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Special Issue "Metabolic Engineering of Plants"

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Plant Sciences".

Deadline for manuscript submissions: closed (31 December 2020).

Special Issue Editors

Prof. Dr. Maria Lourdes Gómez-Gómez
E-Mail Website
Guest Editor
Universidad de Castilla-La Mancha, Ciudad Real, Spain
Interests: apocarotenoids; carotenoids; carotenogenic enzymes; biotechnology; flavonoids; metabolism regulation; pigments
Special Issues and Collections in MDPI journals
Dr. Gianfranco Diretto
E-Mail Website
Guest Editor
ENEA Centro Ricerche Casaccia, Rome, Italy
Interests: apocarotenoids; carotenoids; phenylpropanoids; transcriptomics; metabolomics; systems biology; biological networks

Special Issue Information

Dear Colleagues,

Plants have the ability to use CO2 and the energy derived from the Sun to produce a highly diverse range of compounds. Therefore, plants are the most obvious platform for the sustainable generation of food, chemicals, and valuable materials. Metabolic engineering emerged and developed over the past 20 years as a field in which methodologies for the rational engineering of biological systems have been applied to specific industrial, medical, or scientific problems. Metabolic engineering in plants involves direct and indirect modifications of endogenous pathways to redirect one or more enzymatic reactions in order to produce new compounds or improve/retard the production of known compounds.

This Special Issue will focus on the recent advances and breakthroughs on the metabolic engineering of plants. We will notably focus on the powerful tools for the accurate analysis of plant metabolism, for fast and precise genome engineering, for the modification of specific biochemical reactions or the introduction of new ones, and for the design and creation of tailor-made non-natural enzymes and synthetic metabolic pathways. Successful examples of metabolic engineering strategies using synthetic biology tools will also be included.

Prof. Dr. Maria Lourdes Gómez-Gómez
Dr. Gianfranco Diretto
Guest Editors

Manuscript Submission Information

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

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Keywords

  • synthetic biology
  • secondary metabolites
  • biosynthetic modules
  • isolation of enzyme complexes
  • metabolic engineering
  • pathway flux analyses
  • storage
  • cell compartmentalization
  • photosynthesis
  • pathogen resistance
  • metabolomics
  • modelling
  • crop yield
  • fortification
  • plastids
  • bioenergy

Published Papers (6 papers)

