Bioassays Against Pinewood Nematode: Assessment of a Suitable Dilution Agent and Screening for Bioactive Essential Oils
Abstract
:1. Introduction
2. Results and Discussion
2.1. Assessment of Triton X-100 and Acetone Nematicidal Activity
2.2. Comparative Evaluation of Essential Oils’ Nematicidal Activity Using Triton X-100 or Acetone as Solvent
Species | Dilution agent | 0.25 mg/mL | 0.5 mg/mL | 1 mg/mL |
---|---|---|---|---|
Cymbopogon citratus | Triton X-100 * | 14.98 ± 2.17 | 81.60 ± 1.72 | − |
Acetone | 83.80 ± 1.08 | 89.39 ± 1.18 | − | |
Origanum vulgare | Triton X-100 * | 2.78 ± 0.68 | 3.72 ± 0.56 | 26.61 ± 3.83 |
Acetone | 94.90 ± 1.06 | 98.81 ± 0.51 | 100.00 ± 0.00 | |
Satureja montana 1 | Triton X-100 * | 7.13 ± 1.19 | − | − |
Acetone | 57.60 ± 2.44 | − | − |
Species | l | c | LC100 | |
---|---|---|---|---|
Origanum vulgare * | 0.100 ± 0.0017 a | 1.770 ± 0.095 a | 0.498 ± 0.028 a | |
Ruta graveolens 1 | 0.096 ± 0.0014 a | 1.799 ± 0.052 a | 0.571 ± 0.046 b | |
Ruta graveolens 2 | 0.095 ± 0.0008 a | 1.915 ± 0.135a | 0.663 ± 0.032 c | |
Satureja montana 1 * | 0.099 ± 0.0003 a | 1.946 ± 0.008 a | 0.793 ± 0.002d | |
Satureja montana 2 | 0.089 ± 0.0024 b | 2.832 ± 0.056 b | 0.819 ± 0.007 d | |
Satureja montana 3 | 0.089 ± 0.0013 c | 2.798 ± 0.020 c | 0.828 ± 0.001d |
2.3. Essential Oils Nematicide Activity
Code | Family / Species | Collection place or source b | Date | Plant part c Status | I.P. d | Oil yield(%, v/w) | Mortality e (%) |
---|---|---|---|---|---|---|---|
Anacardiaceae | |||||||
Scm | Schinus molle L. | Évora | 2005 | Leaves, Fresh | H | 0.40 | 1.54 ± 0.47 |
Apiaceae | |||||||
Al | Angelica lignescens Reduron et Danton | Flores (Az) | 2008 | AP (V), Fresh | H | 0.08 | 1.75 ± 0.47 |
Cha | Chaerophyllum azoricum Trelease | Flores (Az) | 2008 | AP (V), Fresh | H | 0.25 | 1.20 ± 0.48 |
Fv1 | Foeniculum vulgare Miller | Graciosa (Az) | 2008 | AP (F), Fresh | H | 0.33 | 6.21 ± 0.71 * |
Fv2 | Foeniculum vulgare Miller | HS | 2008 | Seeds, Dried | H | 5.61 | 8.60 ± 0.81 * |
Fv3 | Foeniculum vulgare Miller | HS | 2008 | Seeds, Dried | H | 6.09 | 9.89 ± 1.71 * |
Fv4 | Foeniculum vulgare Miller | HS | 2008 | Seeds, Dried | H | 5.88 | 6.29 ± 0.91 * |
Fv5 | Foeniculum vulgare Miller | BPGV | 2008 | Seeds, Dried | H | 4.78 | 5.95 ± 0.79 * |
Fv6 | Foeniculum vulgare Miller | HS | 2008 | Seeds, Dried | H | 1.07 | 7.13 ± 0.50 * |
Cupressaceae | |||||||
Cj | Cryptomeria japonica (L. fil.) D. Don. f | Flores (Az) | 2008 | Berries, Fresh | H | 0.41 | 0.79 ± 0.39 |
Jb1 | Juniperus brevifolia (Seub.) Antoine | Flores (Az) | 2008 | Berries, Fresh | H | 0.06 | 0.84 ± 0.16 * |
Jb2 | Juniperus brevifolia (Seub.) Antoine | Flores (Az) | 2008 | AP (V), Fresh | H | 0.45 | 2.56 ± 0.66 |
