Magnaporthe oryzae is the causal agent of rice blast. Glycosylation plays key roles in vegetative growth,development, and infection of M. oryzae. However, several glycosylation-related genes have not been characterize...Magnaporthe oryzae is the causal agent of rice blast. Glycosylation plays key roles in vegetative growth,development, and infection of M. oryzae. However, several glycosylation-related genes have not been characterized.In this study, we identified a Glyco_transf_22 domain-containing protein, MoAlg9, and found that MoAlg9 islocalized to the endoplasmic reticulum(ER). Deletion of MoALG9 significantly affected conidial production, normalappressorium formation, responses to stressors, and pathogenicity of M. oryzae. We also found that the ΔMoalg9mutant was defective in glycogen utilization, appressorial penetration, and invasive growth in host cells. Moreover,we further demonstrated that MoALG9 regulates the transcription of several target genes involved in conidiation,appressorium formation, and cell wall integrity. In addition, we found that the Glyco_transf_22 domain is essentialfor normal MoAlg9 function and localization. We also provide evidence that MoAlg9 is involved in N-glycosylationpathway in M. oryzae. Taken together, these results show that MoAlg9 is important for conidiation, appressoriumformation, maintenance of cell wall integrity, and the pathogenesis of M. oryzae.展开更多
甘露糖基转移酶Alg11是N糖基化途径中的关键酶,催化第4和第5个甘露糖(Man)残基以α-1,2连键的形式转移到底物Man3-GlcNAc2-PP-Dol生成Man5-GlcNAc2-PP-Dol,人类ALG11基因致病突变会导致相关的先天性糖基化疾病ALG11-CDG(Congenital Diso...甘露糖基转移酶Alg11是N糖基化途径中的关键酶,催化第4和第5个甘露糖(Man)残基以α-1,2连键的形式转移到底物Man3-GlcNAc2-PP-Dol生成Man5-GlcNAc2-PP-Dol,人类ALG11基因致病突变会导致相关的先天性糖基化疾病ALG11-CDG(Congenital Disorders of Glycosylation)。目前,对ALG11-CDG的研究更侧重于其临床症状,而患者Alg11突变蛋白的性质还有待探索。作者以研究ALG11-CDG突变蛋白的重组表达及活性为目的,首先在人胚胎肾细胞HEK293和酿酒酵母中分别表达了Alg11野生型及突变体蛋白,发现其表达量存在显著差异,突变体蛋白不能稳定表达。最终在大肠杆菌中成功稳定表达了野生型及ALG11-CDG相关突变蛋白,利用液质联用定量分析了蛋白质的活性,结果显示,突变蛋白的活性存在不同程度的降低,其降低程度与ALG11-CDG疾病的严重程度存在一定的相关性。上述研究结果证明:1)ALG11-CDG相关突变蛋白的不稳定有可能是造成疾病的原因之一;2)体外定量测试大肠杆菌表达的ALG11-CDG相关突变蛋白的活性可以为相应患者的疾病严重程度提供参考。展开更多
To overcome the fast or burst release of hydrophilic drugs from hydrophilic alginate-based carriers,hydrophobic molecule(vinyl acetate,VAc)was grafted on alginate(Alg),which was further used to prepare drug carriers.A...To overcome the fast or burst release of hydrophilic drugs from hydrophilic alginate-based carriers,hydrophobic molecule(vinyl acetate,VAc)was grafted on alginate(Alg),which was further used to prepare drug carriers.Amphiphilic Alg-g-PVAc hydrogel beads were firstly prepared by emulsification/internal gelation technique for the loading of bovine serum albumin(BSA).Then,chitosan was coated on the surface of beads to form novel amphiphilic Alg-g-PVAc/chitosan(Alg-g-PVAc/CS)microcapsules.The BSA-loading amphiphilic Alg-g-PVAc/chitosan(Alg-g-PVAc/CS)microcapsules display similar morphology and size to the hydrophilic alginate/chitosan(AC)microcapsules.However,the drug loading and loading efficiency of BSA in Alg-g-PVAc/CS microcapsules are higher,and the release rate of BSA from Alg-g-PVAc/CS microcapsules is slower.The results demonstrate that the introduction of hydrophobic PVAc on alginate can effectively help retard the release of BSA,and the higher degree of substitution is,the slower the release rate is.In addition,the complex membrane can also be adjusted to delay the release of BSA.As a whole,amphiphilic sodium alginate-vinyl acetate/CS microparticles could be developed for macromolecular drug delivery.展开更多
