A Study on the Use of the Phyto-Courier Technology in Tobacco Leaves Infected by Agrobacterium tumefaciens
Abstract
:1. Introduction
2. Results
2.1. Agroinfiltration Stimulated Stress-Related Genes, but Priming with GS3 Induced Some Significant Changes
2.2. Agroinfiltration Increased the Chloroplasts’ Mean Area and GS3 Prevented This Effect
2.3. Relative Flavonoid Accumulation and Antioxidant Power of the Leaf Metabolite Extracts, Phyto-Courier, and Control Solutions
3. Discussion
4. Materials and Methods
4.1. Experimental Set-Up and Infiltration
4.2. Sampling and RNA Extraction
4.3. RNA Quality Check and Quantification
4.4. Primer Design
4.5. RT-qPCR
4.6. Preparation of Samples for Optical Microscopy
4.7. Preparation of Methanolic Extracts and Quercetin Extraction from the GS3 Phyto-Courier and Q Solution
4.8. Spectrophotometric Measurements
4.9. Data Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Samples | sshLNPs | Trehalose | Quercetin | Hypromellose + Pluronic L61 Vehicle | Total Volume (25 mL PBS Addition) | ||
---|---|---|---|---|---|---|---|
Si-NPs | Hydro-PC | Arginine: Glycine | |||||
B | 25 mL | 50 mL | |||||
GS3 | 4 mg | 16 mg | 4 mg: 2 mg | 4 mg | 25 mg | 25 mL | 50 mL |
Q | 25 mg | 25 mL | 50 mL |
Primer Name | Gene Name | Identifier | Primer Sequence fwd (5′→3′) | Primer Sequence rev (5′→3′) | Size (bp) | Efficiency % |
---|---|---|---|---|---|---|
Genes of interest [34] | ||||||
NB_Chit6 | Chitinase 6 | k58:171433 | GATCGCTGCTTTCTTTGCTC | CGCCTGAAGGACCATTTATC | 80 | 95.69 |
NB_Chn-A | Endochitinase A | k72:280831 | AACCTTCTTGCCACGATGTC | GTGATGACACCAAATCCAG | 91 | 92.96 |
NB_GEβGluc | Glucan endo-1,3-beta-glucosidase | k60:395851 | GCTGCTTGTTGGGAGAAAAC | AGCCTGGACCTATTGAAACC | 102 | 95.17 |
NB_PDRP1 | Pleiotropic drug resistance protein | k80:119278 | AGGTTTCATCGTTCCACGAC | AGGTCTCCGAATTGAGATGC | 111 | 92.55 |
NB_PR-1a | Pathogenesis-related protein 1A | k58:459599 | TCCAACACGAACCGAGTTAC | TTGAGATGTGGGTCGATGAG | 116 | 81.84 |
NB_PR-Q | Acidic endochitinase Q | k78:23474 | CCCCAGGAGCAACATTTAAC | AATGACGCAGTGGAAGATCG | 70 | 91.93 |
NB_RNAP-β | DNA-directed-RNA polymerase subunit beta’ | k58:179615 | TCCTCTTATCCCAATCTGGTG | TTGACGGACACAAACTCTGC | 95 | 88.01 |
NB_VAS | Lipid transfer-like protein | k64:395173 | TAGTCACGGTGGCGATTATG | GCGGTTTGGTGGAATTAAGG | 105 | 91.8 |
NB_WIN2 | Wound-induced protein | k58:223872 | CCGTCAAAGGGTAAACATGG | GATGGAAGAGGGAATCAACG | 144 | 97.23 |
NB_OEE1 | Oxygen-evolving enhancer protein 1 | k80:117138 | CCACATCATTCACGGTCAAG | TGCCATCAGAAGACACTTCG | 135 | 92.6 |
NB_RuBisCO-BP | RuBisCO large subunit-binding protein subunit beta | k64:394932 | TACTGGCTTTTCCGTTCACC | TTGAGCAAGAAGCACTAGC | 103 | 88.00 |
NB_SerThrKin | Serine/threonine-protein kinase | k78:183558 | TACCGATACCGTCCAATTCC | TGCACAGTCATGGTCTTG | 129 | 92.94 |
NB_PSI-N | Photosystem I reaction centre subunit N | k76:226809 | GGCAGCAATGAACTCAAGTG | TGATTGGGAAGCCATAGAGG | 100 | 87.00 |
Reference genes [70] | ||||||
Nb_F-Box | F-box protein | Niben.v0.3.Ctg24993647 (At5g15710) | GGCACTCACAAACGTCTATTTC | ACCTGGGAGGCATCCTGCTTAT | 127 | 100.8 |
Nb_SAND | Sand family protein | Niben.v0.3.Ctg25188435 (At2g28390) | ACCACCAACACCTATGAATGCT | CAGTCTCGCCTCATCTGGGTCA | 83 | 88.27 |
Nb_L23 | 60S ribosomal protein | TC19271 (At2g39460) | AAGGATGCCGTGAAGAAGATGT | GCATCGTAGTCAGGAGTCAACC | 110 | 91.46 |
Nb_UK | Uridylate kinase | EH363935 (At5g26667) | CTAGGAGTATATTGGAAGAGCG | AAAGATACATCGCCTTGCTGAA | 107 | 96.72 |
Nb_GBP | GTP-binding protein | TC20872 (At5g59840) | GGAACTGGATTCGCAACATAGA | GACCCTTGGAAGTTGGCACAGC | 114 | 92.5 |
Nb_RdR6 | Putative RNA-dependent RNA polymerase SDE1 | AY722008 (At3g49500) | TTCAGGAATGTCTTCGAGCG | AGTGATCTAGCAACCCAATGAG | 134 | 93.3 |
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Gutsch, A.; Berni, R.; Hausman, J.-F.; Sutera, F.M.; Dehsorkhi, A.; Torabi-Pour, N.; Saffie-Siebert, S.; Guerriero, G. A Study on the Use of the Phyto-Courier Technology in Tobacco Leaves Infected by Agrobacterium tumefaciens. Int. J. Mol. Sci. 2023, 24, 14153. https://doi.org/10.3390/ijms241814153
Gutsch A, Berni R, Hausman J-F, Sutera FM, Dehsorkhi A, Torabi-Pour N, Saffie-Siebert S, Guerriero G. A Study on the Use of the Phyto-Courier Technology in Tobacco Leaves Infected by Agrobacterium tumefaciens. International Journal of Molecular Sciences. 2023; 24(18):14153. https://doi.org/10.3390/ijms241814153
Chicago/Turabian StyleGutsch, Annelie, Roberto Berni, Jean-Francois Hausman, Flavia Maria Sutera, Ashkan Dehsorkhi, Nissim Torabi-Pour, Suzanne Saffie-Siebert, and Gea Guerriero. 2023. "A Study on the Use of the Phyto-Courier Technology in Tobacco Leaves Infected by Agrobacterium tumefaciens" International Journal of Molecular Sciences 24, no. 18: 14153. https://doi.org/10.3390/ijms241814153