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Article

Phylogenetic Studies and Metabolite Analysis of Sticta Species from Colombia and Chile by Ultra-High Performance Liquid Chromatography-High Resolution-Q-Orbitrap-Mass Spectrometry

by
Laura Albornoz
1,†,
Alfredo Torres-Benítez
2,†,
Miguel Moreno-Palacios
3,
Mario J. Simirgiotis
2,
Saúl A. Montoya-Serrano
4,
Beatriz Sepulveda
5,
Elena Stashenko
6,
Olimpo García-Beltrán
7,8,* and
Carlos Areche
1,*
1
Departamento de Química, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Nuñoa, Santiago 7800024, Chile
2
Instituto de Farmacia, Facultad de Ciencias, Universidad Austral de Chile, Campus Isla Teja, Valdivia 5090000, Chile
3
Laboratorio de Biología Evolutiva de Vertebrados, Departamento de Ciencias Biológicas, Universidad de Los Andes, Bogotá 111711, Colombia
4
Laboratorio de Toxicología, Seccional Tolima, Instituto Nacional de Medicina Legal y Ciencias Forenses, Ibagué 730006, Colombia
5
Departamento de Ciencias Químicas, Universidad Andres Bello, Campus Viña del Mar, Quillota 980, Viña del Mar 2520000, Chile
6
Research Center of Excellence CENIVAM, CIBIMOL, Universidad Industrial de Santander, Building 45, UIS, Carrera 27, Calle 9, Bucaramanga 680002, Colombia
7
Centro Integrativo de Biología y Química Aplicada (CIBQA), Universidad Bernardo O’Higgins, General Gana 1702, Santiago 8370854, Chile
8
Facultad de Ciencias Naturales y Matemáticas, Universidad de Ibagué, Carrera 22 Calle 67, Ibagué 730002, Colombia
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Metabolites 2022, 12(2), 156; https://doi.org/10.3390/metabo12020156
Submission received: 18 January 2022 / Revised: 1 February 2022 / Accepted: 2 February 2022 / Published: 8 February 2022
(This article belongs to the Section Plant Metabolism)
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Abstract

:
Eleven species of lichens of the genus Sticta, ten of which were collected in Colombia (S. pseudosylvatica S. luteocyphellata S. cf. andina S. cf. hypoglabra, S. cordillerana, S. cf. gyalocarpa S. leucoblepharis, S. parahumboldtii S. impressula, S. ocaniensis) and one collected in Chile (S. lineariloba), were analyzed for the first time using hyphenated liquid chromatography with high-resolution mass spectrometry. In the metabolomic analysis, a total of 189 peaks were tentatively detected; the analyses were divided in five (5) groups of compounds comprising lipids, small phenolic compounds, saturated acids, terpenes, and typical phenolic lichen compounds such as depsides, depsidones and anthraquinones. The metabolome profiles of these eleven species are important since some compounds were identified as chemical markers for the fast identification of Sticta lichens for the first time. Finally, the usefulness of chemical compounds in comparison to traditional morphological traits to the study of ancestor–descendant relationships in the genus was assessed. Chemical and morphological consensus trees were not consistent with each other and recovered different relationships between taxa.

Graphical Abstract

1. Introduction

Lichens constitute a mutualistic symbiosis with green algae and/or cyanobacteria [1], and in some cases present a tripartite symbiosis between different mycobionts and photobionts known as photosymbiodemas [2,3,4]. The genus Sticta (Schreb.) Ach. is the most species-diverse group of macrolichens in the family Lobariaceae, with about 120 species, and is characterized by a heteromeric thallus, with wide and rounded or elongated and truncated lobes, sometimes overlapping and rarely polyphilic, with smooth upper surface or with light ribs that may carry isidia or soralia, and the presence of cyphelas on the ventral surface of variable sizes dispersed in the tomentum [5,6,7,8,9,10,11]. In South America, the genus is found in Andean, sub-Andean, and moorland ecosystems, and develops in substrates of bark, soil, wood and rocks [5,6,7,8,9,10,11,12].
Chemical studies of the genus Sticta are scarce. In Sticta and other genera, most compounds are of fungal origin and their chemical variety is related to the environmental conditions in which the species develops [13,14], which is observed in this work. From the earliest reports found for this genus, triterpenes were identified in the species S. billardierii, S. coronata, S. colensoi, and S. favicans [15,16]. The latest reports on species such as S. fuliginosa, S. weigelia, S. caulescens and S. santessonii show the presence of substances such as 7β-acetoxy-22-hydroxy hopane usnic acid, ursolic acid, ergosterol peroxide and β-sitosterol [17]. The S. nylanderiana ethyl-3-formyl-2,4-dihydroxy-5,6-dimethylbenzoate, methyl-2,4-dihydroxy-3,5,6-trimethylbenzoate, orsellinic acid, methyl orsellinate, ethyl orsellinate, lecanoric acid, 4-O-methyl gyrophoric acid and retigeric acid B compounds were isolated and identified [18]. In an unidentified species of the genus Sticta, the compounds stictamides A-C were isolated and identified. In addition, stictamide A acts as an inhibitor of the MMP12 protease (metallopeptidase 12) [19].
Currently, for the identification and elucidation of metabolites in complex extracts, the technique of ultra-high performance liquid chromatography-diode array detection (UHPLC-PDA) coupled to an electrospray ionization tandem mass spectrometer (ESI–MS–MS) [20,21,22,23,24,25,26] is used. The Q-Exactive Focus is a hybrid high-resolution mass spectrometer used to detect and quantify small organic compounds via high-resolution accurate mass spectrometry. This machine combines UHPLC-PDA (ultrahigh pressure liquid chromatography with photodiode array) with an orbitrap, a quadrupole (Q) and a high-resolution collision cell (HRCD), which allows for high resolution diagnostic untargeted metabolomics and accurate determination of fragments [15,16,17,18,19,20,21,22,23,24,25,26,27,28]. This technique has been useful for the chemical study of several lichens of the genera Ramalina, Parmotrema and Usnea [23,24,25,26,27,28,29,30] conducted by our research group and others.
In this research, we analyzed the phytochemical profile of several species of the Sticta genus collected in different geographical regions of Colombia and Chile, based on UHPLC-DAD coupled with high-resolution electrospray ionization tandem mass spectrometry (ESI-MS-MS) for the first time. The eleven (11) lichens of the genus Sticta analyzed were S. pseudosylvatica S. luteocyphellata S. cf. andina S. cf. hypoglabra, S. cordillerana, S. cf. gyalocarpa S. leucoblepharis, S. parahumboldtii S. impressula, S. ocaniensis and S. Lineariloba (Figure 1). Based on the quantity and variety of chemical compounds found in the studied specimens, we complemented the chemical characterization with a maximum parsimony analysis based on the hypothesis that phytochemical compounds in lichens may show a phylogenetic signal, thus proving utility for chemotaxonomy.

2. Results and Discussion

In this study, the metabolome profile of eleven species of Sticta lichens are reported for the first time. Lichen substances have gained considerable attention due to their potential health benefits and possible food nutraceutical or biotechnological applications [31]. Such compounds consist mostly of aliphatic and aromatic substances which proved to have biological and pharmacological activities compared to higher plants, in particular several depsidones and depsides which proved to be antioxidant, cytotoxic and anti-inflammatory agents, among several other bioactivities reported from the genus [31,32,33,34,35]. Their identification by hyphenated mass spectrometry liquid chromatography techniques is highly important for the fast metabolite profiling of these important biodiverse organisms, and the possibility of finding biomarkers that could be of help for their identification.

2.1. Identification of Metabolites in 11 Lichen Species

In this study, eleven species of the genus Sticta were analyzed and Figure 2 shows the chromatograms for the species S. pseudosylvatica and S. lineariloba. In the metabolomic analysis, a total of 189 peaks were detected, 41 of which were unknown compounds, and the analyses were divided in five groups of compounds as explained below (Table 1).

2.1.1. Saturated Organic Acids

Peak 1 was tentatively identified as gluconic acid (C6H11O7), peak 3 as manitol, peak 4 as arabic acid, peak 5 as citric acid and peak 10 as its isomer isocitric acid (C6H7O7), peak 7 as 4-ethyl-2-ethylisophthalic acid (C10H9O4) with peak 2 as its isomer 2-ethylisophthalic acid, peak 15 as the derivative 2-hydroxyisophthalic acid, peak 17 was identified as 4-O-demethylglomellic acid (C24H25O9), peak 30 as 1,5-pentanedicarboxylic acid (C7H11O4) and peak 44 as 2,4-dicarboxy-3-hydroxy-5-methoxytoluene (C10H9O6)

2.1.2. Small Phenolic Compounds

Peaks 14 and 16 were identified as trihydroxy benzalaldehyde (C7H5O4) and 2,4-dihydroxy benzaldehyde (C7H5O3), peak 25 as 5,7-dihydroxy-6-methylphthalide (C9H7O4), peak 39 as metil-2,6-dihidroxibenzoate (C8H7O4), peak 51 as 2,4-dihydroxy benzaldehyde (C7H5O3), peak 53 as 4-ethoxy-3-formyl-2-hydroxy-6-methylbenzoic acid (C11H11O5), peak 95 5,7-dihydroxy-6-methylphthalide derivative (C9H7O3), peak 132 as ethyl 2,4-dihydroxy-6-n-nonylbenzoate (C18H27O4).

