Petal blotch is a prevalent pigmentation pattern observed in the Xibei tree peony(Paeonia rockii), possessing significant aesthetic value and playing a crucial role in the species' reproduction and fitness. Despit...Petal blotch is a prevalent pigmentation pattern observed in the Xibei tree peony(Paeonia rockii), possessing significant aesthetic value and playing a crucial role in the species' reproduction and fitness. Despite years of research, deciphering the molecular mechanisms underlying blotch formation remains challenging. As is well known, floral pigmentation is frequently associated with the familiar R2R3-MYB transcription factors. The key MYB anthocyanin activators of P. rockii ‘Shu Sheng Peng Mo' were previously reported in our preceding study. In this study, we identified and characterized three R2R3-MYBs, Pr MYBi1, Pr MYBi2, and Pr MYBi3, which belong to subgroup 4(SG4) and play repressor roles in anthocyanin biosynthesis. A quantitative real-time PCR(q RT-PCR) assay indicated that the expression of Pr MYBi1 and Pr MYBi3 gradually increased during flowering development and was substantially up-regulated in non-blotch compared to blotch. Yeast one-hybrid and dualluciferase assays demonstrated that Pr MYBi(1-3) directly target the anthocyanin structural genes and repress their transcription. The genetic transformation of tobacco demonstrated that the overexpression of Pr MYBi(1-3) decreased anthocyanin accumulation in flowers, with Pr MYBi1 serving as the most effective repressor. Our results revealed that SG4 R2R3-MYBs negatively regulate the anthocyanin pathway in P.rockii conservatively, and we provide the definite members. These findings will advance future research to unravel the mystery of blotch pattern formation.展开更多
Camptotheca acuminata produces camptothecin(CPT),a monoterpene indole alkaloid(MIA)that is widely used in the treatment of lung,colorectal,cervical,and ovarian cancers.Its biosynthesis pathway has attracted significan...Camptotheca acuminata produces camptothecin(CPT),a monoterpene indole alkaloid(MIA)that is widely used in the treatment of lung,colorectal,cervical,and ovarian cancers.Its biosynthesis pathway has attracted significant attention,but the regulation of CPT biosynthesis by the APETALA2/ethylene-responsive factor(AP2/ERF)transcription factors(TFs)remains unclear.In this study,a systematic analysis of the AP2/ERF TFs family in C.acuminata was performed,including phylogeny,gene structure,conserved motifs,and gene expression profiles in different tissues and organs(immature bark,cotyledons,young flower,immature fruit,mature fruit,mature leaf,roots,upper stem,and lower stem)of C.acuminata.A total of 198 AP2/ERF genes were identified and divided into five relatively conserved subfamilies,including AP2(26 genes),DREB(61 genes),ERF(92 genes),RAV(18 genes),and Soloist(one gene).The combination of gene expression patterns in different C.acuminata tissues and organs,the phylogenetic tree,the co-expression analysis with biosynthetic genes,and the analysis of promoter sequences of key enzymes genes involved in CPT biosynthesis pathways revealed that eight AP2/ERF TFs in C.acuminata might be involved in CPT synthesis regulation,which exhibit relatively high expression levels in the upper stem or immature bark.Among these,four genes(Cac AP2/ERF123,Cac AP2/ERF125,Cac AP2/ERF126,and Cac AP2/ERF127)belong to the ERF–B2 subgroup;two genes(Cac AP2/ERF149 and Cac AP2/ERF152)belong to the ERF–B3 subgroup;and two more genes(Cac AP2/ERF095 and Cac AP2/ERF096)belong to the DREB–A6 subgroup.These results provide a foundation for future functional characterization of the AP2/ERF genes to enhance the biosynthesis of CPT compounds of C.acuminata.展开更多
