Development of breeder chip for gene detection and molecular-assisted selection by target sequencing in wheat.

Wheat is an essential food crop and its high and stable yield is suffering from great challenges due to the limitations of current breeding technology and various stresses. Accelerating molecularly assisted stress-resistance breeding is critical. Through a meta-analysis of published loci in wheat over the last two decades, we selected 60 loci with main breeding objectives, high heritability, and reliable genotyping, such as stress resistance, yield, plant height, and resistance to spike germination. Then, using genotyping by target sequencing (GBTS) technology, we developed a liquid phase chip based on 101 functional or closely linked markers. The genotyping of 42 loci was confirmed in an extensive collection of Chinese wheat cultivars, indicating that the chip can be used in molecular-assisted selection (MAS) for target breeding goals. Besides, we can perform the preliminary parentage analysis with the genotype data. The most significant contribution of this work lies in translating a large number of molecular markers into a viable chip and providing reliable genotypes. Breeders can quickly screen germplasm resources, parental breeding materials, and intermediate materials for the presence of excellent allelic variants using the genotyping data by this chip, which is high throughput, convenient, reliable, and cost-efficient. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-023-01359-3.

Reduction of Rhizoctonia cerealis Infection on Wheat Through Host- and Spray-Induced Gene Silencing of an Orphan Secreted Gene.

Rhizoctonia cerealis is a soilborne fungus that can cause sharp eyespot in wheat, resulting in massive yield losses found in many countries. Due to the lack of resistant cultivars, fungicides have been widely used to control this pathogen. However, chemical control is not environmentally friendly and is costly. Meanwhile, the lack of genetic transformation tools has hindered the functional characterization of virulence genes. In this study, we attempted to characterize the function of virulence genes by two transient methods, host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS), which use RNA interference to suppress the pathogenic development. We identified ten secretory orphan genes from the genome. After silencing these ten genes, only the RcOSP1 knocked-down plant significantly inhibited the growth of R. cerealis. We then described RcOSP1 as an effector that could impair wheat biological processes and suppress pathogen-associated molecular pattern-triggered immunity in the infection process. These findings confirm that HIGS and SIGS can be practical tools for researching R. cerealis virulence genes. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Black carp IKKε collaborates with IRF3 in the antiviral signaling.

Interferon regulatory factor 3 (IRF3) is activated by IκB kinase ε (IKKε) and Tank-binding kinase 1 (TBK1), which plays a crucial role in the interferon signaling in vertebrates. However, the regulation of teleost IRF3 by IKKε remains largely unknown. In this study, the IRF3 homologue (bcIRF3) of black carp (Mylopharyngodon piceus) has been cloned and characterized. The transcription of bcIRF3 was detected to increase in host cells in response to different stimuli. bcIRF3 distributed predominantly in the cytosolic area; however, translocated into nuclei after virus infection. bcIRF3 showed IFN-inducing ability in reporter assay and EPC cells expressing bcIRF3 showed enhanced antiviral ability against both grass carp reovirus (GCRV) and spring viremia of carp virus (SVCV). Moreover, knockdown of bcIRF3 reduced the antiviral ability of the host cells, and the transcription of antiviral-related cytokines was obviously lower in bcIRF3-deficient host cells than that of control cells. The data of reporter assay and plaque assay demonstrated that bcIKKε obviously enhanced bcIRF3-mediated IFN production and antiviral activity. Immunofluorescent staining and co-immunoprecipitation assay revealed that bcIKKε interacted with bcIRF3. It was interesting that the nuclear translocation of bcIRF3 and bcIKKε was enhanced by each other when these two molecules were co-expressed in the cells, however, the protein levels of bcIRF3 and bcIKKε were decreased mutually. Thus, our data support the conclusion that bcIKKε interacts with bcIRF3 and enhances bcIRF3-mediated antiviral signaling during host innate immune activation.

Effects of high-intensity interval exercise on arterial stiffness in individuals at risk for cardiovascular disease: a meta-analysis.

