Abstract
Fluorescent glucose derivatives are valuable tools as glucose analogs in plant research to explore metabolic pathways, study enzyme activity, and investigate cellular processes related to glucose metabolism and sugar transport. They allow visualization and tracking of glucose uptake, its utilization, and distribution within plant cells and tissues. This study investigates the phenotypic and metabolic impact of the exogenously fed glucose derivative, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose) (2-NBDG) on the fibers of Gossypium hirsutum (Upland cotton) ovule in vitro cultures. The presence of 2-NBDG in the culture medium did not lead to macroscopic morphological alterations in ovule and fiber development or to the acquisition of fluorescence or yellow coloration. Confocal laser scanning microscope imaging and chromatographic analysis of cotton ovules’ outer rim cross-sections showed that the 2-NBDG is transported from the extracellular space and accumulated inside some outer integument cells, epidermal cells, and fertilized epidermal cells (fibers), but is not incorporated into the cell walls. Untargeted metabolic profiling of the fibers revealed significant changes in the relative levels of metabolites involved in glycolysis and upregulation of alternative energy-related pathways. To provide biochemical and structural evidence for the observed downregulation of glycolysis pathways in the fibers containing 2-NBDG, kinetics analysis and docking simulations were performed on hexokinase from G. hirsutum (GhHxk). Notably, the catalytic activity of heterologously expressed recombinant active GhHxk exhibited a five-fold decrease in reaction rates compared to D-glucose. Furthermore, GhHxk exhibited a linear kinetic behavior in the presence of 2-NBDG instead of the Michaelis-Menten kinetics found for D-glucose. Docking simulations suggested that 2-NBDG interacts with a distinct binding site of GhHxk9, possibly inducing a conformational change. These results highlight the importance of considering fluorescent glucose derivatives as ready-to-use analogs for tracking glucose-related biological processes. However, a direct comparison between their mode of action and its extrapolation into biochemical considerations should go beyond microscopic inspection and include complementary analytical techniques.
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Claims 10
Claim · model-written summary
2-NBDG in the culture medium caused no macroscopic morphological alterations in cotton ovule and fiber development.
- Taxon
- Gossypium hirsutum
Gossypium hirsutumt0 - Treatment
- 2-NBDG in the culture mediume1
- Outcome
- macroscopic morphological alterations in ovule and fiber developmente1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“The presence of 2-NBDG in the culture medium did not lead to macroscopic morphological alterations in ovule and fiber development or to the acquisition of fluorescence or yellow coloration”
t0“Gossypium hirsutum”
Claim · model-written summary
2-NBDG in the culture medium did not lead to fluorescence or yellow coloration of the cotton ovules.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Treatment
- 2-NBDG in the culture mediume1
- Outcome
- acquisition of fluorescence or yellow coloratione1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“The presence of 2-NBDG in the culture medium did not lead to macroscopic morphological alterations in ovule and fiber development or to the acquisition of fluorescence or yellow coloration”
t0“Gossypium hirsutum”
Claim · model-written summary
2-NBDG was transported from the extracellular space and accumulated inside some outer integument cells, epidermal cells and fertilized epidermal cells (fibers).
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Treatment
- 2-NBDGe1
- Outcome
- accumulated inside some outer integument cells, epidermal cells, and fertilized epidermal cells (fibers)e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“the 2-NBDG is transported from the extracellular space and accumulated inside some outer integument cells, epidermal cells, and fertilized epidermal cells (fibers), but is not incorporated into the cell walls”
t0“Gossypium hirsutum”
Claim · model-written summary
2-NBDG was not incorporated into the cell walls.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Treatment
- 2-NBDGe1
- Outcome
- not incorporated into the cell wallse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“the 2-NBDG is transported from the extracellular space and accumulated inside some outer integument cells, epidermal cells, and fertilized epidermal cells (fibers), but is not incorporated into the cell walls”
t0“Gossypium hirsutum”
Claim · model-written summary
Untargeted metabolic profiling of the fibers revealed significant changes in relative levels of glycolysis metabolites.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Organ / stage
- fiberse1
- Outcome
- relative levels of metabolites involved in glycolysise1
- Direction
- significant changese1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“Untargeted metabolic profiling of the fibers revealed significant changes in the relative levels of metabolites involved in glycolysis and upregulation of alternative energy-related pathways”
t0“Gossypium hirsutum”
Claim · model-written summary
Metabolic profiling of the fibers revealed upregulation of alternative energy-related pathways.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Organ / stage
- fiberse1
- Outcome
- alternative energy-related pathwayse1
- Direction
- upregulatione1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“Untargeted metabolic profiling of the fibers revealed significant changes in the relative levels of metabolites involved in glycolysis and upregulation of alternative energy-related pathways”
t0“Gossypium hirsutum”
Claim · model-written summary
Recombinant GhHxk showed a five-fold decrease in reaction rate with 2-NBDG compared to D-glucose.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Gene
- GhHxke1
- Outcome
- reaction ratese1
- Direction
- decreasee1
- Value
- five-folde1
- Comparator
- D-glucosee1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“the catalytic activity of heterologously expressed recombinant active GhHxk exhibited a five-fold decrease in reaction rates compared to D-glucose”
t0“Gossypium hirsutum”
Claim · model-written summary
GhHxk showed linear kinetics with 2-NBDG instead of the Michaelis-Menten kinetics seen with D-glucose.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Treatment
- 2-NBDGe1
- Gene
- GhHxke1
- Outcome
- linear kinetic behaviore1
- Comparator
- D-glucosee1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“GhHxk exhibited a linear kinetic behavior in the presence of 2-NBDG instead of the Michaelis-Menten kinetics found for D-glucose”
t0“Gossypium hirsutum”
Claim · model-written summary
Docking simulations suggested that 2-NBDG interacts with a distinct binding site of GhHxk9, possibly inducing a conformational change.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Treatment
- 2-NBDGe1
- Gene
- GhHxk9e1
- Outcome
- distinct binding sitee1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“Docking simulations suggested that 2-NBDG interacts with a distinct binding site of GhHxk9, possibly inducing a conformational change”
t0“Gossypium hirsutum”
Claim · model-written summary
The authors recommend that comparing the mode of action of fluorescent glucose derivatives should go beyond microscopic inspection and include complementary analytical techniques.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Outcome
- complementary analytical techniquese1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“a direct comparison between their mode of action and its extrapolation into biochemical considerations should go beyond microscopic inspection and include complementary analytical techniques”
t0“Gossypium hirsutum”
The same questions in other papers
in vitro · quality: grain · Gossypium hirsutum (cotton)Positive and negative results
5 in the opposite direction
- positive
- positive
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2 papers more in this group
See all 6 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.