-
Frontiers in Microbiology 2024The plant secondary metabolites (PSMs), as important plant resistance indicators, are important targets for screening plant insect resistance breeding. In this study, we...
BACKGROUND
The plant secondary metabolites (PSMs), as important plant resistance indicators, are important targets for screening plant insect resistance breeding. In this study, we aimed to investigate whether the population of (ZC) is affected by different varieties of PSMs content. At the same time, the structure and function of the gut microbiome of ZC were also analyzed in relation to different pecan varieties.
METHODS
We counted the populations of ZC larvae in four pecan varieties and determined the content of four types of PSMs. The structure and function of the larval gut microbiota were studied in connection to the number of larvae and the content of PSMs. The relationships were investigated between larval number, larval gut microbiota, and PSM content.
RESULTS
We found that the tannins, total phenolics, and total saponins of 4 various pecans PSMs stifled the development of the ZC larval population. The PSMs can significantly affect the diversity and abundance of the larval gut microbiota. Enrichment of ASV46 ( sp.), ASV994 ( sp.), ASV743 ( sp.), ASV709 ( sp.), ASV671 ( sp.), ASV599 ( sp.), ASV575 ( sp.), and ASV27 ( sp.) in the gut of larvae fed on high-resistance cultivars was positively associated with their tannin, total saponin, and total phenolic content. The results of the gut microbiome functional prediction for larvae fed highly resistant pecan varieties showed that the enriched pathways in the gut were related to the breakdown of hazardous chemicals.
CONCLUSION
Our findings provide further evidence that pecan PSMs influence the structure and function of the gut microbiota, which in turn affects the population stability of ZC. The study's findings can serve as a theoretical foundation for further work on selecting ZC-resistant cultivars and developing green management technology for ZC.
PubMed: 38680914
DOI: 10.3389/fmicb.2024.1379488 -
International Journal of Molecular... Apr 2024As global climate change continues, drought episodes have become increasingly frequent. Studying plant stress tolerance is urgently needed to ensure food security. The...
As global climate change continues, drought episodes have become increasingly frequent. Studying plant stress tolerance is urgently needed to ensure food security. The common ice plant is one of the model halophyte plants for plant stress biology research. This study aimed to investigate the functions of a newly discovered transcription factor, Homeobox 7 (HB7), from the ice plant in response to drought stress. An efficient -mediated transformation method was established in the ice plant, where ectopic expression may be sustained for four weeks. The overexpression (OE) plants displayed drought tolerance, and the activities of redox enzymes and chlorophyll content in the OE plants were higher than the wild type. Quantitative proteomics revealed 1910 and 495 proteins significantly changed in the OE leaves compared to the wild type under the control and drought conditions, respectively. Most increased proteins were involved in the tricarboxylic acid cycle, photosynthesis, glycolysis, pyruvate metabolism, and oxidative phosphorylation pathways. Some were found to participate in abscisic acid signaling or response. Furthermore, the abscisic acid levels increased in the OE compared with the wild type. HB7 was revealed to bind to the promoter motifs of Early Responsive to Dehydration genes and abscisic acid-responsive genes, and protein-protein interaction analysis revealed candidate proteins responsive to stresses and hormones (e.g., abscisic acid). To conclude, may contribute to enhance plant drought tolerance through abscisic acid signaling.
Topics: Abscisic Acid; Signal Transduction; Droughts; Gene Expression Regulation, Plant; Transcription Factors; Stress, Physiological; Plant Proteins; Plants, Genetically Modified; Proteomics; Homeodomain Proteins; Drought Resistance
PubMed: 38674154
DOI: 10.3390/ijms25084569 -
Life (Basel, Switzerland) Apr 2024Quorum sensing (QS) controls the virulence of This study aims to determine the anti-QS activity of aspirin alone and in combination with chitosan to reach maximum...
