Fusarium pseudograminearum is a devastating pathogen that causes Fusarium crown rot(FCR)in wheat and poses a significant threat to wheat production in terms of grain yield and quality.However,the mechanism by which F....Fusarium pseudograminearum is a devastating pathogen that causes Fusarium crown rot(FCR)in wheat and poses a significant threat to wheat production in terms of grain yield and quality.However,the mechanism by which F.pseudograminearum infects wheat remains unclear.In this study,we aimed to elucidate these mechanisms by constructing a T-DNA insertion mutant library for the highly virulent strain WZ-8A of F.pseudograminearum.By screening this mutant library,we identified nine independent mutants that displayed impaired pathogenesis in barley leaves.Among these mutants,one possessed a disruption in the gene FpRCO1 that is an ortholog of Saccharomyces cerevisiae RCO1,encoding essential component of the Rpd3S histone deacetylase complex in F.pseudograminearum.To further investigate the role of FpRCO1 in F.pseudograminearum,we employed a split-marker approach to knock out FpRCO1 in F.pseudograminearum WZ-8A.FpRCO1 deletion mutants exhibit reduced vegetative growth,conidium production,and virulence in wheat coleoptiles and barley leaves,whereas the complementary strain restores these phenotypes.Moreover,under stress conditions,the FpRCO1 deletion mutants exhibited increased sensitivity to NaCl,sorbitol,and SDS,but possessed reduced sensitivity to H_(2)O_(2)compared to these characteristics in the wild-type strain.RNA-seq analysis revealed that deletion of FpRCO1 affected gene expression(particularly the downregulation of TRI gene expression),thus resulting in significantly reduced deoxynivalenol(DON)production.In summary,our findings highlight the pivotal role of FpRCO1 in regulating vegetative growth and development,asexual reproduction,DON production,and pathogenicity of F.pseudograminearum.This study provides valuable insights into the molecular mechanisms underlying F.pseudograminearum infection in wheat and may pave the way for the development of novel strategies to combat this devastating disease.展开更多
Hyphal fusion(anastomosis)is a common process serving many important functions at various developmental stages in the life cycle of ascomycetous fungi.However,the biological roles and molecular mechanisms in plant pat...Hyphal fusion(anastomosis)is a common process serving many important functions at various developmental stages in the life cycle of ascomycetous fungi.However,the biological roles and molecular mechanisms in plant pathogenic fungi were widely unknown.In this study,a hyphal fusion protein FpHam-2 was screened from a T-DNA insertion mutant library of Fusarium pseudograminearum,and FpHam-2 interacts with another 2 hyphal fusion protein homologues FpHam-3 and FpHam-4.Each of these 3 genes deletion mutant revealed in similar defective phenotypes compared with the WT and complemented strains,including reduction in growth rate,defects in hyphal fusion and conidiation,more sensitive for cell membrane,cell wall and oxidative stress responses,and decreased in virulence.The yeast two-hybrid assay was used to identify that FpHam-2 interacts with 3 autophagy-related proteins,including FpAtg3,FpAtg28 and FpAtg33.Furthermore,FpHam-2-deletion mutant showed decreased accumulation of autophagic bodies in hypha.In conclusion,FpHam-2,FpHam-3 and FpHam-4 have an essential role for hyphal fusion and regulating the growth,conidiation and virulence in F.pseudograminearum.展开更多
Objective The aim of the study was to explore the difference between immune cell subsets during the incubation of cytokine-induced kill cells (CIKs) from patients with and without hepatitis B virus (HBV). Methods ...Objective The aim of the study was to explore the difference between immune cell subsets during the incubation of cytokine-induced kill cells (CIKs) from patients with and without hepatitis B virus (HBV). Methods Peripheral blood samples were extracted from 50 tumor patients, and were divided into two groups according to the presence or absence of HBV. The proliferation rate and activity of CIK cells were examined based on counts on days 1, 5, 7, 9, 11, 13, and 15 of culture. Additionally, the CD3+, CD4+, CD8+, CD3+CD8+, C+)3+CD4+, and CD3+CD56+ T cell populations were analyzed by flow cytometry on days 5, 7, 10, 13, and 15 of culture. Results