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Engineering 2024年第4期35卷 201-213页

作者： Guangjin Li Zhanquan Zhang Yong Chen Tong Chen Boqiang Li Shiping Tian Key Laboratory of Plant Resources The Innovative Academy of Seed DesignInstitute of BotanyChinese Academy of SciencesBeijing 100093China University of Chinese Academy of Sciences Beijing 100049China

Light is a fundamental environmental factor for living organisms on earth—not only as a primary energy source but also as an informational signal.In fungi,light can be used as an indicator for both time and space to control important physiological and morphological responses.Botrytis cinerea(B.cinerea)is a devastating phytopathogenic fungus that exploits light cues to optimize virulence and the balance between conidiation and sclerotia development,thereby improving its dispersal and survival in ecosystems.However,the components and mechanisms underlying these processes remain obscure.Here,we identify a novel light-signaling component in B.cinerea,BcCfaS,which encodes a putative cyclopropane fatty-acyl-phospholipid synthase.BcCfaS is strongly induced by light at the transcriptional level and plays a crucial role in regulating photomorphogenesis.Deletion of BcCfaS results in reduced vegetative growth,altered colony morphology,impaired sclerotial development,and enhanced conidiation in a lightdependent manner.Moreover,the mutant exhibits serious defects in stress response and virulence on the host.Based on a lipidomics analysis,a number of previously unknown fungal lipids and many BcCfaS-regulated lipids are identified in B.cinerea,including several novel phospholipids and fatty acids.Importantly,we find that BcCfaS controls conidiation and sclerotial development by positively regulating methyl jasmonate(MeJA)synthesis to activate the transcription of light-signaling components,revealing for the first time the metabolic base of photomorphogenesis in fungi.Thus,we propose that BcCfaS serves as an integration node for light and lipid metabolism,thereby providing a regulatory mechanism by which fungi adapt their development to a changing light environment.These new findings provide an important target for antifungal design to prevent and control fungal disease.

关键词： Light Botrytis cinerea Cyclopropane fatty-acyl-phospholipid synthase Lipid metabolism Photomorphogenesis Virulence

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Protective Effect of Naringenin on Acute Myocardial Ischemia-reperfusion Injury in Rats

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Medicinal Plant 2024年第3期15卷 50-52页

作者： Xia ZHANG Ping ZHOU Juan LI Zhaojun XIANG Qianqian LUO Qing DENG Department of Neurosurgery Taihe Hospital of Shiyan CityHubei University of MedicineShiyan 442000China Gynecology and Obstetrics Taihe HospitalHubei University of MedicineShiyan 442000China

[Objectives]To investigate the protective mechanism of naringenin on acute myocardial ischemia-reperfusion injury(AMI-RI)in Sprague-Dawley(SD)rats.[Methods]A total of 32 SD rats with AMI-RI model construction were randomly divided into AMI-RI model control group and citrus pigment A/B/C groups(n=8).The naringenin A,B,and C groups were administrated 20,40 and 80 mg/(kg•d)for 10 d.The AMI group served as the negative control and was not treated.At the conclusion of the treatment regimen,a sample of intraventricular blood was collected for the purpose of measuring lactate dehydrogenase(LDH),glutathione peroxidase(GLH-PX),nitric oxide(NO),and superoxide dismutase(SOD)levels.Additionally,myocardial tissue was identified within the ischemic region.The content of malondialdehyde(MDA)was determined by inducing nitric oxide synthase(iNOS)and endodermal nitric oxide synthase(eNOS)positive cells in the left anterior descending coronary artery.[Results]Following citrus treatment,the contents of GLH-PX and SOD in ventricular blood of the citrus B group were found to be significantly elevated,while the contents of NO and LDH in myocardial MDA and ventricle were observed to be significantly reduced.The number of eNOS-positive cells was significantly increased,while the number of iNOS-positive cells was significantly decreased.The difference was statistically significant when compared with the AMI-RI group(P<0.05).The changes observed in the above indicators in the citrus C group were more pronounced than those observed in the citrus B group.The difference between the citrus C and the B group was statistically significant(P<0.05),indicating that this effect is concentration dependent.[Conclusions]In addition to its ability to inhibit myocardial lipid peroxidation during AMI-RI by increasing SOD activity,naringenin may also affect the synthesis and release of NO by regulating eNOS and iNOS,thereby achieving protection against AMI-RI.One effect is enhanced as the dose of the drug increases.

