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Analysis of the Interface between Primary and Secondary Metabolism in Catharanthus roseus Cell Cultures Using ^13C-Stable Isotope Feeding and Coupled Mass Spectrometry

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molecular plant 2013年第2期6卷 581-584页

作者： Carla Antonio Natali R. Mustafa Sonia Osorio Takayuki Tohge Patrick Giavalbco Lothar Willmitzer Heiko Rischer Kirsi-Marj Oksman-Caldentey Robert Verpoorte Alisdair R. Fernie Central Metabolism Group Max Planck Institute of Molecular Plant Physiology Am Muehlenberg 1 D-14476 Potsdam-Golm Germany Natural Products Laboratory Institute of Biology Leiden University Einsteinweg 55 2300 RA Leiden The Netherlands present address: Departamento de Biologia molecular y Bioquimica Universidad de M.~laga 29071 M=~laga Spain Genes and Small Molecules Group Max Planck Institute of MolecularPlant Physiology Am Muehlenberg 1 D-14476 Potsdam-Golm Germany VTT Technical Research Centre of Finland PO Box 1000 F1-02044 VTT (Espoo) Finland

Dear Editor, The tropical plant Madagascar periwinkle Catharanthus roseus （L.） *** is a rich source of plant-derived medicinal ter-penoid indole alkaloids （TIAs）, including the anti-hypertensive ajmalicine, the sedative compound serpentine, and the anti-cancer drugs vinblastine and vincristine. However, the latter two compounds are produced in C. roseus plants only in very low amounts. Elicitors such as hormones （e.g. jasmonates or salicylic acid） activate plant natural defense responses, includ-ing increased secondary metabolite production （EI-Sayed and Verpoorte, 2007; Lackman et al.. 2011）.

关键词： GAS-CHROMATOGRAPHY BIOSYNTHESIS ARABIDOPSIS

Responses of <i>Marsilea minuta</i>L. to Cadmium Stress and Assessment of Some Oxidative Biomarkers

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American Journal of plant Sciences 2014年第10期5卷 1467-1476页

作者： Kingsuk Das Chiranjib Mandal Nirmalya Ghosh Narottam Dey Malay Kumar Adak Department of biotechnology Visva Bharati Santiniketan India plant physiology and plant molecular Biology Research Unit Department of Botany University of Kalyani Kalyani India

In a hydroponic based experiment, the Cd toxicity is monitored with some cellular responses of Marsilea plant. Initially, plants were grown under varying concentrations (0, 50, 100 and 200 μM of Cd) of cadmium (Cd) with supplementation of 2 mM spermidine (Spd). The oxidative stress developed by Cd overaccumulation was measured with fall in Relative Growth Rate (RGR) by 27.11% to 59.83% growth reduction over control under varying Cd treatments. The retrieval of RGR was recovered by 1.59 folds as compared to the highest concentration of Cd (200 μM) when plants were fed with Spd. A concomitant degradation of chlorophyll was recorded in dose-dependant manner, however, the retrieval was not much pronounced with Spd. On the contrary, the non-oxidant thiol had borne more clarity with ongoing Cd concentration and appeared to be 40.51% increase maximally for GSH: GSSG at the highest concentration of Cd. Spd has minimized the ratio by 27.4%. The recovery of osmotic turgidity was indexed with a sharp rise in glycine betaine by 3.86 folds maximum at the highest concentration of Cd over control which declined by 30.9% with Spd. Another cellular response of treated plants was more evident from their isozymic profiles with regard to superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX). The intensity of protein expression was significantly variable but not in band numbers as evident from Cd treated plants. In vitro enzyme assay of catalase showed as declining trend within the limit of 33.13% to 43.22% which was reported by 1.45 folds when Spd was applied. Therefore, from the present study, the cellular responses of Marsilea plant which showed compatibility for their expression with Cd toxicity could be hypothesized as a case of bioindication.

关键词： Cadmium Toxicity Marsilea minuta Bioindicators

Anterograde and Retrograde Regulation of Nuclear Genes Encoding Mitochondrial Proteins during Growth, Development, and Stress

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molecular plant 2014年第7期7卷 1075-1093页

作者： Sophia Ng Inge De Clercqc Olivier Van Akena Simon R. Lawd Aneta Ivanovad Patrick Willems Estelle Giraud Frank Van Breusegem James Wheland ARC Centre of Excellence in plant Energy Biology University of Western Australia Australia Joint Research Laboratory in Genomics and Nutriomics College of Life Sciences Zhejiang University Hangzhou 310058 RR. China Department of plant Systems Biology VIB and Department of Plant Biotechnology and Bioinformatics Ghent UniversityTechnologiepark 927 B-9052 Gent Belgium Department of Botany ARC Centre of Excellence in Plant Energy Biology School of Life Science La Trobe University Bundoora 3086Victoria Australia Department of Medical Protein Research and Department of Biochemistry 9000 Ghent Belgium present address: Illumina ANZ 1 International Court Scoresby Victoria 3179 Australia

