J. Blanco Salas 1,2 , T. Ruiz Téllez 3 , F. M. Vázquez Pardo 2 , M. A. Cases Capdevilla 4 , M. J. Pérez-Alonso 5 , D. García Alonso 2 1 FOTEX, Badajoz; 2 Grupo HABITAT, Instituto de Investigaciones Agrarias Finca La Orden-Valdesequera, CICYTEX, Gobierno de Extremadura, Autovía A-5 Km 372, 06187 Guadajira, Badajoz, España; 3 Grupo de Investigación en Biología de la Conservación. Facultad de Ciencias. Universidad de Extremadura. Avda. de Elvas s/n, 06071 Badajoz, España. (truiz@unex.es); 4 Departamento de Medio Ambiente, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, 28071. Madrid, España; 5 Universidad Complutense. Facultad de Biología. Departamento de Biología Vegetal, Botánica. 28040. Madrid, España. CULTIVATED vs WILD POPULATION: THE CASE OF THE ESSENTIAL OIL OF THYMUS MASTICHINA (L.) L. AND T. ZYGIS S.L. IN EXTREMADURA (SPAIN) This work is partially based on data from Blanco J. (2005) Contribución al conocimiento del los recursos fitogenéticos de Extremadura: el caso de los Tomillos. Tesis Doctoral. Universidad de Extremadura. Acknowledgments to INIA (RF00-019-C2-2) and J. Sanz (C.S.I.C.). Introduction  Many of the productions obtained from native Thymus species (thymes) in the Iberian Peninsula are collected in wild populations.  However, legislation limits such collections, and industry demands homogeneous quality raw materials. Cultivation of useful line plants for the global market has been made necessary. Material and Methods • Juvenile’s transplant of 4 populations of T. mastichina and 4 populations of T. zygis, were performed, for an experimental crop with drip irrigation at the “La Orden” Agricultural Research Center, Badajoz (Spain). • Individuals were collected in wild populations and transplanted to pots and finally to the soil, where they were grown for 1 year. The following 2 years they were mowed twice, at flowering and fruiting season. At the same time, the same mowing was made at the source (wild) populations. • Essential oil was extracted by hydrodistillation. For each sample 2 extractions were performed: one from the whole plant (WP) and one from flowers and leaves (FL). Essential oil extracted was quantified to estimate the yield of essential oil and then used to determine the components and their percentages relative to the total. Gas chromatography (GC) and gas chromatography - mass spectrometry (GC-MS) were used. Results were compared by statistical analysis. Results and Discussion  Wild and cultivated T. mastichina , at fruiting stage, have a significant different yield (for both WP and FL materials), and a different chemical composition.  Wild and cultivated T. zygis have a different yield at flowering stage (just for WP material) but differences in its chemical composition are found only at fruiting stage.  