BACKGROUND
Quorum sensing (QS) controls the virulence of This study aims to determine the anti-QS activity of aspirin alone and in combination with chitosan to reach maximum inhibition. We tested ten virulent (. ) isolates and screened for N-acyl homoserine lactone (AHL) production using as a biosensor. isolates were treated with sub-minimum inhibitory concentrations (MICs) of aspirin and chitosan-aspirin. We used broth microdilution and checkerboard titration methods to determine the MICs and the synergistic effect of these two compounds, respectively. Real-time polymerase chain reaction (PCR) was used to estimate the anti-QS activity of the aspirin-chitosan combination on the expression of and genes.
RESULTS
Aspirin decreased the motility and production of AHLs, pyocyanin, and biofilm. Chitosan potentiated the inhibitory effect of aspirin. The chitosan-aspirin combination inhibited and gene expression in PAO1 (ATCC 15692) by 7.12- and 0.92-fold, respectively. In clinical isolates, the expression of and was decreased by 1.76 × 10- and 1.63 × 10-fold, respectively. Molecular docking analysis revealed that aspirin could fit into the active sites of the QS synthases and with a high binding affinity, causing conformational changes that resulted in their inhibition.
CONCLUSIONS
The chitosan-aspirin combination provides new insights into treating virulent and resistant
PubMed: 38672752
DOI: 10.3390/life14040481 -
Methods and Protocols Mar 2024Additional Affiliation(s): Updated Affiliation [...].
Additional Affiliation(s): Updated Affiliation [...].
PubMed: 38668143
DOI: 10.3390/mps7020029 -
Frontiers in Microbiology 2024It is essential to consider a practical antibody test to successfully implement marker vaccines and validate vaccination efficacy against classical swine fever virus...
BACKGROUND
It is essential to consider a practical antibody test to successfully implement marker vaccines and validate vaccination efficacy against classical swine fever virus (CSFV). The test should include a serological antibody assay, combined with a tool for differentiating infected from vaccinated animals (DIVA). The immunochromatographic test strip (ICS) has been exclusively designed for detecting CSFV E2 antibodies while lacking in detecting E antibodies, which can be employed and satisfy DIVA strategy. This study developed a novel ICS for detecting CSFV E2/E dual-antibody. The effectiveness of ICS in evaluating the DIVA capability of two novel chimeric pestivirus vaccine candidates was assessed.
METHODS
Recombinant E2 or E protein was transiently expressed in the plant using . ICS was subsequently assembled, and goat anti-rabbit IgG and recombinant CSFV E2 or E protein were plated onto the nitrocellulose membrane as control and test lines, respectively. The sensitivity and specificity of ICS were evaluated using sera with different neutralizing antibody titers or positive for antibodies against CSFV and other pestiviruses. The coincidence rates for detecting E2 and E antibodies between ICS and commercial enzyme-linked immunosorbent assay (ELISA) kits were also computed. ICS performance for DIVA capability was evaluated using sera from pigs vaccinated with conventional vaccine or chimeric vaccine candidates.
RESULTS
E2 and E proteins were successfully expressed in -produced recombinant proteins. ICS demonstrated high sensitivity in identifying CSFV E2 and E antibodies, even at the low neutralizing antibody titers. No cross-reactivity with antibodies from other pestiviruses was confirmed using ICS. There were high agreement rates of 93.0 and 96.5% between ICS and two commercial ELISA kits for E2 antibody testing. ICS also achieved strong coincidence rates of 92.9 and 89.3% with two ELISA kits for E antibody detection. ICS confirmed the absence of CSFV E-specific antibodies in sera from pigs vaccinated with chimeric vaccine candidates.
CONCLUSION
E2 and E proteins derived from the plant showed great potential and can be used to engineer a CSFV E2/E dual-antibody ICS. The ICS was also highly sensitive and specific for detecting CSFV E2 and E antibodies. Significantly, ICS can fulfill the DIVA concept by incorporating chimeric vaccine candidates.
