Nonhost Disease Resistance in Pea: Chitosan’s Suggested Role in DNA Minor Groove Actions Relative to Phytoalexin-Eliciting Anti-Cancer Compounds
Abstract
:1. Introduction
1.1. AT-Rich DNA Sequences in Pre-Promoter Region of Pea HMG A Gene
- TCAATTAAAA AATCAATTTA TTTATTTCAT TTCATAAATA TATTCATAA
- AATTAAATAC AATGAGTAGA ATTTCAAAC TCTCAATAAATTTTAGT
1.2. Sequences Used in Crystal Modeling of Drug/DNA Minor Groove Complexes
- Netropsin: d(CGCAATTCGCG)
- Distamycin: d(CGCAAATTTGCG)
- Hoechst 33258: d(CGCGAATTCGCG)
1.3. DNA Minor Groove Compounds Evaluated as Pisatin Elicitors Include Chitosan, Chromomycin A3, Distamycin, Hoecsht 33258, Netropsin, and Methyl Blue
1.4. Production of PR Gene Proteins and Pisatin
1.5. Natural Components
1.6. Effects of Chitosan-Oligomer and Pathogen Challenge on Animal Cells
1.7. Chitosan Signaling in Pea Tissue
2. Results and Discussion
2.1. Chromomycin
2.2. Other DNA Minor Groove Localizing Compounds
2.3. DNA-Fragmentation Assciated with Netropsin Treatment
2.4. The Specifics of Netropsin/DNA Dodecamer Attachment
2.5. Signal Reception in the Hypothesized Role of Host Chromatin
3. Experimental Section
3.1. Elicitor Compound Sources
3.2. Plant and Pathogen
3.3. Elicitor Treatments and Pisatin Quantization
3.4. DNA Damage Assessment
4. Conclusions
Funding
Acknowledgments
Conflicts of Interest
References
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Pre-Treatment | Secondary Trmt in 30 min | Pisatin µg/g fr. wt. 22 h | Hypersensitive Response at 24 h |
---|---|---|---|
Netropsin 0.5 mg/mL | Fsph spores | 40.8 ± 5.0 | +++ |
Netropsin 0.5 mg/mL | water | 26.3 ± 1.6 | +++ |
Netropsin 0.25 mg/mL | Fsph spores | 22.6 ± 2.5 | +++ |
Netropsin 0.25 mg/mL | water | 2.1 ± 0.1 | +++ |
Netropsin 0.125 mg/mL | Fsph spores | 36.1 ± 24.0 | ++ |
Netropsin 0.125 mg/mL | water | 2.7 ± 1.6 | ++ |
Water | Fsph spores | 34.7± 8.2 | ++ |
Pre-Treatment 20 µL/Pod Half | Secondary Trmt in 30 min 20 µL/Pod Half | Pisatin µg/g fr. wt. 24 h | Hyper. Response 24 h |
---|---|---|---|
Netropsin 0.5 mg/mL | Fsph spores (3 × 106/mL) | 56.7 ± 3.8 | ++ |
Water | Fsph spores (3 × 106/mL) | 101.3 ± 22.7 | ++ |
Netropsin 0.5 mg/mL | Fsph spores (7.5 × 105/mL) | 39.0 ± 3.9 | +++ |
Water | Fsph spores (7.5 × 105/mL) | 57.0 ± 15.8 | +++ |
Netropsin 0.5 mg/mL | Fsph spores (3.7 × 105/mL) | 43.8 ± 2.6 | + |
Water | Fsph spores (3.7 × 105/mL) | 35.3 ± 6.1 | ++ |
Netropsin 0.5 mg/mL | Fsph spores (1.8 × 105/mL) | 24.2 ± 5.0 | ++ |
Water | Fsph spores (1.8 × 105/mL) | 52.6 ± 10.1 | ++ |
Netropsin 0.5 mg/mL | |||
Fsph spores 3 × 106/mL) | -------------------------------- | 48.9 ± 7.0 | +++ |
Water | -------------------------------- | --------------- | + |
Netropsin 0.5 mg/mL | |||
Chitosan 0.5 mg/mL | ------------------------------- | 10.7 ± 4.9 | ++ |
Secondary Trmt in 30 min | Pisatin µg/g fr. wt. 24 h | Hypersensitivity Response 24 h | |
---|---|---|---|
Chitosan 0.2 mg/mL | Netropsin 1 mg/mL | 50.2 ± 3.2 | +++ |
Water | Netropsin 1 mg/mL | 22.5 ± 0.9 | +++ |
Chitosan 0.2 mg/mL | Netropsin 0.5 mg/mL | 38.7 ± 0.9 | +++ |
Water | Netropsin 0.5 mg/mL | 13.7 ± 0.1 | ++ |
Chitosan 0.2 mg/mL | Netropsin 0.25 mg/mL | 27.2 ± 3.4 | ++ |
Water | Netropsin 0.25 mg/mL | 14.2 ± 5.2 | +++ |
Chitosan 0.2 mg/mL | Netropsin 0.12 mg/mL | 9.8 ± 1.3 | ++ |
Water | Netropsin 0.12 mg/mL | ---------- | + |
Fsph spores (2.4 × 106) | ---------------------------- | 244.4 ± 27.4 | +++ |
Water | ---------------------------- | ---------------- | Light |
Chitosan 0.2 mg/mL | ---------------------------- | 0.9 ± 9.27 | ++ |