Objective: The purpose of this meta-analysis was to investigate the effect of high-intensity interval training (HIIT) on arterial stiffness (AS) and vascular function in persons at high risk of cardiovascular disease (CVD). Methods: We conducted a comprehensive search of randomized controlled trials (RCTs) published in electronic databases (PubMed, Web of Science, Cochrane, Embase, and Ebsco) since their inception through October 2023 to evaluate the effect of HIIT on AS and vascular function in persons at high risk for CVD. The weighted mean difference (WMD) and 95% confidence intervals (95% CI) were calculated, and heterogeneity was assessed using the I2 test. Results: This study included 661 participants from 16 studies. HIIT significantly reduced pulse wave velocity (PWV) in persons at high risk for CVD [weighted mean difference (WMD), -0.62; 95% CI, -0.86--0.38; P < 0.00001]. Subgroup analysis showed that the PWV improvement effect was better when the HIIT program was performed 2-3 times per week and the duration was controlled within 40 min [2-3 times, -0.67; 95% CI, -0.93--0.41; P < 0.00001; time of duration, ≤40 min, -0.66; 95% CI, -0.91--0.41; P < 0.00001]. HIIT significantly reduced systolic blood pressure (SBP, -5.43; 95% CI, -8.82--2.04; P = 0.002), diastolic blood pressure (DPB, -2.96; 95% CI, -4.88--1.04; P = 0.002), and resting heart rate (RHR, -4.35; 95% CI, -7.04--1.66; P = 0.002), but had no significant effect on augmentation index (AIX, -2.14; 95% CI, -6.77-2.50; P = 0.37). Conclusion: HIIT can improve PWV in high-risk individuals with CVD and reduce SBP, DBP, and RHR, but has no significant effect on AIX. HIIT can effectively improve AS and vascular function and can be recommended as an effective method to improve AS in high-risk persons with CVD. Systematic Review Registration: https://www.crd.york.ac.uk/PROSPERO/, identifier: CRD42023471593.

Theoretical and Experimental Investigation on the Flexural Behaviour of Prestressed NC-UHPC Composite Beams.

To investigate the normal section strength and cracking bending moment of normal concrete-ultra-high-performance concrete (NC-UHPC) composite beams, calculation formulas were established considering the tensile strength of UHPC based on the current railway bridge design code. Using the railway T-beam as a template, prestressed NC-UHPC composite beams with different NC layer heights were built. A static bending test was performed, the pressure of the steel strand and the deflection and strain of the beam were measured, and the evolution of cracks in each beam was observed. The calculation formulas of the normal section strength and cracking bending moment of NC-UHPC composite beam were verified by the test. The results showed that the type of strain was similar to load-deflection curves with increasing load; the bending failure process of the NC-UHPC composite beam showed four obvious stages: elasticity, uniform cracking, crack development, and yield. Cracks in the beam started to appear at stage II, developed rapidly at stage III, and stopped emerging at stage IV. The calculation formulas for the normal section strength and the cracking bending moment of the NC-UHPC composite beam were in good agreement with the test values. Normal concrete with a compressive strength of 80 MPa can replace UHPC for the design of NC-UHPC composite beams.

Identification of Long Intergenic Noncoding RNAs in Rhizoctonia cerealis following Inoculation of Wheat.

Wheat sharp eyespot caused by Rhizoctonia cerealis is primarily a severe threat to worldwide wheat production. Currently, there are no resistant wheat cultivars, and the use of fungicides is the primary method for controlling this disease. Elucidating the mechanisms of R. cerealis pathogenicity can accelerate the pace of the control of this disease. Long intergenic noncoding RNAs (lincRNAs) that function in plant-pathogen interactions might provide a new perspective. We systematically analyzed lincRNAs and identified a total of 1,319 lincRNAs in R. cerealis. We found that lincRNAs are involved in various biological processes, as shown by differential expression analysis and weighted correlation network analysis (WGCNA). Next, one of nine hub lincRNAs in the blue module that was related to infection and growth processes, MSTRG.4380.1, was verified to reduce R. cerealis virulence on wheat by a host-induced gene silencing (HIGS) assay. Following that, RNA sequencing (RNA-Seq) analysis revealed that the significantly downregulated genes in the MSTRG.4380.1 knockdown lines were associated mainly with infection-related processes, including hydrolase, transmembrane transporter, and energy metabolism activities. Additionally, 23 novel microRNAs (miRNAs) were discovered during small RNA (sRNA) sequencing (sRNA-Seq) analysis of MSTRG.4380.1 knockdown, and target prediction of miRNAs suggested that MSTRG.4380.1 does not act as a competitive endogenous RNA (ceRNA). This study performed the first genome-wide identification of R. cerealis lincRNAs and miRNAs. It confirmed the involvement of a lincRNA in the infection process, providing new insights into the mechanism of R. cerealis infection and offering a new approach for protecting wheat from R. cerealis. IMPORTANCE Rhizoctonia cerealis, the primary causal agent of wheat sharp eyespot, has caused significant losses in worldwide wheat production. Since no resistant wheat cultivars exist, chemical control is the primary method. However, this approach is environmentally unfriendly and costly. RNA interference (RNAi)-mediated pathogenicity gene silencing has been proven to reduce the growth of Rhizoctonia and provides a new perspective for disease control. Recent studies have shown that lincRNAs are involved in various biological processes across species, such as biotic and abiotic stresses. Therefore, verifying the function of lincRNAs in R. cerealis is beneficial for understanding the infection mechanism. In this study, we reveal that lincRNAs could contribute to the virulence of R. cerealis, which provides new insights into controlling this pathogen.