2.1.3. Typical Lichenic Phenolic Compounds (Depsides, Depsidones and Anthraquinones)

Several compounds were identified as the typical types of compounds occurring in lichens in our Sticta species. Peak 20, with a deprotonated molecule at m/z: 413.1569, was identified as grayanic acid (C23H25O7), peak 23 was identified as atranol (C8H7O3) [25], peak 32 as didechlorolecideoidin (C17 H13O7) [36] showing diagnostic fragments at m/z: 209.0456; 285.0776; 151.0396; 179.0347 and 123.0443, peak 34 was identified as orsellinic acid (C8H7O4), peak 37 as nor 8′-methylconstictic acid (C21H19O11), showing a diagnostic daughter fragment at m/z: 209.0455, peak 40 as hypostictic acid isomer (C19H15O8), peak 42 as fumarprotocetraric acid derivative (C17H11O6), peak 52 as consalizinic acid derivative I (C19H13O11, with diagnostic daughter ions at m/z: 373.0573; 387.0373; 225.0406 and 177.0193, and peak 55 as consalizinic acid derivative II, with ions at m/z: 401.0524; 417.0474 and 373.0574), peak 56 and 57 as an cynodontin, citreorosein isomer or consalizinic acid derivative I isomer, respectively, peak 59 as 1,4,5,6,8-pentahydroxy-3-ethylanthraquinone (C15H9O7), peak 62, with an ion at m/z: 403.0681 was identified as a haemathamnolic acid isomer (C19H15O10) and peak 64 as constictic acid (C19H13O10), peak 63 as a fumarprotocetraric acid derivative (C17H11O6), peak 65 as hypostictic acid isomer (C19H15O8), producing daughter ions at m/z: 327.0883, 195.0664 and 179.0347 peak 66 as terphenylquinones thelephoric acid (C18H7O8) and peak 67 as methylstictic acid (C20H15O9, ions at 371.0779 and 193.0504). Peak 68 was tentatively identified as 8′-metilconstictic acid isomer (C21H19O11), and peak 69 as protocetraric acid (C18H13O9) [29], peak 70 as hypoconstictic acid (C19H15O9), peak 77 as menegazziaic acid (C18H13O9) [37] with ionic fragments at m/z: 311.0570; 255.0666; 329.0679, peak 78 as norstictic acid (C18H11O9 ions at m/z: 327.0526; 151.0396 and 123.0444), peak 80 as the antioxidant agent physodalic acid [35] (C20H15O10, MS2 peaks at m/z: 359.0417; 315.0520; 343.0832; 387.0367; 373.0573 and 401.0525), peak 82 as derivative methyl 8-hydroxy-4-O-demethylbarbatate (C19H19O9), peak 83 as 12,13,15-trihydroxy-9-octadecenoic acid (C18H33O5), peak 84 as the cytotoxic compound haemoventosin (C15H11O7) [38], with ions at 259.0619, 231.0667, 189.0560, peak 86 as conhypoprotocetraric acid or convirensic acid (C18H15O8) [29], peak 87 as 4-O-dimethylbaeomycesic acid (C18H15O8, 181.0714; 163.0397 and 137.0236), a methyl derivative of baeomycesic acid [39], peak 88 as orsellinic acid isomer (C8H7O4, 123.0440; 149.0235), peak 89 as lecanoric acid (C16H13O7, 167.0345; 123.0443; 149.0238) [23], peak 90 as constictic acid isomer (C19H13O10), peak 92 with a deprotonated molecule at m/z: 235.0615 and daughter ion at m/z: 181.0504 was identified as 2-methyl-5-hydroxy-6-hydroxymethyl-7-methoxychromone (C12H11O5), peak 96 was identified as criptostictic acid derivative (C18H11O8), peak 105 as stictic acid (C19H13O9) [25], peak 106 as the typical lichen anthraquinone parietin (C16H11O5), while peak 108 was identified as evernic acid isomer (C17H15O7), peak 109 as hypoconstictic acid and peak 110 as cryptostictic acid [40] (C19H15O9, diagnostic daughter ions at m/z: 267.0661; 343.0825, 311.05067 and 239.0710). peak 113 as salazinic acid isomer (C18H11O1), peak 117 with a deprotonated molecule at m/z: 323.0556 was identified as pulvinic acid derivative I (C18H11O6), producing ions at m/z: 133.0286; 117.0335. Peak 119, with a parent ion at m/z: 345.0989, was identified as 4-O-demethylbarbatic acid (C18H17O7) [29], peak 121 as methyl orsellinate [37] and peak 123 as gyrophoric acid. Peak 125, producing fragments at m/z: 313.0723; 135.0444 and 179.0348, was identified as hyposalazinic acid (C18H13O8) [37], peak 127 as an isomer of orsellinic acid [23], peak 124 as galbinic acid (C20H13O11, 403.0681; 371.0417; 401.0524; 327.0518 and 149.0239) [37], peak 129 as norstictic acid (C18H11O9, 27.0517; 227.0716; 151.0390; 243.0297) [1,2], peak 133 as evernic acid (C17H15O7) [25], peak 131 as loxodinol isomer (C25H29O9), peak 136 as the dibenzophenone strepsilin (C15H9O5), peak 146 as squamatic acid and peak 147 as the depsone picrolichenic acid (C25H29O7), peak 144 and 145 as hydroxytetracosapentaenoic acid derivative (C24H37O3) and hydroxytrioxotricosanoic acid (C23H39O6), respectively. Peak 150, with an ion at m/z: 457.2244, was identified as 2,2′-di-O-methylanziaic acid (C26H33O7) [41], peak 154 was determined to be pulvinic acid, (C18H11O5, ions at m/z: 117.0338; 263.0713), peak 164 as a pulvinic acid derivative (C19H13O5), peak 164 as a pulvinic acid derivative of 321.0770 (C19H13O5), and peak 165 as another isomer of pulvinic acid (C18H11O5), while peak 155 was identified as 4-O-demethylbarbatic acid (C18H17O7, diagnostic ions at m/z: 123.0443; 137.0237; 181.0502). Peak 156 was determined as soromic acid (ions at m/z: 313.0726; 181.0502; 179.0347; 327.0520; 269.0826 and 285.0776), peak 157 as methylgyrophoric acid (C25H21O10, diagnostic ions at m/z: 149.0238; 123.0442; 167.0346 and 317.0671), and peak 158 as evernic acid isomer. Similarly, peak 159 was identified as anthraquinones skyrin (C30H17O10), peak 160 as angardianic acid (C19H35O4) [42], peak 166, with an ion at m/z: 551.1197, was identified as furfuric acid isomer (C28H23O12, producing fragments at m/z: 371.0784; 193.0504; 179.0347; 207.0297 and 193.0504), peak 172 as barbatic acid (C19H19O7) [25], peak 174 as thamnolic acid, peak 175 as orsenillic acid derivative II (C8H7O4), peak 177 as lobaric acid (C25H27O8 with ions at m/z: 411.1824; 367.1811), peak 180 as hypothamnolic acid (C19H17O10 with ions at m/z: 209.0456; 181.0499) [43], peak 184 as usnic acid (C18H15O7) [25], peak 185 as either nephromopsic acid or roccellaric acid (C19H33O4) [44], and finally, peak 187 was identified as the cytotoxic compound perlatolic acid [45], peak 188 as the antibacterial compound caperatic acid [46] and peak 189 as atranorin.

2.1.4. Terpenes

Peak 112 was identified as retigeric acid B (C30H45O6), while peak 111 with an ion at m/z: 515.3025 was tentatively identified as a retigeric acid derivative (C30H43O7).

2.1.5. Lipids

Oxylipins polyunsaturated fatty acids are an important dietary compounds, and can be found in edible fruits by HPLC orbitrap mass spectrometry [47] and also can be found in useful plants [48] and lichens [49]. In this study, several fatty acids including saturated fats and oxylipins were found using this technique in Sticta lichens. Peak 33, with a parent ion at m/z: 555.3047, was identified as decahydroxyoxopentacosanoic acid (C25H47O13); peak 43, with a parent ion at m/z: 187.0977, was determined to be 4,5-dihydroxy-2-nonenoic acid (C9H15O4); peak 72 as 12,13,15-trihydroxy-9-octadecenoic acid (C18H15O5), while peaks 91 and 94 were determined as pentahydroxytetracosanoic acid (C24H47O7) and heptahydroxytrioxooctadecanoic acid (C18H29O12), respectively. In the same manner, peaks 102–104 were identified as heptahydroxytetraoxoicosanoic acid (C20H31O13), tetrahydroxytricosanoic acid (C23H45O6), and tetrahydroxytrioxoundecanoic acid (C11H15O9), respectively. Peak 115 was assigned as 9,10-dihydroxyoctadecatrienoic acid (C18H29O4) and peak 118 as 9,10,12 trihydroxytriacontaheptaenoic acid; peak 122 as eptahydroxyetraoxoicosanoic acid (C20H31O13); peak 126 as hydroxytetracosapentaenoic acid (C24H37O3); and peak 128 as dihydroxyoctadecenoic acid (C18H33O4). Peak 130 was tentatively identified as dihydroxyoctadec-6-enoic acid (C18H33O4); peak 134 as a protocetraric acid isomer (C18H13O9) [29]; peak 139 as hexahydroxytrioxooctacosatrienoic acid (C28H43O11); peak 140 as nonahydroxyoctacosatetraenoic acid (C28H47O11); peak 142 as norsolorinic acid (C20H17O7); peak 148 as heptahydroxydioxohexacosanoic acid (C26H47O11); peak 151 as dihydroxytetracosahexaenoic acid (C24H35O4); and peak 152 as hydroxyoctadecadienoic acid (C18H31O3). In the same manner, peak 161 and 162 were attributed to pentadecatetraenoic acid and 9-hydroxyoctadecatrienoic acid, respectively. Finally, peak 171 was identified as trihydroxyheptacosa pentaenoic acid; peak 173 as hydroxytrioxodocosanoic acid; and peak 183 as dihydroxyicosahexaenoic acid (C20H27O4).
In this study, we worked on 11 species of the genus Sticta from Colombia and Chile. It should be noted that the species were collected in different ecosystems and environmental conditions in South America. The analyses includes 189 compounds, 41 of which had not yet been identified, of which the most representative are gluconic acid (1), citric acid (5), 2-Ethylisophthalic acid (13), orsellinic acid (34), lecanoric acid (89), stictic acid (105), parietin (106), gyrophoric acid (123) and usnic acid (184) (Figure 3). It should be noted that none of the identified and unidentified compounds are present simultaneously in all 11 species.