关键词： Rat Naringenin Acute myocardial ischemia-reperfusion Lipid peroxidation Inducible/endothelial nitric oxide synthase

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Molecular cloning, characterization, and promoter analysis of the isochorismate synthase （AalCS1） gene from Artemisia annua

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2017年第8期18卷 662-673页

作者： Lu-yao WANG Ying ZHANG Xue-qing FU Ting-ting ZHANG Jia-wei MA Li-da ZHANG Hong-mei QIAN Ke-xuan TANG Shan LI Jing-ya ZHAO Plant Bioteehnology Research Center Fudan-SJTU-Nottingham Plant Biotechnology R&D Center School of Agriculture and Biology Shanghai Jiao Tong University Shanghai 200240 China School of Bioscience and Bioengineering South China University of Technology Guangzhou 510006 China

Isochorismate synthase （ICS） is a crucial enzyme in the salicylic acid （SA） synthesis pathway. Thefull-length complementary DNA （cDNA） sequence of the ICS gene was isolated from Artemisia annua L. The gene,named AalCS1, contained a 1710-bp open reading frame, which encoded a protein with 570 amino acids. Bioinfor-matics and comparative study revealed that the polypeptide protein of AalCS1 had high homology with ICSs from otherplant species. Southern blot analysis suggested that AalCS1 might be a single-copy gene. Analysis of the 1470-bppromoter of AalCS1 identified distinct cis-acting regulatory elements, including TC-rich repeats, MYB binding site（MBS）, and TCA-elements. An analysis of AalCS1 transcript levels in multifarious tissues of A. annua using quantita-tive real-time polymerase chain reaction （qRT-PCR） showed that old leaves had the highest transcription levels.AalCS1 was up-regulated under wounding, drought, salinity, and SA treatments. This was corroborated by thepresence of the predicted cis-acting elements in the promoter region of AalCSI. Overexpressing transgenic plants andRNA interference transgenic lines of AalCSl were generated and their expression was compared. High-performanceliquid chromatography （HPLC） results from leaf tissue of transgenic A. annua showed an increase in artemisinincontent in the overexpressing plants. These results confirm that AalCS1 is involved in the isochorismate pathway.

关键词： Salicylic acid Artemisia annua L. Quantitative real-time polymerase chain reaction (qRT-PCR) Isochorismate synthase

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Drosophila CTP synthase can form distinct substrate-and product-bound filaments

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Journal of Genetics and Genomics 2019年第11期46卷 537-545页

作者： Xian Zhou Chen-Jun Guo Huan-Huan Hu Jiale Zhong Qianqian Sun Dandan Liu Shuang Zhou Chia Chun Chang Ji-Long Liu School of Life Science and Technology ShanghaiTech UniversityShanghai201210China Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai200031China University of Chinese Academy of Sciences Beijing100049China iHuman Institute ShanghaiTech UniversityShanghai201210China

Intracellular compartmentation is a key strategy for the functioning of a cell.In 2010,several studies revealed that the metabolic enzyme CTP synthase(CTPS)can form filamentous structures termed cytoophidia in prokaryotic and eukaryotic cells.However,recent structural studies showed that CTPS only forms inactive product-bound filaments in bacteria while forming active substrate-bound filaments in eukaryotic cells.In this study,using negative staining and cryo-electron microscopy,we demonstrate that Drosophila CTPS,whether in substrate-bound or product-bound form,can form filaments.Our results challenge the previous model and indicate that substrate-bound and product-bound filaments can coexist in the same species.We speculate that the ability to switch between active and inactive cytoophidia in the same cells provides an additional layer of metabolic regulation.

关键词： CTP synthase Drosophila Cytoophidium Cryo-EM

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Trehalose phosphate synthase 5-dependent trehalose metabolism modulates basal defense responses in Arabidopsis thaliana

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Journal of Integrative Plant Biology 2019年第4期61卷 509-527页

作者： Xuelan Wang Yan Du Diqiu Yu Key Laboratory of Tropical Plant Resources and Sustainable Use Xishuangbanna Tropical Botanical Garden Chinese Academy of Sciences University of Chinese Academy of Sciences

Despite the recent discovery that trehalose synthesis is important for plant development and abiotic stress tolerance, the effects of trehalose on biotic stress responses remain relatively unknown. In this study, we demonstrate that TREHALOSE PHOSPHATE synthase 5(TPS5)-dependent trehalose metabolism regulates Arabidopsis thaliana defenses against pathogens(necrotrophic Botrytis cinerea and biotrophic Pseudomonas syringae).Pathogen infection increased trehalose levels and upregulated TPS5 expression. Application of exogenous trehalose significantly improved plant defenses against B. cinerea, but increased the susceptibility of plants to P. syringae. We demonstrate that elevated trehalose biosynthesis, in transgenic plants over-expressing TPS5, also increased the susceptibility to P. syringae, but decreased the disease elcit r symptoms caused by B. cinerea. The knockout of TPS5 prevented the accumulation of trehalose and enhanced defense responses against P. syringae. Additionally, we observed that a TPS5-interacting protein(multiprotein bridging factor 1c) was required for induced expression of TPS5 during pathogen infections. Furthermore, we show that trehalose promotes P. syringae growth and disease development, via a mechanism involving suppression of the plant defense gene, Pathogenesis-Related Protein 1. These findings provide insight into the function of TPS5-dependent trehalose metabolism in plant basal defense responses.

