Role of BraRGL1 in regulation of Brassica rapa bolting and flowering

Gibberellin(GA)plays a major role in controlling Brassica rapa stalk development.As an essential negative regulator of GA signal transduction,DELLA proteins may exert significant effects on stalk development.However,the regulatory mechanisms underlying this regulation remain unclear.In this study,we report highly efficient and inheritable mutagenesis using the CRISPR/Cas9 gene editing system in BraPDS(phytoene desaturase)and BraRGL1(key DELLA protein)genes.We observed a loss-of-function mutation in BraRGL1 due to two amino acids in GRAS domain.The flower bud differentiation and bolting time of BraRGL1 mutants were significantly advanced.The expression of GA-regulatory protein(BraGASA6),flowering related genes(BraSOC1,BraLFY),expansion protein(BraEXPA11)and xyloglucan endotransferase(BraXTH3)genes was also significantly upregulated in these mutants.BraRGL1-overexpressing plants displayed the contrasting phenotypes.BraRGL1 mutants were more sensitive to GA signaling.BraRGL1 interacted with BraSOC1,and the interaction intensity decreased after GA3 treatment.In addition,BraRGL1 inhibited the transcription-activation ability of BraSOC1 for BraXTH3 and BraLFY genes,but the presence of GA3 enhanced the activation ability of BraSOC1,suggesting that the BraRGL1-BraSOC1 module regulates bolting and flowering of B.rapa through GA signal transduction.Thus,we hypothesized that BraRGL1 is degraded,and BraSOC1 is released in the presence of GA3,which promotes the expression of BraXTH3 and BraLFY,thereby inducing stalk development in B.rapa.Further,the BraRGL1-M mutant promoted the flower bud differentiation without affecting the stalk quality.Thus,BraRGL1 can serve as a valuable target for the molecular breeding of early maturing varieties.

The Systemic Immune Inflammatory Index Predicts No-Reflow Phenomenon after Primary Percutaneous Coronary Intervention in Older Patients with STEMI

Purpose:Coronary no-reflow phenomenon(NRP),a common adverse complication in patients with ST-segment eleva-tion myocardial infarction(STEMI)treated by percutaneous coronary intervention(PCI),is associated with poor patient prognosis.In this study,the correlation between the systemic immune-inflammation index(SII)and NRP in older patients with STEMI was studied,to provide a basis for early identification of high-risk patients and improve their prognosis.Materials and methods:Between January 2017 and June 2020,578 older patients with acute STEMI admitted to the Department of Cardiology of Hebei General Hospital for direct PCI treatment were selected for this retrospective study.Patients were divided into an NRP group and normal-flow group according to whether NRP occurred during the operation.Clinical data and the examination indexes of the two groups were collected.Logistic regression was used to analyze the independent predictors of NRP,and the receiver operating characteristic curve was used to further analyze the ability of SII to predict NRP in older patients with STEMI.Results:Multivariate logistic analysis indicated that hypertension(OR=2.048,95%CI:1.252–3.352,P=0.004),lymphocyte count(OR=0.571,95%CI:0.368–0.885,P=0.012),platelet count(OR=1.009,95%CI:1.005–1.013,P<0.001),hemoglobin(OR=1.015,95%CI:1.003–1.028,P=0.018),multivessel disease(OR=2.237,95%CI:1.407–3.558,P=0.001),and SII≥1814(OR=3.799,95%CI:2.190–6.593,P<0.001)were independent predictors of NRP after primary PCI in older patients with STEMI.Receiver operating characteristic curve analysis demonstrated that SII had a high predictive value for NRP(AUC=0.738;95%CI:0.686–0.790),with the best cut-off value of 1814,a sensitivity of 52.85%and a specificity of 85.71%.Conclusion:For older patients with STEMI undergoing primary PCI,SII is a valid predictor of NRP.