2.1.6. Distance and Phylogenetic Analysis

We found that 69/189 (37%) chemical characters and 7/16 (44%) morphological characters were parsimony informative. Optimally retained trees had a minimum parsimony score of 256 (chemical compounds) and 87 (morphology). The chemical compounds tree showed higher consistency and retention indexes and lower homoplasy (CI = 0.966, RI = 0.786) than the morphology tree (CI = 0.738., RI = 0.531).
In general, maximum-parsimony strict consensus trees from morphological and chemical characters were not consistent with each other and did not recover the same relationship between taxa (Figure 4), whereas the morphological tree recovers most of the evolutionary relationships (positions in phylogeny) documented in the published molecular phylogeny of Colombian Sticta [50] and the chemical-compound tree mirrors the geographic clusters of collected samples exactly.
In the last two decades, the study of secondary metabolites in lichens has represented an input for the determination of specimens in different complex groups, through their intervention in taxonomic keys. These compounds are mostly aromatic derivatives such as depsides, depsidones, dibenzofurans, dibenzoquinones and usnic acid among others that derive from the biochemical pathways generated by malonic, mevalonic and shikimic acids [51]. The morphological data used in the present analysis of phylogenetic relationships within species of Sticta demonstrate the relevance of morphological traits in lichen taxonomy. Nevertheless, the chemical characters offer the possibility of an alternative comparison, independent from the morphology-based classification system [52]. In this study, the analysis of chemical traits recovered more geographic than ancestor-descendant relationships among taxa. These results enrich the discussion of the role of the local environment on lichen adaptation through the actions of natural selection on biochemical pathways.
In other studies, groupings based on chemical compounds such as the case of S. cf ocaniensis, S. cf. pseudolobaria, and S. canariensis, were consistent with the known molecular phylogeny of Sticta, as was the case with S. pulmonarioides and S. cf. weigelia [53]. For the genus Cetrelia, an assessment of the composition of secondary metabolites allowed for a confirmation of the presence of species only reported in America in Europe, as in the case of C. chicitae. Metabolite composition has also facilitated the confirmation of new species for the genus by chemical fingerprinting, which contrasts the phenotypic plasticity of some morphological characters used for identification [54]. In the genus Psoroma, an analysis of the distribution of secondary metabolites has validated the presence of chemical markers unique to the group, and the presumed description of subgenera by taxa heterogeneity, causing spatial segregation [55].
On the other hand, in the genus Cladonia, there is evidence of the use of chemotaxonomic methods to determine and differentiate phylogenetically related species (C. arbuscula, C. borealis, C. chlorophaea, C. coccifera, C. coniocraea, C. cornuta, C. fimbriata, C. mitis, C. monomorpha, C. pyxidate, C. rangiferina, C. stellaris, and C. stygia), which contain chemical markers exclusive to the group [56]. In this way, the morphological–anatomical data are complemented, and discriminatory characters are provided to distinguish the species. In the genus Blastenia, reduced chemotypes are also reported in some lineages with particular genetic characteristics and distribution [57].
Currently, the process of chemotaxonomic discrimination analysis in lichen groups requires reinforcement with complementary techniques, such as the use of pigments derived from anthraquinone-type compounds in specimens of the family Teloschistaceae (Pyrenodesmia sensu lato) [58] and optical-sensor profiles for metabolic profiling in species of the genera Cladonia, Stereocaulon, Lichina, Collema and Peltigera [59]. In addition, advances in analytical chemistry and mass spectrometry have allowed for a greater specificity in the elaboration of bioactive compounds profiles, which, together with modern DNA-sequencing techniques and the extension of morphological descriptions as a “polyphasic approach”, provide objectivity in the delimitation of lichen species [60,61]. However, due to the wide variation in lichen chemotypes, the use of new compounds such as fatty acids is proposed in chemotaxonomy and phylogeny analyses, and in building an understanding of molecular-complex communication and compound biosynthetic pathways [62].

3. Materials and Methods

3.1. Chemicals

Ultrapure water was obtained from a water purification system brand Millipore (Milli-Q Merck Millipore, Santiago, Chile). Analytical reagents were all purchased from Sigma Aldrich Co. (Santiago, Chile). Ethanol, Methanol, formic acid, acetone, and acetonitrile were of chromatographic grade for HPLC analysis. Analytical lichen standards (purity: 98% by HPLC) were purchased from Sigma-Aldrich Chemical Company (Santiago, Chile).

3.2. Lichen Material

The lichen specimens S. pseudosylvatica Moncada & Suárez (35 g) and S. luteocyphellata Moncada & Lücking (28 g) were collected by Olimpo García Beltrán and Alfredo Torres Benítez in 2018 in Villahermosa, in the department of Tolima-Colombia, at the farm La Estrella (5°02′48.63″ N–75°07′37.98″ W). The species S. cf. andina Moncada & Lücking (31 g), S. cf. hypoglabra Moncada & Lücking (42 g), S. cordillerana Gyeln (37 g), S. cf. gyalocarpa (Nyl.) (29 g), S. leucoblepharis (Nyl.) Tuck. & Mont (34 g). y S. parahumboldtii Moncada & Lücking (40 g) were collected by Alfredo Torres Benítez and Emmanuel Campos in 2018 in the Semillas de Agua Civil Society Nature Reserve in the Anaime páramo (4°15′18.09″ N–73°33′23.27″ W) and the species S. impressula (Nyl.) Zahlbr (29 g) and S. ocaniensis (33 g) Moncada & Simijaca were collected by Alfredo Torres Benítez and María Rivera Montalvo in 2017 in the “Combéima river basin”, Ibagué-Tolima, Colombia (4°36′02.35″ N–75°19′50.45″ W). All voucher specimens were deposited in the herbarium of Universidad Distrital Francisco José de Caldas (Colombia) and Prof. Alejandra Suárez Corredor confirmed their identity.

3.3. Preparation of the Sample for Analyses

Fresh samples were weighed and frozen for two days at −80 °C. Then, the samples were taken to a freeze–evaporation system (Model 7670541 FreeZone 2.5 Liter Labconco Freeze Dry Systems) and all the water contained in the original product was removed by freeze–evaporation cycles. A total of 3 g of each dried lichen was macerated with methanol (3 times, 30 mL each time, 3 days/extraction). The solutions were concentrated to obtain 11 mg of extract from S. pseudosylvatica; 9 mg S. luteocyphellata; 14 mg S. cf. andina; 12 mg S. cf. hypoglabra; 10 mg S. cordillerana; 9 mg S. cf. gyalocarpa; 13 mg S. leucoblepharis; 13 mg S. parahumboldtii; 8 mg S. impressula and 9 mg S. ocaniensis, respectively. Then, the lichen extracts were processed individually for HPLC-MS analyses (redissolved in methanol at a concentration of 1 mg/mL for the analyses).

3.4. Instrument

A Thermo Scientific Ultimate 3000 UHPLC with a PDA (photodiode array detector) detector controlled by Chromeleon 7.2 Software (Thermo Fisher Scientific, Waltham, MA, USA) in conjunction with a Thermo high resolution Q-Exactive focus mass spectrometer (Thermo, Bremen, Germany) were used for analysis. The chromatographic system was coupled to the MS using a type II heated electrospray ionization source. Nitrogen obtained (purity >99.999%) from a nitrogen generator (Genius NM32LA, Peak Scientific, Billerica, MA, USA) was employed as both the collision and damping gas. Mass calibration for Orbitrap was performed once a day, in both negative and positive modes, to ensure working mass 5 ppm of accuracy. Sodium dodecyl sulfate, caffeine, N-butylamine, buspirone hydrochloride, and taurocholic acid sodium salt (Sigma Aldrich, Saint Louis, MO, USA) plus Ultramark 1621 (Alpha Aezar, Stevensville, MI, USA), a phosphazine fluorinated solution, was the standard mixture used to calibrate the mass spectrometer. These compounds were dissolved in a mixture of acetic acid, acetonitrile, water, and methanol (Merck, Darmstadt, Germany) and were infused using a Chemyx Fusion 100 syringe pump, XCalibur 2.3 software and Trace Finder 3.2 (Thermo Fisher Scientific, San José, CA, USA), which were used for control and data processing. Q Exactive 2.0 SP 2 from Thermo Fisher Scientific was used to control the mass spectrometer. The lichens extracts were individually redissolved in methanol (at a concentration of 1 mg/mL), each solution was filtered (PTFE filter, Merck) and then 10 microliters were injected in the UHPLC instrument for UHPLC-MS analysis. XCalibur 2.3 software (Thermo Fisher Scientific, Bremen, Germany) and Trace Finder 3.2 (Thermo Fisher Scientific, San José, CA, USA) were used for UHPLC control and data processing, respectively. Q Exactive 2.0 SP 2 from Thermo Fisher Scientific was used to control the mass spectrometer.

3.5. LC Parameters

Liquid chromatography on a UHPLC C-18 column (Acclaim, 150 mm × 4.6 mm ID, 2.5 μm, Thermo Fisher Scientific, Bremen, Germany) was performed as reported previously. The mobile phases were 1% formic aqueous solution, (A) methanol 1% formic acid (B) and acetonitrile 1% formic acid (C). The gradient program time were as follows: 0.00 min, 18 B, 75 C; 5.00 min, 18 B, 75 C; 15.00 min, 40 B, 60 C; 20.00 min. B, 100 C; and 12 min for column equilibration at starting conditions.

