Gene pyramiding of ZmGLK36 and ZmGDIα-hel for rough dwarf disease resistance in maize.

Maize rough dwarf disease (MRDD) caused by pathogenic viruses in the genus Fijivirus in the family Reoviridae is one of the most destructive diseases in maize. The pyramiding of effective resistance genes into maize varieties is a potential approach to reduce the damage resulting from the disease. Two major quantitative trait loci (QTLs) (qMrdd2 and qMrdd8) have been previously identified. The resistance genes ZmGLK36 and ZmGDIα-hel have also been cloned with the functional markers Indel-26 and IDP25K, respectively. In this study, ZmGLK36 and ZmGDIα-hel were introgressed to improve MRDD resistance of maize lines (Zheng58, Chang7-2, B73, Mo17, and their derived hybrids Zhengdan958 and B73 × Mo17) via marker-assisted selection (MAS). The converted lines and their derived hybrids, carrying one or two genes, were evaluated for MRDD resistance using artificial inoculation methods. The double-gene pyramiding lines and their derived hybrids exhibited increased resistance to MRDD compared to the monogenic lines and the respective hybrids. The genetic backgrounds of the converted lines were highly similar (90.85-98.58%) to the recurrent parents. In addition, agronomic trait evaluation demonstrated that pyramiding lines with one or two genes and their derived hybrids were not significantly different from the recurrent parents and their hybrids under nonpathogenic stress, including period traits (tasseling, pollen shedding, and silking), yield traits (ear length, grain weight per ear and 100-kernel weight) and quality traits (protein and starch content). There were differences in plant architecture traits between the improved lines and their hybrids. This study illustrated the successful development of gene pyramiding for improving MRDD resistance by advancing the breeding process. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-024-01466-9.

Genome-Wide Identification, Phylogeny and Expression Analysis of Subtilisin (SBT) Gene Family under Wheat Biotic and Abiotic Stress.

The subtilisin-like protease (SBT) family is widely known for its role in stress resistance to a number of stressors in different plant species, but is rarely studied in wheat. Subtilisin-like serine proteases (SBTs) are serine proteolytic enzymes that hydrolyze proteins into small peptides, which bind to receptors as signal molecules or ligands and participate in signal transduction. In this study, we identified 255 putative SBT genes from the wheat reference genome and then divided these into seven clades. Subsequently, we performed syntenic relation analysis, exon-intron organization, motif composition, and cis-element analysis. Further, expression analysis based on RNA-seq and tissue-specific expression patterns revealed that TaSBT gene family expression has multiple intrinsic functions during various abiotic and biotic stresses. Analysis of RNA-seq expression assays and further validation through qRT PCR suggested that some of the TaSBT genes have significant changes in expression levels during Pst interaction. TaSBT7, TaSBT26, TaSBT102, and TaSBT193 genes showed increasing expression levels during compatible and non-compatible interactions, while the expression levels of TaSBT111 and TaSBT213 showed a decreasing trend, indicating that these members of the wheat SBT gene family may have a role in wheat's defense against pathogens. In conclusion, these results expand our understanding of the SBT gene family, and provide a valuable reference for future research on the stress resistance function and comprehensive data of wheat SBT members.

KLF4 Affects Acute Renal Allograft Injury via Binding to MicroRNA-155-5p Promoter to Regulate ERRFI1.

Kruppel-like factor 4 (KLF4) owns the promising potential in treating kidney injury, which inevitably occurs during renal allograft. Given that, this research targets to unveil KLF4-oriented mechanism from microRNA-155-5p/ERBB receptor feedback inhibitor 1 (miR-155-5p/ERRFI1) axis in acute renal allograft injury. Mice were injected with miR-155-5p-related sequences before acute renal allograft modeling. Afterwards, serum inflammation, along with oxidative stress, renal tubular injury, and apoptosis in renal tissues were detected. HK-2 cells were processed by hypoxia/reoxygenation (H/R) and transfected with miR-155-5p- or ERRFI1-related sequences, after which cell proliferation and apoptosis were measured. KLF4, miR-155-5p, and ERRFI1 expressions and their interaction were tested. KLF4 and miR-155-5p levels were enhanced, and ERRFI1 level was repressed in mice after acute renal allograft and in H/R-treated HK-2 cells. KLF4 bound to the promoter of miR-155-5p. Depleting miR-155-5p reduced serum inflammation and attenuated oxidative stress, renal tubular injury, and apoptosis in mice with acute renal allograft injury. Downregulating miR-155-5p facilitated proliferation and repressed apoptosis of H/R-treated HK-2 cells. miR-155-5p targeted ERRFI1. Knocking down ERRFI1 antagonized the effects of downregulated miR-155-5p on acute renal allograft injury, as well as on H/R-treated HK-2 cell proliferation and apoptosis. A summary displays that silencing KLF4 suppresses miR-155-5p to attenuate acute renal allograft injury by upregulating ERRFI1, which provides a way to control acute renal allograft injury.

