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3. A redundant transcription factor network steers spatiotemporal Arabidopsis triterpene synthesis

Author

European Commission, Ghent University, Flemish Government, Research Foundation - Flanders, Swiss National Science Foundation, China Scholarship Council, John Innes Foundation, Nguyen, Trang Hieu [0000-0003-1051-8068], Thiers, Louis [0000-0002-8650-508X], Van Moerkercke, Alex [0000-0002-6393-4754], Bai, Yuechen [0000-0002-2314-8660], Fernández-Calvo, Patricia [0000-0002-1576-9651], Minne, Max [0000-0003-3089-2284], Depuydt, Thomas [0000-0001-5889-3962], Colinas, Maite [0000-0001-7053-2983], Verstaen, Kevin [0000-0003-3582-0298], Van Isterdael, Gert [0000-0001-6626-1316], Nützmann, Hans-Wilhelm [0000-0002-6855-7556], Osbourn, Anne [0000-0003-2195-5810], Saeys, Yvan [0000-0002-0415-1506], De Rybel, Bert [0000-0002-9551-042X], Vandepoele, Klaas [0000-0003-4790-2725], Ritter, Andrés [0000-0001-7011-6824], Goossens, Alain [0000-0002-1599-551X], Nguyen, Trang Hieu, Thiers, Louis, Van Moerkercke, Alex, Bai, Yuechen, Fernández-Calvo, Patricia, Minne, Max, Depuydt, Thomas, Colinas, Maite, Verstaen, Kevin, Van Isterdael, Gert, Nützmann, Hans-Wilhelm, Osbourn, Anne, Saeys, Yvan, De Rybel, Bert, Vandepoele, Klaas, Ritter, Andrés, Goossens, Alain, European Commission, Ghent University, Flemish Government, Research Foundation - Flanders, Swiss National Science Foundation, China Scholarship Council, John Innes Foundation, Nguyen, Trang Hieu [0000-0003-1051-8068], Thiers, Louis [0000-0002-8650-508X], Van Moerkercke, Alex [0000-0002-6393-4754], Bai, Yuechen [0000-0002-2314-8660], Fernández-Calvo, Patricia [0000-0002-1576-9651], Minne, Max [0000-0003-3089-2284], Depuydt, Thomas [0000-0001-5889-3962], Colinas, Maite [0000-0001-7053-2983], Verstaen, Kevin [0000-0003-3582-0298], Van Isterdael, Gert [0000-0001-6626-1316], Nützmann, Hans-Wilhelm [0000-0002-6855-7556], Osbourn, Anne [0000-0003-2195-5810], Saeys, Yvan [0000-0002-0415-1506], De Rybel, Bert [0000-0002-9551-042X], Vandepoele, Klaas [0000-0003-4790-2725], Ritter, Andrés [0000-0001-7011-6824], Goossens, Alain [0000-0002-1599-551X], Nguyen, Trang Hieu, Thiers, Louis, Van Moerkercke, Alex, Bai, Yuechen, Fernández-Calvo, Patricia, Minne, Max, Depuydt, Thomas, Colinas, Maite, Verstaen, Kevin, Van Isterdael, Gert, Nützmann, Hans-Wilhelm, Osbourn, Anne, Saeys, Yvan, De Rybel, Bert, Vandepoele, Klaas, Ritter, Andrés, and Goossens, Alain

Abstract

Plant specialized metabolites modulate developmental and ecological functions and comprise many therapeutic and other high-value compounds. However, the mechanisms determining their cell-specific expression remain unknown. Here we describe the transcriptional regulatory network that underlies cell-specific biosynthesis of triterpenes in Arabidopsis thaliana root tips. Expression of thalianol and marneral biosynthesis pathway genes depends on the phytohormone jasmonate and is limited to outer tissues. We show that this is promoted by the activity of redundant bHLH-type transcription factors from two distinct clades and coactivated by homeodomain factors. Conversely, the DOF-type transcription factor DAG1 and other regulators prevent expression of the triterpene pathway genes in inner tissues. We thus show how precise expression of triterpene biosynthesis genes is determined by a robust network of transactivators, coactivators and counteracting repressors.