3.6. MS Parameters

The HESI parameters were as follows: sheath gas-flow rate of 75 units; aux. gas unit flow rate of 20; capillary temperature of 400 °C; aux gas heater temperature of 500 °C; spray voltage of 2500 V (for ESI−); and S lens RF level of 30. Full scan data in both the positive and negative modes were acquired at a resolving power of 70,000 FWHM (full width half maximum) at m/z 200. For the compounds of interest, a scan range of m/z 100–1000 was chosen; the automatic gain control (AGC) was set at 3 × 106 and the injection time set to 200 ms. Scan-rate was set at 2 scans s−1. External calibration was performed using a calibration solution in the positive and negative modes. For confirmation purposes, a targeted MS/MS analysis was performed using the mass inclusion list, with a 30 s time window, with the Orbitrap spectrometer operating both in the positive and negative mode at 17,500 FWHM (m/z 200). The AGC target was set to 2 × 105, with the max. injection time of 20 ms. The precursor ions were filtered by the quadrupole, which operates at an isolation window of m/z 2. The fore vacuum, high vacuum and ultrahigh vacuum were maintained at approximately 2 mbar, from 105 and below 1010 mbar, respectively. Collision energy (HCD cell) was operated at 30 kv. Detection was based on calculated exact mass and on retention time of target compounds, as shown in Table 1. The mass tolerance window was set to 5 ppm for the two modes for most compounds.

3.7. Similarity and Phylogenetic Analyses

We carried out a phylogenetic study to analyze if the chemical compounds found in Sticta specimens recovered a phylogenetic signal consistent with the current taxonomic relationships in the genera. First, we built two character-state matrices. One matrix included the 189 chemical compounds reported in this paper, and the other encompassed 16 morphological traits (Suppl. Tables S1 and S2). Then, the compounds were coded as binary characters (presence/absence), whereas morphological traits were coded as multistate characters. All character states receive the same weight and were set as unordered. Next, exploratory Neighbor-Joining distance trees were built. After that, we used a maximum parsimony phylogenetic approach to perform an exhaustive search of optimal trees. A maximum of 100 trees were retained after evaluating ca. 34 million trees per matrix. Trees were unrooted given the absence of descriptions of chemical compounds and morphological traits for potential outgroups. Then, the consistency and retention indexes were calculated. Finally, we obtained the strict consensus of optimal trees. All the procedures were performed in PAUP 4a168 for mac.

4. Conclusions

Eleven lichens of the Sticta genera from two different country zones were phytochemically investigated. More scientific data on chemistry is presented for these interesting lichens that can significantly increase the knowledge and potential for sustainable applications and industrial interest. This valuable natural-product biomass has potential applications in food, medicine, biotechnology, pharmaceuticals, and cosmetics, with many possible applications from food-conserving agents to anticancer biomaterials. The morphological data used in the present analysis of phylogenetic relationships within genus of Sticta demonstrate the relevance of morphological traits in lichen taxonomy. Nevertheless, chemical characters for chemotaxonomic studies offer the possibility of an alternative comparison, independent from the morphology-based classification system. In this study, chemical traits’ analysis recovered more geographic than ancestor-descendant relationships among taxa, and these results enriched the discussion of the role of the local environment on lichen adaptation through natural selection acting on biochemical pathways.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/metabo12020156/s1, Table S1: Description of character and character states used in the present study of Sticta, Table S2: Character state matrix of morphological traits used to recover phylogenetic relationships in species of Sticta.

Author Contributions

M.J.S., O.G.-B., C.A. conceived and designed the experiments; A.T.-B. performed lichens characterization, L.A., B.S., S.A.M.S. and E.S. performed the LC MS experiments and organized the data of HPLC/MS, M.M.-P. performed the calculations of phylogenetic studies and their chemotaxonomic relationships. All authors wrote the paper, and read and approved the final manuscript. All authors have read and agreed to the published version of the manuscript.

Funding

O.G.-B. Thank funding from the Ministry of Science, Technology and Innovation, the Ministry of Education, the Ministry of Industry, Commerce and Tourism, and ICETEX, Programme Ecosistema Científico-Colombia Científica, from the Francisco José de Caldas Fund, Grand RC-FP44842-212-2018; M.J.S. received financial support from Fondecyt, Chile (Grant 1220075) and INACh RT 16-17, while C.A. received funding from Fondecyt Regular 1190314.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data is contained within the article or Supplementary Material, but raw Thermo HPLC profiles of the plant or other data can be available on author’s request.

Conflicts of Interest

The authors declare no conflict of interest. The funding sponsors had no role in the design of the study; in the collection, analyses, interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

