The Sesquiterpene Synthase PtTPS5 Produces (1S,5S,7R,10R)-Guaia-4(15)-en-11-ol and (1S,7R,10R)-Guaia-4-en-11-ol in Oomycete-Infected Poplar Roots
Abstract
:1. Introduction
2. Results
2.1. P. cactorum Infection Induces the Accumulation of Terpenes, Aromatic Compounds, and Fatty Acids in P. trichocarpa Roots
2.2. Transcriptome Analysis of Infected and Non-Infected Poplar Roots Revealed a Sesquiterpene Synthase Gene PtTPS5 that Is highly Induced upon P. cactorum Infection
2.3. PtTPS5 Produces (1S,5S,7R,10R)-guaia-4(15)-en-11-ol and (1S,7R,10R)-guaia-4-en-11-ol as Major Products
2.4. The Accumulation of (1S,5S,7R,10R)-guaia-4(15)-en-11-ol, (1S,7R,10R)-guaia-4-en-11-ol, and Hedycaryol in P. cactorum-Infected and Non-Infected Roots Matches the Expression of PtTPS5
3. Discussion
4. Materials and Methods
4.1. Biological Material
4.2. Phytophthora Cactorum Treatment
4.3. Hexane Extraction of Root Tissue and GC-MS/GC-FID Analysis
4.4. Methanol Extraction of Root Tissue and HPLC-UV, LC-MS/MS Analysis of Methanol Extracts
4.5. RNA Extraction and Reverse Transcription
4.6. Heterologous Expression of PtTPS5 and Enzyme Assays
4.7. RNA Sequencing and RT-qPCR Analysis
4.8. Compound Isolation and Structure Elucidation
4.9. Statistical Analysis
4.10. Accession Numbers
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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Compound | Uninfected Roots | Infected Roots | t-Value/T-Value | p-Value |
---|---|---|---|---|
Aromatic compounds | ||||
Benzylalcohol # | 0.22 ± 0.20 | 0.77 ± 0.20 | 48.00 (WR) | 0.038 * |
Salicylaldehyde # | 4.02 ± 1.00 | 4.93 ± 1.79 | 0.02 (ST) | 0.988 |
2-Phenylethanol # | 0.11 ± 0.04 | 0.74 ± 0.23 | 39.00 (WR) | <0.001 *** |
Benzyl salicylate | n.q. | n.q. | - | - |
Terpenes | ||||
Limonene # | n.q | n.q | - | - |
1,8-Cineole # | n.q | n.q | - | - |
α-Terpineol # | 0.10 ± 0.03 | 0.44 ± 0.05 | 5.26 (ST) | <0.001 *** |
Elemol | 0.01 ± 0.01 | 0.27 ± 0.09 | 40.50 (WR) | 0.002 ** |
Guaia-4(15)-en-11-ol # + Guaia-4-en-11-ol # | 0.04 ± 0.04 | 4.31 ± 1.35 | 36.00 (WR) | <0.001 *** |
Fatty acids/aldehydes | ||||
(E)-4-Nonenal | n.q. | n.q. | - | - |
Myristaldehyde | 3.16 ± 0.34 | 8.53 ± 2.17 | 3.13 (ST) | 0.007 ** |
Myristic acid # | 0.40 ± 0.08 | 6.13 ± 1.59 | 8.19 (ST) | <0.001 *** |
Pentadecanoic acid # | 1.51 ± 0.30 | 4.49 ± 0.51 | 38.00 (WR) | <0.001 *** |
Palmitic acid # | 24.53 ± 3.57 | 72.04 ± 6.13 | 36.00 (WR) | <0.001 *** |
Oleic acid # | 5.04 ± 0.92 | 7.34 ± 1.09 | 1.51 (ST) | 0.154 |
Stearic acid # | 1.65 ±0.19 | 5.29 ± 0.71 | 6.39 (ST) | <0.001 *** |
Others | ||||
1-Hexanol | 0.14 ± 0.01 | 0.17 ± 0.01 | 12.00 (WR) | 0.038 * |
Unidentified compound | traces | 1.64 ± 0.88 | 40.00 (WR) | 0.002 ** |