The patterns of variation of chemical profiles here described, should be considered to implement strategic plans to promote cultivation of these two etnobotanical species in the region. Córdoba, España 17 - 21 de noviembre, 2014 Objective: This study aims to know if the cultivation of two etnobotanical species, very popular in Spain, Thymus mastichina (L.) L. and T. zygis s.l., influences yields and chemical composition of their essential oils. Table 1. Chemical composition (% values range for 2-3 years of study) of the main components of the essential oil of 4 wild populations (PW) of T. zygis s.l. and their cultivated homologues (PC). P1 = Badajoz; P2 = Guadajira; P3= Los Santos de Maimona; P4 = Solana de los Barros . RI = retention index according to n-paraffins on DB-1 column; RTM= Retention time in GLC-MS; t = trace (< 0.1%); R= yield; WP = whole plant; FL = flowers and leaves. Table 2. Chemical composition (% values range for 2-3 years of study) of the main components of the essential oil of 4 wild populations (PW) of T. mastichina and their cultivated homologues (PC). P1 = Alía; P2 = La Garganta; P3= El Portanchito; P4 = Villafranca de los Barros . RI = retention index according to n- paraffins on DB-1 column; RTM= Retention time in GLC-MS; t = trace (< 0.1%); R= yield; WP = whole plant; FL = flowers and leaves. Compound RI RTM P1W P1C P2W P2C P3W P3C P4W P4C P1W P1C P2W P2C P3W P3C P4W P4C camphene 949 5,32 0,5-1,3 1,1-1,4 0,9-1,3 1,2-2,0 1,2-2,7 1,0-1,2 1,2-2,0 0,8-0,9 0,8-1,1 0,9-2,0 1,8-2,8 2,4-3,0 1,7-2,4 2,0-2,8 1,9-2,5 1,6-1,9 β-pinene 985 5,76 0,4-0,5 0,4-0,9 0,4-1,7 1,4-1,7 1,1-2,0 0,6-0,8 1,3-3,0 0,5-0,7 0,3-1,0 0,3-1,1 3,2-6,2 4,3-4,4 2,6-4,4 2,6-3,0 2,6-4,0 1,7-2,2 myrcene 991 5,86 1,8-3,9 2,5-3,1 1,7-2,1 1,3-1,8 2,2-4,2 2,0-2,2 2,1-2,4 1,5-1,9 1,2-2,4 1,6-4,4 1,8-2,3 2,2-2,7 1,0-2,2 2,6-3,2 1,9-2,4 2,3-3,1 p -cymene 1021 6,53 8,8-23,8 18,5-19,3 22,5-31,3 18,7-19,9 16,22,9 12,3-12,7 17,7-25,8 12,2-14,5 23,8-30,7 21,6-31,2 20,5-43,5 26,1-28,0 34,9-40,1 22,6-35,9 37,8-42,4 19,7-32,5 (Z )-β-ocimene 1038 6,91 t t-0,8 0,9-6,5 0,9-1,4 5,0-21,7 4,0-4,2 1,2-1,8 0,3-0,7 t-0,8 t-0,3 1,2-6,9 2,3-3,2 3,2-5,8 6,1-7,2 1,2-4,6 1,5-4,3 γ-terpinene 1060 7,17 9,6-12,9 9,8-11,0 5,8-10,7 9,0-10,1 8,0-10,7 9,3-9,5 7,8-13,7 8,8-9,8 3,6-4,9 4,7-16,4 1,9-14,1 7,7-8,7 2,2-3,0 9,7-11,2 2,9-4,1 5,8-10,5 linalool 1102 7,93 1,0-2,6 1,1-1,7 7,4-18,4 10,8-12,3 10,2-17,6 8,0-8,8 5,8-7,1 9,2,9,8 1,4-2,2 1-2,6 11,2-12,6 14,1-16,0 10,8-12,2 6,7-7,3 4,1-5,4 5,6-10,6 borneol 1172 9,44 0,3-0,9 0,4-0,6 1,1-1,8 0,7-0,9 0,3-1,0 0,5-0,7 0,9-1,6 t-0,8 0,6-1,4 t-0,5 2,9-4,3 3,6-3,8 2,2-5,7 2,4-2,6 1,6-1,7 0,9-1,1 terpinen-4-ol 1216 9,68 0,6-1,7 1,2-1,8 2,3-5,4 1,8-2,2 1,0-2,3 0,8,1,1 1,7-3,7 0,7-1,5 0,7-1,2 0,8-1,9 5,2-7,5 6,0-6,3 t-5,1 3,1-3,8 2,7-5,5 2,3-4,4 thymol 1290 12,01 41,9-74,0 48,5-49,3 32,4-35-8 48,0-49,1 22,4-35,4 52,1-53,8 40,0-49,5 54,0-58,3 50,7-62,2 28,2-65,8 13,9-28,9 22,9-23,3 18,8-27,0 22,5-32,6 27,2-31,5 32,6-42,3 carvacrol 1298 12,23 1,0-4,43 5,5-7,8 1,4-1,6 1,4-1,8 1,2-1,6 1,99-2,3 1,9-2,5 1,9-3,2 1,0-3,2 1,5-4,3 1,1-1,2 1,0-1,3 1,9-2,0 1,7-1,8 2,3-3,6 2,2-2,5 WP 1,7-3,6 4,2 0,9-1,4 1,7 1,0-1,1 3,3 1,3-1,7 3,6 0,7-2,1 0,9-1,4 0,5-0,8 0,6-0,9 0,4-0,8 0,6-1,5 0,9-1,0 1,1-2,3 FL 3,3-5,2 7,4 1,4-2,2 2,2 1,9-2,3 4,2 2,3-2,8 4,9 1,5-5,0 1,9-2,4 1,0-1,4 0,9-1,1 0,8-1,4 1,1-2,1 1,2-1,9 1,4-2,4 R (mL/100gr) fruiting stage flowering stage Table 3. Kruskal-Wallis test significance levels for the comparison of the yields between wild and cultivated population of T. zygis y T. mastichina in flowering and fruiting stage for whole plant to flower (WP) and leaves (FL). ns = no significance; * = p < 0.05; ** = p < 0.01; *** = p< 0.001. Table 4. Kruskal-Wallis