evolutionary achievement of almost all terrestrial plants, lignin biosynthesis is essential for various mechanical and physiological processes. Possible effects of plant cell wall lignification on large-scale vegetation distribution are, however, not yet fully understood. Here, we present double-stained, wood anatomical stem measurements of 207 perennial herbs (Potentilla pamirica Wolf), which were collected between 5550 and 5850 m asl on the north-western Tibetan Plateau in Ladakh, India. We also measured changes in situ root zone and surface air temperatures along the sampling gradient and applied piecewise structural equation models to assess direct and indirect relationships between the age and size of plants, the degree of cell wall lignification in their stems, and the elevation at which they were growing. Based on the world's highest-occurring vascular plants, the Pamir Cinquefoils, we demonstrate that the amount of lignin in the secondary cell walls decreases significantly with increasing elevation (r = -0.73; p < 0.01). Since elevation is a proxy for temperature, our findings suggest a thermal constrain on lignin biosynthesis at the cold range limit of woody plant growth.

The mechanism of arsenic behaviour in the soil-plant system and its interaction with biogenic macroelements of plants under conditions of toxic stress

ABSTRACT The mobility of arsenic compounds in the soil-plant system and its interaction with biogenic macroelements K, P, Ca, Mg, Si, S, Na in plants under conditions of toxic stress have been studied. This is illustrated by the wheatgrass Elytrigia repens. It has been established that the root system is responsible for protecting the aerial part of wheatgrass from excess arsenic, and it is of great importance for phytostabilisation of arsenic pollution therefore indicating the increased tolerance of this plant. The intensity of As accumulation in plants depends on its speciation in the soil. Antagonism of the main essential elements (K, P) to the accumulation of As in wheatgrass shoots is observed. It can lead to disruption of metabolic processes in plant cells. It has been shown that changes in the K+/Na + values may lead to violation of osmotic processes in the aerial plant organs and contribute to their degradation not only under salt stress but also under the influence of higher concentrations of As in the plants. In areas intensively contaminated with arsenic, a decrease in the K/Na value in the above-ground organs of wheatgrass can be used as a criterion for the degree of phytotoxicity.

Drought Tolerance in Plants: Physiological and Molecular Responses

Drought, a significant environmental challenge, presents a substantial risk to worldwide agriculture and the security of food supplies. In response, plants can perceive stimuli from their environment and activate defense pathways via various modulating networks to cope with stress. Drought tolerance, a multifaceted attribute, can be dissected into distinct contributing mechanisms and factors. Osmotic stress, dehydration stress, dysfunction of plasma and endosome membranes, loss of cellular turgidity, inhibition of metabolite synthesis, cellular energy depletion, impaired chloroplast function, and oxidative stress are among the most critical consequences of drought on plant cells. Understanding the intricate interplay of these physiological and molecular responses provides insights into the adaptive strategies plants employ to navigate through drought stress. Plant cells express various mechanisms to withstand and reverse the cellular effects of drought stress. These mechanisms include osmotic adjustment to preserve cellular turgor, synthesis of protective proteins like dehydrins, and triggering antioxidant systems to counterbalance oxidative stress. A better understanding of drought tolerance is crucial for devising specific methods to improve crop resilience and promote sustainable agricultural practices in environments with limited water resources. This review explores the physiological and molecular responses employed by plants to address the challenges of drought stress.

Chemical inhibition of stomatal differentiation by perturbation of the master-regulatory bHLH heterodimer via an ACT-Like domain

Selective perturbation of protein interactions with chemical compounds enables dissection and control of developmental processes. Differentiation of stomata, cellular valves vital for plant growth and survival, is specified by the basic-helix-loop-helix (bHLH) heterodimers. Harnessing a new amination reaction, we here report a synthesis, derivatization, target identification, and mode of action of an atypical doubly-sulfonylated imidazolone, Stomidazolone, which triggers stomatal stem cell arrest. Our forward chemical genetics followed by biophysical analyses elucidates that Stomidazolone directly binds to the C-terminal ACT-Like (ACTL) domain of MUTE, a master regulator of stomatal differentiation, and perturbs its heterodimerization with a partner bHLH, SCREAM in vitro and in plant cells. On the other hand, Stomidazolone analogs that are biologically inactive do not bind to MUTE or disrupt the SCREAM-MUTE heterodimers. Guided by structural docking modeling, we rationally design MUTE with reduced Stomidazolone binding. These engineered MUTE proteins are fully functional and confer Stomidazolone resistance in vivo. Our study identifies doubly-sulfonylated imidazolone as a direct inhibitor of the stomatal master regulator, further expanding the chemical space for perturbing bHLH-ACTL proteins to manipulate plant development.