Proliferation over a 15-day period was higher in the HBV-positive group than in the negative group (280-fold vs. 180-fold increase, respectively), but there was no significant difference between the two groups at each time point. The frequencies of CD3+, CD8+ T, CD3+CD8+, and CD3+CD56+T cells increased over time, while those of CD4+ and CD3+CD4+ T cells decreased over time, and these changes were greater in the positive group than in the negative group. The differences in CD8+ T cells and CD3+CD4+ T cells between the two groups were significant (P 〈 0.05). Conclusion The proliferative capacity of CIK cells was higher for patients in the HBV-positive group than those in the HBV-negative group, and immune cell subsets were more favorable in the HBV-positive group than the neaative arouD.展开更多
As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri(Chai-Hu) is commonly used for the treatment of influenza,which is derived from the dried root...As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri(Chai-Hu) is commonly used for the treatment of influenza,which is derived from the dried roots of Bupleurum chinense DC.and Bupleurum scorzonerifolium Willd.Among of diverse chemical components,saikosaponins are the key active components of the herb medicine.In the present study,we established a method of high performance liquid chromatography(HPLC) coupled with evaporative light scattering detection(ELSD) for simultaneous determination of saikosaponin a,c and d in root,stem,leaf and flower of Bupleurum chinense(B chinense) collected from different areas of Shanxi Province,China.The results from 16 samples of root,stem,leaf and flower of B chinense demonstrated that the total contents of the three saikosaponins in the root of B chinense collected from Dongshan Taiyuan,Xishan,Tianlongshan and Pangquangou were 4.26 mg/g,3.22 mg/g,4.23 mg/g and 3.05 mg/g,respectively.However,there was scarcely any saikosaponins in the stem,leaf and flower of B chinense collected from above-mentioned areas.The method of HPLC coupled with ELSD was suitable for quality control of Radix Bupleuri.The result also confirmed that the root of B chinense was the best medicinal part.展开更多
基金supported by grants from the National Natural Science Foundation of China(31901835)the Science and Technology Planning Project of Henan Province of China(212102110145)the International(Regional)Cooperation and Exchange Program of the National Natural Science Foundation of China(31961143018).
文摘Fusarium pseudograminearum is a devastating pathogen that causes Fusarium crown rot(FCR)in wheat and poses a significant threat to wheat production in terms of grain yield and quality.However,the mechanism by which F.pseudograminearum infects wheat remains unclear.In this study,we aimed to elucidate these mechanisms by constructing a T-DNA insertion mutant library for the highly virulent strain WZ-8A of F.pseudograminearum.By screening this mutant library,we identified nine independent mutants that displayed impaired pathogenesis in barley leaves.Among these mutants,one possessed a disruption in the gene FpRCO1 that is an ortholog of Saccharomyces cerevisiae RCO1,encoding essential component of the Rpd3S histone deacetylase complex in F.pseudograminearum.To further investigate the role of FpRCO1 in F.pseudograminearum,we employed a split-marker approach to knock out FpRCO1 in F.pseudograminearum WZ-8A.FpRCO1 deletion mutants exhibit reduced vegetative growth,conidium production,and virulence in wheat coleoptiles and barley leaves,whereas the complementary strain restores these phenotypes.Moreover,under stress conditions,the FpRCO1 deletion mutants exhibited increased sensitivity to NaCl,sorbitol,and SDS,but possessed reduced sensitivity to H_(2)O_(2)compared to these characteristics in the wild-type strain.RNA-seq analysis revealed that deletion of FpRCO1 affected gene expression(particularly the downregulation of TRI gene expression),thus resulting in significantly reduced deoxynivalenol(DON)production.In summary,our findings highlight the pivotal role of FpRCO1 in regulating vegetative growth and development,asexual reproduction,DON production,and pathogenicity of F.pseudograminearum.This study provides valuable insights into the molecular mechanisms underlying F.pseudograminearum infection in wheat and may pave the way for the development of novel strategies to combat this devastating disease.