References

  1. Nash, T. Lichen Biology, 2nd ed.; Cambridge University Press: Cambridge, MA, USA, 2008. [Google Scholar]
  2. Armaleo, D.; Clerc’, P. Lichen chimeras: DNA analysis suggests that one fungus forms twp morphotypes. Exp. Mycol. 1991, 15, 1–10. [Google Scholar] [CrossRef]
  3. Sanders, W.B. Composite lichen thalli of sticta sp. from Brazil, with morphologically similar lobes containing either a chlorobiont or a cyanobiont layer. Symbiosis 2001, 31, 47–55. [Google Scholar]
  4. Henskens, F.L.; Green, T.G.A.; Wilkins, A. Cyanolichens can have both cyanobacteria and green algae in a common layer as major contributors to photosynthesis. Ann. Bot. 2012, 110, 555–563. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  5. Galloway, D.J. Studies on the lichen genus Sticta (Schreber) Ach.: I. Southern South American species. Lichenol 1994, 26, 223–282. [Google Scholar] [CrossRef]
  6. Galloway, D.J. Studies on the lichen genus Sticta (Schreber) Ach.: V*. Australian species. Bryophyt. Divers. Evol. 1998, 15, 117–160. [Google Scholar] [CrossRef]
  7. Galloway, D.J. Studies on the lichen genus Sticta (Schreber) Ach. IV. New Zealand species. Lichenologist 1997, 29, 105–168. [Google Scholar] [CrossRef]
  8. Moncada, B.; Aguirre, J.; Lücking, R. Ecogeografía del género Sticta (ascomycota liquenizados: Lobariaceae) en Colombia. Rev. Biol. Trop. 2014, 62, 266–281. [Google Scholar] [CrossRef] [Green Version]
  9. Moncada, B.; Luecking, R.K.; Lumbsch, H.T. Rewriting the evolutionary history of the lichen genus Sticta (Ascomycota: Peltigeraceae subfam. Lobarioideae) in the Hawaiian Islands. Plant Fungal Syst. 2020, 65, 95–119. [Google Scholar] [CrossRef]
  10. Moncada, B.; Lücking, R. Ten new species of Sticta and counting: Colombia as a hot spot for unrecognized diversification in a conspicuous macrolichen genus. Phytotaxa 2012, 74, 1–29. [Google Scholar] [CrossRef]
  11. Moncada, B.; Mercado-Díaz, J.A.; Magain, N.; Hodkinson, B.P.; Smith, C.W.; Bungartz, F.; Pérez-Pérez, R.E.; Gumboski, E.; Sérusiaux, E.; Lumbsch, H.T.; et al. Phylogenetic diversity of two geographically overlapping lichens: Isolation by distance, environment, or fragmentation? J. Biogeogr. 2021, 48, 676–689. [Google Scholar] [CrossRef]
  12. Mercado-Díaz, J.A.; Lücking, R.; Moncada, B.; Widhelm, T.J.; Lumbsch, H.T. Elucidating species richness in lichen fungi: The genus Sticta (Ascomycota: Peltigeraceae) in Puerto Rico. Taxon 2020, 69, 851–891. [Google Scholar] [CrossRef]
  13. Sepahvand, A.; Studzińska-Sroka, E.; Ramak, P.; Karimian, V. Usnea sp.: Antimicrobial potential, bioactive compounds, ethnopharmacological uses and other pharmacological properties; a review article. J. Ethnopharmacol. 2021, 268, 113656. [Google Scholar] [CrossRef] [PubMed]
  14. Calcott, M.J.; Ackerley, D.F.; Knight, A.; Keyzers, R.A.; Owen, J.G. Secondary metabolism in the lichen symbiosis. Chem. Soc. Rev. 2018, 47, 1730–1760. [Google Scholar] [CrossRef] [PubMed]
  15. Corbett, B.R.E.; Young, H.J. Lichens and fungi. Part III. Structural elucidation of 15α, 22-dihydroxyhopane from Sticta billardierii Del. Chem. Soc. C Org. 1966, 1564–1567. [Google Scholar] [CrossRef]
  16. Chin, W.J.; Corbett, R.E.; Heng, C.K.; Wilkins, A.L. Lichens and fungi. Part XI. Isolation and structural elucidation of a new group of triterpenes from Sticta coronata, S. colensoi, and S. flavicans. J. Chem. Soc. Perkin Trans. 1 1973, 14, 1437–1446. [Google Scholar] [CrossRef]
  17. Piovano, M.; Chamy, M.C.; Garbarino, J.A.; Quilhot, W. Secondary metabolites in the genus Sticta (lichens). Biochem. Syst. Ecol. 2000, 28, 589–590. [Google Scholar] [CrossRef]
  18. Zhang, H.J.; Guo, H.F.; Lou, H.X. Secondary metabolites from the Chinese lichen Sticta nylanderiana A. Z. Biochem. Syst. Ecol. 2006, 34, 760–762. [Google Scholar] [CrossRef]
  19. Liang, Z.; Sorribas, A.; Sulzmaier, F.J.; Jiménez, J.I.; Wang, X.; Sauvage, T.; Yoshida, W.Y.; Wang, G.; Ramos, J.W.; Williams, P.G. Stictamides A-C, MMP12 inhibitors containing 4-amino-3-hydroxy-5- phenylpentanoic acid subunits. J. Org. Chem. 2011, 76, 3635–3643. [Google Scholar] [CrossRef] [Green Version]
  20. Le Pogam, P.; Schinkovitz, A.; Legouin, B.; Le Lamer, A.C.; Boustie, J.; Richomme, P. Matrix-free UV-laser desorption ionization mass spectrometry as a versatile approach for accelerating dereplication studies on lichens. Anal. Chem. 2015, 87, 10421–10428. [Google Scholar] [CrossRef] [Green Version]
  21. Musharraf, S.G.; Kanwal, N.; Thadhani, V.M.; Choudhary, M.I. Rapid identification of lichen compounds based on the structure-fragmentation relationship using ESI-MS/MS analysis. Anal. Methods 2015, 7, 6066–6076. [Google Scholar] [CrossRef]
  22. Simirgiotis, M.J.; Quispe, C.; Areche, C.; Sepúlveda, B. Phenolic compounds in chilean mistletoe (quintral, Tristerix tetrandus) analyzed by UHPLC-Q/Orbitrap/MS/MS and its antioxidant properties. Molecules 2016, 21, 245. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  23. Cornejo, A.; Salgado, F.; Caballero, J.; Vargas, R.; Simirgiotis, M.; Areche, C. Secondary metabolites in Ramalina terebrata detected by UHPLC/ESI/MS/MS and identification of parietin as tau protein inhibitor. Int. J. Mol. Sci. 2016, 17, 1303. [Google Scholar] [CrossRef] [Green Version]
  24. Castro, O.N.; Benites, J.; Rodilla, J.; Santiago, J.C.; Simirgiotis, M.; Sepulveda, B.; Areche, C. Metabolomic analysis of the lichen Everniopsis trulla using ultra high performance liquid chromatography-quadrupole-orbitrap mass spectrometry (UHPLC-Q-OT-MS). Chromatographia 2017, 80, 967–973. [Google Scholar] [CrossRef]
  25. Torres-Benítez, A.; Rivera-Montalvo, M.; Sepúlveda, B.; Castro, O.N.; Nagles, E.; Simirgiotis, M.J.; Garciá-Beltrán, O.; Areche, C. Metabolomic analysis of two Parmotrema lichens: P. robustum (Degel.) Hale and P. andinum (Mull. rg.) hale using UHPLC-ESI-OT-MS-MS. Molecules 2017, 22, 1861. [Google Scholar] [CrossRef] [Green Version]
  26. Salgado, F.; Albornoz, L.; Cortéz, C.; Stashenko, E.; Urrea-Vallejo, K.; Nagles, E.; Galicia-Virviescas, C.; Cornejo, A.; Ardiles, A.; Simirgiotis, M.; et al. Secondary metabolite profiling of species of the genus Usnea by UHPLC-ESI-OT-MS-MS. Molecules 2018, 23, 54. [Google Scholar] [CrossRef] [Green Version]
  27. Jiménez-González, A.; Quispe, C.; Bórquez, J.; Sepúlveda, B.; Riveros, F.; Areche, C.; Nagles, E.; García-Beltrán, O.; Simirgiotis, M.J. UHPLC-ESI-ORBITRAP-MS analysis of the native Mapuche medicinal plant palo negro (Leptocarpha rivularis DC.–Asteraceae) and evaluation of its antioxidant and cholinesterase inhibitory properties. J. Enzyme Inhib. Med. Chem. 2018, 23, 54. [Google Scholar] [CrossRef] [PubMed]
  28. Areche, C.; Fernandez-Burgos, R.; Terrones, T.C.D.; Simirgiotis, M.; García-Beltrán, O.; Borquez, J.; Sepulveda, B. Mulinum crassifolium Phil; two new mulinanes, gastroprotective activity and metabolomic analysis by UHPLC-orbitrap mass spectrometry. Molecules 2019, 24, 1673. [Google Scholar] [CrossRef] [Green Version]
  29. Parrot, D.; Jan, S.; Baert, N.; Guyot, S.; Tomasi, S. Comparative metabolite profiling and chemical study of Ramalina siliquosa complex using LC-ESI-MS/MS approach. Phytochemistry 2013, 89, 114–124. [Google Scholar] [CrossRef]
  30. Ly, H.D.; Vo, T.N.; Duong, T.H.; Nguyen, T.D.; Nguyen, K.P.P. A new depside and two new diphenyl ether compounds from the lichen Ramalina farinacea (L.). Ach. Phytochem. Lett. 2015, 11, 146–150. [Google Scholar] [CrossRef]
  31. Alam, M.A.; Khatoon, R.; Huda, S.; Ahmad, N.; Sharma, P.K. Biotechnological Applications of Lichens. In Lichen-Derived Products; John Wiley and Sons: Hoboken, NJ, USA, 2020; pp. 203–219. [Google Scholar]
  32. Nguyen, T.T.; Nallapaty, S.; Rao, G.S.N.K.; Koneru, S.T.; Annam, S.S.P.; Tatipamula, V.B. Evaluating the in vitro activity of depsidones from Usnea subfloridana Stirton as key enzymes involved in inflammation and gout. Pharm. Sci. 2021, 27, 291–296. [Google Scholar] [CrossRef]
  33. Zhao, Y.; Wang, M.; Xu, B. A comprehensive review on secondary metabolites and health-promoting effects of edible lichen. J. Funct. Foods 2020, 80, 104283. [Google Scholar] [CrossRef]
  34. Nguyen, T.T.H.; Dinh, M.H.; Chi, H.T.; Wang, S.L.; Nguyen, Q.V.; Tran, T.D.; Nguyen, A.D. Antioxidant and cytotoxic activity of lichens collected from Bidoup Nui Ba National Park, Vietnam. Res. Chem. Intermed. 2019, 45, 33–49. [Google Scholar] [CrossRef]
  35. Aoussar, N.; Laasri, F.E.; Bourhia, M.; Manoljovic, N.; Mhand, R.A.; Rhallabi, N.; Ullah, R.; Shahat, A.A.; Noman, O.M.; Nasr, F.A.; et al. Phytochemical analysis, cytotoxic, antioxidant, and antibacterial activities of lichens. Evid. Based Complement. Altern. Med. 2020, 2020, 1–11. [Google Scholar] [CrossRef] [PubMed]
  36. Elix, J.; Elix, J.; Venables, D.; Venables, D.; Lumbsch, H.; Lumbsch, H.; Brako, L.; Brako, L. Further new metabolites from lichens. Aust. J. Chem. 1994, 47, 1619. [Google Scholar] [CrossRef]
  37. Sultana, N.; Afolayan, A.J. A new depsidone and antibacterial activities of compounds from Usnea undulata Stirton. J. Asian Nat. Prod. Res. 2011, 13, 1158–1164. [Google Scholar] [CrossRef]
  38. Le Pogam, P.; Le Lamer, A.C.; Siva, B.; Legouin, B.; Bondon, A.; Graton, J.; Jacquemin, D.; Rouaud, I.; Ferron, S.; Obermayer, W.; et al. Minor pyranonaphthoquinones from the apothecia of the lichen Ophioparma ventosa. J. Nat. Prod. 2016, 79, 1005–1011. [Google Scholar] [CrossRef]
  39. Goel, M.; Dureja, P.; Rani, A.; Uniyal, P.L.; Laatsch, H. Isolation, characterization and antifungal activity of major constituents of the himalayan Lichen Parmelia reticulata tayl. J. Agric. Food Chem. 2011, 59, 2299–2307. [Google Scholar] [CrossRef]
  40. Asplund, J.; Gauslaa, Y. Content of secondary compounds depends on thallus size in the foliose lichen Lobaria pulmonaria. Lichenologist 2007, 39, 273–278. [Google Scholar] [CrossRef]
  41. Elix, J.A.; Jariangprasert, S.; Archer, A.W. New Pertusaria (lichenized Ascomycota) from Australia and Thailand. Telopea 2008, 12, 263–272. [Google Scholar] [CrossRef]
  42. Brodo, I.M.; Tønsberg, T. Opegrapha halophila (Opegraphaceae), a new lichen species from coastal British Columbia, Canada, and Alaska, USA. Bryologist 2019, 122, 457–462. [Google Scholar] [CrossRef]
  43. Xiang, W.J.; Wang, Q.Q.; Ma, L.; Hu, L.H. β-Orcinol-type depsides from the lichen Thamnolia vermicularis. Nat. Prod. Res. 2013, 27, 804–808. [Google Scholar] [CrossRef] [PubMed]
  44. Nallasivam, J.L.; Fernandes, R.A. A protecting-group-free synthesis of (+)-nephrosteranic, (+)-protolichesterinic, (+)-nephrosterinic, (+)-phaseolinic, (+)-rocellaric acids and (+)-methylenolactocin. Org. Biomol. Chem. 2017, 15, 708–716. [Google Scholar] [CrossRef] [PubMed]
