Identification of genes with differential transcript abundance(GDTA)in seedless mutants may enhance understanding of seedless citrus development.Transcriptome analysis was conducted at three time points during early f...Identification of genes with differential transcript abundance(GDTA)in seedless mutants may enhance understanding of seedless citrus development.Transcriptome analysis was conducted at three time points during early fruit development(Phase 1)of three seedy citrus genotypes:Fallglo(Bower citrus hybrid(Citrus reticulata×C.reticulata×C.paradisi)×Temple(C.reticulata×C.sinensis)),grapefruit(C.paradisi),Pineapple sweet orange(C.sinensis),and their seedless mutants.Seed abortion in seedless mutants was observed at 26 days post anthesis(Time point 2).Affymetrix transcriptomic analysis revealed 359 to 1077 probe sets with differential transcript abundance in the comparison of seedless versus seedy fruits for each citrus genotypes and time points.The GDTA identified by 18 microarray probe sets were validated by qPCR.Hierarchical clustering analysis revealed a range of GDTA associated with development,hormone and protein metabolism,all of which may reflect genes associated with seedless fruit development.There were 14,9 and 12 genes found exhibiting similar abundance ratios in all three seedless versus seedy genotype comparisons at time point 1,2 and 3,respectively.Among those genes were genes coding for an aspartic protease and a cysteine protease,which may play important roles in seedless fruit development.New insights into seedless citrus fruit development may contribute to biotech approaches to create seedless cultivars.展开更多
SUMO PROTEASE RELATED TO FERTILITY 1(SPF1)and SPF2 are responsible for deSUMOylation of SUMO-conjugated protein substrates and for maintaining protein SUMOylation homeostasis.To date,the role of SUMO proteases in fatt...SUMO PROTEASE RELATED TO FERTILITY 1(SPF1)and SPF2 are responsible for deSUMOylation of SUMO-conjugated protein substrates and for maintaining protein SUMOylation homeostasis.To date,the role of SUMO proteases in fatty acid biosynthesis and lipid accumulation remains unclear.Here,we demonstrate that the Arabidopsis thaliana mutants spf1-1,spf2-1,and spf1-1 spf2-1 exhibit increased seed size and elevated seed oil content(SOC).We further show that SPF1 and SPF2 interact with WRINKLED1(WRI1),a master regulator of the transcriptional control of plant oil synthesis.Genetic analyses indicate that the spf1-1 wri1-3 and spf2-1 wri1-3 double mutants,as well as the spf1-1 spf2-1 wri1-3 triple mutant,phenocopy wri1-3 and display severe seed shriveling,indicating that SPF1 and SPF2 act upstream of WRI1.WRI1 was identified as a SUMO1 substrate with two conserved SUMOylation sites,lysine 257(K257)and K266,in cruciferous plants,with K257 acting as the dominant site required for seed oil synthesis.SUMOylation enhances WRI1 stability,whereas SPF1and SPF2mediated deSUMOylation promotes WRI1 degradation.In spf1-1,spf2-1,and spf1-1 spf2-1 mutants,the abundance of SUMOylated WRI1 increases during seed development and correlates with elevated seed oil accumulation.Together,these results indicate that SPF1 and SPF2 negatively regulate oil synthesis by deSUMOylating WRI1,establishing a dynamic SUMOylation and deSUMOylation switch centered on the SPF1/SPF2-WRI1 module that fine-tunes seed development and oil synthesis.展开更多