Geraniaceae | |||||||
Pg1 | Pelargonium graveolens L’Hér. | Lisboa | 2009 | AP (V), Fresh | H | 0.19 | 74.79 ± 2.56 * |
Lamiaceae | |||||||
Mo | Melissa officinalis L. | HS | 2009 | AP (F), Fresh | H | 0.04 | 99.30 ± 0.54 * |
Ma | Mentha aquatica L. | HS | 2009 | AP (F), Dried | H | 0.90 | 7.77 ± 0.83 * |
Mc1 | Mentha cervina L. | Beja | 2005 | AP (F), Fresh | H | 2.00 | 93.56 ± 1.07 * |
Mc2 | Mentha cervina L. | HS | 2009 | AP (V), Dried | H | 2.12 | 92.57 ± 1.48 * |
Ms | Mentha spicata L. | Beja | 2009 | AP (V), Fresh | H | 0.25 | 47.36 ± 2.22 * |
Nc | Nepeta cataria L. | HS | 2009 | AP (F), Fresh | H | 0.18 | 22.03 ± 2.66 * |
Ov | Origanum vulgare L. a | Évora | 2007 | AP (F), Fresh | H | 1.70 | 100.00 ± 0.00 * |
Ro1 | Rosmarinus officinalis L. | HS | 2009 | Leaves, Dried | H | 1.95 | 2.55 ± 0.84 |
Ro2 | Rosmarinus officinalis L. | Lisboa | 2009 | AP (V), Fresh | H | 0.64 | 0.40 ± 0.41 |
Ro3 | Rosmarinus officinalis L. | Lisboa | 2009 | AP (F), Fresh | H | 1.14 | 2.30 ± 0.52 |
So1 | Salvia officinalis L. | Lisboa | 2009 | AP (V), Fresh | H | 0.54 | 1.06 ± 0.40 |
So2 | Salvia officinalis L. | Lisboa | 2009 | AP (V), Fresh | H | 0.71 | 0.07 ± 0.43 |
Sm1 | Satureja montana L.a | HS | 2008 | Leaves, Dried | H | 1.60 | 100.00 ± 0.00 * |
Sm2 | Satureja montana L. | HS | 2009 | AP (V), Dried | H | 0.55 | 100.00 ± 0.00 * |
Sm3 | Satureja montana L. | Beja | 2009 | AP (F), Fresh | D-E | − | 100.00 ± 0.00 * |
Tc1 | Thymus caespititius Brot. | Madeira | 2006 | AP (F), Fresh | D-E | − | 6.06 ± 0.62 * |
Tc2 | Thymus caespititius Brot. | S. Jorge (Az) | 2007 | AP (F), Fresh | D-E | − | 97.01 ± 0.98 * |
Tc3 | Thymus caespititius Brot. | Flores (Az) | 2008 | AP (F), Fresh | H | 0.06 | 94.63 ± 1.30 * |
Tc4 | Thymus caespititius Brot. | Corvo (Az) | 2008 | AP (F), Fresh | H | 0.22 | 99.44 ± 0.26 * |
Tc5 | Thymus caespititius Brot. | Gerês | 2008 | AP (F), Fresh | H | 0.35 | 51.61 ± 3.60 * |
Tc6 | Thymus caespititius Brot. | Graciosa (Az) | 2008 | AP (F), Fresh | H | 0.38 | 58.21 ± 2.19 * |
Tca | Thymus camphoratus Hoffmans. & Link | Faro | 2008 | AP (F), Fresh | H | 0.21 | 3.30 ± 0.59 * |
Tvl | Thymus villosus ssp. lusitanicus (Boiss.) Coutinho | Leiria | 2008 | AP (F), Fresh | H | 1.25 | 66.85 ± 3.44 * |
Tzs | Thymus zygis ssp. sylvestris (Hoffmans. & Link) Coutinho | Leiria | 2008 | AP (F), Fresh | H | 0.23 | 24.25 ± 3.18 * |
Lauraceae | |||||||
Cc | Cinnamomum camphora (L.) T. Nees & C.H. Eberm. | Coimbra | 2009 | Branches without leaves, Dried | H | 0.55 | 1.56 ± 0.16 * |
La | Laurus azorica (Seub.) J. Franco | Flores (Az) | 2008 | AP (V), Fresh | H | 0.25 | 2.17 ± 0.66 |
Lnc1 | Laurus novocanariensis Rivas Mart., Lousã, Fern. Prieto, E. Díaz, J.C. Costa & C. Aguiar | Porto da Cruz, Madeira | 2009 | Branches, Fresh | H | 0.42 | 2.22 ± 0.39 * |