  45. Bogo, D.; Honda, N.K.; Alcantara, G.B.; Brandão, L.F.G.; Aléssio, G.F.; Guimarães, R.D.C.A.; Matos, M.D.F.C. Cytotoxic activity of compounds from lichens of the cerrado biome in Brazil. Orbital 2020, 12, 7–16. [Google Scholar] [CrossRef]
  46. Dieu, A.; Mambu, L.; Champavier, Y.; Chaleix, V.; Sol, V.; Gloaguen, V.; Millot, M. Antibacterial activity of the lichens Usnea florida and Flavoparmelia caperata (Parmeliaceae). Nat. Prod. Res. 2020, 34, 3358–3362. [Google Scholar] [CrossRef] [PubMed]
  47. Simirgiotis, M.J.; Ramirez, J.E.; Schmeda, H.G.; Kennelly, E.J. Bioactive coumarins and HPLC-PDA-ESI-ToF-MS metabolic profiling of edible queule fruits (Gomortega keule), an endangered endemic Chilean species. Food Res. Int. 2013, 54, 532–543. [Google Scholar] [CrossRef]
  48. Brahmi-Chendouh, N.; Piccolella, S.; Nigro, E.; Hamri-Zeghichi, S.; Madani, K.; Daniele, A.; Pacifico, S. Urtica dioica L. leaf chemical composition: A never-ending disclosure by means of HR-MS/MS techniques. J. Pharm. Biomed. Anal. 2021, 195, 113892. [Google Scholar] [CrossRef]
  49. Gouveia-Figueira, S.; Danielsson, K.; Fowler, C.J. Changes in proportions of linoleic acid-derived oxylipins in oral lichen planus. Acta Derm. Venereol. 2019, 99, 1051–1052. [Google Scholar] [CrossRef]
  50. Moncada, B.; Lücking, R.; Suárez, A. Molecular phylogeny of the genus Sticta (lichenized Ascomycota: Lobariaceae) in Colombia. Fungal Divers. 2014, 64, 205–231. [Google Scholar] [CrossRef]
  51. Swofford, D.L. PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods); Version 4; Sinauer Associates: Sunderland, MA, USA, 2003. [Google Scholar]
  52. Culberson, W.L. Chemosystematics and Ecology of Lichen-Forming Fungi. Annu. Rev. Ecol. Syst 1970, 1, 153–170. [Google Scholar] [CrossRef]
  53. Mittermeier, V.K.; Schmitt, N.; Volk, L.P.M.; Suárez, J.P.; Beck, A.; Eisenreich, W. Metabolic profiling of alpine and ecuadorian lichens. Molecules 2015, 20, 18047–18065. [Google Scholar] [CrossRef]
  54. Farkas, E.; Biró, B.; Varga, N.; Sinigla, M.; Lőkös, L. Analysis of lichen secondary chemistry doubled the number of Cetrelia W.L. Culb. & C.F. Culb. species (Parmeliaceae, lichenised Ascomycota) in Hungary. Cryptogamie. Mycologie 2021, 42, 1–16. [Google Scholar] [CrossRef]
  55. Quilhot, W.; Piovano, M.; Arancibia, H. Studies on Chilean Lichens, XII. Chemotaxonomy of the Genus Psoroma. J. Nat. Prod. 1989, 52, 191–192. [Google Scholar] [CrossRef]
  56. Konoreva, L.; Prokopiev, I.; Frolov, I.; Chesnokov, S.; Rozhina, S.; Poryadina, L.; Shavarda, A. Metabolite profiling of the Cladonia lichens using gas chromatography-mass spectrometry. Biochem. Syst. Ecol. 2019, 85, 3–12. [Google Scholar] [CrossRef]
  57. Vondrák, J.; Frolov, I.; Košnar, J.; Arup, U.; Veselská, T.; Halıcı, G.; Malíček, J.; Søchting, U. Substrate switches, phenotypic innovations and allopatric speciation formed taxonomic diversity within the lichen genus Blastenia. J. Syst. Evol. 2020, 58, 295–330. [Google Scholar] [CrossRef]
  58. Frolov, I.; Vondrák, J.; Košnar, J.; Arup, U. Phylogenetic relationships within Pyrenodesmia sensu lato and the role of pigments in its taxonomic interpretation. J. Syst. Evol. 2021, 59, 454–474. [Google Scholar] [CrossRef]
  59. Le Corvec, M.; Boussard-Plédel, C.; Charpentier, F.; Fatih, N.; Le Dare, B.; Massart, F.; Rojas, F.; Tariel, H.; Loréal, O.; Bureau, B.; et al. Chemotaxonomic discrimination of lichen species using an infrared chalcogenide fibre optic sensor: A useful tool for on-field biosourcing. RSC Adv. 2016, 6, 108187–108195. [Google Scholar] [CrossRef]
  60. Frisvad, J.C.; Andersen, B.; Thrane, U. The use of secondary metabolite profiling in chemotaxonomy of filamentous fungi. Mycol. Res. 2008, 112, 231–240. [Google Scholar] [CrossRef]
  61. Xu, M.; Heidmarsson, S.; Olafsdottir, E.S.; Buonfiglio, R.; Kogej, T.; Omarsdottir, S. Secondary metabolites from cetrarioid lichens: Chemotaxonomy, biological activities and pharmaceutical potential. Phytomedicine 2016, 23, 441–459. [Google Scholar] [CrossRef]
  62. Vu, T.H.; Catheline, D.; Delmail, D.; Boustie, J.; Legrand, P.; Lohezic-Le Devehat, F. Gas chromatographic analysis to compare the fatty acid composition of fifteen lichen species, with a focus on Stereocaulon. Lichenologist 2016, 48, 323–337. [Google Scholar] [CrossRef]
Metabolites 12 00156 g001 550
Figure 1. (A) S. pseudosylvatica (Colombia); (B) S. lineariloba (Chile).
Figure 1. (A) S. pseudosylvatica (Colombia); (B) S. lineariloba (Chile).
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Figure 2. Chromatogram of the species S. pseudosylvatica and S. lineariloba.
Figure 2. Chromatogram of the species S. pseudosylvatica and S. lineariloba.
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Figure 3. Structures of some representative compounds found in Sticta lichens.
Figure 3. Structures of some representative compounds found in Sticta lichens.
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Figure 4. Neighbor joining trees (A,B) and unrooted strict consensus trees from maximum parsimony analysis (C,D) with morphological traits (A,C) and chemical compounds (B,D) in some Colombian species of Sticta. Colored lines represent localities from collected specimens. Colored circles represent their position in the molecular phylogeny of Moncada et al. (2014); grayish colors in the color scale refer to more basal taxa, whereas bluish colors are more recent taxa. Although S. lineariloba occurs in Colombia, the specimen in this study was collected in Valdivia (Chile). Independent of the grouping method, trees built based on chemical compounds recovered the geographic relationships, while trees built with morphological traits resemble the molecular phylogeny of Colombian Sticta.
Figure 4. Neighbor joining trees (A,B) and unrooted strict consensus trees from maximum parsimony analysis (C,D) with morphological traits (A,C) and chemical compounds (B,D) in some Colombian species of Sticta. Colored lines represent localities from collected specimens. Colored circles represent their position in the molecular phylogeny of Moncada et al. (2014); grayish colors in the color scale refer to more basal taxa, whereas bluish colors are more recent taxa. Although S. lineariloba occurs in Colombia, the specimen in this study was collected in Valdivia (Chile). Independent of the grouping method, trees built based on chemical compounds recovered the geographic relationships, while trees built with morphological traits resemble the molecular phylogeny of Colombian Sticta.
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Table
Table 1. Identification of metabolites in 11 species of the genus Sticta from Colombia and Chile.
Table 1. Identification of metabolites in 11 species of the genus Sticta from Colombia and Chile.
PeakTentative Identification[M-H]-Retention Time (min)Theoretical Mass (m/z)Measured Mass (m/z)Accuracy (ppm)Metabolite Type **MS Ions (m/z)Lichen Species *
1Gluconic AcidC6H11O71.28195.0509195.05052.0Acid165.0401;2; 3; 7; 9; 10; 11
2UnknownC11H5O5N1.28231.0184231.0195−4.8----6
3ManitolC6H13O61.31181.0712181.0714−1.1Carbohidrate181.0717; 163.06067; 9; 10; 11
4Arabic acidC5H9O61.29165.0399165.0401−1.2Acid147.0293; 113.0237; 129.01966
5Citric AcidC6H7O71.38191.0196191.01922.0Acid111.00801; 2; 3; 4; 5; 6; 7; 8; 9; 11
6UnknownC15H5O3N21.38261.0289261.0300−4.2-----1; 7; 8; 10
74-ethyl-2-Ethylisophthalic acid C10H9O41.44193.0504193.05011.6A133.02881; 2; 3; 4; 6
8UnknownC8H10O5N1.56200.0563200.05592.0---1; 3; 4; 6
9UnknownC15H5O3N21.64261.0247261.0278−11.8----7; 8; 10
10Isocitric AcidC6H7O71.77191.0195191.01921.6Acid111.0079;1; 2; 3; 4; 6; 7
11UnknownC7H9O71.91205.0352205.03482.0-187.0245; 173.0087; 121.11312
12UnknownC7H7O62.24187.0246187.02431.6-125.0237; 137.2503;2
132-Ethylisophthalic acidC10H9O42.85193.0505193.05012.0A161.0240; 133.02901; 2; 3; 4; 6; 7; 8; 9; 10
14Trihydroxy benzaledehydeC7H5O44.81153.0188153.0189−0.7A137.02381; 2; 3; 4
152-Hydroxyisophthalic acidC8H5O57.63181.0137181.0141−2.2A137.02384
162,4-dihydroxy benzaldehydeC7H5O38.02137.0238137.0239−0.7A121.02891; 2; 3; 4
174-O-Demethylglomellic acid C24H25O98.65457.1476457.1499−5.0d---5; 8
18UnknowC22H23O78.77399.1444399.14137.8----8
19UnknowC7H11O58.86175.0606175.0611−2.9----5
20Grayanic acid C23H25O78.91413.1600413.15697.5d181.065038
21UnknownC13H16O7N8.98298.0940298.09274.4-175.0609; 283.0210; 202.0696;5
22UnknownC11H9O79.33253.0361253.03485.1-137.0603; 181,0505; 249.3808; 173.4203;5
23Atranol C8H7O39.44151.0395151.0396−0.7A123.0445; 135.04453
24UnknownC24H23 O89.48439.1371439.1393−5.0-151.0397; 105.0948; 121.4871;5
255,7-Dihydroxy-6-methylphthalide C9H7O49.64179.0344179.0347−1.6A135.0444; 107.04944; 5; 7; 8; 10
26UnknownC16H15O109.73367.0665367.06397.1----1; 4
27UnknownC30H27O13N10.03609.1475609.1482−1.1----1; 4; 6
28UnknowC18H15O410.31295.0970295.093511.8----9
29UnknowC17H13O611.31313.0712313.0724−3.8-----1; 2; 3; 4; 5
301,5-Pentanedicarboxylic acidC7H11O410.56159.0657159.0660−1.9L115.07585
31UnknownC29H25O13N10.54595.1316595.1326−1.7----3
32Didechlorolecideoidin C17 H13O710.64329.0661329.0676−4.6D209.0456; 285.0776; 151.0396; 179.0347; 123.04433
33Decahydroxyoxopentacosanoic acidC25H47O1310.71555.3017555.3047−5.4L---8
34Orsellinic acid C8H7O411.04167.0347167.03441.8A123.04422; 3; 4; 5; 6
35UnknowC21H19O1211.04463.0877463.0893−3.4----3
36UnknowC10H9O511.07209.0450209.0458−3.8----5; 6
37Nor 8′-methylconstictic acidC21H19O1111.11447.0927447.0942−3.4d209.04552
38UnknowC17H13O611.20313.0712313.0720−2.5----7
39Metil-2,6-dihidroxibenzoate C8H7O411.21167.0344167.0346−1.1A109.0287; 137.02389
40Hypostictic acid isomerC19H15O811.36371.0778371.0782−1.1D195.0665; 327.0885; 341.0679; 179.03471; 3; 4; 7
41UnknowC19H16O9 N11.69402.0825402.0841−3.98----11
42Fumarprotocetraric acid derivativeC17H11O611,70311.0556311.0564−2.5d---7; 10
434,5-Dihydroxy-2-nonenoic acid C9H15O412.11187.0974187.0977−1.6L171.1025; 143.10721; 2; 3; 4; 9; 10
442,4-Dicarboxy-3-hydroxy-5-methoxytoluene C10H9O612.14225.0407225.03993.5A181.0503; 167.0346; 149.02404
45UnknownC17H13O612.16313.0724313.0727−1.0----4
46UnknownC18H15O712.37343.0818343.0826−2.3----7; 9
47UnknownC21H17O1212.40461.0737461.07203.7----6
48UnknownC20H17O812.52385.0939385.09234.1----1; 2; 3
49UnknowC10H9O412.81193.0501193.0502−0.5----1; 2; 3; 4; 5; 6; 7; 8; 9; 10
50UnknowC27H45O613.04465.3216465.3231−3.2----3
512,4-dihydroxy benzaldehydeC7H5O313.07137.0237137.0239−1.5A121.02885
52Consalizinic acid derivative IC19H13O1113.36417.0458417.0474−3.8D373.0573; 387.0373; 225.0406, 177.019311
534-Ethoxy-3-formyl-2-hydroxy-6-methylbenzoic acid C11H11O513.41223.0614223.06063.6A177.0190; 133.0296;1; 2; 3; 4; 7; 9; 10
54UnknowC20H15O813.47383.0767383.0781−3.6----1
55Consalizinic acid derivative IIC20H17O1113,47433.0771433.0787−3.7D401.0524; 417.0474; 373.057411
56Cynodontin or Citreorosein isomer, C15H9O613.71285.0399285.0410−3.9Anthraquinone151.0396; 137.02372
