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Evaluation of Essential Plant Oils for the Control of Plasmopara Viticola A

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Evaluation of Essential Plant Oils for the Control of Plasmopara Viticola A This article was downloaded by: [CRA] On: 21 February 2014, At: 03:10 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK Journal of Essential Oil Research Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tjeo20 Evaluation of essential plant oils for the control of Plasmopara viticola A. La Torrea, C. Mandalàa, L. Pezzaa, F. Caradoniaa & V. Battagliaa a Consiglio per la Ricerca e la sperimentazione in Agricoltura - Centro di ricerca per la patologia vegetale, Rome, Italy Published online: 20 Feb 2014. To cite this article: A. La Torre, C. Mandalà, L. Pezza, F. Caradonia & V. Battaglia , Journal of Essential Oil Research (2014): Evaluation of essential plant oils for the control of Plasmopara viticola, Journal of Essential Oil Research, DOI: 10.1080/10412905.2014.889049 To link to this article: http://dx.doi.org/10.1080/10412905.2014.889049 PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http:// www.tandfonline.com/page/terms-and-conditions Journal of Essential Oil Research, 2014 http://dx.doi.org/10.1080/10412905.2014.889049 RESEARCH ARTICLE Evaluation of essential plant oils for the control of Plasmopara viticola A. La Torre*, C. Mandalà, L. Pezza, F. Caradonia and V. Battaglia Consiglio per la Ricerca e la sperimentazione in Agricoltura - Centro di ricerca per la patologia vegetale, Rome, Italy (Received 15 October 2012; accepted 24 January 2014) The aim of this study was to test the effectiveness of various concentrations of tea tree oil and clove oil against Plasmopara viticola. In vitro tests entailed assessing the development of P. viticola on leaf disks and calculating sporangial germination. In a field trial, formulated products containing 23.8% tea tree oil (BM-608) and 15% clove oil (Sporatec) were tested in an organic vineyard to control downy mildew. The efficacy of pure and formulated essential oils was compared with a water control and a reference product (Airone, containing copper oxychloride 50% and copper hydroxide 50%). Laboratory experiments revealed the inhibitory activity of essential oils, although the effectiveness was lower than that of the reference product. The field data demonstrated that BM-608 and Sporatec were able to control the development of downy mildew although they were not as effective as the reference product. This study suggests the application of essential oils in order to reduce the use of synthetic pesticides in agriculture, in accordance with European laws, and to avoid the environmental pollution of copper in organic farming. Keywords: natural products; tea tree oil; clove oil; grape downy mildew; organic farming 1. Introduction alcohols (11). Although the composition is variable, it Awareness of the importance of environmental protec- is regulated by an international standard for ‘Oil of tion and the food safety has increased noticeably over Melaleuca – terpinen-4-ol type’, which sets the maxima the last years. According to EU legislation on the and/or minima of fourteen components of the oil, sustainable use of pesticides (Directive 2009/128/EC), which were selected for a variety of reasons, including member states should take all necessary measures to biological activities. According to ISO 4730:2004, the reduce the risks and impacts of pesticide use on human major components are as follows: terpinen-4-ol health and the environment. In addition, Commission (≥30%), γ-terpinen (10–28%), α-terpinen (5–13%) and Regulation (EC) No. 473/2002 aimed to reduce the 1.8-cineole (≤15%) (8, 10, 12). annual amount of copper used in organic farming. TTO has been found to have some of the strongest Since the use of copper as a fungicide may have antimicrobial properties ever discovered in a plant (13). long-term consequences due to its accumulation in the The antimicrobial activity is attributed mainly to terpi- Downloaded by [CRA] at 03:10 21 February 2014 soil, research efforts should be increased in order to nen-4-ol, which is the major component of the oil (11, find appropriate alternative solutions. 