Lnc2 | Laurus novocanariensis Rivas Mart., Lousã, Fern. Prieto, E. Díaz, J.C. Costa & C. Aguiar | Porto da Cruz, Madeira | 2009 | Branches, Fresh | H | 0.48 | 2.80 ± 0.34 * |
Lnc3 | Laurus novocanariensis Rivas Mart., Lousã, Fern. Prieto, E. Díaz, J.C. Costa & C. Aguiar | Ribeiro Frio, Madeira | 2009 | Branches, Fresh | H | 0.39 | 2.66 ± 0.75 |
Lnc4 | Laurus novocanariensis Rivas Mart., Lousã, Fern. Prieto, E. Díaz, J.C. Costa & C. Aguiar | Ribeiro Frio, Madeira | 2009 | Branches, Fresh | H | 0.64 | 2.91 ± 0.44 * |
Lnc5 | Laurus novocanariensis Rivas Mart., Lousã, Fern. Prieto, E. Díaz, J.C. Costa & C. Aguiar | S. Vicente, Madeira | 2000 | Leaves, Fresh | H | 0.30 | 4.46 ± 0.54 * |
Myrtaceae | |||||||
Eg | Eucalyptus globulus Labill. | Lisbon | 2009 | AP (F), Fresh | H | 2.15 | 4.14 ± 0.85 * |
Pittosporaceae | |||||||
Pu1 | Pittosporum undulatum Vent. | Graciosa (Az) | 2008 | Berries, Fresh | H | 0.21 | 1.22 ± 0.34 |
Pu2 | Pittosporum undulatum Vent. | Graciosa (Az) | 2008 | Leaves, Fresh | H | 0.08 | 1.46 ± 0.44 |
Poaceae | |||||||
Cyc | Cymbopogon citratus (DC) Stapf. a | Faro | 2008 | Leaves, Fresh | H | 0.80 | 98.86 ± 0.32 * |
Rutaceae | |||||||
Ca | Citrus aurantium L. | Évora | 2009 | Leaves, Fresh | H | 0.31 | 26.59 ± 1.47 * |
Rg1 | Ruta graveolens L. | Évora | 2009 | AP (V), Fresh | H | 2.60 | 100.00 ± 0.00 * |
Rg2 | Ruta graveolens L. | HS | 2009 | AP (F), Dried | H | 0.90 | 100.00 ± 0.00 * |
Verbenaceae | |||||||
Lc | Lippia citriodora Kunth | HS | 2009 | AP (V), Dried | H | 0.19 | 54.63 ± 3.53 * |
2.4. Chemical Profile of Essential Oils
Lamiaceae | Rutaceae | ||||
---|---|---|---|---|---|
Compounds | RI a | Sm2 | Sm3 | Rg1 | Rg2 |
2-Methyloctane | 887 | − | − | t b | t |
Tricyclene | 921 | t | t | − | − |
α-Thujene | 924 | 0.3 | 2.4 | − | − |
α-Pinene | 930 | 1.6 | 2.3 | − | − |
Camphene | 938 | 1.6 | 0.1 | − | − |
1-Octen-3-ol | 961 | t | t | − | − |
β-Pinene | 963 | 0.2 | 1.2 | − | − |
n-Octanal | 973 | − | − | t | t |
β-Myrcene | 975 | t | 2.7 | − | − |
α-Phellandrene | 995 | t | 0.4 | − | − |
δ-3-Carene | 1000 | t | 0.1 | − | − |
α-Terpinene | 1002 | 0.3 | 4.1 | − | − |
p-Cymene | 1003 | 20.3 | 8.1 | − | − |
1,8-Cineole | 1005 | t | t | − | − |
β-Phellandrene | 1005 | t | 0.1 | − | − |
Limonene | 1009 | 0.6 | 0.5 | − | − |
cis-β-Ocimene | 1017 | t | t | − | − |
γ-Terpinene | 1035 | 4.3 | 41.1 | − | − |
trans-Sabinene hydrate | 1037 | t | t | − | − |
2-Nonanone | 1058 | − | − | t | t |
2,5-Dimethyl styrene | 1059 | t | t | − | − |
Terpinolene | 1064 | 0.4 | t | − | − |
cis-Sabinene hydrate | 1066 | t | t | − | − |
n-Nonanal | 1073 | − | − | t | t |
Linalool | 1074 | t | t | − | − |
Geigerene isomer | 1116 | − | − | t | t |
