Phenolic profile and antioxidant activity from wild and in vitro cultivated Rhynchostele rossii (Orchidaceae)

Nieves del Socorro Martínez Cruz, Juan Luis Monribot Villanueva, Yolanda Cocotle Ronzón, Rosa Angélica Gutiérrez Sánchez, José Antonio Guerrero Analco


Background and Aims: Rhynchostele rossii is an orchid native to Mexico known as gallinitas (little chickens) that is threatened due to overexploitation for ornamental purposes, as a consequence of which it is necessary to realize efforts for its conservation. To date there are no reports of phytochemical studies of this orchid, although it is well known that species of the Orchidaceae family are a good source of bioactive and nutraceutical compounds (e. g. vanillin). Therefore, the main objective of this research was to establish a R. rossii in vitro germination protocol for propagation and determination of phenolic compounds that contribute to its phytochemical knowledge.
Methods: A specimen of a wild plant and some seedlings obtained by in vitro culture were dried, milled and their components extracted with MeOH; anti-free radical activity (DPPH), total phenols and flavonoids were determined by spectrophotometric methods, and individual phenolic compounds were identified and quantified by liquid chromatography coupled to mass spectrometry (LC-MS).
Key results: The root of the wild plant showed the highest content of total phenols and flavonoids with 121.60 mg GAE g-1 and 108.73 mg CE g-1, respectively, and the best anti-free radical activity with an IC50 53.63 μg ml-1. Extracts from seedlings obtained by in vitro culture also produced phenolic compounds, showing a total phenolic and flavonoid content of 37.35 mg GAE g-1 and 0.16 mg CE g-1, respectively. One coumarin (scopoletin) , three cinnamic acids (4-coumaric acid, ferulic acid and trans-cinnamic acid), three benzoic acids (vanillic acid, vanillin and 4-hydroxybenzoic acid) and three flavonoids (quercetin-3-D-galactoside, quercetin-3-glucoside and kaempferide) were identified and quantified by LC-MS.
Conclusions: This study demonstrates that R. rossii is a potential source of antioxidant metabolites that can be obtained by in vitro culture, without harming the wild specimens.

Palabras clave

biotechnology, orchid in vitro propagation, secondary metabolites

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Atefipour, N., M. Dianat, M. Badavi and A. Sarkaki. 2016. Ameliorative effect of vanillic acid on serum bilirubin, chronotropic and dromotropic properties in the cholestasis induced model rats. Electron Physician 8(5): 2410-2415. DOI:

Baltazar-Bernal, O., J. Zavala Ruiz, F. Solís Zatonelli, J. Pérez Sato and O. Sánchez Eugenio. 2014. Sendero interpretativo de orquídeas y bromelias en Tepexilotla, Chocamán, Veracruz. Revista Mexicana de Ciencias Agrícolas 5(9): 1687-1699. DOI:

Bami, E., O. B. Ozakpinar, Z. N. Ozdemir-Kumral, K. Köroglu, F. Ercan, Z. Cirakli, T. Sekerler, F. V. Izzettin, M. Sancar and B. Okuyan. 2017. Protective effect of ferulic acid on cisplatin induced nephrotoxicity in rats. Environmental Toxicology Pharmacology 54: 105-111. DOI:

Bhattacharyya, P., S. Kumaria, R. Diengdoh and P. Tandon. 2014. Genetic stability and phytochemical analysis of the in vitro regenerated plants of Dendrobium nobile Lindl., an endangered medicinal orchid. Meta Gene 2: 489-504. DOI:

Bhatnagar, M., N. Sarkar, N. Gandharv, O. Apang, S. Singh and S. Ghosal. 2017. Evaluation of antimycobacterial, leishmanicidal and antibacterial activity of three medicinal orchids of Arunachal Pradesh, India. BMC Complementary and Alternative Medicine 17(379). DOI:

Brand-Williams, W., M. E. Cuvelier and C. Berset. 1995. Use of a free radical method to evaluate antioxidant activity. LWT-Food Science and Technology 28(1): 25-30. DOI:

Cano Asseleih, L. M., R. A. Menchaca García and J. Y. S. Ruiz Cruz. 2015. Ethnobotany, pharmacology and chemistry of medicinal orchids from Veracruz. Journal of Agricultural Science and Technology 5: 745-754. DOI:

Castañeda-Zárate, M., J. Viccon-Esquivel, S. Ramos-Castro and R. Solano-Gómez. 2012. Registros nuevos de Orchidaceae para Veracruz, México. Revista Mexicana de Biodiversidad 83(1): 281-284. DOI:

Castillo-Pérez, L. J., D. Martínez-Soto, J. J. Maldonado-Miranda, A. J. Alonso-Castro and C. Carranza-Álvarez. 2019. The endemic orchids of Mexico: a review. Biologia 74(1): 1-13. DOI:

Chand, M. B., M. R. Paudel and B. Pant. 2016. The antioxidant activity of selected wild orchids of Nepal. Journal of Coastal Life Medicine 4(9): 731-736.

