Overexpression of Ricinus communis L. malate synthase enhances seed tolerance to abiotic stress during germination

Valdinei Carvalho Brito, Catherine P. de Almeida, Rhaíssa R. Barbosa, Maria G.A. Carosio, Antônio G. Ferreira, Luzimar G. Fernandez, Renato D. de Castro, Henk Hilhorst, Wilco Ligterink, Paulo Roberto Ribeiro*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Ricinus communis L. seeds can germinate at high temperatures, but further development of the seedlings is negatively affected. This mainly caused by impairment of energy-generating pathways when seeds are germinated at 35 °C. Ricinus communis malate synthase (RcMLS) is a key responsive gene in lipid mobilization and gluconeogenesis and as such might have a role in sustaining successful seed germination and seedling growth. Herein, we raised the question whether RcMLS might be involved in the biochemical and molecular mechanisms required for R. communis seed germination under unfavourable environmental conditions. For that, we used a robust approach that encompassed bioinformatics analysis, transgenic Arabidopsis thaliana (L.) Heynh seeds overexpressing RcMLS, along with phenotypical characterization of seed germination under abiotic stress. The phylogenetic tree revealed important evolutionary relationship amongst MLS sequences from R. communis and from other crop species/model plants. Overexpression of RcMLS enhanced A. thaliana seed germination under high temperature and salt stress. For example, wild-type A. thaliana Columbia seeds (Col-0) showed 37 % of maximum germination at 35 °C, whereas A. thaliana seeds overexpressing RcMLS showed up to 71%. When salt stress was applied (75 mM NaCl), maximum germination of Col-0 seeds reached 37%, whereas for A. thaliana seeds overexpressing RcMLS it reached up to 93%. Nuclear Magnetic Resonance (NMR) and Gas Chromatography coupled to Time-Of-Flight Mass Spectrometry (GC-TOF-MS) metabolomics analysis showed a robust metabolic signature of A. thaliana seeds overexpressing RcMLS in response to abiotic stress. They accumulated high levels of Met, Ile, fructose, glucose, and sucrose. Therefore, we suggested that overexpression of RcMLS has modulated the glyoxylate cycle and gluconeogenesis pathway in order to maintain cellular homeostasis under unfavorable environmental conditions. Our results provide important leads into the contribution of RcMLS to the underlying mechanism of R. communis seed germination under adverse environmental conditions. This might be helpful for breeding programs to develop more resistant R. communis cultivars which are more likely to sustain growth and high yield under the severe conditions found in arid and semi-arid areas worldwide.

Original languageEnglish
Article number112110
JournalIndustrial Crops and Products
Volume145
DOIs
Publication statusPublished - Mar 2020

Fingerprint

malate synthase
Ricinus communis
abiotic stress
germination
seeds
Arabidopsis thaliana
seed germination
gluconeogenesis
environmental factors
salt stress
glyoxylate cycle
dry environmental conditions
metabolomics

Keywords

  • Abiotic stress
  • Castor bean
  • Functional characterization
  • Temperature-responsive genes