PubMed: 38666258
DOI: 10.3389/fmicb.2024.1383976 -
STAR Protocols Jun 2024Cellular protein homeostasis is maintained by the disposal of aggregated misfolded proteins. Here, we present a protocol for investigating the involvement of the...
Cellular protein homeostasis is maintained by the disposal of aggregated misfolded proteins. Here, we present a protocol for investigating the involvement of the proteins of interest in misfolded protein degradation via Agrobacterium-mediated transient expression in Nicotiana benthamiana. We describe in detail the steps of misfolded protein design, transient protein expression in N. benthamiana, subsequent total protein extraction, and quantification of misfolded proteins through western blotting. This generalizable system can be used for misfolded proteins derived from various plants or microbes. For complete details on the use and execution of this protocol, please refer to Ai et al..
Topics: Nicotiana; Agrobacterium; Proteolysis; Protein Folding; Plant Proteins; Plants, Genetically Modified
PubMed: 38662545
DOI: 10.1016/j.xpro.2024.103034 -
Heliyon Apr 2024The study aimed to enhance quercetin production in radish by optimizing -mediated transformation. This protocol involved infecting radish seed embryo axis with EHA105...
The study aimed to enhance quercetin production in radish by optimizing -mediated transformation. This protocol involved infecting radish seed embryo axis with EHA105 strain carrying the . Radish seeds were infected with the suspension (0.8 OD) for 30 min, followed by sonication for 60 s and vacuum infiltration for 90 s at 100 mm Hg. A 3-day co-cultivation in Murashige and Skoog medium with 150 μM acetosyringone yielded a transformation efficiency of 59.6% and a transgenic callus induction rate of 32.3%. Transgenic plant and callus lines were confirmed by GUS histochemical assay, PCR, and qRT-PCR. The transgenic lines showed an increased expression of flavonoid pathway genes ( and ) and antioxidant genes ( and ) compared to WT plants. Overexpression of in transgenic callus increased enzyme activity of phenylalanine ammonia lyase, catalase, and ascorbate peroxidase. In half-strength MS medium with 116.8 mM sucrose, the highest growth index (7.63) was achieved after 20 days. In overexpressed callus lines, phenolic content (357.31 mg g dry weight), flavonoid content (463 mg g dry weight), and quercetin content (48.24 mg g dry weight) increased significantly by 9.41-fold. Micro-wounding, sonication, and vacuum infiltration improved transformation in radishes. These high-quercetin-content transgenic callus lines hold promise as valuable sources of flavonoids.
PubMed: 38660267
DOI: 10.1016/j.heliyon.2024.e27053 -
BMC Biotechnology Apr 2024Since the 2000's, plants have been used as bioreactors for the transient production of molecules of interest such as vaccines. To improve protein yield, "amplicon"...
BACKGROUND
Since the 2000's, plants have been used as bioreactors for the transient production of molecules of interest such as vaccines. To improve protein yield, "amplicon" vectors based on plant viruses are used. These viral constructs, engineered to carry the gene of interest replicate strongly once introduced into the plant cell, allowing significant accumulation of the protein. Here, we evaluated the suitability of the monocot-infecting RNA virus Rice yellow mottle virus (RYMV) as an amplicon vector. The promastigote surface antigen (PSA) of the protozoan Leishmania was considered as a protein of interest due to its vaccine properties against canine leishmaniasis.
RESULTS
Since P1 (ORF1) and CP (ORF3) proteins are not strictly necessary for viral replication, ORF1 was deleted and the PSA gene was substituted to ORF3 in the RYMV-based vector. We evaluated its expression in the best described plant bioreactor system, Nicotiana benthamiana which, unlike rice, allows transient transformation by Agrobacterium. Despite not being its natural host, we demonstrated a low level of RYMV-based vector replication in N. benthamiana leaves. Under optimized ratio, we showed that the P19 silencing suppressor in combination with the missing viral CP ORF significantly enhanced RYMV amplicon replication in N. benthamiana. Under these optimized CP/P19 conditions, we showed that the RYMV amplicon replicated autonomously in the infiltrated N. benthamiana cells, but was unable to move out of the infiltrated zones. Finally, we showed that when the RYMV amplicon was expressed under the optimized conditions we set up, it allowed enhanced PSA protein accumulation in N. benthamiana compared to the PSA coding sequence driven by the 35S promoter without amplicon background.
