Abstract
The objective of this study was to evaluate the effects of different natural ventilation systems and explant types on the growth and volatile compound content of Lippia gracilis cultured in vitro. The treatments consisted of four membrane systems (without membrane, with one, two, and four porous membranes) and two explant types (nodal segments with and without a pair of leaves). The evaluation of growth, photosynthetic pigments and chemical analysis of the volatile fraction were performed at 35 days of cultivation in half strength MS basal medium. Natural ventilation systems significantly influenced the in vitro growth and volatile fraction of L. gracilis. Explants with a pair of leaves obtained the best experimental responses. The natural ventilation system with four membranes provided the best growth parameters and leaf area response of L. gracilis explants with leaves. The photosynthetic pigments decreased with an increase in the number of porous membranes in the culture flask. Variations in the number, content, and profile of volatile compounds under the influence of natural ventilation systems were observed. Major constituents such as ρ-cymene, γ-terpinene, thymol, carvacrol, and E-caryophyllene, regardless of experimental conditions, were identified. The highest carvacrol and thymol contents were observed in plantlets grown in culture flasks with four porous membranes. To maximize the content of carvacrol and thymol from the in vitro culture of L. gracilis, explants with a pair of leaves and four porous membranes in culture flasks are recommended for use.
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Abbreviations
- GC/MS:
-
Gas chromatography/mass spectrometry
- NMS:
-
No membrane system
- AMS1:
-
Alternative membrane system with one filter
- AMS2:
-
Alternative membrane system with two filters
- AMS4:
-
Alternative membrane system with four filters
- PTFE:
-
Polytetrafluoroethylene
- MS:
-
Murashige and Skoog
- PCA:
-
Principal component analysis
References
Adams RP (2017) Identification of essential oil components by gas chromatography/mass spectrometry, 5 edn. Texensis Publishing, Texas
Alvarez C, Sáez P, Sáez K, Sánchez-Olate M, Ríos D (2012) Effects of light and ventilation on physiological parameters during in vitro acclimatization of Gevuina avellana mol. Plant Cell Tissue Organ Cult 110(1):93–101. https://doi.org/10.1007/s11240-012-0133-x
Benincasa M (2003) Análise de crescimento de plantas: noções básicas. FUNEP, Jaboticabal, p 42
Chandra S, Bandopadhyay R, Kumar V, Chandra R (2010) Acclimatization of tissue cultured plantlets: from laboratory to land. Biotechnol Lett 32(9):1199–1205. https://doi.org/10.1007/s10529-010-0290-0
Cha-um S, Chanseetis C, Chintakovid W, Pichakum A, Supaibulwatana K (2011) Promoting root induction and growth of in vitro macadamia (Macadamia tetraphylla L.‘Keaau’) plantlets using CO2 enriched photoautotrophic conditions. Plant Cell Tissue Organ Cult 106(3):435. https://doi.org/10.1007/s11240-011-9940-8
Chen C (2015) Application of growth models to evaluate the microenvironmental conditions using tissue culture plantlets of Phalaenopsis Sogo Yukidian ‘V3’. Sci Hortic 191:25–30. https://doi.org/10.1016/j.scienta.2015.05.007
da Silva FFS, Dantas BF (2014) Taxas de crescimento de mudas de quixabeira submetidas a diferentes condições. Scientia Plena 10(9):1–7
de Oliveira Cruz EM, Costa-Junior LM, Pinto JAO, de Alexandria Santos D, de Araujo SA, de Fátima Arrigoni-Blank M, Bacci L, Alves PB, de Holanda Cavalcanti SC, Blank AF (2013) Acaricidal activity of Lippia gracilis essential oil and its major constituents on the tick Rhipicephalus (Boophilus) microplus. Vet Parasitol 195(1–2):198–202. https://doi.org/10.1016/j.vetpar.2012.12.046