Cite this

Brito, V. C., de Almeida, C. P., Barbosa, R. R., Carosio, M. G. A., Ferreira, A. G., Fernandez, L. G., ... Ribeiro, P. R. (2020). Overexpression of Ricinus communis L. malate synthase enhances seed tolerance to abiotic stress during germination. Industrial Crops and Products, 145, [112110]. https://doi.org/10.1016/j.indcrop.2020.112110
Brito, Valdinei Carvalho ; de Almeida, Catherine P. ; Barbosa, Rhaíssa R. ; Carosio, Maria G.A. ; Ferreira, Antônio G. ; Fernandez, Luzimar G. ; de Castro, Renato D. ; Hilhorst, Henk ; Ligterink, Wilco ; Ribeiro, Paulo Roberto. / Overexpression of Ricinus communis L. malate synthase enhances seed tolerance to abiotic stress during germination. In: Industrial Crops and Products. 2020 ; Vol. 145.
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title = "Overexpression of Ricinus communis L. malate synthase enhances seed tolerance to abiotic stress during germination",
abstract = "Ricinus communis L. seeds can germinate at high temperatures, but further development of the seedlings is negatively affected. This mainly caused by impairment of energy-generating pathways when seeds are germinated at 35 °C. Ricinus communis malate synthase (RcMLS) is a key responsive gene in lipid mobilization and gluconeogenesis and as such might have a role in sustaining successful seed germination and seedling growth. Herein, we raised the question whether RcMLS might be involved in the biochemical and molecular mechanisms required for R. communis seed germination under unfavourable environmental conditions. For that, we used a robust approach that encompassed bioinformatics analysis, transgenic Arabidopsis thaliana (L.) Heynh seeds overexpressing RcMLS, along with phenotypical characterization of seed germination under abiotic stress. The phylogenetic tree revealed important evolutionary relationship amongst MLS sequences from R. communis and from other crop species/model plants. Overexpression of RcMLS enhanced A. thaliana seed germination under high temperature and salt stress. For example, wild-type A. thaliana Columbia seeds (Col-0) showed 37 {\%} of maximum germination at 35 °C, whereas A. thaliana seeds overexpressing RcMLS showed up to 71{\%}. When salt stress was applied (75 mM NaCl), maximum germination of Col-0 seeds reached 37{\%}, whereas for A. thaliana seeds overexpressing RcMLS it reached up to 93{\%}. Nuclear Magnetic Resonance (NMR) and Gas Chromatography coupled to Time-Of-Flight Mass Spectrometry (GC-TOF-MS) metabolomics analysis showed a robust metabolic signature of A. thaliana seeds overexpressing RcMLS in response to abiotic stress. They accumulated high levels of Met, Ile, fructose, glucose, and sucrose. Therefore, we suggested that overexpression of RcMLS has modulated the glyoxylate cycle and gluconeogenesis pathway in order to maintain cellular homeostasis under unfavorable environmental conditions. Our results provide important leads into the contribution of RcMLS to the underlying mechanism of R. communis seed germination under adverse environmental conditions. This might be helpful for breeding programs to develop more resistant R. communis cultivars which are more likely to sustain growth and high yield under the severe conditions found in arid and semi-arid areas worldwide.",
keywords = "Abiotic stress, Castor bean, Functional characterization, Temperature-responsive genes",
author = "Brito, {Valdinei Carvalho} and {de Almeida}, {Catherine P.} and Barbosa, {Rha{\'i}ssa R.} and Carosio, {Maria G.A.} and Ferreira, {Ant{\^o}nio G.} and Fernandez, {Luzimar G.} and {de Castro}, {Renato D.} and Henk Hilhorst and Wilco Ligterink and Ribeiro, {Paulo Roberto}",
year = "2020",
month = "3",
doi = "10.1016/j.indcrop.2020.112110",
language = "English",
volume = "145",
journal = "Industrial Crops and Products",
issn = "0926-6690",
publisher = "Elsevier",

}

Brito, VC, de Almeida, CP, Barbosa, RR, Carosio, MGA, Ferreira, AG, Fernandez, LG, de Castro, RD, Hilhorst, H, Ligterink, W & Ribeiro, PR 2020, 'Overexpression of Ricinus communis L. malate synthase enhances seed tolerance to abiotic stress during germination', Industrial Crops and Products, vol. 145, 112110. https://doi.org/10.1016/j.indcrop.2020.112110

Overexpression of Ricinus communis L. malate synthase enhances seed tolerance to abiotic stress during germination. / Brito, Valdinei Carvalho; de Almeida, Catherine P.; Barbosa, Rhaíssa R.; Carosio, Maria G.A.; Ferreira, Antônio G.; Fernandez, Luzimar G.; de Castro, Renato D.; Hilhorst, Henk; Ligterink, Wilco; Ribeiro, Paulo Roberto.

In: Industrial Crops and Products, Vol. 145, 112110, 03.2020.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Overexpression of Ricinus communis L. malate synthase enhances seed tolerance to abiotic stress during germination

AU - Brito, Valdinei Carvalho

AU - de Almeida, Catherine P.

AU - Barbosa, Rhaíssa R.

AU - Carosio, Maria G.A.

AU - Ferreira, Antônio G.

AU - Fernandez, Luzimar G.

AU - de Castro, Renato D.