57Consalizinic acid derivative I isomerC19H13O1113.78417.0458417.0474−3.84D373.0573; 343.0467, 77.0190; 401.052310
58UnknowC30H47O713.79519.3322519.3337−2.9----3
591,4,5,6,8-Pentahydroxy-3-ethylanthraquinone C15H9O713.87301.0348301.0361−4.3Anthraquinone---4
60UnknowC19H15O413.87307.0970307.093910.0----8
61UnknowC14H13O713.99293.0661293.0674−4.4----3
62Haemathamnolic acid isomer C19H15O1014.18403.0665403.0681−3.97D359.0781; 371.0414; 209.045511
63Fumarprotocetraric acid derivativeC17H11O614.78311.0567311.0569−0.6d---1; 2; 3; 7; 10
64Constictic acid C19H13O1014.61401.0509401.0528−4.74D357.0625; 313.0726; 283.0619; 255.0670; 121.028911
65Hypostictic acid isomer C19H15O814.98371.0781371.07673.8D327.0883; 195.0664; 179.03475
66Thelephoric acid C18H7O814.81351.0141351.0154−3.7Terphenylquinones---3
67Methylstictic acidC20H15O915.26399.0716399.0728−3.0D371.0779; 193.05042; 7
68Nor 8′-metilconstictic acid C21H19O1115.28447.0927447.0944−3.80D401.0524; 209.045511
69Protocetraric acid C18H13O917.17373.0560373.0574−3.75D355.0468; 329.0674; 311.0568; 227.0352; 267.0669; 285.077711
70Hypoconstictic acid C19H15O917.27387.0716387.0729−3.4D267.0673; 311.0552; 149.0238; 343.0827;167.03453
71UnknowC14H13O617.84277.0712277.0724−4.3---3; 6
7212,13,15-Trihydroxy-9-octadecenoic acidC18H15O518.11329.2328329.2340−3.6L285.1716;2; 6
73UnknowC20H13O818.19381.0610381.0626−4.2----3; 4
74Salazinic acidC18H11O1018.21387.0352387.0368−4.13D343.0468; 269.0458; 241.0507; 325.0365; 299.056911
75UnknownC10H9O418.45193.0505193.05012.1----3; 4
76UnknownC30H47O718.61519.3322519.33190.6---3
77Menegazziaic acid C18H13O918.79373.0560373.0575−4.0D311,0570; 255.0666; 329.06793
78Norstictic acidC18H11O918.86371.0403371.0417−3.8D327.0526; 151.0396; 123.0444;3
79UnknowC22H19O1018.88443.0978443.0996−4.1----3
80Physodalic acidC20H15O1018.99415.0665415.0681−3.85D359.0417; 315.0520; 343.0832; 387,0367; 373.0573; 401.052511
81UnknowC26H19O1019.00491.0978491.0997−3.9----3
82Derivative methyl 8-hydroxy-4-0-demethylbarbatateC19H19O919.05391.1045391.10294.1d359.07883
8312,13,15-Trihydroxy-9-octadecenoic acidC18H33O519.05329.2328329.2336−2.4299L---7
84HaemoventosinC15H11O719.16303,0519303.05054.7Naphthaquinone259.0619; 231.0667; 189.0560;3
85α-acetilconstictic acid derivative IC21H17O1119.22445.0771445.0786−3.3D415.0680; 371.0780; 427.0676; 343.0830;193.0504, 401.052211
86Conhypoprotocetraric acid or Convirensic acid C18H15O819.25359.0781359.07673.9D344.0545; 302.04423
874-O-dimethylbaemycesic acid C18H15O819.27359.0781359.07673.9d181.0714; 163.0397; 137.02361; 2; 3;5; 6
88Orsellinic acid IsomerC8H7O419.45167.0344167.0346−1.1974A123.0440; 149.02359; 10
89Lecanoric acidC16H13O719.51317.0661317.06710.6d167.0345; 123.0443; 149.0238;1; 2; 3; 4; 5; 6
90Constictic acid isomerC19H13O1019.56401.0509401.0524−3.74D357,0626; 313.0726; 343.0831; 255.062211
91Pentahydroxytetracosanoic acidC24H47O719.67447.3322447.3336−3.1L---1; 3; 7; 9; 10
922-Methyl-5-hydroxy-6-hydroxymethyl-7-MethoxychromoneC12H11O519.73235.0606235.0615−3.8C181.05043
93UnknownC20H17O819.79385.0939385.09234.1----1; 4; 5; 10
94Heptahydroxytrioxooctadecanoic acidC18H29O1219.86437.1664437.16454.3L---1; 4; 5; 6; 7; 9; 10
955,7-Dihydroxy-6-methylphthalide derivative C9H7O319.88163.0395163.03921.8A119.049210
96Criptostictic acid derivativeC18H11O820.04355.0454355.0462−2.2D133.0288; 239.0715; 311.0572; 179.0345;7
97UnknowC18H17O620.08329.1025329.1032−2.1----9
98UnknowC20H15O820.16383.0767383.0775−2.0----9; 10
99UnknowC21H19O920.18415.1045415.10293.9----3; 5; 9; 10
100UnknowC28H23O1120.12535.1240535.1257−3.1----3
101UnknowC15H13O320.13241.0872241.0874−0.8-----1; 2
102Heptahydroxytetraoxoicosanoic acid C20H31O1321.19479.1765479.17463.9L---7
103Tetrahydroxytricosanoic acid C23H45O620.26417.3232417.32163.9L403.30731; 3; 4; 7
104Tetrahydroxytrioxoundecanoic acid C11H15O920.30291.0716291.06995.8L---8
105Stictic acid C19H13O920.34385.0560385.0576−4.1D341.0674; 357.0622; 297.0774; 313.0721; 193.0504; 269.082611
106ParietinC16H11O520.39283.0606283.0617−3.9Antraquinone179.03451; 2; 6
107UnknowC24H47O11N220.39539.3157539.3180−4.3----3
108Evernic acid isomerC17H15O720.46331.0818331.0830−3.6d167.0347; 123.0447; 149.02401
109Hypoconstictic acidC19H15O920.50387.0716387.0732−4.1D149.0238; 343.0836; 167.03454
110Cryptostictic acidC19H15O920.50387.0716387.0725−2.3D267,0661; 343,0825; 311.05067; 239.07107; 8
111Retigeric acid derivativeC30H43O720.51515.3009515.3025−3.1Triterpene---3
112Retigeric acid BC30H45O620.56501.3216501.3236−4.0Triterpene---3
113Salazinic acid isomerC18H11O1020.58387.0352387.0368−4.13D343.0468; 299.056511
114UnknownC23H22O10N20.68472.1244472.1259−3.2------6
1159,10-dihydroxyoctadecatrienoic acidC18H29O420.69309.2081309.20664.9L291.19751; 2; 4
116UnknowC17H13O620.84313.0712313.0727−4.8-----1; 2; 6
117Pulvinic acid derivative IC18H11O620.97323.0556323.05560.0Pulvinic acid y derivates133.0286; 117.033510
1189,10,12 Trihydroxytriacontaheptaenoic acidC30H45O520.99485.3284485.32673.5L---3
1194-0-Demethylbarbatic acidC18H17O720.99345.0974345.0989−4.3d181.0505; 163.0396; 137.06034
120UnknowC24H23O10N21.02485.1322485.13190.6----1; 5
121Methyl orsellinateC9H9O421.05181.0502181.05010.5A163.03891
122Heptahydroxyetraoxoicosanoic acidC20H31O1321.17479.1752479.1765−2.7L---1; 2; 3; 4; 5; 6; 9
123Gyrophoric AcidC24H19O1021.25467.0991467.09782.8d167.0346; 317.0673; 123.0445; 149.0238;1; 3; 4; 6; 11
124Galbinic acidC20H13O1121.27429.0458429.0474−3.73D403.0681; 371.0417; 401.0524; 327.0518; 149.023911
125Hyposalazinic acid C18H13O821.28357.0610357.0623−3.6D313.0723; 135.0444; 179.03481
126Hydroxytetracosapentaenoic acidC24H37O321.42373.2743373.27430.0L---10
127Orsellinic acid isomerC8H7O421.47167.0347167.03441.8A149.0239; 123.04431; 4; 6
128Dihydroxyoctadecenoic acidC18H33O421.48313.2390313.2395−1.6L----2; 5; 6
129Norstictic acidC18H11O921.64371.0403371.0417−3.77D27.0517; 227.0716; 151.0390; 243.029711
130Dihydroxyoctadec-6-enoic acidC18H33O421.58313.2379313.23790.0L---10
131Loxodinol isomerC25H29O921.64473.1812473.1818−1.2DE429.19199
132EthyI 2,4-dihydroxy-6-n-nonylbenzoate C18H27O421.65307.1909307.1922−4.2A263.16591; 2; 3; 4
133Evernic AcidC17H15O721.81331.0828331.08183.0d167.0345; 123.0444; 149,0238;1; 2; 3; 4; 5
134Protocetraric acid IsomerC18H13O921.85373.0560373.0573−3.48D355.0460; 329.0674; 285.0780, 311.0567; 255.,067211
135UnknownC22H22O8N21.83428.1360428.13453.5-----6
136StrepsilinC15H9O521.89269.0450269.0462−4.5DBF225.05542; 5
137UnknownC30H29O421.97453.2066453.20611.1----3
138UnknownC18H11O622.01323.0556323.0570−4.33----11
139Hexahydroxytrioxooctacosatrienoic acidC28H43O1122.12555.2805555.2841−6.4832L---9
140Nonahydroxyoctacosatetraenoic acidC28 H47O1122.26559.3124559.3132−1.4L---2
141UnknowC28H41O9N222.26549.2849549.28126.8----3
142Norsolorinic acidC20H17O722.44369.0974369.0989−4.06----11
143UnknowC25H33O1322.46541.1921541.19092.2----1; 2; 3; 6; 10
144Hydroxytetracosapentaenoic acid derivativeC24H37O322.61373.2743373.27410.5----10
145Hydroxytrioxotricosanoic acidC23H39O622.53411.2747411.2757−2.4L---8
146Squamatic acidC19H17O922.89389.0873389.0886−3.3d343.0836; 163.0396; 193.0139; 149.0238; 121.02861; 3; 4
147Picrolichenic acidC25H29O722.72441.1913441.1926−3.0Depsones---1
148Heptahydroxydioxohexacosanoic acidC26H47O1122.76535.3118535.3134−3.0L---6
149UnknowC30H45O422.83469.3319469.3335−3.4----3
1502,2′-Di-O-methylanziaic acidC26H33O722.85457.2226457.2244−3.9d413.2345;4
151Dihydroxytetracosahexaenoic acidC24H35O422.85387.2535387.2552−4.4L---5
152Hydroxyoctadecadienoic acidC18H31O322.90295.2273295.22730.0L---10
153Orsellinic acid IsomerC8H7O422.92167.0344167.0348−2.3A149.0240; 123.044511
154Pulvinic acidC18H11O522.98307.0606307.0613−2.2Pulvinic acid y derivates117.0338; 263.07139
1554-0-Demethylbarbatic acid C18H17O723.02345.0974345.0986−3.5d123.0443; 137.0237; 181.05021
156Psoromic acidC18H13O823.06357.0610357.0626−4.4D313.0726; 181.0502; 179.0347; 327.0520; 269.0826; 285.077611
157Methylgyrophoric acidC25H21O1023.15481.1135481.1147−2.5d149.0238; 123.0442; 167.0346; 317.06711; 4
158Evernic acid isomerC17H15O723.22331.0818331.0832−4.2d149.0239; 123.0443; 167.0346; 105.033711
159SkyrinC30H17O1023.28537.0822537.0840−3.4Anthraquinones----3
160Angardianic acidC19H35O423.36327.2543327.2547−1.2Acids283.2649; 309.20812; 4
161Pentadecatetraenoic acidC15H21O223.38233.1542233.1545−1.2L---9; 10
1629-hydroxyoctadecatrienoic acidC18 H29O323.45293.2117293.2130−4.4L277.21806
163UnknowC18H15O723.53343.0818343.0824−1.7----9
164Pulvinic acid derivative IIC19H13O523.68321.0763321.0770−2.1Pulvinic acid y derivates117.03379; 10; 11
165Pulvinic acidC18H11O523.77307.0606307.0620−4.5Pulvinic acid y derivates263.0720; 117.033911
166Furfuric acid isomerC28H23O1223.82551.1190551.1197−1.2D371.0784; 193.0504; 179.0347; 207.0297; 193.05048
167UnknowC26H47O5N223.82467.3485467.3492−3,9----3
168UnknowC30H27O623.89483.1808483.1820−2,5----1
169UnknowC30H25O624.01481.1651481.1663−2,5----1
170UnknowC15H13O324.02241.0872241.08720,0----1
171Trihydroxyheptacosa pentaenoic acidC27H43O524.05447.3110447.3127−3.8L---8
172 Barbatic Acid C19H19O724.26359.1141359.11312.8d137.0603; 163.0396; 181.05091; 4
173Hydroxytrioxodocosanoic acidC22H37O624.29397.2590397.2601−2.8L---8
174Thamnolic acid isomerC19H15O1124.41419.0614419.0630−3.8d375.0730; 167.0344; 209.0455; 181.050311
175Orsenillic acid derivated IIC8H7O424.73167.0344167.0347−1.80-149.0239; 1230444;11
176UnknowC26H33O824.73473.2190473.21753.2----2
177Lobaric acidC25H27O824.81455.1706455.1718−2.6D411.1824; 367.18111
178UnknowC22H27O724.97403.1770403.17573.2- 2
179UnknowC30H41O825.27529.2819529.28013.4----4
180Hypothamnolic acidC19H17O1025.49405.0822405.0832−2.5d209.0456; 181.04991
181UnknowC25H11O725.53423.0505423.04971.9----4
182Pulvinic acid derivative IIIC19H13O525.67321.0763321.0777−4.3Pulvinic acid y derivates117.033811
183Dihydroxyicosahexaenoic acidC20H27O426.02331.1909331.1925−4.8L-----11
184Usnic acidC18H15O726.05343.0818343.0831−3.8DBF231.0658; 328.0585; 259.06043; 4; 6; 7; 8
185Nephromopsic acid orRoccellaric acidC19H33O426.32325.2392325.23794.0Acids281.24943; 4
186UnknowC28H25O5N26.87455.1733455.17232.20----11
187Perlatolic acidC25H31O726.98443.2070443.2078−1.8d205.0867; 179.1073; 223.09737; 8
188Caperatic acidC21H37O728.14401.2539401.2549−2.4Acids255.23278
189AtranorinC19H17O829.64373.0923373.0937−3.75d177.0192; 163.03979; 11
* Identified by addition experiments with a genuine compound. A = Aromatic compound; L = Lipid; D = depsidone; d = depside; DE = diphenilether; DBF = dibenzofurane. C = Chromone. ** 1 = S. cf. andinaa; 2 = S. cf. hypoglabra; 3 = S. cordillerana; 4 = S. cf. gyalocarpa; 5 = S. leucoblepharis; 6 = S. parahumboldtii; 7 = S. impressula; 8 = S. ocaniesnsis; 9 = S. speudosylvatica; 10 = S. luteocyphelata; 11 = S. lineariloba.
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MDPI and ACS Style