Background:We previously reported that activation of the cell cycle in human-induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs)enhances their remuscularization capacity after human cardiac muscle patch tr...Background:We previously reported that activation of the cell cycle in human-induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs)enhances their remuscularization capacity after human cardiac muscle patch transplantation in infarcted mouse hearts.Herein,we sought to identify the effect of magnesium lithospermate B(MLB)on hiPSC-CMs during myocardial repair using a myocardial infarction(MI)mouse model.Methods:In C57BL/6 mice,MI was surgically induced by ligating the left anterior descending coronary artery.The mice were randomly divided into five groups(n=10 per group);a MI group(treated with phosphate-buffered saline only),a hiPSC-CMs group,a MLB group,a hiPSC-CMs+MLB group,and a Sham operation group.Cardiac function and MLB therapeutic efficacy were evaluated by echocardiography and histochemical staining 4 weeks after surgery.To identify the associated mechanism,nuclear factor(NF)-κB p65 and intercellular cell adhesion molecule-1(ICAM1)signals,cell adhesion ability,generation of reactive oxygen species,and rates of apoptosis were detected in human umbilical vein endothelial cells(HUVECs)and hiPSC-CMs.Results:After 4 weeks of transplantation,the number of cells that engrafted in the hiPSC-CMs+MLB group was about five times higher than those in the hiPSC-CMs group.Additionally,MLB treatment significantly reduced tohoku hospital pediatrics-1(THP-1)cell adhesion,ICAM1 expression,NF-κB nuclear translocation,reactive oxygen species production,NF-κB p65 phosphorylation,and cell apoptosis in HUVECs cultured under hypoxia.Similarly,treatment with MLB significantly inhibited the apoptosis of hiPSC-CMs via enhancing signal transducer and activator of transcription 3(STAT3)phosphorylation and B-cell lymphoma-2(BCL2)expression,promoting STAT3 nuclear translocation,and downregulating BCL2-Associated X,dual specificity phosphatase 2(DUSP2),and cleaved-caspase-3 expression under hypoxia.Furthermore,MLB significantly suppressed the production of malondialdehyde and lactate dehydrogenase and the reduction in glutathione content induced by hypoxia in both HUVECs and hiPSC-CMs in vitro.Conclusions:MLB significantly enhanced the potential of hiPSC-CMs in repairing injured myocardium by improving endothelial cell function via the NF-κB/ICAM1 pathway and inhibiting hiPSC-CMs apoptosis via the DUSP2/STAT3 pathway.展开更多
基金This project was conducted using USDA/ARS base funding.
文摘Identification of genes with differential transcript abundance(GDTA)in seedless mutants may enhance understanding of seedless citrus development.Transcriptome analysis was conducted at three time points during early fruit development(Phase 1)of three seedy citrus genotypes:Fallglo(Bower citrus hybrid(Citrus reticulata×C.reticulata×C.paradisi)×Temple(C.reticulata×C.sinensis)),grapefruit(C.paradisi),Pineapple sweet orange(C.sinensis),and their seedless mutants.Seed abortion in seedless mutants was observed at 26 days post anthesis(Time point 2).Affymetrix transcriptomic analysis revealed 359 to 1077 probe sets with differential transcript abundance in the comparison of seedless versus seedy fruits for each citrus genotypes and time points.The GDTA identified by 18 microarray probe sets were validated by qPCR.Hierarchical clustering analysis revealed a range of GDTA associated with development,hormone and protein metabolism,all of which may reflect genes associated with seedless fruit development.There were 14,9 and 12 genes found exhibiting similar abundance ratios in all three seedless versus seedy genotype comparisons at time point 1,2 and 3,respectively.Among those genes were genes coding for an aspartic protease and a cysteine protease,which may play important roles in seedless fruit development.New insights into seedless citrus fruit development may contribute to biotech approaches to create seedless cultivars.