AU - Hilhorst, Henk

AU - Ligterink, Wilco

AU - Ribeiro, Paulo Roberto

PY - 2020/3

Y1 - 2020/3

N2 - Ricinus communis L. seeds can germinate at high temperatures, but further development of the seedlings is negatively affected. This mainly caused by impairment of energy-generating pathways when seeds are germinated at 35 °C. Ricinus communis malate synthase (RcMLS) is a key responsive gene in lipid mobilization and gluconeogenesis and as such might have a role in sustaining successful seed germination and seedling growth. Herein, we raised the question whether RcMLS might be involved in the biochemical and molecular mechanisms required for R. communis seed germination under unfavourable environmental conditions. For that, we used a robust approach that encompassed bioinformatics analysis, transgenic Arabidopsis thaliana (L.) Heynh seeds overexpressing RcMLS, along with phenotypical characterization of seed germination under abiotic stress. The phylogenetic tree revealed important evolutionary relationship amongst MLS sequences from R. communis and from other crop species/model plants. Overexpression of RcMLS enhanced A. thaliana seed germination under high temperature and salt stress. For example, wild-type A. thaliana Columbia seeds (Col-0) showed 37 % of maximum germination at 35 °C, whereas A. thaliana seeds overexpressing RcMLS showed up to 71%. When salt stress was applied (75 mM NaCl), maximum germination of Col-0 seeds reached 37%, whereas for A. thaliana seeds overexpressing RcMLS it reached up to 93%. Nuclear Magnetic Resonance (NMR) and Gas Chromatography coupled to Time-Of-Flight Mass Spectrometry (GC-TOF-MS) metabolomics analysis showed a robust metabolic signature of A. thaliana seeds overexpressing RcMLS in response to abiotic stress. They accumulated high levels of Met, Ile, fructose, glucose, and sucrose. Therefore, we suggested that overexpression of RcMLS has modulated the glyoxylate cycle and gluconeogenesis pathway in order to maintain cellular homeostasis under unfavorable environmental conditions. Our results provide important leads into the contribution of RcMLS to the underlying mechanism of R. communis seed germination under adverse environmental conditions. This might be helpful for breeding programs to develop more resistant R. communis cultivars which are more likely to sustain growth and high yield under the severe conditions found in arid and semi-arid areas worldwide.

AB - Ricinus communis L. seeds can germinate at high temperatures, but further development of the seedlings is negatively affected. This mainly caused by impairment of energy-generating pathways when seeds are germinated at 35 °C. Ricinus communis malate synthase (RcMLS) is a key responsive gene in lipid mobilization and gluconeogenesis and as such might have a role in sustaining successful seed germination and seedling growth. Herein, we raised the question whether RcMLS might be involved in the biochemical and molecular mechanisms required for R. communis seed germination under unfavourable environmental conditions. For that, we used a robust approach that encompassed bioinformatics analysis, transgenic Arabidopsis thaliana (L.) Heynh seeds overexpressing RcMLS, along with phenotypical characterization of seed germination under abiotic stress. The phylogenetic tree revealed important evolutionary relationship amongst MLS sequences from R. communis and from other crop species/model plants. Overexpression of RcMLS enhanced A. thaliana seed germination under high temperature and salt stress. For example, wild-type A. thaliana Columbia seeds (Col-0) showed 37 % of maximum germination at 35 °C, whereas A. thaliana seeds overexpressing RcMLS showed up to 71%. When salt stress was applied (75 mM NaCl), maximum germination of Col-0 seeds reached 37%, whereas for A. thaliana seeds overexpressing RcMLS it reached up to 93%. Nuclear Magnetic Resonance (NMR) and Gas Chromatography coupled to Time-Of-Flight Mass Spectrometry (GC-TOF-MS) metabolomics analysis showed a robust metabolic signature of A. thaliana seeds overexpressing RcMLS in response to abiotic stress. They accumulated high levels of Met, Ile, fructose, glucose, and sucrose. Therefore, we suggested that overexpression of RcMLS has modulated the glyoxylate cycle and gluconeogenesis pathway in order to maintain cellular homeostasis under unfavorable environmental conditions. Our results provide important leads into the contribution of RcMLS to the underlying mechanism of R. communis seed germination under adverse environmental conditions. This might be helpful for breeding programs to develop more resistant R. communis cultivars which are more likely to sustain growth and high yield under the severe conditions found in arid and semi-arid areas worldwide.

KW - Abiotic stress

KW - Castor bean

KW - Functional characterization

KW - Temperature-responsive genes

U2 - 10.1016/j.indcrop.2020.112110

DO - 10.1016/j.indcrop.2020.112110

M3 - Article

VL - 145

JO - Industrial Crops and Products

JF - Industrial Crops and Products

SN - 0926-6690

M1 - 112110

ER -