Geigerene | 1121 | − | − | 0.5 | 0.1 |
Borneol | 1134 | 3.9 | 0.1 | − | − |
Terpinen-4-ol | 1148 | 2.3 | 0.2 | − | − |
α-Terpineol | 1159 | t | t | − | − |
2-Decanone | 1166 | − | − | t | t |
Carvacrol methyl ether | 1224 | 3.7 | t | − | − |
2-Undecanone | 1275 | − | − | 94.4 | 92.8 |
Thymol | 1275 | 15.2 | t | − | − |
Carvacrol | 1286 | 40 | 35.3 | − | − |
β-Bourbonene | 1379 | t | t | − | − |
2-Dodecanone c | 1389 | − | − | t | t |
β-Caryophyllene | 1414 | 2.6 | 1.1 | − | − |
β-Copaene | 1426 | t | t | − | − |
Aromadendrene | 1428 | 0.3 | t | − | − |
α-Humulene | 1447 | t | t | − | − |
2-Tridecanone | 1479 | − | − | t | t |
β-Bisabolene | 1500 | 0.1 | t | − | − |
trans-Calamenene | 1505 | 0.1 | t | − | − |
δ-Cadinene | 1505 | 0.1 | 0.1 | − | − |
β-Caryophyllene oxide | 1561 | 0.6 | t | − | − |
UI Rg d | 1626 | − | − | 5.1 | 7.1 |
% of identification | 98.6 | 99.9 | 94.9 | 92.9 | |
Grouped components | |||||
Monoterpene hydrocarbons | 29.6 | 63.1 | − | − | |
Oxygen-containing monoterpenes | 65.2 | 35.6 | − | − | |
Sesquiterpene hydrocarbons | 3.2 | 1.2 | − | − | |
Oxygen-containing sesquiterpenes | 0.6 | − | − | − | |
C13 compounds | − | − | 0.5 | 0.1 | |
Others | − | − | 94.4 | 92.8 |
3. Experimental
3.1. Plant Material
3.2. Essential Oils and Volatiles Extraction
3.3. Rearing and Collection of Nematodes
3.4. Bioassays
3.5. Determination of Essential Oils Composition
3.5.1. Gas Chromatography (GC)
3.5.2. Gas Chromatography-Mass Spectrometry (GC-MS)
3.6. Data Analysis
4. Conclusions
Acknowledgements
- Samples Availability: No pure compounds were used. Samples of the tested essential oils are available from the authors.
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Barbosa, P.; Faria, J.M.S.; Mendes, M.D.; Dias, L.S.; Tinoco, M.T.; Barroso, J.G.; Pedro, L.G.; Figueiredo, A.C.; Mota, M. Bioassays Against Pinewood Nematode: Assessment of a Suitable Dilution Agent and Screening for Bioactive Essential Oils. Molecules 2012, 17, 12312-12329. https://doi.org/10.3390/molecules171012312
Barbosa P, Faria JMS, Mendes MD, Dias LS, Tinoco MT, Barroso JG, Pedro LG, Figueiredo AC, Mota M. Bioassays Against Pinewood Nematode: Assessment of a Suitable Dilution Agent and Screening for Bioactive Essential Oils. Molecules. 2012; 17(10):12312-12329. https://doi.org/10.3390/molecules171012312
Chicago/Turabian StyleBarbosa, Pedro, Jorge M. S. Faria, Marta D. Mendes, Luís Silva Dias, Maria Teresa Tinoco, José G. Barroso, Luis G. Pedro, Ana Cristina Figueiredo, and Manuel Mota. 2012. "Bioassays Against Pinewood Nematode: Assessment of a Suitable Dilution Agent and Screening for Bioactive Essential Oils" Molecules 17, no. 10: 12312-12329. https://doi.org/10.3390/molecules171012312