Alternaria Mycotoxins: An Overview of Toxicity, Metabolism, and Analysis in Food.

The genus Alternaria is widely distributed in the environment. Numerous species of the genus Alternaria can produce a variety of toxic secondary metabolites, called Alternaria mycotoxins. In this review, natural occurrence, toxicity, metabolism, and analytical methods are introduced. The contamination of these toxins in foodstuffs is ubiquitous, and most of these metabolites present genotoxic and cytotoxic effects. Moreover, Alternaria toxins are mainly hydroxylated to catechol metabolites and combined with sulfate and glucuronic acid in in vitro arrays. A more detailed summary of the metabolism of Alternaria toxins is presented in this work. To effectively detect and determine the mycotoxins in food, analytical methods with high sensitivity and good accuracy are also reviewed. This review will guide the formulation of maximum residue limit standards in the future, covering both toxicity and metabolic mechanism of Alternaria toxins.

Cytokinin dehydrogenase: a genetic target for yield improvement in wheat.

The plant hormone group, the cytokinins, is implicated in both qualitative and quantitative components of yield. Cytokinins have opposing actions in shoot and root growth-actions shown to involve cytokinin dehydrogenase (CKX), the enzyme that inactivates cytokinin. We revise and provide unambiguous names for the CKX gene family members in wheat, based on the most recently released wheat genome database, IWGSC RefSeq v1.0 & v2.0. We review expression data of CKX gene family members in wheat, revealing tissue-specific gene family member expression as well as sub-genome-specific expression. Manipulation of CKX in cereals shows clear impacts on yield, root growth and orientation, and Zn nutrition, but this also emphasizes the necessity to unlink promotive effects on grain yield from negative effects of cytokinin on root growth and uptake of mineral nutrients, particularly Zn and Fe. Wheat is the most widely grown cereal crop globally, yet is under-research compared with rice and maize. We highlight gaps in our knowledge of the involvement of CKX for wheat. We also highlight the necessity for accurate analysis of endogenous cytokinins, acknowledging why this is challenging, and provide examples where inadequate analyses of endogenous cytokinins have led to unjustified conclusions. We acknowledge that the allohexaploid nature of bread wheat poses challenges in terms of uncovering useful mutations. However, we predict TILLING followed by whole-exome sequencing will uncover informative mutations and we indicate the potential for stacking mutations within the three genomes to modify yield components. We model a wheat ideotype based on CKX manipulation.

Identification and expression of genes associated with the abscission layer controlling seed shattering in Lolium perenne.

Perennial ryegrass (Lolium perenne) is one of the most important pasture grasses in the world. However, seed production is negatively impacted by the seed shattering (shedding) nature of this species. Recently, genes involved in the seed shattering process have been isolated and functionally characterized in several crop species. The aim of this study was to identify the genes playing critical roles in the seed shattering process in perennial ryegrass. DNA sequences of genes involved in seed shattering in the Poaceae were used to identify and isolate target genes in perennial ryegrass using a comparative genomics strategy. The candidate seed shattering genes were identified using an 'in-house' perennial ryegrass transcriptome database. The relative expression levels of the candidate ryegrass shattering genes were determined using RT-qPCR during different floret and seed developmental stages. Histological analysis of the abscission layer was also conducted. Homologues of seed shattering genes were identified and isolated from perennial ryegrass, and the relative gene expression results suggested that several genes, including LpqSH1 and LpSH1, might have a role in abscission layer formation during seed development. In addition, lignification of the abscission layer may play an important role in the abscission process. A genetic model for seed shattering in perennial ryegrass is suggested and may be useful for directing gene editing towards the production of a reduced-shattering ryegrass.

A mouse model of interstitial pneumonitis induced by murine cytomegalovirus infection after allogeneic skin transplantation.

We investigated the effect of murine cytomegalovirus (MCMV) on interstitial pneumonia in transplant recipients in an experimental skin allograft model. Skin transplantation between C57BL/6J and BALB/c mice was performed in the presence or absence of cyclosporin A treatment. Flow cytometry showed that the number of CD4(+) and CD8(+) cells and the level of IFN- γ decreased significantly in the groups treated with cyclosporin A. We either mock-infected or infected the mice with MCMV by intranasal administration and monitored pathophysiological behavior and body weight. The infected mice were sacrificed at different days postinfection for histology, immunohistochemistry, and molecular biological evaluations. Interstitial pneumonitis was observed in positive control groups as well as in experimental group that received cyclosporin A, a skin transplant, and infected with the highest dose of virus (10(5) PFU). Transmission electronic microscopy demonstrated the presence of herpes virus particles. MCMV DNA and glycoprotein B were demonstrated in the epithelial cells of the lung tissue in those animals by in situ hybridization and immunohistochemistry, respectively. Our data demonstrated the establishment of a mouse model of interstitial pneumonitis via MCMV infection after allogeneic skin transplantation.

