Asian soybean rust,caused by Phakopsora pachyrhizi,is a devastating fungal disease threatening global soybean production,particularly in tropical regions where chemical control is increasingly unsustainable.This study...Asian soybean rust,caused by Phakopsora pachyrhizi,is a devastating fungal disease threatening global soybean production,particularly in tropical regions where chemical control is increasingly unsustainable.This study employed cutting-edge 4D-DIA proteomics to investigate molecular defense mechanisms in resistant(SX6907)and susceptible(Tianlong 1)soybean cultivars during early infection(12 hpi and 3 dpi).We identified 12,852 proteins,with 1,510 differentially expressed proteins(DEPs)revealing genotype-specific responses.Resistant plants exhibited sustained upregulation of immune receptors(CRKs,LRR-RLKs),MAPK signaling components,and cell wall reinforcement proteins(peroxidases,XTHs),alongside dynamic modulation of calcium signaling and ROS homeostasis.These patterns suggest key pathways enriched in resistance may include phenylpropanoid biosynthesis,isoflavonoid production,and ER stress responses,while susceptible plants showed suppression of photosynthesis and defense pathways.Weighted Protein Co-expression Network Analysis(WPCNA)highlighted co-expression modules linked to resistance,potentially including NLR-mediated effector-triggered immunity.Crucially,DIR proteins and organelle-specific defense hubs(e.g.,chloroplasts,nuclei)were implicated in rust resistance.Validation by qPCR confirmed concordance for 84%of tested DEPs.Our findings provide a protein-level blueprint of soybean rust resistance,identifying candidate targets for marker-assisted breeding and genetic engineering to develop durable resistant varieties,reducing reliance on fungicides.展开更多
The increase in human population has led to imminent pressures to develop new edible proteins with decreased environmental footprints.The most promising approach involves the production of single cell protein(SCP)from...The increase in human population has led to imminent pressures to develop new edible proteins with decreased environmental footprints.The most promising approach involves the production of single cell protein(SCP)from yeasts,which have been utilized in a wide variety of foods for thousands of years.In this study,102 yeast strains isolated from traditional fermented pork(Nanx Wudl)were investigated for their potential as SCP producer for the first time.Based on preliminary screening,Saccharomyces cerevisiae Y70 and Candida parapsilosis H5Y13,both showing high protein content and excellent growth capability,were selected for further analysis via 4D-DIA proteomics technology.Proteomic analysis indicated that the oxidative metabolism pathways,including TCA cycle,oxidative phosphorylation and pentose phosphate pathway,may have a significant impact on global protein synthesis and production.This study provides useful information for selecting SCP-producing yeast from Chinese fermented meat products and contribute to a deeper understanding of the underlying metabolic mechanisms behind global protein synthesis in yeast.Furthermore,these findings also provide potential molecular targets for genetic engineering modifications in yeast,aimed at constructing highly efficient cell factories for protein production.展开更多
In plants,heteromorphic self-incompatibility(HetSI)is a strategy for avoiding self-pollination and promoting outcrossing,and during this process,numerous protein-protein interaction events occur between the pistil and...In plants,heteromorphic self-incompatibility(HetSI)is a strategy for avoiding self-pollination and promoting outcrossing,and during this process,numerous protein-protein interaction events occur between the pistil and pollen.Previous studies in Primula and Fagopyrum that focused on HetSI systems have provided interesting insights;however,the molecular mechanism underlying HetSI remains largely unknown.In this study,we profiled the proteome of Plumbago auriculata stigmas before and after self-incompatible(SI)and self-compatible(SC)pollination.Comparative analyses were conducted by 4D-DIA(Four-dimensional data independent acquisition),a promising technology that increases the sensitivity and reduces the spectral complexity of proteomic analysis by adding a fourth dimension,ion mobility.The results revealed 33387 peptides and 5311 proteins in all samples.The pathways in which the differentially expressed proteins(DEPs)identified in the P×P(Pin style self-pollinated with pin pollen)vs.PS(Pin style)and