基金supported by the grants from the National Natural Science Foundation of China(U2004140)the Henan Provincial Science and Technology Major Project,China(221100110100)。
文摘Hyphal fusion(anastomosis)is a common process serving many important functions at various developmental stages in the life cycle of ascomycetous fungi.However,the biological roles and molecular mechanisms in plant pathogenic fungi were widely unknown.In this study,a hyphal fusion protein FpHam-2 was screened from a T-DNA insertion mutant library of Fusarium pseudograminearum,and FpHam-2 interacts with another 2 hyphal fusion protein homologues FpHam-3 and FpHam-4.Each of these 3 genes deletion mutant revealed in similar defective phenotypes compared with the WT and complemented strains,including reduction in growth rate,defects in hyphal fusion and conidiation,more sensitive for cell membrane,cell wall and oxidative stress responses,and decreased in virulence.The yeast two-hybrid assay was used to identify that FpHam-2 interacts with 3 autophagy-related proteins,including FpAtg3,FpAtg28 and FpAtg33.Furthermore,FpHam-2-deletion mutant showed decreased accumulation of autophagic bodies in hypha.In conclusion,FpHam-2,FpHam-3 and FpHam-4 have an essential role for hyphal fusion and regulating the growth,conidiation and virulence in F.pseudograminearum.
文摘Objective The aim of the study was to explore the difference between immune cell subsets during the incubation of cytokine-induced kill cells (CIKs) from patients with and without hepatitis B virus (HBV). Methods Peripheral blood samples were extracted from 50 tumor patients, and were divided into two groups according to the presence or absence of HBV. The proliferation rate and activity of CIK cells were examined based on counts on days 1, 5, 7, 9, 11, 13, and 15 of culture. Additionally, the CD3+, CD4+, CD8+, CD3+CD8+, C+)3+CD4+, and CD3+CD56+ T cell populations were analyzed by flow cytometry on days 5, 7, 10, 13, and 15 of culture. Results Proliferation over a 15-day period was higher in the HBV-positive group than in the negative group (280-fold vs. 180-fold increase, respectively), but there was no significant difference between the two groups at each time point. The frequencies of CD3+, CD8+ T, CD3+CD8+, and CD3+CD56+T cells increased over time, while those of CD4+ and CD3+CD4+ T cells decreased over time, and these changes were greater in the positive group than in the negative group. The differences in CD8+ T cells and CD3+CD4+ T cells between the two groups were significant (P 〈 0.05). Conclusion The proliferative capacity of CIK cells was higher for patients in the HBV-positive group than those in the HBV-negative group, and immune cell subsets were more favorable in the HBV-positive group than the neaative arouD.
基金Shanxi Educational Committee(Grant No.20111113)Shanxi Science and Technology Department(Grant No.2016ZD0201)
文摘As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri(Chai-Hu) is commonly used for the treatment of influenza,which is derived from the dried roots of Bupleurum chinense DC.and Bupleurum scorzonerifolium Willd.Among of diverse chemical components,saikosaponins are the key active components of the herb medicine.In the present study,we established a method of high performance liquid chromatography(HPLC) coupled with evaporative light scattering detection(ELSD) for simultaneous determination of saikosaponin a,c and d in root,stem,leaf and flower of Bupleurum chinense(B chinense) collected from different areas of Shanxi Province,China.The results from 16 samples of root,stem,leaf and flower of B chinense demonstrated that the total contents of the three saikosaponins in the root of B chinense collected from Dongshan Taiyuan,Xishan,Tianlongshan and Pangquangou were 4.26 mg/g,3.22 mg/g,4.23 mg/g and 3.05 mg/g,respectively.However,there was scarcely any saikosaponins in the stem,leaf and flower of B chinense collected from above-mentioned areas.The method of HPLC coupled with ELSD was suitable for quality control of Radix Bupleuri.The result also confirmed that the root of B chinense was the best medicinal part.