14). TTO is effective as an antiseptic, fungicide and A large number of investigations have focused on bactericide (15). It has been re-evaluated in recent years natural products able to reduce or replace copper (1–7). as an alternative agent to antibiotics (10, 16). The mode However, copper is still essential for the cultivation of of action of this oil, which has been studied in bacteria a wide range of plants, especially for the control of and fungi, involves both the loss of membrane integrity Plasmopara viticola (Berk. & M.A. Curtis) Berl. & De and permeability, accompanied by the release of Toni on grapevine. These considerations led us to carry intracellular material and the inhibition of cellular out research in order to control grape downy mildew in respiration, with the consequent inability to maintain an environmentally friendly way by identifying the homeostasis associated with changes in cell morphol- effectiveness of essential oils: tea tree oil and clove oil. ogy (10, 11, 17). Tea tree oil (TTO) is obtained by the steam distilla- Cloves (Syzygium aromaticum (L.) Merr. & L.M. tion or hydro-distillation of leaves and terminal Perry, syn. Eugenia aromaticum or Eugenia caryophyl- branches of Australian bush Melaleuca alternifolia lata) are the aromatic dried flower buds of the tree (Maiden & Betche) Cheel (Myrtaceae) (8–10). It is a family Myrtaceae (18, 19). Clove oil (CO) is a steam complex mixture of terpene hydrocarbons, mainly distillation product obtained from clove leaves or buds, monoterpenes, sesquiterpenes and their associated which contain phenylpropanoides such as eugenol, *Corresponding author. Email: [email protected] © 2014 Taylor & Francis 2 A. La Torre et al. thymol, carvacrol and cinnamaldehyde (19). The major Candia located near Rome, Italy. Infected grapevine component is eugenol (84-88%) and further distillation leaves showing the symptoms of the disease were used of CO produces a refined product containing almost pure as the source of sporangial inoculum of oomycete. eugenol (i.e. > 95%) (8). Several studies have demon- strated the potent antifungal (20–22), antiviral (19) and antibacterial effects of cloves (23–26). Cloves are used 2.1.3. Leaf disk assay and inoculation with P. viticola in Ayurveda, Chinese medicine and Western herbalism Leaf disks were prepared according to Boso and fl (27). Cloves are antimutagenic (28), anti-in ammatory Kassemeyer (31). Untreated leaves of Vitis vinifera L. (29), antioxidant (19), antiulcerogenic (24), antithrom- cv. Malvasia di Candia, were used to obtain 18-mm botic (18) and antiparasitic (30). The main antifungal diameter leaf disks, which were punched out with a action of phenolic compounds, such as eugenol, appears cork borer. Ten leaf disks, obtained from ten healthy to be exerted on the cellular membrane (11, 17). leaves, were prepared for each treatment and, after TTO and CO have recently been included in EU surface sterilization, were immersed for 20 minutes in Regulation No. 540/2011 (the list of active substances solutions of different concentrations (ranging from approved for use in plant protection products) and are 0.0125% to 1%) of the tested products. The immersion authorized for use in many European countries. To provided a uniform distribution of products on all parts date, commercial formulations have not been authorized of the leaf disk. In order to overcome limited solubility against P. viticola. TTO has been authorized as a in aqueous media, E-TTO and T-4-ol were emulsified fungicide to control early blight on potato, and with 0.05% Tween 20. Distilled sterile water was used powdery mildew on carrot, herbs, cucumber, water- as a control. A control solution containing 0.05% melon, tomato, pepper and ornamentals, and CO has Tween 20 was also considered in order to examine the been authorized for post-harvest use on apples and effects of the solubilizing agent. A standard (Airone, pears against Gloesporium spp. and Penicillium spp.. marketed by ISAGRO S.p.A. – copper oxychloride fi Our research thus aimed to evaluate the ef cacy of 50% and copper hydroxide 50%) was used as a these essential oils for the control of P. viticola, a key positive control in the dosage prescribed on the label. pathogen in grapes. The activity of pure and formulated Leaf disks were dried aseptically and then placed in essential oils was evaluated both in vitro and in vivo. Petri dishes (90 mm) containing 1% water agar and 30 mg/L benzimidazole. The leaf disks were placed 2. Experimental with the abaxial side up and after 24 hours each disk 2.1. Laboratory experiments was administered one drop of 30 μL inoculum by means of a micropipette.
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