Mitochondrial biogenesis and function in plants require the expression of over 1000 nuclear genes encoding mitochondrial proteins （NGEMPs）. The expression of these genes is regulated by tissue-specific, developmental, internal, and external stimuli that result in a dynamic organelle involved in both metabolic and a variety of signaling processes. Although the metabolic and biosynthetic machinery of mitochondria is relatively well understood, the factors that regu- late these processes and the various signaling pathways involved are only beginning to be identified at a molecular level. The molecular components of anterograde （nuclear to mitochondrial） and retrograde （mitochondrial to nuclear） signaling pathways that regulate the expression of NGEMPs interact with chloroplast-, growth-, and stress-signaling pathways in the cell at a variety of levels, with common components involved in transmission and execution of these signals. This positions mitochondria as important hubs for signaling in the cell, not only in direct signaling of mitochondrial function per se, but also in sensing and/or integrating a variety of other internal and external signals. This integrates and optimizes growth with energy metabolism and stress responses, which is required in both photosynthetic and non-photosynthetic cells.

关键词： mitochondria mitochondrial retrograde regulation (MRR) organellar crosstalk signaling.

Rewilding staple crops for the lost halophytism:Toward sustainability and profitability of agricultural production systems

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molecular plant 2022年第1期15卷 45-64页

作者： Nishtha Rawat Silas Wungrampha Sneh L.Singla-Pareek Min Yu Sergey Shabala Ashwani Pareek Stress physiology and molecular Biology Lab School of Life SciencesJawaharlal Nehru UniversityNew Delhi 110067India plant Stress Biology International Centre for Genetic Engineering and BiotechnologyNew Delhi 110067India International Research Centre for Environmental Membrane Biology Foshan UniversityFoshan 528000China Tasmanian Institute for Agriculture University of TasmaniaHobart Tas 7001Australia National Agri-Food biotechnology institute Mohali 140306India

Abiotic stress tolerance has been weakened during the domestication of all major staple *** salinity is a major environmental constraint that impacts over half of the world population;however,given the increasing reliance on irrigation and the lack of available freshwater,agriculture in the 21st century will increasingly become ***,global food security is critically dependent on the ability of plant breeders to create high-yielding staple crop varieties that will incorporate salinity tolerance traits and ac-count for future climate ***,we have argued that the current agricultural practices and reliance on crops that exclude salt from uptake is counterproductive and environmentally unsustainable,and thus called for a need for a major shift in a breeding paradigm to incorporate some halophytic traits that were present in wild relatives but were lost in modern crops during *** this review,we provide a comprehensive physiological and molecular analysis of the key traits conferring crop halophy-tism,such as vacuolar Na+sequestration,ROS desensitization,succulence,metabolic photosynthetic switch,and salt deposition in trichomes,and discuss the strategies for incorporating them into elite germ-plasm,to address a pressing issue of boosting plant salinity tolerance.

关键词： halophyte salinity salt sequestration tissue tolerance trichomes water use efficiency

Cadmium Accumulation in <i>Marsilea minuta</i>Linn. and Its Antioxidative Responses

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American Journal of plant Sciences 2013年第2期4卷 365-371页

作者： Kingsuk Das Chiranjib Mandal Nirmalya Ghosh Narottam Dey Malay Kumar Adak Department of biotechnology Visva Bharati Visvavidyalaya Santiniketan India plant physiology and plant molecular Biology Research Unit Department of Botany University of Kalyani Kalyani India

The present study was carried out to investigate the extent of cadmium (Cd) accumulation with its possible impact on physiological and biochemical basis of heavy metal tolerance in Marsilea minuta Linn. Cd salt (0 μM, 50 μM and 100 μM) was allowed to absorb by the plants for prolong days in hydroponic culture and a significant deterioration of the plant biomass was recorded. However, roots absorbed more metals than the leaves. plants recorded a significant rise of superoxide (O-2) and hydrogen peroxide (H2O2). A noticeable amount of protein oxidation and lipid peroxidation were in proportionate to Cd accumulation. Anthocyanin and flavonoid content were decreased as compared to control condition. Superoxide dismutase (SOD), guaiacol peroxidase (GPX) and glutathione reductase (GR) contributed their antioxidative functions according to the Cd doses. The expression of GR was also evident from its activity staining in gel. So, it may suggest that antioxidative enzymes are up regulated and likely to be responsible for tolerance to Cd induced oxidative stress in Marsilea minuta Linn.