test significance levels for the comparison of the main component of the essential oil composition between wild and cultivated population of T. zygis y T. mastichina in flowering and fruiting stage. ns = no significative; * = p< 0.05; ** = p < 0.01; *** = p < 0.001. Figure 2. Experimental cultivation of the 4 populations of T. zygis. In the rafts, mowed material for the present study. Figure 1. Harvesting cultivated populations of T. mastichina for the present study. Compound RI RTM P1W P1C P2W P2C P3W P3C P4W P4C P1W P1C P2W P2C P3W P3C P4W P4C Camphene 933 5,32 2,7-3,8 3,2-3,6 2,8-3,1 3,6-3,9 3,3-4,3 3,2-3,8 3,3-4,2 3,0-3,3 2,6-2,8 3,9-4,1 3,2-4,2 4,4-4,6 4,1-4,4 4,6-4,8 3,5-4,4 4,5-4,6 Sabinene 949 5,66 2,1-2,5 2,4-2,6 2,3-2,4 2,5-2,9 2,6-3,1 3,0-3,4 2,6-3,4 2,2-3,1 1,3-1,7 3,2-3,4 1,4-2,1 3,3-3,6 1,6-1,9 3,9-4,2 1,6-2,3 3,9-4,2 β-pinene 970 5,76 0,2-0,8 t-0,8 1,4-1,6 1,4-1,9 0,6-1,5 0,8-0,9 t-0,1 t-0,3 0,3-0,4 0,2-0,6 1,6-2,3 1,7-2,2 0,8-1,6 0,6-0,9 t-0,4 0,2-0,3 myrcene 978 5,86 5,1-7,2 6,3-6,9 4,7-5,7 5,9-6,5 6,2-7,2 6,2-7,3 6,2-8,1 6,4-6,7 4,7-5,7 7,9-8,7 5,2-6,6 7,6-7,8 6,3-7,7 8,6-9,0 6,9-8,0 8,8-9,3 limonene 1031 6,60 0,9-1,1 0,8-1,3 0,7-0,9 1,1-1,3 1,0-1,1 0,7-1,3 0,5-0,8 0,9-1,1 1,5-2,0 1,8-1,9 0,8-1,0 0,9-0,9 0,7-1,1 0,9-1,1 0,7-0,8 0,9-1,0 1,8 cineol 1039 6,70 73,3-75,1 72,2-75,1 70,7-70,9 64,0-68,3 71,2-73,5 65,5-68,7 73,7-77,3 69,9-72,1 75,5-78,0 69,3-70,6 68,8-73,2 63,4-65,7 70,0-75,5 65,5-68,7 72,0-73,5 69,3-71,7 linalool 1102 7,91 0,8-2,1 1,8-2,2 2,9-3,5 2,9-3,5 1,3-2,7 3,0-3,9 0,7-2,4 2,6-2,9 0,9-1,0 0,4-0,5 2,0-2,2 1,4-2,0 0,7-1,9 0,8-1,1 0,9-2,0 0,8-0,9 trans-dihydro-α-terp 1165 9,42 2,3-3,6 2,1-2,3 4,1-4,6 4,0-4,6 3,1-3,8 3,3-3,6 1,6-1,9 1,9-2,2 1,8-2,4 2,2-2,9 4,1-5,0 3,0-4,8 3,0-4,1 2,8-3,2 2,1-2,5 2,0-2,2 4-terpineol 1178 9,66 0,8-0,9 0,6-0,9 0,4-0,9 0,7-0,8 0,4-0,9 0,6-0,8 0,7-0,9 0,5-0,8 1,1-1,5 0,7-1,0 1,0-1,3 0,7-1,0 1,0-1,1 0,6-0,7 1,2-1,4 0,7-0,7 α-terpineol 1193 9,93 4,4-5,5 5,0-5,4 2,5-3,2 3,5-3,9 4,3-5,9 5,5-6,1 4,2-5,2 6,0-6,2 4,9-5,5 6,4-6,4 2,7-3,5 2,9-3,5 4,3-6,0 6,6-7,1 5,3-6,2 5,9-6,5 WP 3,1-3,2 2,8 1,1-2,1 2,9 2,7-2,8 4,0 3,6-3,7 3,4 1,0-2,3 4,1-7,0 1,3-2,0 1,9-3,6 1,9-2,2 2,7-6,7 2,1-2,2 2,8-5,6 FL 3,6-5,3 4,9 2,9-3,6 5 5,6-7,0 7,61 4,6-6,3 6,3 2,3-4,9 9,8-11,1 1,7-3,4 5,4-5,9 6,4-9,3 10,0-12,2 4,5-6,3 8,6-12,0 R (mL/100gr) fruiting stage flowering stage flowering stage fruiting stage camphene ns 0.052 ns 0.280 β-pinene ns 0.225 ns 0.487 myrcene ns 0.363 ** 0.006 p -cymene ns 0.090 * 0.031 ( Z )-β-ocimene ns 0.224 ns 0.876 γ-terpinene ns 0.628 ** 0.002 linalool ns 1.000 ns 1.000 borneol ns 0.052 ns 0.440 terpinen-4-ol ns 0.225 ns 0.939 thymol ns 0.052 ns 0.537 carvacrol ns 0.115 ns 0.787 Camphene ns 0.514 * 0.013 Sabinene ns 0.214 *** 0.000 β-pinene ns 0.598 ns 0.934 myrcene ns 0.433 *** 0.001 limonene ns 0,598 ns 0,514 1,8 cineol ns 0.192 ** 0.005 linalool ns 0.090 ns 0.098 trans-dihydro- α-terpineol ns 1.000 ns 0.457 4-terpineol ns 0.191 *** 0.001 α-terpineol ns 0.240 ns 0.058 wild / cultivated Thymus zygis Thymus mastichina T. mastichina T. zygis (s.l.) n=12 n=12 ns 0.495 *0.041 n=16 n=16 ns 0.275 ns 0.090 n=16 n=16 ** 0.006 ns 0.248 n=19 n=20 ** 0.004 ns 0.537 flowering stage WP FL fruiting stage WP FL View publication stats View publication stats