Sesquiterpenoids play an import role in the direct or indirect defense of plants.Farnesyl pyrophosphate synthases(FPSs)catalyze the biosynthesis of farnesyl pyrophosphate,which is a key precursor of farnesol and(E)-β...Sesquiterpenoids play an import role in the direct or indirect defense of plants.Farnesyl pyrophosphate synthases(FPSs)catalyze the biosynthesis of farnesyl pyrophosphate,which is a key precursor of farnesol and(E)-β-farnesene.In the current study,two FPS genes in Gossypium hirsutum,GhFPS1 and GhFPS2,were heterologously cloned and functionally characterized in a greenhouse setting.The open reading frames for full-length GhFPS1 and GhFPS2 were each 1029 nucleotides,and encoded two proteins of 342 amino acids with molecular weights of 39.4 kDa.The deduced amino acid sequences of GhFPS1–2 showed high identity to FPSs of other plants.Quantitative real-time PCR analysis revealed that GhFPS1 and GhFPS2 were highly expressed in G.hirsutum leaves,and were upregulated in methyl jasmonate(MeJA)-,methyl salicylate(MeSA)-and aphid infestation-treated cotton plants.The recombinant proteins of either GhFPS1 or GhFPS2 plus calf intestinal alkaline phosphatase could convert geranyl diphosphate(GPP)or isopentenyl diphosphate(IPP)to one major product,farnesol.Moreover,in electrophysiological response and Y-tube olfactometer assays,farnesol showed obvious attractiveness to female Aphidius gifuensis,which is an important parasitic wasp of aphids.Our findings suggest that two GhFPSs are involved in farnesol biosynthesis and they play a crucial role in indirect defense of cotton against aphid infestation.展开更多
Zanthoxylum bungeanum is an economically important crop worldwide due to its high content of aroma-producing monoterpenoids,and development of varieties with enhanced flavor and overall quality is a crucial research a...Zanthoxylum bungeanum is an economically important crop worldwide due to its high content of aroma-producing monoterpenoids,and development of varieties with enhanced flavor and overall quality is a crucial research area.However,the transcriptional regulatory mechanisms underlying monoterpenoid synthesis in Z.bungeanum remain unclear,hindering these breeding efforts.In this study,RNA sequencing,gas chromatography–mass spectrometry,and other molecular biology techniques were used to identify the underlying transcriptional regulation mechanisms.Two transcription factors,ZbbHLH2 and ZbERF6,were identified as key regulators of monoterpenoid synthesis in Z.bungeanum that upregulate various monoterpenoid synthesis-associated genes and are novel transcriptional activators of ZbIDI,which encodes the rate-limiting enzyme in plant monoterpenoid synthesis.Functional analysis revealed that the expression of three genes[1]modulates monoterpenoid accumulation in Z.bungeanum peel.These findings provide novel insights into the metabolic regulatory network of monoterpenoid synthesis in Z.bungeanum peel,offer potential strategies for the biofortification of specific monoterpenoids,and will promote the development of Z.bungeanum germplasm for targeted breeding and quality improvement.展开更多
2-Oxoglutarate(2OG)-dependent dioxygenases(2-ODDs)are omnipresent iron-containing non-heme enzymes that catalyze various oxidation-reduction reactions in plant growth and development,nucleic acid modification and seco...2-Oxoglutarate(2OG)-dependent dioxygenases(2-ODDs)are omnipresent iron-containing non-heme enzymes that catalyze various oxidation-reduction reactions in plant growth and development,nucleic acid modification and secondary metabolism.We systematically summarized recent research on the oxidative modifications of plant 2-ODDs and related enzymes,their vital importance in the biosynthesis of plant special metabolites,and their catalytic specificity/flexibility,and discussed the potential of 2-ODD as a new approach for the identification of pivotal genes and the elucidation of biosynthetic pathway.展开更多
