Phytochemical variability between Colombian accessions of Lippia alba (Mill.) N.E. Brown Variabilidad fitoquímica entre accesiones colombianas de Lippia alba (Mill.) N.E. Brown Variabilidade fitoquímica entre acessos colombianos de Lippia alba (Mill.) N.E. Brown José Omar Cardona Montoya1 & Jaime Eduardo Muñoz Flórez2 1Ingeniero Agrónomo, Doctor en Ciencias Agropecuarias. 2Ingeniero Agrónomo, Especialista en Matemática Avanzada, Doctor en Ciencias Agrarias. 1 Docente-investigador Universidad del Pacífico-Buenaventura, Valle. Colombia. 2Docente-investigador Universidad Nacional de Colombia-Sede Palmira. Colombia. [email protected], [email protected] Abstract Resumen The physiological fluctuation of organic constituents La fluctuación fisiológica de constituyentes orgánicos about its environmental response has two compo- alrededor de su respuesta ambiental, tiene dos com- nents, intra-inter individual variation. This study as- ponentes, variación intra e interindividual. Este estudio sessed the components of biological variation based evaluó los componentes de variación biológica con on a matrix of presence-absence, generating preci- base en una matriz de presencia-ausencia, generando sion values inter-intra individual/population based valores de precisión inter-intra poblacional/individual a on a sample of 58 accessions of L. alba. Chemical partir de una muestra de 58 accesiones de L. alba. La intra-individual variability was high (WI=2.06 [98%]) variabilidad química intra-individual fue alta (WI=2.06 and the inter-population, was low (AP=0.033), with [98%]) y la inter-poblacional, fue baja (AP=0.033), con FST=0.023 p(rand>data=0.07) and 2% proportion. FST=0.023 p(rand>data=0.07) y proporción 2%. No There were no high values of Estimates of variance se observaron valores altos de estimativos de vari- [Est.Var.] leading to establish chemical variability anza [Est.Var.] que conduzcan a establecer diversidad among individuals and populations. The results gen- genética-DG- entre poblaciones e individuos. Los re- erated by the test point to the presence of two sub- sultados generados por la prueba apuntan a la pres- populations, defined each by local environmental encia de dos sub-poblaciones, delimitadas cada una conditions. por condiciones ambientales locales. Keywords: WARDMLM.SAS, phytochemical screen- Palabras clave: WARDMLM.SAS, tamizaje fito- ing, Verbenaceae, intra/interindividual variation químico, Verbenaceae, variación intra/interindividual 39 Revista de Investigación Agraria y Ambiental – Volumen 7 Número 2 – julio - diciembre de 2016 – ISSN 2145-6097 Resumo (AP=0.033), com FST=0.023 p(rand>data=0.07) e pro- A flutuação fisiológica dos constituintes orgânicos ao porção 2%. Não foram observados valores altos de redor de sua resposta ambiental têm dois componen- Estimativos de Variância [Est.Var.] que conduzem a tes, variação intra e inter individual. Neste estudo, fo- estabelecer Diversidade Genética-DG- entre popula- ram avaliados os componentes da variação biológica ções e indivíduos. Os resultados gerados pelo teste baseado em uma matriz de presença – ausência, ge- apontam á presença de dois subpopulações, delimi- rando valores de precisão inter e intrapopulacional/ tadas (cada uma) por condições ambientais locais. individual partindo de uma amostra de 58 acessos de L. alba. A variabilidade química intraindividual foi Palavras-chave: WARDMLM.SAS, peneiramento fi- alta (WI=2.06 [98%]) e a interpopulacional, foi baixa toquímico, Verbenaceae, variação intra/interindividual Introduction Plant essential oils and secondary metabolites. content of carvone, between 30% and 35% (Castro Essential oils are liquid oily aromatic fragrant [some- et al., 2002), turning it into a substitute for other es- times semi-liquid or solid], obtained from plant ma- sential oils that contain this same component. An- terial [flowers, buds, seeds, leaves, branches, bark, other important chemical component of L. alba is herbs, Woods, fruits and roots] (UNCTAD, 2005). limonene, making up more than 25% of essential oil Take advantage mainly in the food industry as fla- (UNCTAD, 2005). The main compounds of the es- voring, the perfume-fragrances industry, and the sential oil observed in leaves are: terpinene, p-cy- pharmaceutical, to add taste and/or smell drugs mene, caryophylene, myrcene, geraniol, neral and (UNCTAD, 2005). Plant secondary metabolites pro- (Castro et al., 2002.; Jarvis et al., 2006; Blanco et duction is genetic, performing functions of attrac- al., 2007; Celis et al., 2007; Mesa et al., 2009; Esco- tion-repulsion (Viccini et al., 2004). However, its ex- bar et al., 2010). pression is determined by pre-existing conditions in their habitat (presence or absence of phytophagous, Chemical variability of essential