CONCLUSION
This work demonstrates that a non-dicot-infecting virus can be used as an amplicon vector for the efficient production of proteins of interest such as PSA in N. benthamiana leaves.
Topics: Nicotiana; Genetic Vectors; Plant Leaves; Animals; Dogs; Antigens, Protozoan; Bioreactors; Plants, Genetically Modified
PubMed: 38658899
DOI: 10.1186/s12896-024-00851-8 -
Sheng Wu Gong Cheng Xue Bao = Chinese... Apr 2024This study aimed to visualize the morphological features and dynamic changes of tomato mitochondria to provide a basis for the study of its mitochondrial functions. In...
This study aimed to visualize the morphological features and dynamic changes of tomato mitochondria to provide a basis for the study of its mitochondrial functions. In this study, transgenic tomatoes expressing mitochondria-localized green fluorescent protein (mitochondria-GFP, Mt-GFP) were obtained by -mediated genetic transformation. The color, hardness, soluble solids, acidity content, respiration rate, and ethylene production of the transgenic Mt-GFP tomato fruits were determined at the stage of mature green, breaker, and 3, 6, 9 days after breaker, while the wild-type tomato fruits were used as a control. As expected, Mt-GFP recombinant protein did not affect the ripening process, but induced the increased acidity of tomato fruits. The accumulations of Mt-GFP protein in tomato leaves and fruits were successfully verified by Western blotting. The morphological characteristics of mitochondria in flower, leaf and fruit cells as well as the dynamic changes of mitochondria in flower cells were clearly observed and studied under confocal laser microscope. The development of transgenic Mt-GFP tomato plants helps the visualization of tomato mitochondria and provides good research materials for the study of mitochondrial function during tomato development and fruit ripening.
Topics: Solanum lycopersicum; Green Fluorescent Proteins; Plants, Genetically Modified; Mitochondria; Mitochondrial Dynamics; Fruit
PubMed: 38658146
DOI: 10.13345/j.cjb.230488 -
Horticulture Research Apr 2024Grafting is a traditional and significant strategy to suppress soil-borne diseases, such as the crown gall disease caused by tumorigenic and . Root exudates and the...
Grafting is a traditional and significant strategy to suppress soil-borne diseases, such as the crown gall disease caused by tumorigenic and . Root exudates and the rhizosphere microbiome play critical roles in controlling crown gall disease, but their roles in suppressing crown gall disease in grafted plants remain unclear. Here, disease-susceptible cherry rootstock 'Gisela 6' and disease-resistant cherry rootstock 'Haiying 1' were grafted onto each other or self-grafted. The effect of their root exudates on the soil microbiome composition and the abundance of pathogenic were studied. Grafting onto the disease-resistant rootstock helped to reduce the abundance of pathogenic , accompanied by altering root exudation, enriching potential beneficial bacteria, and changing soil function. Then, the composition of the root exudates from grafted plants was analyzed and the potential compounds responsible for decreasing pathogenic abundance were identified. Based on quantitative measurement of the concentrations of the compounds and testing the impacts of supplied pure chemicals on abundance and chemotaxis of pathogenic and potential beneficial bacteria, the decreased valine in root exudates of the plant grafted onto resistant rootstock was found to contribute to decreasing abundance, enriching some potential beneficial bacteria and suppressing crown gall disease. This study provides insights into the mechanism whereby grafted plants suppress soil-borne disease.
PubMed: 38645683
DOI: 10.1093/hr/uhae049