关键词： Oxidative Stress Heavy Metals Aquatic Fern

MapMan4: A Refined Protein Classification and Annotation Framework Applicable to Multi-Omics Data Analysis

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molecular plant 2019年第6期12卷 879-892页

作者： Rainer Schwacke Gabriel Y.Ponce-Soto Kirsten Krause Anthony M.Bolger Borjana Arsova Asis Hallab Kristina Gruden Mark Stitt Marie E.Bolger Bjorn Usadel Institute for Bio- and Geosciences (IBG-2: plant Sciences) Forschungszentrum Julich Wilhelm Johnen StraBe Jiilich Germany Department of Arctic and Marine Biology The Arctic University of Norway Biology Building 9037 Tromso Norway Institute for Botany and molecular Genetics BioEconomy Science Center Worringer Weg RWTH Aachen University 52074 Aachen Germany National Institute of Biology Department of Biotechnology and Systems Biology Vedna Pot 111 1000 Ljubljana Slovenia Max Planck Institute for molecular plant physiology Department of Systems Regulation 14476 Potsdam-Golm Germany

Genome sequences from over 200 plant species have already been published, with this number expected to increase rapidly due to advances in sequencing technologies. Once a new genome has been assembled and the genes identified, the functional annotation of their putative translational products, proteins, using ontologies is of key importance as it places the sequencing data in a biological context. Furthermore, to keep pace with rapid production of genome sequences, this functional annotation process must be fully automated. Here we present a redesigned and significantly enhanced MapMan4 framework, together with a revised version of the associated online Mercator annotation tool. Compared with the original MapMan, the new ontology has been expanded almost threefold and enforces stricter assignment rules. This framework was then incorporated into Mercator4, which has been upgraded to reflect current knowledge across the land plant group, providing protein annotations for all embryophytes with a comparably high quality. The annotation process has been optimized to allow a plant genome to be annotated in a matter of minutes. The output results continue to be compatible with the established MapMan desktop application.

关键词： plant genomes Functional annotation MapMan Mercator Transcriptomes

Salt Stress Triggers Phosphorylation of the Arabidopsis Vacuolar K＋ Channel TPK1 by Calcium-Dependent Protein Kinases （CDPKs）

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molecular plant 2013年第4期6卷 1274-1289页

作者： Andreas Latz Norbert Mehlmer Simone Zapf Thomas D. Mueller Bernhard Wurzinger Barbara Pfister Edina Csaszar Rainer Hedrich Markus Teige Dirk Becker Department of molecular plant physiology and Biophysics Julius-von-Sachs-lnstitute University of Wurzburg Julius-von-Sachs-Platz 2 D-97082 Wurzburg Germany Department of Biochemistry and Cell Biology Max F. Perutz Laboratories University of Vienna Dr. Bohrgasse 9 A-1030 Vienna Austria present address: Department of Biology I Botany Munich Center for Integrated Protein Science CiPSM Ludwig-Maximilians-Universitat Munchen D-82152 Planegg-Martinsried Germany College of Science King Saud University Riyadh 11451 Saudi Arabia

14-3-3 proteins play an important role in the regulation of many cellular processes. The Arabidopsis vacuolar two-pore K＋ channel 1 （TPK1） interacts with the 14-3-3 protein GRF6 （GF14-λ）. Upon phosphorylation of the putative binding motif in the N-terminus of TPK1, GRF6 binds to TPK1 and activates the potassium channel. In order to gain a deeper understanding of this 14-3-3-mediated signal transduction, we set out to identify the respective kinases, which regulate the phosphorylation status of the 14-3-3 binding motif in TPK1. Here, we report that the calcium-dependent protein kinases （CDPKs） can phosphorylate and thereby activate the 14-3-3 binding motif in TPK1. Focusing on the stress-activated kinase CPK3, we visualized direct and specific interaction of TPK1 with the kinase at the tonoplast in vivo. In line with its proposed role in K＋ homeostasis, TPK1 phosphorylation was found to be induced by salt stress in planta, and both cpk3 and tpkl mutants displayed salt-sensitive phenotypes. molecular modeling of the TPK1-CPK3 interaction domain provided mechanistic insights into TPK1 stress-regulated phosphorylation responses and pinpointed two arginine residues in the N-terminal 14-3-3 binding motif in TPK1 critical for kinase interaction. Taken together, our studies provide evidence for an essential role of the vacuolar potassium channel TPK1 in salt-stress adaptation as a target of calcium-regulated stress signaling pathways involving Ca2＋, Ca2＋-dependent kinases, and 14-3-3 proteins.

关键词： potassium channel vacuole calcium calcium-dependent kinase 14-3-3 protein salt stress.