Nanofabrication of 50 nm zone plates through e-beam lithography with local proximity effect correction for x-ray imaging

High resolution Fresnel zone plates for nanoscale three-dimensional imaging of materials by both soft and hard x-rays are increasingly needed by the broad applications in nanoscience and nanotechnology.When the outmost zone-width is shrinking down to 50 nm or even below,patterning the zone plates with high aspect ratio by electron beam lithography still remains a challenge because of the proximity effect.The uneven charge distribution in the exposed resist is still frequently observed even after standard proximity effect correction(PEC),because of the large variety in the line width.This work develops a new strategy,nicknamed as local proximity effect correction(LPEC),efficiently modifying the deposited energy over the whole zone plate on the top of proximity effect correction.By this way,50 nm zone plates with the aspect ratio from 4:1 up to 15:1 and the duty cycle close to 0.5 have been fabricated.Their imaging capability in soft(1.3 keV)and hard(9 keV)x-ray,respectively,has been demonstrated in Shanghai Synchrotron Radiation Facility(SSRF)with the resolution of 50 nm.The local proximity effect correction developed in this work should also be generally significant for the generation of zone plates with high resolutions beyond 50 nm.

Identification and Characterization of a LEA-like Gene, CaMF5,Specifically Expressed in the Anthers of Male-fertile Capsicum annuum

To improve the understanding of molecular mechanisms of anther and/or pollen development in Chili pepper, in the present study, fulllength cDNA and DNA sequences of the pollen development-related gene CaMF5 were obtained from the anthers of a Capsicum annuum nuclear male-fertile line. Sequence analysis indicated that the full length of CaMF5 was 747 bp, containing a maximum opening reading frame of 447 bp.Amino acid sequence alignment and phylogenetic analysis revealed that CaMF5 shared approximately 37%–77% homology with a series of uncharacterized or hypothetical proteins and late embryogenesis abundant(LEA) proteins from other plants. However, no LEA structural domain was detected in CaMF5, which indicated that it might be a new type of LEA gene. CaMF5 was only expressed in flower buds at stages 7 and 8 and in open flowers of the male-fertile line, whereas it exhibited no expression in any examined organs of the male-sterile line. In addition, CaMF5 showed the highest transcript abundance in the anthers of the male-fertile line, with no expression being detected in any other examined organs, such as the sepals, petals, pistils, roots, stems, or leaves. Taken together, our results suggest that CaMF5 is an anther-specific gene that might encode a new type of LEA protein related to anther and/or pollen development in C. annuum.

Claroideoglomus etunicatum improved the growth and saline-alkaline tolerance of Potentilla anserina by altering physiological and biochemical properties

To investigate the effects of arbuscular mycorrhizal(AM)fungi on the growth and saline–alkaline tolerance of Potentilla anserina L.,the seedlings were inoculated with Claroideoglomus etunicatum(W.N.Becker&Gerd.)C.Walker&A.Schüßler in pot cultivation.After 90 days of culture,saline–alkaline stress was induced with NaCl and NaHCO_(3)solution according to the main salt components in saline–alkaline soils.Based on the physiological response of P.anserina to the stress in the preliminary experiment,the solution concentrations of 0 mmol/L,75 mmol/L,150 mmol/L,225 mmol/L and 300 mmol/L were treated with stress for 10 days,respectively.The mycorrhizal colonization rate,mycorrhizal dependence,chlorophyll content,malondialdehyde content,antioxidant enzyme activities,osmoregulation substances content and water status were measured.The results showed that with the increase of NaCl and NaHCO_(3)stress concentration,mycorrhizal colonization rate,colonization intensity,arbuscular abundance and vesicle abundance decreased,and reached the lowest value at 300 mmol/L.Strong mycorrhizal dependence was observed after the symbiosis with AM fungus,and the dependence was higher under NaHCO_(3)treatment.Under NaCl and NaHCO_(3)stress,inoculation with AM fungus could increase chlorophyll content,decrease malondialdehyde content,increase activities of superoxide dismutase,peroxidase and catalase,increase contents of proline,soluble sugar and soluble protein,increase tissue relative water content and decrease water saturation deficit.It was concluded that salt–alkali stress inhibited the colonization of AM fungus,but the mycorrhiza still played a positive role in maintaining the normal growth of plants under salt–alkali stress.