Lipid biosynthesis is essential for eukaryotic cells, but the mechanisms of the process in microalgae remain poorly understood. Phosphatidic acid phosphohydrolase or 3-sn-phosphatidate phosphohydrolase(PAP) catalyzes ...Lipid biosynthesis is essential for eukaryotic cells, but the mechanisms of the process in microalgae remain poorly understood. Phosphatidic acid phosphohydrolase or 3-sn-phosphatidate phosphohydrolase(PAP) catalyzes the dephosphorylation of phosphatidic acid to form diacylglycerols and inorganic orthophosphates. This reaction is integral in the synthesis of triacylglycerols. In this study, the mRNA level of the PAP isoform CrPAP2 in a species of Chlamydomonas was found to increase in nitrogen-free conditions. Silencing of the CrPAP2 gene using RNA interference resulted in the decline of lipid content by 2.4%–17.4%. By contrast, over-expression of the CrPAP2 gene resulted in an increase in lipid content by 7.5%–21.8%. These observations indicate that regulation of the CrPAP2 gene can control the lipid content of the algal cells. In vitro CrPAP2 enzyme activity assay indicated that the cloned CrPAP2 gene exhibited biological activities.展开更多
In this study we designed a novel,cost‐efficient and green method for the synthesis of copper nanoparticles(Cu NPs)supported on manganese dioxide(MnO2)NPs,using Centella asiatica L.leaf extract as a naturally‐source...In this study we designed a novel,cost‐efficient and green method for the synthesis of copper nanoparticles(Cu NPs)supported on manganese dioxide(MnO2)NPs,using Centella asiatica L.leaf extract as a naturally‐sourced reducing agent,without stabilizers or surfactants.This synthetic process is environmentally‐friendly and avoids the use of toxic reducing agents.Phenolic hydroxyl groups in the leaf extract are believed to reduce Cu2+in solution to generate Cu NPs that are subsequently stabilized on the MnO2NP surfaces.The resulting Cu/MnO2nanocomposite was fully characterized using X‐ray diffraction,transmission electron microscopy,field emission scanning electron microscopy,energy‐dispersive X‐ray spectroscopy and Fourier transform infrared spectroscopy.This material was found to function as a highly active,efficient and recyclable heterogeneous catalyst for the reduction of Congo red,rhodamine B and methylene blue as well as nitro compounds such as2,4‐dinitrophenylhydrazine and4‐nitrophenol in the presence of NaBH4in aqueous media at ambient temperature.The high stability of the Cu/MnO2nanocomposite also allows the catalyst to be separated and reused several times without any significant loss of activity.?2018,Dalian Institute of Chemical Physics,Chinese Academy of Sciences.Published by Elsevier B.V.All rights reserved.展开更多
YdjC chitooligosaccharide deacetylase homolog(YDJC)has been identified as a susceptibility gene for inflammatory bowel disease(IBD),yet its role in the pathogenesis of IBD,particularly in regulating immune responses i...YdjC chitooligosaccharide deacetylase homolog(YDJC)has been identified as a susceptibility gene for inflammatory bowel disease(IBD),yet its role in the pathogenesis of IBD,particularly in regulating immune responses in the gut mucosa,remains elusive.In this study,we demonstrated that YDJC expression is downregulated in inflamed mucosa,particularly in the CD4^(+)T cells of IBD patients,and that Ydjc deficiency promotes CD4^(+)T-cell proliferation and Th1 cell differentiation,thereby exacerbating acute and chronic colitis in mice.Integrative transcriptomic,proteomic,and metabolomic analyses revealed that Ydjc^(-/-)CD4^(+)T cells exhibit upregulated SREBP2-mediated cholesterol biosynthesis.Consistently,treatment with key enzyme inhibitors targeting cholesterol biosynthesis,including simvastatin,fatostatin,and AAV-sh-Srebf2,markedly suppressed CD4^(+)T-cell proliferation and Th1 cell differentiation,thereby alleviating colitis in Ydjc^(-/-)mice.Mechanistically,YDJC directly deacetylates SREBP2,which further suppresses downstream target gene expression(e.g.,Hmgcr,Hmgcs1,and Cyp51).Therefore,our findings elucidate a novel mechanism whereby YDJC restrains intestinal mucosal inflammation by downregulating SREBP2-driven Th1 cell differentiation,suggesting that targeting YDJC and SREBP2-mediated cholesterol biosynthesis may serve as promising therapeutic strategies for IBD.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.32030095)the Key project at central government level:The ability establishment of sustainable use for valuable Chinese medicine resources(Grant No.2060302).