Albornoz, L.; Torres-Benítez, A.; Moreno-Palacios, M.; Simirgiotis, M.J.; Montoya-Serrano, S.A.; Sepulveda, B.; Stashenko, E.; García-Beltrán, O.; Areche, C. Phylogenetic Studies and Metabolite Analysis of Sticta Species from Colombia and Chile by Ultra-High Performance Liquid Chromatography-High Resolution-Q-Orbitrap-Mass Spectrometry. Metabolites 2022, 12, 156. https://doi.org/10.3390/metabo12020156

AMA Style

Albornoz L, Torres-Benítez A, Moreno-Palacios M, Simirgiotis MJ, Montoya-Serrano SA, Sepulveda B, Stashenko E, García-Beltrán O, Areche C. Phylogenetic Studies and Metabolite Analysis of Sticta Species from Colombia and Chile by Ultra-High Performance Liquid Chromatography-High Resolution-Q-Orbitrap-Mass Spectrometry. Metabolites. 2022; 12(2):156. https://doi.org/10.3390/metabo12020156

Chicago/Turabian Style

Albornoz, Laura, Alfredo Torres-Benítez, Miguel Moreno-Palacios, Mario J. Simirgiotis, Saúl A. Montoya-Serrano, Beatriz Sepulveda, Elena Stashenko, Olimpo García-Beltrán, and Carlos Areche. 2022. "Phylogenetic Studies and Metabolite Analysis of Sticta Species from Colombia and Chile by Ultra-High Performance Liquid Chromatography-High Resolution-Q-Orbitrap-Mass Spectrometry" Metabolites 12, no. 2: 156. https://doi.org/10.3390/metabo12020156

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