dos Santos CP, Pinto JAO, dos Santos CA, Cruz EMO, de Fátima Arrigoni-Blank M, Andrade TM, de Alexandria Santos D, Alves PB, Blank AF (2016) Harvest time and geographical origin affect the essential oil of Lippia gracilis Schauer. Ind Crops Prod 79:205–210. https://doi.org/10.1016/j.indcrop.2015.11.015
Ferraz RP, Bomfim DS, Carvalho NC, Soares MB, da Silva TB, Machado WJ, Prata APN, Costa EV, Moraes VRS, Nogueira PCL (2013) Cytotoxic effect of leaf essential oil of Lippia gracilis Schauer (Verbenaceae). Phytomedicine 20(7):615–621. https://doi.org/10.1016/j.phymed.2013.01.015
Hartikainen K, Nerg A-m, Kivimäenpää M, Kontunen-Soppela S, Mäenpää M, Oksanen E, Rousi M, Holopainen T (2009) Emissions of volatile organic compounds and leaf structural characteristics of European aspen (Populus tremula) grown under elevated ozone and temperature. Tree Physiol 29(9):1163–1173. https://doi.org/10.1093/treephys/tpp033
Holopainen JK (2011) Can forest trees compensate for stress-generated growth losses by induced production of volatile compounds? Tree Physiol 31(12):1356–1377. https://doi.org/10.1093/treephys/tpr111
Iarema L, da Cruz ACF, Saldanha CW, Dias LLC, Vieira RF, de Oliveira EJ, Otoni WC (2012) Photoautotrophic propagation of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen]. Plant Cell Tissue Organ Cult 110(2):227–238. https://doi.org/10.1007/s11240-012-0145-6
Isah T (2015) Adjustments to in vitro culture conditions and associated anomalies in plants. Acta Biol Cracov Bot 57(2):9–28. https://doi.org/10.1515/abcsb-2015-0026
Ivanova M, Van Staden J (2010) Natural ventilation effectively reduces hyperhydricity in shoot cultures of Aloe polyphylla Schönland ex Pillans. Plant Growth Regul 60(2):143–150. https://doi.org/10.1007/s10725-009-9430-8
Lichtenthaler HK, Buschmann C (2001) Chlorophylls and carotenoids: measurement and characterization by UV–VIS spectroscopy. Curr Protoc Food Anal Chem
Lorenzi H, Matos FJ (2002) Plantas medicinais no Brasil: nativas e exóticas
Loreto F, Sharkey TD (1990) A gas-exchange study of photosynthesis and isoprene emission in Quercus rubra L. Planta 182(4):523–531. https://doi.org/10.1007/BF02341027
Loreto F, Ciccioli P, Brancaleoni E, Cecinato A, Frattoni M, Sharkey TD (1996a) Different sources of reduced carbon contribute to form three classes of terpenoid emitted by Quercus ilex L. leaves. Proc Natl Acad Sci 93(18):9966–9969
Loreto F, Ciccioli P, Cecinato A, Brancaleoni E, Frattoni M, Fabozzi C, Tricoli D (1996b) Evidence of the photosynthetic origin of monoterpenes emitted by Quercus ilex L. leaves by 13C labeling. Plant Physiol 110(4):1317–1322
Maia JGS, Taveira FSN, Andrade EHA, Silva MHLd, Zoghbi MdGB (2003) Essential oils of Lippia grandis Schau. Flav Fragr J 18(5):417–420. https://doi.org/10.1002/ffj.1241
Miguel C, Marum L (2011) An epigenetic view of plant cells cultured in vitro: somaclonal variation and beyond. J Exp Bot 62(11):3713–3725. https://doi.org/10.1093/jxb/err155
Mohamed M-H, Alsadon A (2010) Influence of ventilation and sucrose on growth and leaf anatomy of micropropagated potato plantlets. Sci Hortic 123(3):295–300. https://doi.org/10.1016/j.scienta.2009.09.014
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497
Neto RM, Matos FJ, Andrade VS, de Melo MC, Carvalho C, Guimarães SB, Pessoa OD, Silva SL, Silva SF, Vasconcelos PR (2010) The essential oil from Lippia gracilis Schauer, Verbenaceae, in diabetic rats. Rev Bras Farmacogn 20(2):261–266. https://doi.org/10.1590/S0102-695X2010000200021
Niinemets Ü, Hauff K, Bertin N, Tenhunen JD, Steinbrecher R, Seufert G (2002) Monoterpene emissions in relation to foliar photosynthetic and structural variables in Mediterranean evergreen Quercus species. New Phytol 153(2):243–256. https://doi.org/10.1046/j.0028-646X.2001.00323.x
NIST (2008) National Institute of Standards and Technology. In: Chemistry Web Book. http://webbook.nist.gov/chemistry/. Accessed 17 May 2017.