文摘Petal blotch is a prevalent pigmentation pattern observed in the Xibei tree peony(Paeonia rockii), possessing significant aesthetic value and playing a crucial role in the species' reproduction and fitness. Despite years of research, deciphering the molecular mechanisms underlying blotch formation remains challenging. As is well known, floral pigmentation is frequently associated with the familiar R2R3-MYB transcription factors. The key MYB anthocyanin activators of P. rockii ‘Shu Sheng Peng Mo' were previously reported in our preceding study. In this study, we identified and characterized three R2R3-MYBs, Pr MYBi1, Pr MYBi2, and Pr MYBi3, which belong to subgroup 4(SG4) and play repressor roles in anthocyanin biosynthesis. A quantitative real-time PCR(q RT-PCR) assay indicated that the expression of Pr MYBi1 and Pr MYBi3 gradually increased during flowering development and was substantially up-regulated in non-blotch compared to blotch. Yeast one-hybrid and dualluciferase assays demonstrated that Pr MYBi(1-3) directly target the anthocyanin structural genes and repress their transcription. The genetic transformation of tobacco demonstrated that the overexpression of Pr MYBi(1-3) decreased anthocyanin accumulation in flowers, with Pr MYBi1 serving as the most effective repressor. Our results revealed that SG4 R2R3-MYBs negatively regulate the anthocyanin pathway in P.rockii conservatively, and we provide the definite members. These findings will advance future research to unravel the mystery of blotch pattern formation.
基金supported by the National Key R&D Program of China(No.2019YFC1711100)the CAMS Innovation Fund for Medical Sciences(CIFMS,No.2016-I2M-3-016)。
文摘Camptotheca acuminata produces camptothecin(CPT),a monoterpene indole alkaloid(MIA)that is widely used in the treatment of lung,colorectal,cervical,and ovarian cancers.Its biosynthesis pathway has attracted significant attention,but the regulation of CPT biosynthesis by the APETALA2/ethylene-responsive factor(AP2/ERF)transcription factors(TFs)remains unclear.In this study,a systematic analysis of the AP2/ERF TFs family in C.acuminata was performed,including phylogeny,gene structure,conserved motifs,and gene expression profiles in different tissues and organs(immature bark,cotyledons,young flower,immature fruit,mature fruit,mature leaf,roots,upper stem,and lower stem)of C.acuminata.A total of 198 AP2/ERF genes were identified and divided into five relatively conserved subfamilies,including AP2(26 genes),DREB(61 genes),ERF(92 genes),RAV(18 genes),and Soloist(one gene).The combination of gene expression patterns in different C.acuminata tissues and organs,the phylogenetic tree,the co-expression analysis with biosynthetic genes,and the analysis of promoter sequences of key enzymes genes involved in CPT biosynthesis pathways revealed that eight AP2/ERF TFs in C.acuminata might be involved in CPT synthesis regulation,which exhibit relatively high expression levels in the upper stem or immature bark.Among these,four genes(Cac AP2/ERF123,Cac AP2/ERF125,Cac AP2/ERF126,and Cac AP2/ERF127)belong to the ERF–B2 subgroup;two genes(Cac AP2/ERF149 and Cac AP2/ERF152)belong to the ERF–B3 subgroup;and two more genes(Cac AP2/ERF095 and Cac AP2/ERF096)belong to the DREB–A6 subgroup.These results provide a foundation for future functional characterization of the AP2/ERF genes to enhance the biosynthesis of CPT compounds of C.acuminata.
基金This work was supported by the National Natural Science Foundation of China(31772176,31672038 and 31621064)and the National Key Research and Development Program of China(2017YFDO201900 and 2017YFD0200400).