基金supported by the National Natural Science Foundation of China (32202253)the Natural Science Foundation of Anhui Higher Education Institutions, China (KJ2020A0102)the Talent Research Project of Anhui Agricultural University, China (rc342001)。
文摘Magnaporthe oryzae is the causal agent of rice blast. Glycosylation plays key roles in vegetative growth,development, and infection of M. oryzae. However, several glycosylation-related genes have not been characterized.In this study, we identified a Glyco_transf_22 domain-containing protein, MoAlg9, and found that MoAlg9 islocalized to the endoplasmic reticulum(ER). Deletion of MoALG9 significantly affected conidial production, normalappressorium formation, responses to stressors, and pathogenicity of M. oryzae. We also found that the ΔMoalg9mutant was defective in glycogen utilization, appressorial penetration, and invasive growth in host cells. Moreover,we further demonstrated that MoALG9 regulates the transcription of several target genes involved in conidiation,appressorium formation, and cell wall integrity. In addition, we found that the Glyco_transf_22 domain is essentialfor normal MoAlg9 function and localization. We also provide evidence that MoAlg9 is involved in N-glycosylationpathway in M. oryzae. Taken together, these results show that MoAlg9 is important for conidiation, appressoriumformation, maintenance of cell wall integrity, and the pathogenesis of M. oryzae.
文摘甘露糖基转移酶Alg11是N糖基化途径中的关键酶,催化第4和第5个甘露糖(Man)残基以α-1,2连键的形式转移到底物Man3-GlcNAc2-PP-Dol生成Man5-GlcNAc2-PP-Dol,人类ALG11基因致病突变会导致相关的先天性糖基化疾病ALG11-CDG(Congenital Disorders of Glycosylation)。目前,对ALG11-CDG的研究更侧重于其临床症状,而患者Alg11突变蛋白的性质还有待探索。作者以研究ALG11-CDG突变蛋白的重组表达及活性为目的,首先在人胚胎肾细胞HEK293和酿酒酵母中分别表达了Alg11野生型及突变体蛋白,发现其表达量存在显著差异,突变体蛋白不能稳定表达。最终在大肠杆菌中成功稳定表达了野生型及ALG11-CDG相关突变蛋白,利用液质联用定量分析了蛋白质的活性,结果显示,突变蛋白的活性存在不同程度的降低,其降低程度与ALG11-CDG疾病的严重程度存在一定的相关性。上述研究结果证明:1)ALG11-CDG相关突变蛋白的不稳定有可能是造成疾病的原因之一;2)体外定量测试大肠杆菌表达的ALG11-CDG相关突变蛋白的活性可以为相应患者的疾病严重程度提供参考。
基金Supported by the National Natural Science Foundation of China(No.21276033)the Open Foundation of the State Key Laboratory of Bioactive Seaweed Substances(Nos.SKL-BASS1711,SKL-BASS1707)the National Undergraduates Innovation and Entrepreneurship Training Program of China(No.201711258000001)
文摘To overcome the fast or burst release of hydrophilic drugs from hydrophilic alginate-based carriers,hydrophobic molecule(vinyl acetate,VAc)was grafted on alginate(Alg),which was further used to prepare drug carriers.Amphiphilic Alg-g-PVAc hydrogel beads were firstly prepared by emulsification/internal gelation technique for the loading of bovine serum albumin(BSA).Then,chitosan was coated on the surface of beads to form novel amphiphilic Alg-g-PVAc/chitosan(Alg-g-PVAc/CS)microcapsules.The BSA-loading amphiphilic Alg-g-PVAc/chitosan(Alg-g-PVAc/CS)microcapsules display similar morphology and size to the hydrophilic alginate/chitosan(AC)microcapsules.However,the drug loading and loading efficiency of BSA in Alg-g-PVAc/CS microcapsules are higher,and the release rate of BSA from Alg-g-PVAc/CS microcapsules is slower.The results demonstrate that the introduction of hydrophobic PVAc on alginate can effectively help retard the release of BSA,and the higher degree of substitution is,the slower the release rate is.In addition,the complex membrane can also be adjusted to delay the release of BSA.As a whole,amphiphilic sodium alginate-vinyl acetate/CS microparticles could be developed for macromolecular drug delivery.