Compound | Uninfected Roots | Infected Roots | t-Value/T-Value | p-Value |
---|---|---|---|---|
Salicin | 41.87 ± 16.03 | 77.22 ± 34.06 | 19.00 (WR) | 0.336 |
Salicin-7-sulfate | 2.08 ± 0.26 | 3.39 ± 0.39 | 2.52 (ST) | 0.026 * |
Salirepin | 14.50 ± 2.39 | 20.62 ± 2.79 | 1.53 (ST) | 0.151 |
Salirepin-7-sulfate | 0.30 ± 0.03 | 0.42 ± 0.05 | 1.77 (ST) | 0.1 |
Salicortin | 368.31 ± 172.48 | 204.06 ± 70.32 | 2.25 (ST) | 0.056 |
Tremulacin | 3.16 ± 1.27 | 2.49 ± 1.98 | 0.28 (ST) | 0.785 |
Homaloside D | 16.77 ± 9.28 | 7.78 ± 3.04 | 33.00 (WR) | 0.596 |
C | Guaia-4(15)-en-11-ol (1) | Guaia-4-en-11-ol (2) | ||
---|---|---|---|---|
13C | 1H | 13C | 1H | |
1 | 52.34 (CH) | 1.18 (m) | 55.60 (CH) | 2.28 (m) |
2 | 32.42 (CH2) | 1.76 (m, Hα) 1.00 (m, Hβ) | 30.42 (CH2) | 1.92 (m) 1.51 (m) |
3 | 32.96 (CH2) | 2.32 (ddm, J = 15.9, 7.8, Hβ) 2.19 (m, Hα) | 36.60 (CH2) | 2.23 (m) 2.13 (m) |
4 | 159.08 (Cq) | – | 131.74 (Cq) | – |
5 | 46.33 (CH) | 2.15 (m) | 138.62 (Cq) | – |
6 | 34.56 (CH2) | 1.88 (ddd, J = 13.9, 9.2, 6.1, Hβ) 1.54 (ddd, J = 13.9, 11.3, 8.0, Hα) | 30.37 (CH2) | 2.67 (d, J = 15.2) 2.00 (m) |
7 | 49.56 (CH) | 1.41 (m) | 49.30 (CH) | 1.36 (m) |
8 | 26.71 (CH2) | 1.74 (m, Hα) 1.10 (m, Hβ) | 31.51 (CH2) | 1.90 (m) 1.00 (m) |
9 | 40.29 (CH2) | 1.77 (m, Hβ) 0.96 (m, Hα) | 40.26 (CH2) | 1.78 (m) 1.10 (m) |
10 | 42.32 (CH) | 1.06 (m) | 39.37 (CH) | 1.33 (m) |
11 | 73.26 (Cq) | – | 72.82 (Cq) | – |
12 | 27.75 (CH3) | 1.03 (s) | 26.89 (CH3) | 1.03 (s) |
13 | 25.78 (CH3) | 1.01 (s) | 26.66 (CH3) | 1.02 (s) |
14 | 21.58 (CH3) | 0.87 (d, J = 6.5) | 22.01 (CH3) | 0.93 (d, J = 6.6) |
15 | 104.24 (CH2) | 5.00 (m, HE) 4.91 (br, HZ) | 14.60 (CH3) | 1.63 (br s) |
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Lackus, N.D.; Morawetz, J.; Xu, H.; Gershenzon, J.; Dickschat, J.S.; Köllner, T.G. The Sesquiterpene Synthase PtTPS5 Produces (1S,5S,7R,10R)-Guaia-4(15)-en-11-ol and (1S,7R,10R)-Guaia-4-en-11-ol in Oomycete-Infected Poplar Roots. Molecules 2021, 26, 555. https://doi.org/10.3390/molecules26030555
Lackus ND, Morawetz J, Xu H, Gershenzon J, Dickschat JS, Köllner TG. The Sesquiterpene Synthase PtTPS5 Produces (1S,5S,7R,10R)-Guaia-4(15)-en-11-ol and (1S,7R,10R)-Guaia-4-en-11-ol in Oomycete-Infected Poplar Roots. Molecules. 2021; 26(3):555. https://doi.org/10.3390/molecules26030555
Chicago/Turabian StyleLackus, Nathalie D., Jennifer Morawetz, Houchao Xu, Jonathan Gershenzon, Jeroen S. Dickschat, and Tobias G. Köllner. 2021. "The Sesquiterpene Synthase PtTPS5 Produces (1S,5S,7R,10R)-Guaia-4(15)-en-11-ol and (1S,7R,10R)-Guaia-4-en-11-ol in Oomycete-Infected Poplar Roots" Molecules 26, no. 3: 555. https://doi.org/10.3390/molecules26030555