文摘Sesquiterpenoids play an import role in the direct or indirect defense of plants.Farnesyl pyrophosphate synthases(FPSs)catalyze the biosynthesis of farnesyl pyrophosphate,which is a key precursor of farnesol and(E)-β-farnesene.In the current study,two FPS genes in Gossypium hirsutum,GhFPS1 and GhFPS2,were heterologously cloned and functionally characterized in a greenhouse setting.The open reading frames for full-length GhFPS1 and GhFPS2 were each 1029 nucleotides,and encoded two proteins of 342 amino acids with molecular weights of 39.4 kDa.The deduced amino acid sequences of GhFPS1–2 showed high identity to FPSs of other plants.Quantitative real-time PCR analysis revealed that GhFPS1 and GhFPS2 were highly expressed in G.hirsutum leaves,and were upregulated in methyl jasmonate(MeJA)-,methyl salicylate(MeSA)-and aphid infestation-treated cotton plants.The recombinant proteins of either GhFPS1 or GhFPS2 plus calf intestinal alkaline phosphatase could convert geranyl diphosphate(GPP)or isopentenyl diphosphate(IPP)to one major product,farnesol.Moreover,in electrophysiological response and Y-tube olfactometer assays,farnesol showed obvious attractiveness to female Aphidius gifuensis,which is an important parasitic wasp of aphids.Our findings suggest that two GhFPSs are involved in farnesol biosynthesis and they play a crucial role in indirect defense of cotton against aphid infestation.
基金supported by the National Natural Science Foundation of China(31872706)the National Key Research and Development Program of China(2019 YFD1000603).
文摘Zanthoxylum bungeanum is an economically important crop worldwide due to its high content of aroma-producing monoterpenoids,and development of varieties with enhanced flavor and overall quality is a crucial research area.However,the transcriptional regulatory mechanisms underlying monoterpenoid synthesis in Z.bungeanum remain unclear,hindering these breeding efforts.In this study,RNA sequencing,gas chromatography–mass spectrometry,and other molecular biology techniques were used to identify the underlying transcriptional regulation mechanisms.Two transcription factors,ZbbHLH2 and ZbERF6,were identified as key regulators of monoterpenoid synthesis in Z.bungeanum that upregulate various monoterpenoid synthesis-associated genes and are novel transcriptional activators of ZbIDI,which encodes the rate-limiting enzyme in plant monoterpenoid synthesis.Functional analysis revealed that the expression of three genes[1]modulates monoterpenoid accumulation in Z.bungeanum peel.These findings provide novel insights into the metabolic regulatory network of monoterpenoid synthesis in Z.bungeanum peel,offer potential strategies for the biofortification of specific monoterpenoids,and will promote the development of Z.bungeanum germplasm for targeted breeding and quality improvement.
基金This work was supported by the National Key R&D Program of China(2020YFA0908000)the National Natural Science Foundation of China(81773830)+1 种基金the Key Project at central government level:The ability establishment of sustainable use for valuable Chinese medicine resources(2060302-1806-03)National Program for Special Support of Eminent Professionals.
文摘2-Oxoglutarate(2OG)-dependent dioxygenases(2-ODDs)are omnipresent iron-containing non-heme enzymes that catalyze various oxidation-reduction reactions in plant growth and development,nucleic acid modification and secondary metabolism.We systematically summarized recent research on the oxidative modifications of plant 2-ODDs and related enzymes,their vital importance in the biosynthesis of plant special metabolites,and their catalytic specificity/flexibility,and discussed the potential of 2-ODD as a new approach for the identification of pivotal genes and the elucidation of biosynthetic pathway.