A plasma membrane nucleotide-binding leucinerich repeat receptor mediates the recognition of the Ralstonia pseudosolanacearum effector RipY in Nicotiana benthamiana

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plant Communications 2023年第6期4卷 313-328页

作者： Boyoung Kim Wenjia Yu Haseong Kim Qian Dong Sera Choi Maxim Prokchorchick Alberto PMacho Kee Hoon Sohn Cécile Segonzac Department of Agriculture Forestry and BioresourcesSeoul National UniversitySeoul 08826Republic of Korea plant Immunity Research Center Seoul National UniversitySeoul 08826Republic of Korea Shanghai Center for plant Stress Biology CAS Center for Excellence in Molecular Plant SciencesChinese Academy of SciencesShanghai 201602China Department of Life Sciences Pohang University of Science and TechnologyPohang 37673Republic of Korea plant Genomics and Breeding Institute Seoul National UniversitySeoul 08826Republic of Korea Research Institute of Agriculture and Life Sciences Seoul National UniversitySeoul 08826Republic of Korea Department of Agricultural biotechnology Seoul National UniversitySeoul 08826Republic of Korea present address:Institute of plant and Microbial Biology and Zürich-Basel plant Science Center University of Zürich8008 ZürichSwitzerland The Institute of Crop Science and Resource Conservation(INRES) University of BonnNussallee 953115 BonnGermany

Bacterial wilt disease caused by several Ralstonia species is one of the most destructive diseases in Solanaceae *** a few functional resistance genes against bacterial wilt have been cloned to ***,we showthat the broadly conserved typeⅢsecreted effector RipY is recognized by the Nicotiana benthamiana immune system,leading to cell death induction,induction of defense-related gene expression,and restriction of bacterial pathogen *** a multiplexed virus-induced gene-silencing-based *** nucleotide-binding and leucine-rich repeat receptor(NbNLR)library,we identified a coiled-coil(CC)nucleotide-binding and leucine-rich repeat receptor(CNL)required for recognition of RipY,which we named RESISTANCE TO RALSTONIA SOLANACEARUM RIPY(RRS-Y).Genetic complementation assays in RRS-Y-silenced plants and stable rrs-y knockout mutants demonstrated that RRS-Y is sufficient to activate RipY-induced cell death andRipY-induced immunity to Ralstonia ***-Y function is dependent on the phosphate-binding loop motif of the nucleotide-binding domain but independent of the characterized signaling components ENHANCED DISEASE SUSCEPTIBILITY 1,ACTIVATED DISEASE RESISTANCE 1,and N REQUIREMENT GENE 1 and the NLR helpers NB-LRR REQUIRED FOR HR-ASSOCIATED CELL DEATH-2,-3,and-4 in *** further show that RRS-Y localization at the plasma membrane is mediated by two cysteine residues in the CC domain and is required for RipY *** also broadly recognizes RipY homologs across Ralstonia ***,we show that the C-terminal region of RipY is indispensable for RRS-Y ***,our findings provide an additional effector/receptor pair system to deepen our understanding of CNL activation in plants.

关键词： typeⅢsecreted effector plant immune system nucleotide-binding leucine-rich repeat receptor virusinduced gene silencing

Systematic optimization of the yeast cell factory for sustainable and high efficiency production of bioactive ginsenoside compound K

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Synthetic and Systems biotechnology 2021年第2期6卷 69-76页

作者： Pingping Wang Jiali Wang Guoping Zhao Xing Yan Zhihua Zhou CAS-Key Laboratory of Synthetic Biology CAS Center for Excellence in Molecular Plant SciencesInstitute of Plant Physiology and EcologyChinese Academy of Sciences300 Fenglin RdShanghai200032China School of Life Sciences Henan UniversityKaifeng475001China

Ginsenoside Compound K(CK)has been recognized as a major functional component that is absorbed into the systemic circulation after oral administration of *** demonstrates diverse bioactivities.A phase I clinical study indicated that CK was a potential candidate for arthritis ***,a phase II clinical study was suspended because of the high cost associated with the present CK manufacturing approach,which is based on the traditional planting-extracting-biotransforming *** previously elucidated the complete CK biosynthetic pathway and realized for the first time de novo biosynthesis of CK from glucose by engineered ***,CK production was not sufficient for industrial ***,we systematically engineered Saccharomyces cerevisiae to achieve high titer production of CK from glucose using a previously constructed protopanaxadiol(PPD)-producing chassis,optimizing UGTPg1 expression,improving UDP-glucose biosynthesis,and tuning down UDP-glucose *** final engineered yeast strain produced CK with a titer of 5.74 g/L in fed-batch fermentation,which represents the highest CK production in microbes reported to *** scaled-up,this high titer de novo microbial biosynthesis platform will enable a robust and stable supply of CK,thus facilitating study and medical application of CK.

关键词： Compound K UDP-Glucose Glycosylation efficiency Cell factory Synthetic biology