关键词： Trehalose phosphate synthase Arabidopsis thaliana^FA

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Post-translational modifications of prostaglandin-endoperoxide synthase 2 in colorectal cancer:An update

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World Journal of Gastroenterology 2018年第48期24卷 5454-5461页

作者： Rafael I Jaén Patricia Prieto Marta Casado Paloma Martín-Sanz Lisardo Boscá Department of Metabolism and Physiopathology of Inflammatory Diseases Instituto de Investigaciones Biomédicas Alberto Sols (CSIC-UAM) Department of Biomedicine Instituto de Biomedicina de Valencia (CSIC) Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares y Hepáticas y DigestivasISCⅢ Unidad Asociada IIBMULPGC Universidad de las Palmas de Gran Canaria (ULPGC)

The biosynthesis of prostanoids is involved in both physiological and pathological processes. The expression of prostaglandin-endoperoxide synthase 2(PTGS2; also known as COX-2) has been traditionally associated to the onset of several pathologies, from inflammation to cardiovascular, gastrointestinal and oncologic events. For this reason, the search of selective PTGS2 inhibitors has been a focus for therapeutic interventions. In addition to the classic non-steroidal anti-inflammatory drugs, selective and specific PTGS2 inhibitors, termed coxibs, have been generated and widely used. PTGS2 activity is less restrictive in terms of substrate specificity than the homeostatic counterpart PTGS1, and it accounts for the elevated prostanoid synthesis that accompanies several pathologies. The main regulation of PTGS2 occurs at the transcription level. In addition to this, the stability of the mRNA is finely regulated through the interaction with several cytoplasmic elements, ranging from specificmicroR NAs to proteins that control mR NA degradation. Moreover, the protein has been recognized to be the substrate for several post-translational modifications that affect both the enzyme activity and the targeting for degradation via proteasomal and non-proteasomal mechanisms. Among these modifications, phosphorylation, glycosylation and covalent modifications by reactive lipidic intermediates and by free radicals associated to the proinflammatory condition appear to be the main changes. Identification of these post-translational modifications is relevant to better understand the role of PTGS2 in several pathologies and to establish a correct analysis of the potential function of this protein in diseases progress. Finally, these modifications can be used as biomarkers to establish correlations with other parameters, including the immunomodulation dependent on molecular pathological epidemiology determinants, which may provide a better frame for potential therapeutic interventions.

关键词： Prostaglandins Prostaglandin-endoperoxide synthase 2 Post-translational modifications Glycosylation Colorectal cancer Inflammation

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Calcium Receptor and Nitric Oxide synthase Expression in Circular Muscle of Lower Esophagus from Patients with Achalasia

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Chinese Medical Journal 2018年第23期131卷 2882-2885页

作者： Yang Gao Jun-Feng Liu Xin He Xin-Bo Liu Ling-Ling Zhang Lian-Mei Zhao Chao Zhang Department of Thoracic Surgery The Fourth Hospital of Hebei Medical UniversityShijiazhuangHebei 050011China Graduate School Hebei Medical UniversityShijiazhuangHebei 050017China Department of Pathology The Fourth Hospital of Hebei Medical UniversityShijiazhuangHebei 050011China Research Center The Fourth Hospital of Hebei Medical UniversityShijiazhuangHebei 050011China

To the Editor:Achalasia is a primary esophageal motility disorder.[l] The etiology ofachalasia remains unknown,but it is associated with degeneration,reduction,or absence of neurons in the esophageal muscular layer (A... 详细信息

关键词： Nitric Oxide synthase Expression Achalasia

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Cloning,expression,purification and bioinformatic analysis of 2-methylcitrate synthase from Mycobacterium tuberculosis

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Asian Pacific Journal of Tropical Medicine 2015年第1期8卷 19-23页

作者： Kasamy Eniyan Urmi Bajpai Department of Biomedical Science Acharya Narendra Dev College(University of Delhi)