Synchrotron-Based Data-Constrained Modeling Analysis of Microscopic Mineral Distributions in Limestone

Three dimensional (3D) microscopic distributions of dolomite and calcite in a limestone sample have been analyzed with a data-constrained modeling (DCM) technique using synchrotron radiation-based multi-energy X-ray computed tomography (CT) data as constraints. In order to optimize the experimental parameters, X-ray CT simulations and DCM analysis of a numerical phantom consisting of calcite (CaCO3) and dolomite (CaMg(CO3)2) have been used to investigate the effects on the predicted results in relation to noise, X-ray energy and sample-to-detector distance (SDD). The simulation results indicate that the optimal X-ray energies are 25 and 35 keVs, and the SDD is 10 mm. The high resolution 3D distributions of mineral phases of a natural limestone have been obtained. The results are useful for quantitative understanding of mineral, porosity, and physical property distributions in relation to oil and gas reservoirs hosted in carbonate rocks, which account for more than half of the world’s conventional hydrocarbon resources. The case studied is also instructive for the applicability of the DCM methods for other types of composite materials with modest atomic number contrasts between the mineral phases.

Acute and Sub-Acute Toxicological Evaluations of Bioactive Alkaloidal Extract from Melodinus henryi and Their Main Chemical Constituents

Melodinus henryi is a good source of terpenoid indole alkaloids,and traditionally used as a folk medicine in the treatment of meningitis and fracture.In order to further exploit their potential uses,its anti-inflammatory and immunosuppressive activi-ties,safety evaluations and chemical profiles have been illustrated.Compared to the crude methanol extract from M.henryi and its non-alkaloidal fraction,the total alkaloidal fraction(MHTA)had the strongest anti-inflammatory and immunosup-pressive activities.In the acute oral toxicity assay,the half lethal dose(LD50)of MHTA was more than 2000 mg/kg.The sub-acute toxicity assay for consecutive 28 days exhibited MHTA at a lower concentrations of less than 500 mg/kg might be regarded as safe,and might damage spleen,liver,kidney,and heart when the dose is higher than 1000 mg/kg.In addition,a phytochemical investigation on MHTA led to the isolation of 15 monoterpenoid indole alkaloids.Thus,in regard with the potent side effects of MHTA,it should be used with caution in the development of phytomedicine.

Isolation and species diversity of arbuscular mycorrhizal fungi in the rhizosphere of Puccinellia tenuiflora of Songnen saline-alkaline grassland,China

Salinization has led to the deterioration of the ecological environment,affected the growth of plants,and hindered the development of agriculture and forestry.Arbuscular mycorrhizal(AM)fungi,as important soil microorganisms,play significant physiological and ecological roles in promoting plant nutrient absorption and improving soil structure.Puccinellia tenuiflora(Turcz.)Scribn.et Merr.in Songnen saline-alkaline grassland was selected as the research object to observe AM fungal colonization of the roots and explore the species and diversity of AM fungi in symbiotic association with P.tenuiflora.This study showed that AM fungi colonized in P.tenuiflora roots and formed a typical Arum-type mycorrhizal structure.A significant correlation was observed between vesicular abundance and the colonization intensity of mycorrhiza.Isolation and identification revealed 40 species of AM fungi in the rhizosphere of P.tenuiflora,belonging to 14 genera,of which two species could not be identified.The richness of the genus Glomus was the highest,accounting for 30%of the total species.Funneliformis mosseae and Rhizophagus intraradices were isolated from all the samples and were the species with the widest distribution in the rhizosphere of P.tenuiflora.Correlation analysis showed that pH only had a significant impact on the distribution of a few species,such as Glomus pustulatum,Diversispora spurca,Glomus aggregatum,Rhizophagus clarum,and Acaulospora foveata.The present study provides a theoretical basis for further exploring the resources of AM fungi in saline-alkaline soil.

Roles and Molecular Mechanisms of Biomarkers in Hepatocellular Carcinoma with Microvascular Invasion: A Review

Hepatocellular carcinoma(HCC)being a leading cause of cancer-related death,has high associated mortality and recurrence rates.It has been of great necessity and urgency to find effective HCC diagnosis and treatment measures.Studies have shown that microvascular invasion(MVI)is an independent risk factor for poor prognosis after hepatectomy.The abnormal expression of biomacromolecules such as circ-RNAs,lncRNAs,STIP1,and PD-L1 in HCC patients is strongly correlated with MVI.Deregulation of several markers mentioned in this review affects the proliferation,invasion,metastasis,EMT,and anti-apoptotic processes of HCC cells through multiple complex mechanisms.Therefore,these biomarkers may have an important clinical role and serve as promising interventional targets for HCC.In this review,we provide a comprehensive overview on the functions and regulatory mechanisms of MVI-related biomarkers in HCC.