Published
2023

7. A redundant transcription factor network steers spatiotemporal Arabidopsistriterpene synthesis

Author

Nguyen, Trang Hieu, Thiers, Louis, Van Moerkercke, Alex, Bai, Yuechen, Fernández-Calvo, Patricia, Minne, Max, Depuydt, Thomas, Colinas, Maite, Verstaen, Kevin, Van Isterdael, Gert, Nützmann, Hans-Wilhelm, Osbourn, Anne, Saeys, Yvan, De Rybel, Bert, Vandepoele, Klaas, Ritter, Andrés, and Goossens, Alain

Abstract

Plant specialized metabolites modulate developmental and ecological functions and comprise many therapeutic and other high-value compounds. However, the mechanisms determining their cell-specific expression remain unknown. Here we describe the transcriptional regulatory network that underlies cell-specific biosynthesis of triterpenes in Arabidopsis thalianaroot tips. Expression of thalianol and marneral biosynthesis pathway genes depends on the phytohormone jasmonate and is limited to outer tissues. We show that this is promoted by the activity of redundant bHLH-type transcription factors from two distinct clades and coactivated by homeodomain factors. Conversely, the DOF-type transcription factor DAG1 and other regulators prevent expression of the triterpene pathway genes in inner tissues. We thus show how precise expression of triterpene biosynthesis genes is determined by a robust network of transactivators, coactivators and counteracting repressors.

Published
2023
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8. Modulation of Arabidopsis root growth by specialized triterpenes

Author

Bai, Yuechen, primary, Fernández‐Calvo, Patricia, additional, Ritter, Andrés, additional, Huang, Ancheng C., additional, Morales‐Herrera, Stefania, additional, Bicalho, Keylla U., additional, Karady, Michal, additional, Pauwels, Laurens, additional, Buyst, Dieter, additional, Njo, Maria, additional, Ljung, Karen, additional, Martins, José C., additional, Vanneste, Steffen, additional, Beeckman, Tom, additional, Osbourn, Anne, additional, Goossens, Alain, additional, and Pollier, Jacob, additional

Published
2021
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9. The transcription factor emission of methyl anthranilate 1 stimulates emission of methyl anthranilate from hairy roots

Author

Pollier, Jacob, De Geyter, Nathan, Moses, Tessa, Boachon, Benoit, Franco‐Zorrilla, José M., Bai, Yuechen, Lacchini, Elia, Gholami, Azra, Vanden Bossche, Robin, Werck Reichhart, Daniele, Goormachtig, Sofie, Goossens, Alain, Institut de biologie moléculaire des plantes (IBMP), and Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)

Subjects
[SDV.BV]Life Sciences [q-bio]/Vegetal Biology, ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2019

10. The MYB transcription factor emission of methyl anthranilate 1 stimulates emission of methyl anthranilate from Medicago truncatula hairy roots

Author

Agency for Innovation by Science and Technology (Belgium), China Scholarship Council, Ministry of Science, Research, and Technology (Iran), Research Foundation - Flanders, Pollier, Jacob, Geyter, Nathan De, Moses, Tessa, Boachon, Benoît, Franco-Zorrilla, José Manuel, Bai, Yuechen, Lacchini, Elia, Gholami, Azra, Vanden Bossche, Robin, Werck-Reichhart, Danièle, Goormachtig, Sofie, Goossens, Alain, Agency for Innovation by Science and Technology (Belgium), China Scholarship Council, Ministry of Science, Research, and Technology (Iran), Research Foundation - Flanders, Pollier, Jacob, Geyter, Nathan De, Moses, Tessa, Boachon, Benoît, Franco-Zorrilla, José Manuel, Bai, Yuechen, Lacchini, Elia, Gholami, Azra, Vanden Bossche, Robin, Werck-Reichhart, Danièle, Goormachtig, Sofie, and Goossens, Alain