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Research

Communication
Development of Atropa belladonna L. Plants with High-Yield Hyoscyamine and without Its Derivatives Using the CRISPR/Cas9 System
Int. J. Mol. Sci. 2021, 22(4), 1731; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22041731 - 09 Feb 2021
Viewed by 579
Abstract
Atropa belladonna L. is one of the most important herbal plants that produces hyoscyamine or atropine, and it also produces anisodamine and scopolamine. However, the in planta hyoscyamine content is very low, and it is difficult and expensive to independently separate hyoscyamine from [...] Read more.
Atropa belladonna L. is one of the most important herbal plants that produces hyoscyamine or atropine, and it also produces anisodamine and scopolamine. However, the in planta hyoscyamine content is very low, and it is difficult and expensive to independently separate hyoscyamine from the tropane alkaloids in A. belladonna. Therefore, it is vital to develop A. belladonna plants with high yields of hyoscyamine, and without anisodamine and scopolamine. In this study, we generated A. belladonna plants without anisodamine and scopolamine, via the CRISPR/Cas9-based disruption of hyoscyamine 6β-hydroxylase (AbH6H), for the first time. Hyoscyamine production was significantly elevated, while neither anisodamine nor scopolamine were produced, in the A. belladonna plants with homozygous mutations in AbH6H. In summary, new varieties of A. belladonna with high yields of hyoscyamine and without anisodamine and scopolamine have great potential applicability in producing hyoscyamine at a low cost. Full article
(This article belongs to the Special Issue Metabolic Engineering of Plants)
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Article
Metabolic Engineering of the Native Monoterpene Pathway in Spearmint for Production of Heterologous Monoterpenes Reveals Complex Metabolism and Pathway Interactions
Int. J. Mol. Sci. 2020, 21(17), 6164; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21176164 - 26 Aug 2020
Cited by 1 | Viewed by 880
Abstract
Spearmint produces and stores large amounts of monoterpenes, mainly limonene and carvone, in glandular trichomes and is the major natural source of these compounds. Towards producing heterologous monoterpenes in spearmint, we first reduced the flux into the native limonene pathway by knocking down [...] Read more.
Spearmint produces and stores large amounts of monoterpenes, mainly limonene and carvone, in glandular trichomes and is the major natural source of these compounds. Towards producing heterologous monoterpenes in spearmint, we first reduced the flux into the native limonene pathway by knocking down the expression of limonene synthase (MsLS) by RNAi method. The MsLS RNAi lines exhibited a huge reduction in the synthesis of limonene and carvone. Detailed GC-MS and LC-MS analysis revealed that MsLS RNAi plants also showed an increase in sesquiterpene, phytosterols, fatty acids, flavonoids, and phenolic metabolites, suggesting an interaction between the MEP, MVA shikimate and fatty acid pathways in spearmint. Three different heterologous monoterpene synthases namely, linalool synthase and myrcene synthase from Picea abies and geraniol synthase from Cananga odorata were cloned and introduced independently into the MsLS RNAi mutant background. The expression of these heterologous terpene synthases resulted mainly in production of monoterpene derivatives. Of all the introduced monoterpenes geraniol showed the maximum number of derivatives. Our results provide new insights into MEP pathway interactions and regulation and reveals the existence of mechanisms for complex metabolism of monoterpenes in spearmint. Full article
(This article belongs to the Special Issue Metabolic Engineering of Plants)
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Article
Plastid Glycerol-3-phosphate Acyltransferase Enhanced Plant Growth and Prokaryotic Glycerolipid Synthesis in Brassica napus
Int. J. Mol. Sci. 2020, 21(15), 5325; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21155325 - 27 Jul 2020
Viewed by 596
Abstract
Plastid-localized glycerol-3-phosphate acyltransferase (ATS1) catalyzes the first-step reaction in glycerolipid assembly through transferring an acyl moiety to glycerol-3-phosphate (G3P) to generate lysophosphatidic acid (LPA), an intermediate in lipid metabolism. The effect of ATS1 overexpression on glycerolipid metabolism and growth remained to be elucidated [...] Read more.
Plastid-localized glycerol-3-phosphate acyltransferase (ATS1) catalyzes the first-step reaction in glycerolipid assembly through transferring an acyl moiety to glycerol-3-phosphate (G3P) to generate lysophosphatidic acid (LPA), an intermediate in lipid metabolism. The effect of ATS1 overexpression on glycerolipid metabolism and growth remained to be elucidated in plants, particularly oil crop plants. Here, we found that overexpression of BnATS1 from Brassica napus enhanced plant growth and prokaryotic glycerolipid biosynthesis. BnATS1 is localized in chloroplasts and an in vitro assay showed that BnATS1 had acylation activity toward glycerol 3-phosphate to produce LPA. Lipid profiling showed that overexpression of BnATS1 led to increases in multiple glycerolipids including phosphatidylglycerol (PG), monogalactosyldiacylglycerol (MGDG), phosphatidylcholine (PC), and phosphatidylinositol (PI), with increased polyunsaturated fatty acids. Moreover, increased MGDG was attributed to the elevation of 34:6- and 34:5-MGDG, which were derived from the prokaryotic pathway. These results suggest that BnATS1 promotes accumulation of polyunsaturated fatty acids in cellular membranes, thus enhances plant growth under low-temperature conditions in Brassica napus. Full article
(This article belongs to the Special Issue Metabolic Engineering of Plants)
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Article
A Pyrimidin-Like Plant Activator Stimulates Plant Disease Resistance and Promotes the Synthesis of Primary Metabolites
Int. J. Mol. Sci. 2020, 21(8), 2705; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21082705 - 14 Apr 2020
Cited by 1 | Viewed by 872
Abstract
Plant activators are chemicals that induce plant defense responses to various pathogens. Here, we reported a new potential plant activator, 6-(methoxymethyl)-2-[5-(trifluoromethyl)-2-pyridyl] pyrimidin-4-ol, named PPA2 (pyrimidin-type plant activator 2). Unlike the traditional commercial plant activator benzothiadiazole S-methyl ester (BTH), PPA2 was fully soluble [...] Read more.