基金supported by the National Natural Science Foundation of China(Nos.30960032 and 31000117)the Major Technology Project of Hainan(No.ZDZX2013023-1)+2 种基金the National Nonprofit Institute Research Grants(Nos.CATAS-ITBB 110507 and CATAS-ITBB130305)the Fundamental Scientific Research Funds for Chinese Academy of Tropical Agricultural Sciences(No.1630052013009)the Natural Science Foundation of Hainan Province(No.313077),China
文摘Lipid biosynthesis is essential for eukaryotic cells, but the mechanisms of the process in microalgae remain poorly understood. Phosphatidic acid phosphohydrolase or 3-sn-phosphatidate phosphohydrolase(PAP) catalyzes the dephosphorylation of phosphatidic acid to form diacylglycerols and inorganic orthophosphates. This reaction is integral in the synthesis of triacylglycerols. In this study, the mRNA level of the PAP isoform CrPAP2 in a species of Chlamydomonas was found to increase in nitrogen-free conditions. Silencing of the CrPAP2 gene using RNA interference resulted in the decline of lipid content by 2.4%–17.4%. By contrast, over-expression of the CrPAP2 gene resulted in an increase in lipid content by 7.5%–21.8%. These observations indicate that regulation of the CrPAP2 gene can control the lipid content of the algal cells. In vitro CrPAP2 enzyme activity assay indicated that the cloned CrPAP2 gene exhibited biological activities.
文摘In this study we designed a novel,cost‐efficient and green method for the synthesis of copper nanoparticles(Cu NPs)supported on manganese dioxide(MnO2)NPs,using Centella asiatica L.leaf extract as a naturally‐sourced reducing agent,without stabilizers or surfactants.This synthetic process is environmentally‐friendly and avoids the use of toxic reducing agents.Phenolic hydroxyl groups in the leaf extract are believed to reduce Cu2+in solution to generate Cu NPs that are subsequently stabilized on the MnO2NP surfaces.The resulting Cu/MnO2nanocomposite was fully characterized using X‐ray diffraction,transmission electron microscopy,field emission scanning electron microscopy,energy‐dispersive X‐ray spectroscopy and Fourier transform infrared spectroscopy.This material was found to function as a highly active,efficient and recyclable heterogeneous catalyst for the reduction of Congo red,rhodamine B and methylene blue as well as nitro compounds such as2,4‐dinitrophenylhydrazine and4‐nitrophenol in the presence of NaBH4in aqueous media at ambient temperature.The high stability of the Cu/MnO2nanocomposite also allows the catalyst to be separated and reused several times without any significant loss of activity.?2018,Dalian Institute of Chemical Physics,Chinese Academy of Sciences.Published by Elsevier B.V.All rights reserved.
基金financial support of the National Natural Science Foundation of China(82370532,82341219)Shanghai Hospital Development Center Foundation(SHDC12022118)。
文摘YdjC chitooligosaccharide deacetylase homolog(YDJC)has been identified as a susceptibility gene for inflammatory bowel disease(IBD),yet its role in the pathogenesis of IBD,particularly in regulating immune responses in the gut mucosa,remains elusive.In this study,we demonstrated that YDJC expression is downregulated in inflamed mucosa,particularly in the CD4^(+)T cells of IBD patients,and that Ydjc deficiency promotes CD4^(+)T-cell proliferation and Th1 cell differentiation,thereby exacerbating acute and chronic colitis in mice.Integrative transcriptomic,proteomic,and metabolomic analyses revealed that Ydjc^(-/-)CD4^(+)T cells exhibit upregulated SREBP2-mediated cholesterol biosynthesis.Consistently,treatment with key enzyme inhibitors targeting cholesterol biosynthesis,including simvastatin,fatostatin,and AAV-sh-Srebf2,markedly suppressed CD4^(+)T-cell proliferation and Th1 cell differentiation,thereby alleviating colitis in Ydjc^(-/-)mice.Mechanistically,YDJC directly deacetylates SREBP2,which further suppresses downstream target gene expression(e.g.,Hmgcr,Hmgcs1,and Cyp51).Therefore,our findings elucidate a novel mechanism whereby YDJC restrains intestinal mucosal inflammation by downregulating SREBP2-driven Th1 cell differentiation,suggesting that targeting YDJC and SREBP2-mediated cholesterol biosynthesis may serve as promising therapeutic strategies for IBD.