T×T(Thrum style self-pollinated with thrum pollen)vs.TS(Thrum style)comparisons were significantly enriched were biosynthesis of secondary metabolites and pentose and glucuronate interconversions.In the P×T(Pin style cross-pollinated with thrum pollen)vs.PS and T×P(Thrum style cross-pollinated with pin pollen)vs.TS comparison,the top three pathways were biosynthesis of secondary metabolites,pentose and glucuronate interconversions,and phenylpropanoid biosynthesis.The phenylpropanoid biosynthesis,cutin,suberine and wax biosynthesis,and flavonoid biosynthesis pathways were enriched in the P×T vs.P×P comparison,and starch and sucrose metabolism,glycerophospholipid metabolism,and alpha-linolenic acid metabolism were abundant in the T×T vs.T×P comparison.The enriched pathways between PS and TS were the biosynthesis of secondary metabolites,phenylpropanoid biosynthesis,and pentose and glucuronate interconversion.Self-incompatibility protein S1(SI S1),Mitogen-activated protein kinase 3/4(MPK3/4),Mitogen-activated protein kinase kinase 2/3(M2K2/3),Exocyst complex component EXO70A1(E70A1)and Thioredoxin H1/2(TRXH1/2)were found to be HetSI-related candidates,and O-fucosyltransferase 23(OFT23),3-ketoacyl-CoA synthase 6(KCS6),Receptor-like protein kinase FERONIA(FERON),Fimbrin-5(FIMB5),Pollen-specific leucine-rich repeat extensin-like protein 4(PLRX4),Transcription initiation factor IIB-2(TF2B2)and Pectinesterase 1(AL11A),etc.,were identified as other regulatory transducers.These findings combined with our morphological and reactive oxygen species(ROS)intensity analyses indicate that P.auriculata has typical dry-stigmas and that the HetSI mechanism might differ between the pin and thrum.SI S1 might be the key factor in HetSI,and ROS are overexpressed during SC pollination to rapidly activate the mitogen-activated protein kinase(MAPK)-mediated phosphorylation of E70A1 to maintain stigma receptivity in plants with HetSI.展开更多
文摘为探讨经典名方散偏汤对慢性偏头痛的作用机制,该研究采用4D-DIA蛋白质组学技术分析其对慢性偏头痛模型大鼠的干预影响,并对关键差异蛋白进行实验验证。首先将SD雄性大鼠随机分组,反复注射硝酸甘油制备慢性偏头痛模型。连续灌胃7 d后,采用大鼠痛苦面容量表(RGS)评估药效。取三叉神经节进行4D-DIA相对定量蛋白质组学检测,筛选各组差异蛋白,结合多个数据库进行基因本体论(Gene Ontology,GO)功能和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析;使用STRING数据库及Cytoscape软件进行蛋白-蛋白互作(PPI)网络分析,并对关键差异表达蛋白TRPV1进行蛋白免疫印迹法(Western blot)检测。结果显示,空白组与模型组间有517个差异表达蛋白,模型组与散偏汤中剂量组间有221个差异表达蛋白。GO功能和KEGG通路富集分析显示,差异表达蛋白主要与炎症反应、伤害性感受刺激、甘油三酯代谢、免疫调节等相关,主要集中在炎症相关TRP通路、AMPK信号通路、PI3K-Akt信号通路、TGF-β信号通路等。PPI网络显示,IGF、TOP2A、APOA1、CDK1、TTN、RYR1、CSRP3等靶蛋白具有较高的度值(degree)。Western blot结果显示,与模型组比较,散偏汤中、高剂量组TRPV1表达水平差异有统计学意义(P<0.05)。综上,散偏汤可能通过调节TRP、AMPK、PI3K-Akt等炎症相关通路治疗慢性偏头痛,其对TRPV1蛋白的调节发挥了重要作用,并对伤害性刺激感受、脂代谢和免疫反应等具有潜在的调节作用。
基金supported by the National Key Research and Development Program of China(2022YFF1001504)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-ASTIP-2022-OCRI)+1 种基金the Basic research fund of Oil Crop Research Institute,Chinese Academy of Agricultural Sciences(1610172022002)Hubei Province Technological Innovation Program(2024BBB003).
文摘Asian soybean rust,caused by Phakopsora pachyrhizi,is a devastating fungal disease threatening global soybean production,particularly in tropical regions where chemical control is increasingly unsustainable.This study employed cutting-edge 4D-DIA proteomics to investigate molecular defense mechanisms in resistant(SX6907)and susceptible(Tianlong 1)soybean cultivars during early infection(12 hpi and 3 dpi).We identified 12,852 proteins,with 1,510 differentially expressed proteins(DEPs)revealing genotype-specific responses.Resistant plants exhibited sustained upregulation of immune receptors(CRKs,LRR-RLKs),MAPK signaling components,and cell wall reinforcement proteins(peroxidases,XTHs),alongside dynamic modulation of calcium signaling and ROS homeostasis.These patterns suggest key pathways enriched in resistance may include phenylpropanoid biosynthesis,isoflavonoid production,and ER stress responses,while susceptible plants showed suppression of photosynthesis and defense pathways.Weighted Protein Co-expression Network Analysis(WPCNA)highlighted co-expression modules linked to resistance,potentially including NLR-mediated effector-triggered immunity.Crucially,DIR proteins and organelle-specific defense hubs(e.g.,chloroplasts,nuclei)were implicated in rust resistance.Validation by qPCR confirmed concordance for 84%of tested DEPs.Our findings provide a protein-level blueprint of soybean rust resistance,identifying candidate targets for marker-assisted breeding and genetic engineering to develop durable resistant varieties,reducing reliance on fungicides.