Gene co-expression analysis reveals conserved and distinct gene networks between resistant and susceptible Lens ervoides challenged by hemibiotrophic and necrotrophic pathogens

As field crops are likely to be challenged by multiple pathogens during their development, the investigation of broad-spectrum resistance in the host is of great interest for crop genetic enhancement. In this study, we attempted to address this question by adopting a weighed gene co-expression approach to study the temporal transcriptome dynamics of resistant and susceptible recombinant inbred lines (RILs) derived from an intraspecific Len ervoides cross during the infection process with either the necrotrophic pathogens Ascochyta lentis or Stemphylium botryosum, or the hemibiotrophic pathogen Colletotrichum lentis. By comparing networks of resistant and susceptible RILs, seven network module pairs were found to possess high correlation coefficients (R > 0.70) and large number of overlapping genes (n > 100). The conserved co-regulation of genes in these network module pairs were involved in plant cell wall synthesis, cell division, cytoskeleton organization, and protein ubiquitin related processes and appeared to be common disease responses against these pathogens. On the other hand, we also identified eight modules with low correlation between resistance and susceptibility networks. Among those, a stronger gene co-expression in R-genes and small RNA processes in the resistant hosts may be enhancing L. ervoides resistance against A. lentis, C. lentis, and S. botryosum, whereas the higher level of synergistic regulation in the synthesis of arginine and glutamine and phospholipid and glycerophospholipids in the susceptible hosts may contribute to increased susceptibility in L. ervoides.

Visualizing tomato spotted wilt virus protein localization: Cross-kingdom comparisons of protein-protein interactions.

Tomato spotted wilt virus (TSWV) is an orthotospovirus that infects both plants and insect vectors. Understanding the protein localization and interactions is crucial for unraveling the infection cycle and host-virus interactions. We investigated and compared the localization of TSWV proteins. A change in localization over time was associated with the viral proteins that did not contain signal peptides and transmembrane domains such as N, NSs and NSm, however, this only occurred in the plant cells, not in the insect cells. The localization between plants and insects otherwise was consistent indicating a similar mechanism is utilized by the virus in both types of cells. We also tested the localization of the proteins during an active plant infection using free RFP as a marker to highlight the nucleus and cytoplasm. Voids in the cytoplasm were shown only during infection and N, NSs, NSm and to lesser extent, GN and GC, were surrounding these areas suggesting it may be a site of replication or morphogenesis. Furthermore, we tested the interactions of viral proteins using both bimolecular fluorescence complementation (BiFC) and membrane-based yeast two-hybrid (MbY2H) assays. These revealed self-interactions of NSm, N, GN, GC, and NSs. We also identified interactions between different TSWV proteins, indicating their roles and host interactions, such as between NSs and GC and N and GC which may be necessary during the replication and assembly processes respectively. This research expands our knowledge of TSWV infection and elaborates on the intricate relationships between viral proteins, cellular dynamics, and host responses.

Volatilization and cleaning recovery of Pb, Se and Hg in acid sludge of copper smelting system during vacuum distillation

The high lead acid sludge, containing a small amount of selenium and mercury, is a hazardous waste product of the copper smelting system. Improper disposal of this waste can cause serious harm to the environment. In this study, the method of vacuum distillation was proposed for treating the high lead acid sludge. The study aimed to investigate the volatilization rule of selenium, mercury, and lead in the acid sludge during vacuum distillation, as well as to purify PbSe and metallic lead. The results indicated that under 5 Pa, Hg and Se commence volatilizing at 300 ℃, the content of Hg in volatile substances remains relatively constant after reaching 400 ℃, demonstrating nearly complete volatilization at this temperature with a ratio of 92.93 %. Similarly, selenium almost completely volatilizes at 800 ℃ with a ratio of 96.74 %, while lead does so at around 1000 ℃ with a ratio of approximately 99.27 %. Mercury at 400 ℃ can be recovered as mercury selenide. Lead selenide can be achieved at 800 ℃, while lead sulfate can be decomposed and purified at 1000 ℃ to obtain metallic lead with a purity higher than 99 %. The method can properly treat and recover mercury, selenium, lead and other elements in the acid sludge with short process and no pollution, which provides a new cleaning approach for high lead acid sludge.