Objective:To clone,express and purify 2—methylcitrale synthase(Rv1131) gene of Mycobacterium tuberculosis(M.tuberculosis) and to study its structural characteristics using various bioinformatics tools.Methods:Rv1131 gene was amplified In polymerase chain reaction using M.tuberculosis H37 Rv genomic DNA and cloned into pGEM-T easy vector and sequenced.The gene was sub-cloned in pET28 c vector,expressed in Escherichia coli RL22(E.coli BL21)(DE3) cells and the recombinant protein was identified by Western blotting.The protein was purified using Nickel affinity chromatography and the structural characteristics like sub-cellular localization.presence of transmembrane helices and secondary structure of the protein were predicted by bioinformaties tools.Tertiary structure of the protein and phylogenetic analysis was aiso established by in silico analysis.Results:The expression of the recombinant protein(Rv1131) was confirmed by western blotting using anti-HIS antibodies and the protein was purified from the soluble fraction.In silico analysis showed that the protein contains no signal peptide and transmembrane helices,Active site prediction showed that the protein has histidine and aspartie acid residues at 242.281 & 332 positions respectively.Phylogenetic analysis showed100%homology with major mycobacterial species.Secondary structure predicts 2-methyleitrate synthase contain 51.9%;alpha-helk.8.7%extended strand and 39.4%random coils.Tertiary structure of the protein was also established.Conclusions:The enzyme 2-methylcitrate synthase from M.tuberculosis H37 Rv has been successfully expressed and purified.The purified protein will further be utilized to develop assay methods lor screening new inhibitors.

关键词： Mycobacterium tuberculosis 2-methylcitrate synthase Propionyl-CoA Expression Purification

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Homologous and heterologous interactions between catalytic and regulatory subunits of Escherichia coli acetohydroxyacid synthaseⅠand Ⅲ

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Science China Chemistry 2009年第9期52卷 1362-1371页

作者： NIU CongWei, FENG Wei, ZHOU YanFei, WEN Xin & XI Zhen State Key Laboratory of Elemento-Organic Chemistry and Department of Chemical Biology, Nankai University, Tianjin 300071, China State Key Laboratory of Elemento-Organic Chemistry and Department of Chemical Biology Nankai University Tianjin China

Homologous and heterologous interactions between acetohydroxyacid synthase (AHAS) I and Ⅲ from E. coli have been studied by surface plasmon resonance (SPR). The catalytic and regulatory subunits association for AHAS I (KD = 1.13 × 10-7 M) was stronger than that for AHAS Ⅲ (KD = 5.29 × 10-7 M). A strong heterologous association between regulatory and catalytic subunits and heterologous activation of catalytic subunits were observed. SPR results combined with enzyme kinetics indicate that the reconstituted heterologous enzymes had similar kinetic properties as homologous enzymes, implying that the regulatory subunit of AHAS I could be replaced by the regulatory subunit of AHAS Ⅲ and vice versa. This work may be useful to further understandings of the mechanism of regulation of AHAS.

关键词： acetohydroxyacid synthase subunit interaction surface plasmon resonance

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Suppression of starch synthase I(SSI) by RNA interference alters starch biosynthesis and amylopectin chain distribution in rice plants subjected to high temperature

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The Crop Journal 2019年第5期7卷 573-586页

作者： Qian Zhao Xiaoxia Du Zhanyu Han Yu Ye Gang Pan Muhammad-Asad-Ullah Asad Qifa Zhou Fangmin Cheng Institute of Crop Science College of Agriculture and BiotechnologyZhejiang UniversityHangzhou 310058ZhejiangChina Jiangsu Collaborative Innovation Center for Modern Crop Production Nanjing 210095JiangsuChina

Based on known cDNAs of rice starch synthase isoforms,we constructed dsRNA interference vectors for starch synthase I(SSI)to produce transgenic plants containing starch with a moderately high amylose content.We investigated the effect of SSI suppression on grain quality traits,starch biosynthesis,and amylopectin chain distribution in rice plants exposed to two different temperature regimes.The activities and transcripts of BEs,DBEs,and other SS isoforms were further investigated to clarify the effect of SSI suppression on these key enzymes and their specific isoforms under different temperature treatments.Suppression of SSI by RNAi altered grain starch component and amylopectin chain distribution,but it exerted only a slight effect on total starch content(%)and accumulation amount(mg kernel?1)and on starch granule morphology and particle size distribution.Under normal temperature(NT),insignificant differences in kernel weight,chalky kernel proportion,chalky degree,and starch granule morphology between SSI-RNAi line and its wild type(WT)were observed.However,amylose content(AC)level and granule-bound starch synthase(GBSS)activity in rice endosperms were markedly increased by SSI-RNAi suppression.The chalky kernel proportion and chalky degree of SSIRNAi lines were significantly higher than those of WT under high temperature(HT)exposure at filling stage.Inhibition of SSI by RNAi affected amylopectin chain distribution and raised starch gelatinization temperature(GT)in two ways:directly from the SSI deficiency itself and indirectly by reducing BEIIb amounts in an SSI-deficient background.The deficiency of SSI expression led to an alteration in the susceptibility of grain chalkiness occurrence and starch gelatinization temperature to HT exposure,owing to a pleiotropic effect of SSI deficiency on the expression of other genes associated with starch biosynthesis.

关键词： Rice Starch synthase I RNA interference Grain quality Amylopectin High temperature

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