Chase, M. W., K. M. Cameron, J. V. Freudenstein, A. M. Pridgeon, G. Salazar, C. van den Berg and A. Schuiteman. 2015. An updated classification of Orchidaceae. Botanical Journal of the Linnean Society 177(2): 151-174. DOI:

Chen, Y., U. M. Goodale, X.-L. Fan and J.-Y. Gao. 2015. Asymbiotic seed germination and in vitro seedling development of Paphiopedilum spicerianum: an orchid with an extremely small population in China. Global Ecology and Conservation 3: 367-378. DOI:

Chugh, S., S. Guha and I. U. Rao. 2009. Micropropagation of orchids: A review on the potential of different explants. Scientia Horticulturae 122(4): 507-520. DOI:

Cruz-Higareda, J. B., B. S. Luna-Rosales and A. Barba-Álvarez. 2015. A novel seed baiting technique for the epiphytic orchid Rhynchostele cervantesii, a means to acquire mycorrhizal fungi from protocorms. Lankesteriana 15(1): 67-76. DOI:

Di Ferdinando, M., C. Brunetti, G. Agati and M. Tattini. 2014. Multiple functions of polyphenols in plants inhabiting unfavorable Mediterranean areas. Environmental and Experimental Botany 103: 107-116. DOI:

Ding, Z., Y. Dai, H. Hao, R. Pan, X. Yao and Z. Wang. 2008. Anti-inflammatory effects of scopoletin and underlying mechanisms. Pharmaceutical Biology 46(12): 854-860. DOI:

Divakaran, M. and K. N. Babu. 2009. Micropropagation and in vitro conservation of vanilla (Vanilla planifolia Andrews). In: Jain, S. M. and P. K. Saxena (eds.). Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants. Humana Press. Totowa NJ, USA. Pp. 129-138. DOI:

Divakaran, M., K. N. Babu and K. V. Peter. 2016. Protocols for biotechnological interventions in improvement of vanilla (Vanilla planifolia Andrews). In: Jain, S. (ed.). Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants. Humana Press. New York, USA. Pp. 47-63. DOI:

Espinosa-Leal, C. A., J. F. Treviño-Neávez, R. A. Garza-Padrón, M. J. Verde-Star, C. Rivas-Morales and M. E. Morales-Rubio. 2015. Contenido de fenoles totales y actividad anti-radical de extractos metanólicos de la planta silvestre y cultivada in vitro de Leucophyllum frutescens. Revista Mexicana de Ciencias Farmacéuticas 46(3): 52-56.

Fay, M. F. 2018. Orchid conservation: how can we meet the challenges in the twenty-first century? Botanical Studies 59: 16. DOI:

Ganesan, K. and B. Xu. 2017. A critical review on polyphenols and health benefits of black soybeans. Nutrients 9(5): 455. DOI:

Giri, L., P. Dhyani, S. Rawat, I. D. Bhatt, S. K. Nandi, R. S. Rawal and V. Pande. 2012. In vitro production of phenolic compounds and antioxidant activity in callus suspension cultures of Habenaria edgeworthii: A rare Himalayan medicinal orchid. Industrial Crops and Products 39: 1-6. DOI:

Guerriero, G., R. Berni, J. A. Muñoz-Sánchez, F. Apone, E. M. Abdel-Salam, A. A. Qahtan, A. A. Alatar, C. Cantini, G. Cai, J-F. Hausman, K. S. Siddiqui, S. M. T. Hernández-Sotomayor and M. Faisal. 2018. Production of plant secondary metabolites: examples, tips and suggestions for biotechnologists. Genes 9(6): 309. DOI:

Hinsley, A., H. J. de Boer, M. F. Fay, S. W. Gale, L. M. Gardiner, R. S. Gunasekara, P. Kumar, S. Masters, D. Metusala, D. L. Roberts, S. Veldman, S. Wong and J. Phelps. 2018. A review of the trade in orchids and its implications for conservation. Botanical Journal of the Linnean Society 186(4): 435-455. DOI:

Hussain, M. S., S. Fareed, S. Ansari, M. A. Rahman, I. Z. Ahmad and M. Saeed. 2012. Current approaches toward production of secondary plant metabolites. Journal of Pharmacy and Bioallied Sciences 4(1): 10-20. DOI:

Ješvnik, T. and Z. Luthar. 2015. Successful disinfection protocol for orchid seeds and influence of gelling agent on germination and growth. Acta Agriculturae Slovenica 105: 95-102. DOI:

Jiménez-Peña, N., L. I. Trejo-Téllez and P. Juárez-López. 2018. Concentración de macronutrimentos de tres especies silvestres de Rhynchostele en su hábitat natural. Revista Mexicana de Ciencias Agrícolas 9(5): 971-980. DOI:

Juárez-Trujillo, N., J. L. Monribot-Villanueva, M. Alvarado-Olivares, G. Luna-Solano, J. A. Guerrero-Analco and M. Jiménez-Fernández. 2018. Phenolic profile and antioxidant properties of pulp and seeds of Randia monantha Benth. Industrial Crops and Products 124: 53-58. DOI:

Khamchatra, N., K. W. Dixon, S. Tantiwiwat and J. Piapukiew. 2016. Symbiotic seed germination of an endangered epiphytic slipper orchid, Paphiopedilum villosum (Lindl.) Stein. from Thailand. South African Journal of Botany 104: 76-81. DOI:

Lin, D., M. Xiao, J. Zhao, Z. Li, B. Xing, X. Li, M. Kong, L. Li, Q. Zhang, Y. Liu, H. Chen, W. Qin, H. Wu and S. Chen. 2016. An overview of plant phenolic compounds and their importance in human nutrition and management of type 2 diabetes. Molecules 21(10): 1374. DOI:

Luceri, C., F. Guglielmi, M. Lodovici, L. Giannini, L. Messerini and P. Dolara. 2004. Plant phenolic 4-coumaric acid protects against intestinal inflammation in rats. Scandinavian Journal of Gastrienterology 39(11): 1128-1133.

Menchaca-García, R. and D. Moreno-Martínez. 2010. Rhynchostele rossii (Lindl) Soto Arenas & Salazar. Especie en notable peligro. In: Gómez-Pompa, A., T. Krömer and R. Castro-Cortés (eds.). Atlas de la flora de Veracruz: un patrimonio natural en riesgo. EDIMPLAS. Xalapa, México. Pp. 481-482.

Minh, T. N., P. T. Tuyen, D. T. Khang, N. V. Quan, P. T. T. Ha, N. T. Quan, Y. Andriana, X. Fan, T. M. Van, T. D. Khanh and T. D. Xuan. 2017. Potential use of plant waste from the moth orchid (Phalaenopsis sogo yukidian V3) as an antioxidant source. Foods 6(10): 85. DOI:

Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15(3): 473-497. DOI:

Murthy, H. N., E. Lee and K. Paek. 2014. Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plan Cell Tissue and Organ Culture 118: 1-16. DOI:

Naik, P. M. and J. M. Al-Khayri. 2016. Impact of abiotic elicitors on in vitro production of plant secondary metabolites: A review. Journal of Advanced Research in Biotechnology 1(2): 7. DOI:

Pandey, K. B. and S. I. Rizvi. 2009. Plant polyphenols as dietary antioxidants in human health and disease. Oxidative Medicine and Cellular Longevity 2(5): 270-278. DOI:

Pant, B. 2013. Medicinal orchids and their uses: Tissue culture a potential alternative for conservation. African Journal of Plant Science 7(10): 448-467. DOI:

Paul, S., S. Kumaria and P. Tandon. 2012. An effective nutrient medium for asymbiotic seed germination and large-scale in vitro regeneration of Dendrobium hookerianum, a threatened orchid of northeast India. AoB Plants 2012(1): plr032. DOI:

Pereira, D. M., P. Valentão, J. A. Pereira and P. B. Andrade. 2009. Phenolics: From Chemistry to Biology. Molecules 14(6): 2220-2211. DOI:

Petruk, G., R. Del Giudice, M. M. Rigano and D. M. Monti. 2018. Antioxidants from plants protect against skin photoaging. Oxidative Medicine and cellular Longevity. 2018(2): 1-11, ID 1454936. DOI:

Ramakrishna, A. and G. A. Ravishankar. 2011. Influence of abiotic


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