CRISPR-Cas12a-Based Diagnostics of Wheat Fungal Diseases.

Fusarium head blight (FHB) of wheat, mainly caused by Fusarium graminearum (F. graminearum) infection, reduces crop yield and contaminates grain with mycotoxins. We report a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a-based nucleic acid assay for an early and rapid diagnosis of wheat FHB. Guide RNA (gRNA) was screened for highly specific recognition of polymerase chain reaction (PCR) amplicon of the internal transcribed spacer (ITS) region and the transcription elongation factor 1α (EF1α) of F. graminearum. The trans-activation of Cas12a protein cleaves the single-stranded DNA probes with the terminal fluorophore and quencher groups, thus allowing us to report the presence of ITS and EF1α of F. graminearum. Owing to the dual recognition process through PCR primers and gRNA hybridization, the approach realized specific discrimination of F. graminearum from other pathogenic fungi. It also allowed us to detect as low as 1 fg/µL total DNA from F. graminearum, which is sufficient to diagnose a 4 day F. graminearum infection. CRISPR-Cas12a-based nucleic acid assay promises the molecular diagnosis of crop diseases and broadens the application of CRISPR tools.

PKACα negatively regulates TAK1/IRF7 signaling in black carp Mylopharyngodon piceus.

Protein Kinase A catalytic subunit α (PKACα), plays an important role in the PKA and NF-κB signaling pathway in mammals. However, the function of PKACα in teleost fish remains largely unknown. In this study, PKACα from black carp (bcPKACα) has been cloned and its role in the innate immune antiviral signaling pathway was investigated. The open reading frame of bcPKACα gene contains 1056 nucleotides and the immunofluorescence assay verified that PKACα was mainly distributed in the cytoplasm. The reporter assay showed that bcPKACα expression and co-expression of bcPKACα and black carp TAK1 (bcTAK1) could activate the transcription of NF-κB. However, bcTAK1/bcIRF7-mediated IFN transcription was inhibited by bcPKACα. Knockdown of bcPKACα showed slightly enhanced antiviral activity against spring viremia of carp virus (SVCV) compared with control group. Accordingly, the antiviral activity against SVCV and grass carp reovirus (GCRV) of EPC cells co-expressing bcPKACα, bcTAK1 and bcIRF7 was obviously lower than that of EPC cells co-expressing bcTAK1 and bcIRF7. The similar subcellular distribution and interaction between bcPKACα and bcTAK1 were detected by immunofluorescent staining and co-immunoprecipitation assay separately. The data generated in this study demonstrates that bcPKACα associates with bcTAK1 and positively regulates NF-κB signaling, however, negatively regulates TAK1/IRF7 signaling pathway.

MoS2 nanotubes loaded with TiO2 nanoparticles for enhanced electrocatalytic hydrogen evolution.

Efficient and stable non-precious metal catalysts composed of earth-abundant elements are crucial to the hydrogen evolution reaction (HER) in high-energy conversion efficiency. Herein, TiO2/MoS2-NTs catalyst, in which the MoS2 nanotubes were loaded with TiO2 nanoparticles, have been synthesized via a facile solvothermal and hydrothermal method. The as-prepared TiO2/MoS2-NTs electrocatalyst demonstrated enhanced electrocatalytic hydrogen evolution performance compared with MoS2-NTs. Electrochemical measurements reveal the overpotential and Tafel slope of as-prepared TiO2/MoS2-NTs are -0.21 V and 42 mV dec-1. The HER improvement is proposed to be attributed to the increased edge sites results from the interfaces and synergic effect between TiO2 nanoparticles and MoS2 nanotubes.