Abstract

Plants respond to herbivore or pathogen attacks by activating specific defense programs that include the production of bioactive specialized metabolites to eliminate or deter the attackers. Volatiles play an important role in the interaction of a plant with its environment. Through transcript profiling of jasmonate-elicited Medicago truncatula cells, we identified Emission of Methyl Anthranilate (EMA) 1, a MYB transcription factor that is involved in the emission of the volatile compound methyl anthranilate when expressed in M. truncatula hairy roots, giving them a fruity scent. RNA sequencing (RNA-Seq) analysis of the fragrant roots revealed the upregulation of a methyltransferase that was subsequently characterized to catalyze the O-methylation of anthranilic acid and was hence named M. truncatula anthranilic acid methyl transferase (MtAAMT) 1. Given that direct activation of the MtAAMT1 promoter by EMA1 could not be unambiguously demonstrated, we further probed the RNA-Seq data and identified the repressor protein M. truncatula plant AT-rich sequence and zinc-binding (MtPLATZ) 1. Emission of Methyl Anthranilate 1 binds a tandem repeat of the ACCTAAC motif in the MtPLATZ1 promoter to transactivate gene expression. Overexpression of MtPLATZ1 in transgenic M. truncatula hairy roots led to transcriptional silencing of EMA1, indicating that MtPLATZ1 may be part of a negative feedback loop to control the expression of EMA1. Finally, application of exogenous methyl anthranilate boosted EMA1 and MtAAMT1 expression dramatically, thus also revealing a positive amplification loop. Such positive and negative feedback loops seem to be the norm rather than the exception in the regulation of plant specialized metabolism.

Published
2019

11. The MYB transcription factor Emission of Methyl Anthranilate 1 stimulates emission of methyl anthranilate from Medicago truncatula hairy roots

Author

Pollier, Jacob, primary, De Geyter, Nathan, additional, Moses, Tessa, additional, Boachon, Benoît, additional, Franco‐Zorrilla, José M., additional, Bai, Yuechen, additional, Lacchini, Elia, additional, Gholami, Azra, additional, Vanden Bossche, Robin, additional, Werck‐Reichhart, Danièle, additional, Goormachtig, Sofie, additional, and Goossens, Alain, additional

Published
2019
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14. A redundant transcription factor network steers spatiotemporal Arabidopsis triterpene synthesis

Author

Trang Hieu Nguyen, Louis Thiers, Alex Van Moerkercke, Yuechen Bai, Patricia Fernández-Calvo, Max Minne, Thomas Depuydt, Maite Colinas, Kevin Verstaen, Gert Van Isterdael, Hans-Wilhelm Nützmann, Anne Osbourn, Yvan Saeys, Bert De Rybel, Klaas Vandepoele, Andrés Ritter, Alain Goossens, European Commission, Ghent University, Flemish Government, Research Foundation Flanders, Swiss National Science Foundation, China Scholarship Council, John Innes Foundation, Nguyen, Trang Hieu, Thiers, Louis, Van Moerkercke, Alex, Bai, Yuechen, Fernández-Calvo, Patricia, Minne, Max, Depuydt, Thomas, Colinas, Maite, Verstaen, Kevin, Van Isterdael, Gert, Nützmann, Hans-Wilhelm, Osbourn, Anne, Saeys, Yvan, De Rybel, Bert, Vandepoele, Klaas, Ritter, Andrés, and Goossens, Alain