Treatment (20 µL Volume) | Pisatin µg/g fr. wt. | Hypersensitivity |
---|---|---|
Chromomycin 2.0 mg/mL | 69.9 ± 1.0 | +++ |
Chromomycin 1.0 mg/mL | 133.5 ± 11.2 | ++ |
Chromomycin 0.5 mg/mL | 188.3 ± 3.5 | + |
Chromomycin 0.25 mg/mL | 70.9 ± 33.5 | +++ |
Chromomycin 0.125 mg/mL | 106.1 ± 16.1 | ++ |
Chromomycin 0.06 mg/mL | 77.3 ± 7.9 | +++ |
Chromomycin 0.03 mg/mL | 10.9 ± 10.9 | + |
Chromomycin 0.015 mg/mL | 8.5 ± 7.1 | + |
Water | ---------------- | + |
Fsph spores (4.5 × 106) | 184.7 ± 42.7 | ++ |
Chitosan 1 mg/mL | 8.6 ± 4.6 | +++ |
Netropsin 0.5 mg/mL | 17.8 ± 1.9 | +++ |
Pre-Treatment 20 µL/Pod Half | Secondary Trmt in 30 min 20 µL/Pod Half | Pisatin µg/g fr. wt. 24 h |
---|---|---|
Chromomycin 0.5 mg/mL | Fsph spores (6.4 × 106/mL) | 271.1 ± 23.3 |
Chromomycin 0.5 mg/mL | Water | 111.2 ± 9.2 |
Chromomycin 0.25 mg/mL | Fsph spores (6.4 × 106/mL) | 248.8 ± 10.8 |
Chromomycin 0.25 mg/mL | Water | 109.8 ± 3.8 |
Chromomycin 0.12 mg/mL | Fsph spores (6.4 × 106/mL) | 291.0 ± 9.5 |
Chromomycin 0.12 mg/mL | Water | 58.9 ± 9.9 |
Chromomycin 0.06 mg/mL | Fsph spores (6.4 × 106/mL) | 270.9 ± 17.5 |
Chromomycin 0.06 mg/mL | Water | 53.7 ± 6.3 |
Chromomycin 0.03 mg/mL | Fsph spores (6.4 × 106/mL) | 259.7 ± 28.4 |
Chromomycin 0.03 mg/mL | Water | 78.3 ± 43.2 |
Water | Fsph spores (6.4 × 106/mL) | 210.6 ± 34.9 |
Water | Water | ---------------- |
Pre-Treatment 20 µL/Pod Half | Secondary Trmt in 30 min 20 µL/Pod Half | Pisatin µg/g fr. wt. |
---|---|---|
Chromomycin 0.12 mg/mL | Chitosan 1 mg/mL | 153.0 ± 87.1 |
Chromomycin 0.12 mg/mL | Water | 113.2 ± 22.1 |
Chromomycin 0.06 mg/mL | Chitosan 1 mg/mL | 81.9 ± 1.2 |
Chromomycin 0.06 mg/mL | Water | 29.9 ± 8.9 |
Chromomycin 0.03 mg/mL | Chitosan 1 mg/mL | 33.4 ± 3.5 |
Chromomycin 0.03 mg/mL | Water | 29.7 ± 20.6 |
Chromomycin 0.015 mg/mL | Chitosan 1 mg/mL | 20.6 ± 3.0 |
Chromomycin 0.015 mg/mL | Water | 2.1 ± 0.0 |
Chromomycin 0.007 mg/mL | Chitosan 1 mg/mL | 43.2 ± 22.9 |
Chromomycin 0.007 mg/mL | Water | 2.4 ± 0.5 |
Water | Chitosan 1 mg/mL | 9.4 ± 4.5 |
Water | Water | 0.0 ± 0.0 |
Pre-Treatment | Secondary Treatment 30 min | Pisatin µg/g fr. wt. 24 h |
---|---|---|
Hoechst 33258 1 mg/mL | Fsph spores | 102 ± 7 |
Hoechst 33258 1 mg/mL | Water | 9 ± 1 |
Hoechst 33258 0.5 mg/mL | Fsph spores | 91 ± 7 |
Hoechst 33258 0.5 mg/mL | Water | 3 ± 2 |
Hoechst 33258 0.25 mg/mL | Fsph spores | 116 ± 26 |
Hoechst 33258 0.25 mg/mL | Water | 1 ± 0.6 |
Hoechst 33258 0.125 mg/mL | Fsph spores | 90 ± 5 |
Hoechst 33258 0.125 mg/mL | Water | 2 ± 0.9 |
Water | Fsph spores | 84 ± 21 |
Water | Water | --------- |
Chitosan lactate 1 mg/mL | Fsph spores | 125 ± 62 |
Chitosan lactate | Water | 9 ± 2 |
Sample Availability: Samples of the compounds are not available from the authors. |
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Hadwiger, L.A. Nonhost Disease Resistance in Pea: Chitosan’s Suggested Role in DNA Minor Groove Actions Relative to Phytoalexin-Eliciting Anti-Cancer Compounds. Molecules 2020, 25, 5913. https://doi.org/10.3390/molecules25245913
Hadwiger LA. Nonhost Disease Resistance in Pea: Chitosan’s Suggested Role in DNA Minor Groove Actions Relative to Phytoalexin-Eliciting Anti-Cancer Compounds. Molecules. 2020; 25(24):5913. https://doi.org/10.3390/molecules25245913
Chicago/Turabian StyleHadwiger, Lee A. 2020. "Nonhost Disease Resistance in Pea: Chitosan’s Suggested Role in DNA Minor Groove Actions Relative to Phytoalexin-Eliciting Anti-Cancer Compounds" Molecules 25, no. 24: 5913. https://doi.org/10.3390/molecules25245913