Abstract
ABSTRACT Nanoparticles hold great potential as innovative agrochemicals, offering significant advantages over traditional products like insecticides, herbicides and fertilizers, which often harm soil and human health. This study investigates the effects of free aqueous pomegranate extract (PAE) and PAE encapsulated in CaCO 3 nanocrystals (nanoCaCO 3 @PAE) on cellular uptake and internalization mechanisms in tobacco plants. Our experiments assessed the cytotoxic effect of free PAE on protoplast viability, while the negative effect was reduced in the presence of nanoCaCO 3 @PAE. FM4-64 labelling and secRFP-HDEL-transformed protoplasts showed an indirect increased of endocytic activity when treated with 0.25 mg/mL nanoCaCO 3 @PAE. After optimization of confocal microscope setting, auto-fluorescence of nano-encapsulated extracts were visible in protoplast endosome-like structures. In 3-month-old tobacco seedlings, foliar spraying of PAE or nanoCaCO 3 @PAE showed no visible signs of stress or necrosis 14 days after treatments, as well as chlorophyll and carotenoid content, compared to control seedlings. On the other hand, a significant increase in the amount of soluble phenols in the treated leaves, particularly in those treated with nanoCaCO 3 @PAE compared to their free form was observed. The use of fluorescent carbon nanocrystals, nanoCaCO 3 @FITC, allowed the visualization of the nanoparticles into their final destination, i.e. central vacuole, where the phenols extracted from leaves were not degraded and then probably released from nanoCaCO 3 @PAE due to the acidic nature of this compartment. Through the use of Tyrphostin A23, a specific inhibitor of clathrin-mediated internalization, it has been demonstrated that nanoCaCO 3 @FITC were internalized through clathrin-independent mechanisms.
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Claims 7
Claim · model-written summary
Free PAE reduced protoplast viability; encapsulation reduced that negative effect.
- Taxon
- tobacco
Nicotiana tabacume1 - Treatment
- nanoCaCO 3 @PAEe0
- Outcome
- protoplast viabilitye0
- Direction
- reducede0
- Comparator
- free PAEe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“cytotoxic effect of free PAE on protoplast viability, while the negative effect was reduced in the presence of nanoCaCO 3 @PAE”
e1“in tobacco plants”
Claim · model-written summary
At 0.25 mg/mL nanoCaCO 3 @PAE increased endocytic activity.
- Taxon
- tobaccoNicotiana tabacume1
- Treatment
- 0.25 mg/mL nanoCaCO 3 @PAEe0
- Outcome
- endocytic activitye0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“an indirect increased of endocytic activity when treated with 0.25 mg/mL nanoCaCO 3 @PAE”
e1“in tobacco plants”
Claim · model-written summary
Encapsulated extracts were visible in endosome-like structures.
- Taxon
- tobaccoNicotiana tabacume1
- Outcome
- nano-encapsulated extracts were visible in protoplast endosome-like structurese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“auto-fluorescence of nano-encapsulated extracts were visible in protoplast endosome-like structures”
e1“in tobacco plants”
Claim · model-written summary
Foliar PAE or nanoparticle treatment showed no visible seedling stress or necrosis after 14 days.
- Taxon
- tobaccoNicotiana tabacume1
- Treatment
- foliar spraying of PAE or nanoCaCO 3 @PAEe0
- Outcome
- no visible signs of stress or necrosise0
- Qualifier
- 14 days after treatmentse0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“foliar spraying of PAE or nanoCaCO 3 @PAE showed no visible signs of stress or necrosis 14 days after treatments”
e1“tobacco seedlings”
Claim · model-written summary
Treated tobacco leaves had significantly increased soluble phenols, especially with nanoparticles versus free PAE.
- Taxon
- tobaccoNicotiana tabacume1
- Treatment
- nanoCaCO 3 @PAEe0
- Outcome
- amount of soluble phenols in the treated leavese0
- Direction
- increasee0
- Comparator
- their free forme0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a significant increase in the amount of soluble phenols in the treated leaves, particularly in those treated with nanoCaCO 3 @PAE compared to their free form”
e1“tobacco seedlings”
Claim · model-written summary
Fluorescent nanoparticles reached the central vacuole.
- Taxon
- tobaccoNicotiana tabacume1
- Treatment
- nanoCaCO 3 @FITCe0
- Outcome
- nanoparticles into their final destination, i.e. central vacuolee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“nanoCaCO 3 @FITC, allowed the visualization of the nanoparticles into their final destination, i.e. central vacuole”
e1“tobacco seedlings”
Claim · model-written summary
NanoCaCO 3 @FITC internalization used clathrin-independent mechanisms.
- Taxon
- tobaccoNicotiana tabacume1
- Treatment
- nanoCaCO 3 @FITCe0
- Outcome
- internalized through clathrin-independent mechanismse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“nanoCaCO 3 @FITC were internalized through clathrin-independent mechanisms”
e1“tobacco seedlings”
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