Grazielia intermedia (Spreng.)

de Souza, Tiago J. T., S, Bordignon, ergio A. L., Apel, Miriam A. & Henriques, Amelia T., 2021, Volatile constituents of Eupatorieae (Asteraceae). Compositional multivariate analysis of volatile oils from Southern Brazilian species in the subtribe Disynaphiinae, Phytochemistry (112734) 186, pp. 1-23 : 4-9

Grazielia intermedia (Spreng.) R.M.King & H.Rob. and Grazielia serrata (Spreng.) R.M.King & H.Rob.

are anatomically similar sympatric species. In the same way as other species in this genus, their VOs are mainly composed of hydrocarbon monoterpenes, specifically α- pinene, β- pinene, and limonene. G. intermedia (syn. Eupatorium intermedium , Dimorpholepis intermedia ) is a 1–1.5 m tall, white-flowered, twiggy shrub native from Minas Gerais to Rio Grande do Sul States, Brazil and Uruguay ( King and Robinson, 1987; Matzenbacher, 1979). The VOs in the two samples have similar compositions in the monoterpene fraction, where α- pinene (15.1–19.9%), β- pinene (17.3–24.2%), and limonene (19.7–25.2%) are predominant. However, a compound bearing a caryophyllane nucleus was predominant in the sesquiterpene fractions of both samples (β- caryophyllene, 7.2–8.1%) with higher amounts of bicyclogermacrene (9.4%) among the main compounds in one sample, while in the other, the percentage of this compound was lower (2.3–4.7%). Similar to another Disynaphiinae , Acanthostyles buniifolius , G. intermedia is a host species for Actinote sp. butterflies ( Freitas et al., 2018; Paluch et al., 1999) and bees ( Gonçalves and Melo, 2005), and has been reported to be associated with several other butterfly taxa in the Southern Brazilian Atlantic Rainforest, namely Hesperiidae , Nymphalidae , Papilionidae , Pieridae , and Riodinidae ( Cruz et al., 2012) , as well as the native bee Megachile (Pseudocentron) nudiventris ( Krug et al., 2010) . This species is also important in the reproductive biology of a Long-tailed Reed-Finch Donacospiza albifrons (as is Raulinoreitzia crenulata ) ( Maurício et al., 2013). Tridecapentaynene, germacrene D, bicyclogermacrene, ent-kaurenic acid, spathulenol, and sesquiterpene lactones (germacranolides, melampolides, and guaianolides) have been previously isolated from a Northeastern Brazilian sample (Ferdinand Bohlmann et al., 1981). The chemical composition of the VOs has been reported for inflorescences collected in Parana´State, Southern Brazil with spathulenol, caryophyllene oxide, (E)-β- caryophyllene, germacrene D, bicyclogermacrene, and carotol identified as the major compounds in the supercritical CO 2 or compressed propane extracted oils, while α- pinene (22.6%), β- pinene (14.4%), spathulenol (11.1%), and limonene (7.0%) were the major compounds in the gram-positive antimicrobial hydrodistilled oil ( Czaikoski et al, 2015, 2017). Another sample, collected from the same Parana´area, was composed of α- pinene (22.3%), spathulenol (11.1%), khusimol (10.9%), vetivenic acid (9.2%), β- pinene (5.5%), and limonene (5.7%) ( do Amaral et al., 2018). In contrast to the samples collected in Parana´State, no vetivenic acid, khusimol, or carotol were detected in our samples.

The sample of G. serrata , a 1–1.5 m white-flowered shrub native to Southern Brazil (from Minas Gerais to Rio Grande do Sul), Uruguay, Paraguay, and Northeast Argentina ( King and Robinson, 1987; Matzenbacher, 1979), collected in Gravataí in 2005 also presented VOs similar to others in this genus. The VOs in its leaves and inflorescences were very similar with 76.1 and 77.7% hydrocarbon monoterpenes, respectively. The major compounds were α- pinene (30.2 and 31.5%), β- pinene (17.0 and 13.8%), and limonene (19.4 and 20.9%). The only sesquiterpene among the main compounds in the VOs, as well as in the VOs of G. intermedia , was β- caryophyllene (4.8 and 5.5%). The flower heads of G. serrata have been reported to be associated with Xanthaciura biocellata (Tephritidae) , a fruit fly species associated also with Barrosoa betoniciiformis and Chromolaena congesta , and Xanthaciura chrysura , a species associated with Barrosoa betoniciiformis , Chromolaena hirsuta , and C. laevigata ( Savaris et al., 2015) . The association of this species with Megachile (Moureapis) maculata and Apis mellifera has also been reported ( Weiss, 2008). Southeastern Brazilian butterfly Actinote alalia larvae have also been reported to be associated with this species ( Freitas et al., 2018). Tridecapentaynene, α- pinene, germacrene D, bicyclogermacrene, ent-kaurenic acid, and germacranolides have been previously isolated from a Northeastern Brazilian population ( Bohlmann et al., 1983; Ferdinand Bohlmann et al., 1981).

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The volatile chemistry of the genus Grazielia is a neglected topic. Previously, from 10 species in this genus, only the volatiles in G. intermedia from Parana´State, Brazil have been described ( Czaikoski et al., 2015). In addition, a report on the isolation of volatiles, mainly germacranes, together with other non-volatile compounds in G. serrata have also been published in 1980 (Ferdinand Bohlmann et al., 1981). Herein, we thoroughly described the volatiles in G. serrata , reported a different composition for G. intermedia , and also described the VOs from G. gaudichaudieana and G. nummularia for the first time. None of the samples collected in RS contained compounds bearing either a zizaane or carotane nucleus, as reported for the Parana´sample of G. intermedia . It is noteworthy that all four samples were characterized having very similar compositions, likely reflecting their adaptation to a similar environment.