基金financial support of the National Natural Science Foundation of China(32102016)the Taishan Industrial Experts Program,Beijing Postdoctoral Research Foundation(2323ZZ122).
文摘The increase in human population has led to imminent pressures to develop new edible proteins with decreased environmental footprints.The most promising approach involves the production of single cell protein(SCP)from yeasts,which have been utilized in a wide variety of foods for thousands of years.In this study,102 yeast strains isolated from traditional fermented pork(Nanx Wudl)were investigated for their potential as SCP producer for the first time.Based on preliminary screening,Saccharomyces cerevisiae Y70 and Candida parapsilosis H5Y13,both showing high protein content and excellent growth capability,were selected for further analysis via 4D-DIA proteomics technology.Proteomic analysis indicated that the oxidative metabolism pathways,including TCA cycle,oxidative phosphorylation and pentose phosphate pathway,may have a significant impact on global protein synthesis and production.This study provides useful information for selecting SCP-producing yeast from Chinese fermented meat products and contribute to a deeper understanding of the underlying metabolic mechanisms behind global protein synthesis in yeast.Furthermore,these findings also provide potential molecular targets for genetic engineering modifications in yeast,aimed at constructing highly efficient cell factories for protein production.
基金Scientific Research Foundation of Science&Technology Department of Sichuan Province(2021YJ0497).
文摘In plants,heteromorphic self-incompatibility(HetSI)is a strategy for avoiding self-pollination and promoting outcrossing,and during this process,numerous protein-protein interaction events occur between the pistil and pollen.Previous studies in Primula and Fagopyrum that focused on HetSI systems have provided interesting insights;however,the molecular mechanism underlying HetSI remains largely unknown.In this study,we profiled the proteome of Plumbago auriculata stigmas before and after self-incompatible(SI)and self-compatible(SC)pollination.Comparative analyses were conducted by 4D-DIA(Four-dimensional data independent acquisition),a promising technology that increases the sensitivity and reduces the spectral complexity of proteomic analysis by adding a fourth dimension,ion mobility.The results revealed 33387 peptides and 5311 proteins in all samples.The pathways in which the differentially expressed proteins(DEPs)identified in the P×P(Pin style self-pollinated with pin pollen)vs.PS(Pin style)and T×T(Thrum style self-pollinated with thrum pollen)vs.TS(Thrum style)comparisons were significantly enriched were biosynthesis of secondary metabolites and pentose and glucuronate interconversions.In the P×T(Pin style cross-pollinated with thrum pollen)vs.PS and T×P(Thrum style cross-pollinated with pin pollen)vs.TS comparison,the top three pathways were biosynthesis of secondary metabolites,pentose and glucuronate interconversions,and phenylpropanoid biosynthesis.The phenylpropanoid biosynthesis,cutin,suberine and wax biosynthesis,and flavonoid biosynthesis pathways were enriched in the P×T vs.P×P comparison,and starch and sucrose metabolism,glycerophospholipid metabolism,and alpha-linolenic acid metabolism were abundant in the T×T vs.T×P comparison.The enriched pathways between PS and TS were the biosynthesis of secondary metabolites,phenylpropanoid biosynthesis,and pentose and glucuronate interconversion.Self-incompatibility protein S1(SI S1),Mitogen-activated protein kinase 3/4(MPK3/4),Mitogen-activated protein kinase kinase 2/3(M2K2/3),Exocyst complex component EXO70A1(E70A1)and Thioredoxin H1/2(TRXH1/2)were found to be HetSI-related candidates,and O-fucosyltransferase 23(OFT23),3-ketoacyl-CoA synthase 6(KCS6),Receptor-like protein kinase FERONIA(FERON),Fimbrin-5(FIMB5),Pollen-specific leucine-rich repeat extensin-like protein 4(PLRX4),Transcription initiation factor IIB-2(TF2B2)and Pectinesterase 1(AL11A),etc.,were identified as other regulatory transducers.These findings combined with our morphological and reactive oxygen species(ROS)intensity analyses indicate that P.auriculata has typical dry-stigmas and that the HetSI mechanism might differ between the pin and thrum.SI S1 might be the key factor in HetSI,and ROS are overexpressed during SC pollination to rapidly activate the mitogen-activated protein kinase(MAPK)-mediated phosphorylation of E70A1 to maintain stigma receptivity in plants with HetSI.