基金supported by grants from the National Key Research and Development Plan of China(2021YFF1000103)the National Natural Science Foundation of China(32470339 and 32072095)+1 种基金the Ministry of Education(MOE)of Singapore Tier 2 Program(MOE-T2EP30123-0001 to W.M.)the Asian Institute of Modern Agronomy(AIMA)R&D Funding Program(AIMA-2024-WP2 to W.M.)。
文摘SUMO PROTEASE RELATED TO FERTILITY 1(SPF1)and SPF2 are responsible for deSUMOylation of SUMO-conjugated protein substrates and for maintaining protein SUMOylation homeostasis.To date,the role of SUMO proteases in fatty acid biosynthesis and lipid accumulation remains unclear.Here,we demonstrate that the Arabidopsis thaliana mutants spf1-1,spf2-1,and spf1-1 spf2-1 exhibit increased seed size and elevated seed oil content(SOC).We further show that SPF1 and SPF2 interact with WRINKLED1(WRI1),a master regulator of the transcriptional control of plant oil synthesis.Genetic analyses indicate that the spf1-1 wri1-3 and spf2-1 wri1-3 double mutants,as well as the spf1-1 spf2-1 wri1-3 triple mutant,phenocopy wri1-3 and display severe seed shriveling,indicating that SPF1 and SPF2 act upstream of WRI1.WRI1 was identified as a SUMO1 substrate with two conserved SUMOylation sites,lysine 257(K257)and K266,in cruciferous plants,with K257 acting as the dominant site required for seed oil synthesis.SUMOylation enhances WRI1 stability,whereas SPF1and SPF2mediated deSUMOylation promotes WRI1 degradation.In spf1-1,spf2-1,and spf1-1 spf2-1 mutants,the abundance of SUMOylated WRI1 increases during seed development and correlates with elevated seed oil accumulation.Together,these results indicate that SPF1 and SPF2 negatively regulate oil synthesis by deSUMOylating WRI1,establishing a dynamic SUMOylation and deSUMOylation switch centered on the SPF1/SPF2-WRI1 module that fine-tunes seed development and oil synthesis.
基金supported by the Natural Science Foundation of Hunan Province(Nos.2023JJ30793,2022JJ20088,and 2019JJ50858)the Science and Technology Innovation Program of Hunan Province(No.2021RC2106)+1 种基金the National Natural Science Foundation of China(No.82200323)the Scientific Research Launch Project for new employees of the Second Xiangya Hospital of Central South University.
文摘Background:We previously reported that activation of the cell cycle in human-induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs)enhances their remuscularization capacity after human cardiac muscle patch transplantation in infarcted mouse hearts.Herein,we sought to identify the effect of magnesium lithospermate B(MLB)on hiPSC-CMs during myocardial repair using a myocardial infarction(MI)mouse model.Methods:In C57BL/6 mice,MI was surgically induced by ligating the left anterior descending coronary artery.The mice were randomly divided into five groups(n=10 per group);a MI group(treated with phosphate-buffered saline only),a hiPSC-CMs group,a MLB group,a hiPSC-CMs+MLB group,and a Sham operation group.Cardiac function and MLB therapeutic efficacy were evaluated by echocardiography and histochemical staining 4 weeks after surgery.To identify the associated mechanism,nuclear factor(NF)-κB p65 and intercellular cell adhesion molecule-1(ICAM1)signals,cell adhesion ability,generation of reactive oxygen species,and rates of apoptosis were detected in human umbilical vein endothelial cells(HUVECs)and hiPSC-CMs.Results:After 4 weeks of transplantation,the number of cells that engrafted in the hiPSC-CMs+MLB group was about five times higher than those in the hiPSC-CMs group.Additionally,MLB treatment significantly reduced tohoku hospital pediatrics-1(THP-1)cell adhesion,ICAM1 expression,NF-κB nuclear translocation,reactive oxygen species production,NF-κB p65 phosphorylation,and cell apoptosis in HUVECs cultured under hypoxia.Similarly,treatment with MLB significantly inhibited the apoptosis of hiPSC-CMs via enhancing signal transducer and activator of transcription 3(STAT3)phosphorylation and B-cell lymphoma-2(BCL2)expression,promoting STAT3 nuclear translocation,and downregulating BCL2-Associated X,dual specificity phosphatase 2(DUSP2),and cleaved-caspase-3 expression under hypoxia.Furthermore,MLB significantly suppressed the production of malondialdehyde and lactate dehydrogenase and the reduction in glutathione content induced by hypoxia in both HUVECs and hiPSC-CMs in vitro.Conclusions:MLB significantly enhanced the potential of hiPSC-CMs in repairing injured myocardium by improving endothelial cell function via the NF-κB/ICAM1 pathway and inhibiting hiPSC-CMs apoptosis via the DUSP2/STAT3 pathway.