The role of interleukin-17 in murine cytomegalovirus interstitial pneumonia in mice with skin transplants.

We hypothesized that the T helper (Th)17 response plays an important role in murine cytomegalovirus (MCMV) interstitial pneumonia. BALB/c mice with skin grafts from C57/BJ6 mice were intranasally inoculated with 1.0 × 10(5) PFU MCMV. Lung tissues and skin grafts were histologically evaluated and expression of interleukins (IL)-17, -6 and -8, monocyte chemotactic protein (MCP)-1 and interferon (IFN)-γ in serum and bronchoalveolar lavage (BAL) fluid, intracellular IL-4, -17, and IFN-γ, in spleen lymphocytes were analysed. The levels of IL-17 in the serum and BAL fluid were significantly higher in MCMV-infected mice versus not-infected mice (P = 0.0286 and P = 0.007, respectively) and the BAL levels of IL-17 peaked in 9 days (P = 0.001). The IL-17 level in the BAL was correlated with the grade of lung interstitial inflammation (r = 0.554, P = 0.0144). Serum IFN-γ levels were also higher after infection than that in the not-infected mice (P = 0.0286). IL-17 production increases locally and systemically during MCMV interstitial pneumonia. Neutralization of IL-17 significantly suppressed lung inflammation at day14 as assessed by histology. These findings suggest that IL-17 is important in the pathology of MCMV interstitial pneumonia.

Ultraviolet irradiation induced oxidative stress and response of antioxidant system in an intertidal macroalgae Corallina officinalis L.

The response of the antioxidant defense system of an intertidal macroalgae Corallina officinalis L. to different dosages of UV-B irradiation was investigated. Results showed that superoxide dimutase (SOD) and peroxidase (POX) increased and then maintained at a relatively stable level when subjected to UV-B irradiation. Catalase (CAT) activity under medium dosage of UV-B irradiation (Muv) and high dosage of UV-B irradiation (Huv) treatments were significantly decreased. Ascorbate peroxidase (APX) activity first remained unaltered and then increased in Huv treatment. In addition, the assay on isozymes was carried out using non-denaturing polyacrylamide gel electrophoresis (PAGE). The activities of some SOD isoforms were altered by UV-B. Two new bands (POX V and POX VII) appeared upon exposure to all three UV-B dosages. CAT III activity was increased by low dosage of UV-B irradiation (Luv), whereas CAT III and CAT IV disappeared when the alga was exposed to Muv and Huv. Two bands of APX (APX VI and APX VII) were increased and a new band (APX X) was observed under Huv exposure. H2O2 and thiobarbituric acid reacting substance (TBARS) increased under Muv and Huv treatments. Overall, UV-B protection mechanisms are partly inducible and to a certain extent sufficient to prevent the accumulation of damage in C. officinalis.

Preparation and in vitro, in vivo evaluation of clarithromycin microcapsules.

PURPOSE: To develop and validate a method to prepare clarithromycin (CLM) microcapsules to mask the bitter taste and provide effective treatment, and evaluate the quality of microcapsules in detail, especially the in vitro and in vivo pharmacokinetics behavior. METHODS: CLM microcapsules were prepared using ethyl cellulose as matrix material by an emulsion solvent diffusion method. The physicochemical property, in vitro release study, sensory test and stability test were evaluated. Self-made CLM dry suspension or conventional tablets containing 250 mg of CLM were orally administered with 250 mL of water. The plasma concentration was determined and the pharmacokinetic parameters were calculated by non-compartmental methods. RESULTS: Stable microcapsules could be prepared using ethyl cellulose as matrix material. The quality evaluation of prepared microcapsules was qualified, and the pharmacokinetic parameters of dry suspensions and conventional tablets were as following. Cmax were 1.32±0.62 and 1.40±0.58 µg.ml(-1); Tmax were 3.51±0.54 and 2.01±0.42 h; AUC were 7.65±2.54 and 7.12±2.10 µg.h.ml(-1). CONCLUSION: The preparation method is easy and applicable. The self-made CLM dry suspension containing microcapsules sufficiently alleviate the bitterness of commercial CLM dry suspension, but not decrease the bioavailability and have better effect for delaying drug release in healthy volunteers.

Identification of betacyanin and effects of environmental factors on its accumulation in halophyte Suaeda salsa.

Effects of temperature, light, salinity and developmental phases on the accumulation of red pigments in the C(3) halophyte Suaeda salsa were studied, and the physical and chemical characteristics of the red pigments were also analyzed. The results indicate that: these red pigments are insoluble in organic solvents but free in water, the pigments are red-violet and stable under acidic condition while yellow and unstable under alkaline condition, and they absorb the highest value at wavelength near 538 nm, light suppresses their accumulation and enhances their decomposition. These results suggest that these red pigments are betacyanins. Darkness, low temperatures and high salinity enhance betacyanin accumulation in Suaeda salsa, and darkness in the germination phase is one of the most important environmental factors for the betacyanin accumulation.