Plant activators are chemicals that induce plant defense responses to various pathogens. Here, we reported a new potential plant activator, 6-(methoxymethyl)-2-[5-(trifluoromethyl)-2-pyridyl] pyrimidin-4-ol, named PPA2 (pyrimidin-type plant activator 2). Unlike the traditional commercial plant activator benzothiadiazole S-methyl ester (BTH), PPA2 was fully soluble in water, and it did not inhibit plant growth or root system development in rice (Oryza sativa). PPA2 pretreatment significantly increased plant resistance against bacterial infection in both Arabidopsis and rice, in conjunction with increases in the level of jasmonoyl-isoleucine and 12-oxo-phytodienoic acid. In addition, metabolite profiling indicated that BTH significantly reduced the abundance of various primary metabolites in rice seedlings, including most amino acids, sugars, and organic acids; by contrast, PPA2 promoted their synthesis. Our results thus indicate that PPA2 enhances plant defenses against bacterial infection through the jasmonic acid pathway, and that as a water-soluble compound that can promote the synthesis of primary metabolites it has broad potential applications in agriculture. Full article
(This article belongs to the Special Issue Metabolic Engineering of Plants)
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Article
Tinkering Cis Motifs Jigsaw Puzzle Led to Root-Specific Drought-Inducible Novel Synthetic Promoters
Int. J. Mol. Sci. 2020, 21(4), 1357; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21041357 - 18 Feb 2020
Cited by 5 | Viewed by 1379
Abstract
Following an in-depth transcriptomics-based approach, we first screened out and analyzed (in silico) cis motifs in a group of 63 drought-inducible genes (in soybean). Six novel synthetic promoters (SynP14-SynP19) were designed by concatenating 11 cis motifs, ABF, ABRE, ABRE-Like, CBF, E2F-VARIANT, G-box, GCC-Box, [...] Read more.
Following an in-depth transcriptomics-based approach, we first screened out and analyzed (in silico) cis motifs in a group of 63 drought-inducible genes (in soybean). Six novel synthetic promoters (SynP14-SynP19) were designed by concatenating 11 cis motifs, ABF, ABRE, ABRE-Like, CBF, E2F-VARIANT, G-box, GCC-Box, MYB1, MYB4, RAV1-A, and RAV1-B (in multiple copies and various combination) with a minimal 35s core promoter and a 222 bp synthetic intron sequence. In order to validate their drought-inducibility and root-specificity, the designed synthetic assemblies were transformed in soybean hairy roots to drive GUS gene using pCAMBIA3301. Through GUS histochemical assay (after a 72 h 6% PEG6000 treatment), we noticed higher glucuronidase activity in transgenic hairy roots harboring SynP15, SynP16, and SynP18. Further screening through GUS fluorometric assay flaunted SynP16 as the most appropriate combination of efficient drought-responsive cis motifs. Afterwards, we stably transformed SynP15, SynP16, and SynP18 in Arabidopsis and carried out GUS staining as well as fluorometric assays of the transgenic plants treated with simulated drought stress. Consistently, SynP16 retained higher transcriptional activity in Arabidopsis roots in response to drought. Thus the root-specific drought-inducible synthetic promoters designed using stimulus-specific cis motifs in a definite fashion could be exploited in developing drought tolerance in soybean and other crops as well. Moreover, the rationale of design extends our knowledge of trial-and-error based cis engineering to construct synthetic promoters for transcriptional upgradation against other stresses. Full article
(This article belongs to the Special Issue Metabolic Engineering of Plants)
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Article
Combined Proteomics and Metabolism Analysis Unravels Prominent Roles of Antioxidant System in the Prevention of Alfalfa (Medicago sativa L.) against Salt Stress
Int. J. Mol. Sci. 2020, 21(3), 909; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21030909 - 30 Jan 2020
Cited by 7 | Viewed by 1161
Abstract
Alfalfa is the most extensively cultivated forage legume worldwide, and salinity constitutes the main environmental scourge limiting its growth and productivity. To unravel the potential molecular mechanism involved in salt tolerance in alfalfa, we accomplished a combined analysis of parallel reaction monitoring-based proteomic [...] Read more.
Alfalfa is the most extensively cultivated forage legume worldwide, and salinity constitutes the main environmental scourge limiting its growth and productivity. To unravel the potential molecular mechanism involved in salt tolerance in alfalfa, we accomplished a combined analysis of parallel reaction monitoring-based proteomic technique and targeted metabolism. Based on proteomic analysis, salt stress induced 226 differentially abundant proteins (DAPs). Among them, 118 DAPs related to the antioxidant system, including glutathione metabolism and oxidation-reduction pathways, were significantly up-regulated. Data are available via ProteomeXchange with identifier PXD017166. Overall, 107 determined metabolites revealed that the tricarboxylic acid (TCA) cycle, especially the malate to oxaloacetate conversion step, was strongly stimulated by salt stress. This leads to an up-regulation by about 5 times the ratio of NADPH/NADP+, as well as about 3 to 5 times in the antioxidant enzymes activities, including those of catalase and peroxidase and proline contents. However, the expression levels of DAPs related to the Calvin–Benson–Bassham (CBB) cycle and photorespiration pathway were dramatically inhibited following salt treatment. Consistently, metabolic analysis showed that the metabolite amounts related to carbon assimilation and photorespiration decreased by about 40% after exposure to 200 mM NaCl for 14 d, leading ultimately to a reduction in net photosynthesis by around 30%. Our findings highlighted also the importance of the supplied extra reducing power, thanks to the TCA cycle, in the well-functioning of glutathione to remove and scavenge the reactive oxygen species (ROS) and mitigate subsequently the oxidative deleterious effect of salt on carbon metabolism including the CBB cycle. Full article
(This article belongs to the Special Issue Metabolic Engineering of Plants)
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