Subjects
Plant Science
Abstract

25 Pág., Plant specialized metabolites modulate developmental and ecological functions and comprise many therapeutic and other high-value compounds. However, the mechanisms determining their cell-specific expression remain unknown. Here we describe the transcriptional regulatory network that underlies cell-specific biosynthesis of triterpenes in Arabidopsis thaliana root tips. Expression of thalianol and marneral biosynthesis pathway genes depends on the phytohormone jasmonate and is limited to outer tissues. We show that this is promoted by the activity of redundant bHLH-type transcription factors from two distinct clades and coactivated by homeodomain factors. Conversely, the DOF-type transcription factor DAG1 and other regulators prevent expression of the triterpene pathway genes in inner tissues. We thus show how precise expression of triterpene biosynthesis genes is determined by a robust network of transactivators, coactivators and counteracting repressors., This Article was written in loving memory of A. Van Moerkercke (1979–2021). The authors thank A. Bleys for critically reading the manuscript; D. Gasperini for kindly sharing the ProMYCs:NLS-VENUS reporter lines, and P. Vittorioso for the dag1 mutant, ProDAG1:GUS and DAG1 over-expressing lines; J. R. Wendrich and T. Eekhout for assistance in the launching and analysis of the scRNAseq experiment; and S. Desmet and G. Goeminne from the VIB Metabolomics Core – Ghent for the thalianol profiling. This work was supported by the European Community’s Seventh Framework Program (FP7/2007–2013) under grant agreement 613692-TriForC and H2020 Program under grant agreement 760331-Newcotiana to A.G.; the Special Research Fund from Ghent University to A.G. and A.R. (project BOF18/GOA/013), and M.M. (project BOF20/GOA/012); the Flemish Government (AI Research program) to Y.S.; the Research Foundation Flanders with research project grants to A.G. (G004515N and G008417N) and a postdoctoral fellowship to P.F.-C.; a Swiss National Science Foundation postdoctoral fellowship (P300PA_177831) to M.C.; and a China Scholarship Council PhD scholarship to Y.B. A.O. acknowledges funding support from the John Innes Foundation and the BBSRC Institute Strategic Program Grant ‘Molecules from Nature – Products and Pathways’ (BBS/E/J/000PR9790).

Published
2023

15. [Molecular cloning and prokaryotic expression of phenylalanine ammonia-lyase gene FdPAL from Fagopyrum dibotrys].

Author

Li C, Feng Z, Bai Y, Chen H, Zhao H, and Wu Q

Subjects
Amino Acid Sequence, Base Sequence, Cloning, Molecular, Escherichia coli genetics, Escherichia coli metabolism, Fagopyrum classification, Molecular Sequence Data, Phenylalanine Ammonia-Lyase chemistry, Phenylalanine Ammonia-Lyase metabolism, Phylogeny, Protein Conformation, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Fagopyrum genetics, Phenylalanine Ammonia-Lyase genetics
Abstract

Objective: To clone and characterize the DNA and cDNA sequences of phenylalanine ammonia-lyase gene (PAL) from Fagopyrum dibotrys, and investigate the biological activity of the obtained PAL., Method: Using homology cloning and RT-PCR techniques, the DNA and full-length cDNA sequences of PAL gene were amplified from F. dibotrys. The obtained sequences were analyzed by bioinformatics software. The ORF of PAL gene was cloned into expression vector pET-30b(+) and transformed into Escherichia coli BL21 (DE3) for expression the recombined protein. The catalytic activity of the recombined protein was determined by Spectrophotometer and thin layer chromatography (TLC) methods., Result: The DNA sequence of PAL gene (designated as FdPAL, GenBank accession number: HM628904) was 2 583 bp in size, of which consisted two extrons and a single intron, and the full-length cDNA of FdPAL was 2 169 bp in size, which contained an ORF. The deduced protein of FdPAL contained 722 amino acids with calculated molecular weight (MW) of 78.31 kDa and an isoelectric point (pI) of 5.94. The SDS-PAGE results showed that the molecular weight of recombinant FdPAL protein was 75.37 kDa, which is consistent with the predictions. After 4 hours of induction, the enzymatic specific activity of FdPAL reached the summit, up to 4 386 nmol x g(-1) x min(-1). The reaction products were also identified by TLC, using L-Phe and trans-cinnamic acid as the internal standard., Conclusion: The PAL gene (both DNA sequence and full-length cDNA sequence) was cloned from F. dibotrys, and it has the same classic characters as other PALs in plants. The recombinant FdPAL was efficiently expressed in E. coli and had the activity for catalyzing the conversion from L-phenylalanine to cinnamic acid.

Published
2011

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