oils in L. alba. other plant species, biophysical components, other). The chemical composition of the essential oil of L. These (secondary metabolites), are being actively alba depends on the geographical origin, the condi- used as bioprotectants action _cida [suffix] sustain- tions of culture, age and part of the plant used for able in current systems of agricultural production in extraction, and geobotanical factors (Stashenko et the world (Das & Das, 2005). The determinants of al., 2003). Genotypic determination (Montanari et chemical variability and yield of essential oil (and its al., 2004; Viccini et al., 2004-2006; Dias et al., 2005- complex mix of secondary metabolites) in each spe- 2006; Suárez et al., 2007; Yamamoto et al., 2008; cies are important (Figueiredo et al., 2008). These Martínez et al., 2008), biochemical characterizations includes: (a) physiological variations; (b) environ- (Fischer et al., 2004; Hennebelle et al., 2006) and mental conditions; (c) geographical variations; (d) morphological descriptions (Montanari et al., 2004; factors genetic and evolutionary; (e) socio-cultural Hennebelle, 2007); they make up the bulk of stud- conditions and (f) quantity of plant material (Ricca- ies reported in the species L. alba. Individuals (L. rdi et al., 2000; Jarvis et al., 2006; Figueiredo et al., alba) collected in the northeastern region of Colom- 2008; Palacio & López, 2008). bia, was shown to be a new chemotype, not previ- ously described in the literature, containing carvone Composition of the essential oil of Lippia alba. [40-57%] as a major component in oils, followed In L. alba, secondary metabolites are mainly flavo- by limonene [24-37%], biciclosesquifelandreno [2- noids, produced by epidermal glands in the meso- 22%] piperitenona [1-2%], piperitone [0.8-1.2%] and phyll cells. Essential oil of L. alba has a relative high the β-burboneno [0.6-1.5%]; the component main in 40 Phytochemical variability between Colombian accessions of Lippia alba (Mill.) N.E. Brown volatile fractions was limonene [27-77%], followed by Oliveira et al., (2006) to relate them to their tradition- carvone [14-30%]], biciclosesquifelandreno [1-33%] al uses. The analysis allowed the identification of and the β-burboneno [0.5-6.5%] (Stashenko et al., two chemotypes of L. alba, a myrcene-citral as che- 2003). The analysis of essential oils obtained from motype (15% and 37.1%, respectively) and Lippia leaves of samples of French Guiana, Martinique and alba f. intermedia as a chemotype citral (22.1%). The Guadeloupe, showed seven different ‘chemotypes’ essential oils of both species were active against all (Hennebelle et al., 2006), with a possible connection the tested organisms (bacteria and fungi) by halos of between chemotype and morphotypes (Hennebelle inhibition testing with range from 1.1 to 5.0 cm; prob- et al., 2006). The GxE interaction of 10 genotypes of ably due to its high content of oxygenated monoter- four chemical compositions (chemotype) of L. alba penes (51.0% and 40.1%, respectively), mainly rep- from three regions of the State of São Paulo-Brazil, resented by aldehydes and alcohols. Chemical and had a wide range of leaf production, while essential pharmacological data of L. alba obtained by Oliveira oils performance had a high and uniform genotypic et al., (2006) agreed with the Ethnobotanic survey. determination. No qualitative variation was detected In Colombia, “El Centro de Investigaciones en Bio- for chemical composition and quantitative variation moleculas-CIBIMOL”-from Universidad Industrial was of low magnitude. Linalool and limonene geno- de Santander has made, among others, evaluative types / carvone were invariably more productive for studies on tropical diseases, bioactivity and active performance of oil that the citral genotypes and myr- ingredients of Lippia alba how antimicrobial (Bue- cene/canfor (Yamamoto et al., 2008). no & Stashenko, 2009); antiviral [Ocazionez et al. (2010) in the dengue virus; [Meneses et al., (2009) Lippia alba (Miller) N.E. Brown is characterized by a in the yellow fever virus], antifungal Candida and As- great variability in morphology and chemical compo- pergillus (Montiel et al., 2007), in bacteria [Bueno sition of the essential oil (Oliveira et al., 2006). Tava- et al., (2009) in M. tuberculosis]; as anti-Protozoan res et al. (2005), presented data on the quantitative (Escobar et al., 2010). A complete list of reports variation of the major volatile components of linalol places it in http://tux.uis.edu.co/quimica/ investiga- production in a chemotype of L. alba. The contents cion/centros/ cibimol. of [alpha] - pinene, (Z) - 3-hexenilo and [alpha] - gur- junene was higher in the mother plants cultivated The goal in the present investigation was to deter-