Park S, Moon H, Murthy H, Kim Y (2011) Improved growth and acclimatization of somatic embryo-derived Oplopanax elatus plantlets by ventilated photoautotrophic culture. Biol Plant 55(3):559–562. https://doi.org/10.1007/s10535-011-0125-4
Peña-Ramírez Y, Juárez-Gómez J, González-Rodríguez JA, Robert ML (2012) Tissue culture methods for the clonal propagation and genetic improvement of Spanish red cedar (Cedrela odorata). In: Loyola-Vargas V, Ochoa-Alejo N (eds) Plant cell culture protocols. Methods in molecular biology (methods and protocols), vol 877. Humana Press, Totowa
Pérez-Jiménez M, López-Pérez AJ, Otálora-Alcón G, Marín-Nicolás D, Piñero MC, del Amor FM (2015) A regime of high CO2 concentration improves the acclimatization process and increases plant quality and survival. Plant Cell Tissue Organ Cult 121(3):547–557. https://doi.org/10.1007/s11240-015-0724-4
Possell M, Ryan A, Vickers CE, Mullineaux PM, Hewitt CN (2010) Effects of fosmidomycin on plant photosynthesis as measured by gas exchange and chlorophyll fluorescence. Photosynth Res 104(1):49–59. https://doi.org/10.1007/s11120-009-9504-5
Poulose A, Croteau R (1978) Biosynthesis of aromatic monoterpenes: conversion of γ-terpinene to p-cymene and thymol in Thymus vulgaris L. Arch Biochem Biophys 187(2):307–314. https://doi.org/10.1016/0003-9861(78)90039-5
Saldanha CW, Otoni CG, de Azevedo JLF, Dias LLC, do Rêgo MM, Otoni WC (2012) A low-cost alternative membrane system that promotes growth in nodal cultures of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen]. Plant Cell Tissue Organ Cult 110(3):413–422. https://doi.org/10.1007/s11240-012-0162-5
Saldanha CW, Otoni CG, Rocha DI, Cavatte PC, Detmann KdSC, Tanaka FAO, Dias LLC, DaMatta FM, Otoni WC (2014) CO2-enriched atmosphere and supporting material impact the growth, morphophysiology and ultrastructure of in vitro Brazilian-ginseng [Pfaffia glomerata (Spreng.) Pedersen] plantlets. Plant Cell Tissue Organ Cult 118(1):87–99. https://doi.org/10.1007/s11240-014-0464-x
Sanadze G (1990) The principal scheme of photosynthetic carbon conversion in cells of isoprenereleasing plants. In: Baltscheffsky M (eds) Current research in photosynthesis. Springer, Dordrecht
Sanadze G (2004) Biogenic isoprene (a review). Russ J Plant Physiol 51(6):729–741. https://doi.org/10.1023/B:RUPP.0000047821.63354.a4
Shao M, Czapiewski KV, Heiden AC, Kobel K, Komenda M, Koppmann R, Wildt J (2001) Volatile organic compound emissions from Scots pine: mechanisms and description by algorithms. J Geophys Res 106(D17):20483–20491. https://doi.org/10.1029/2000JD000248
Silva ABD, Lima PP, Oliveira LESD, Moreira AL (2014) In vitro growth and leaf anatomy of Cattleya walkeriana (Gardner, 1839) grown in natural ventilation system. Rev Ceres 61(6):883–890. https://doi.org/10.1590/0034-737X201461060001
Silva ST, Bertolucci SKV, da Cunha SHB, Lazzarini LES, Tavares MC, Pinto JEBP (2017) Effect of light and natural ventilation systems on the growth parameters and carvacrol content in the in vitro cultures of Plectranthus amboinicus (Lour.) Spreng. Plant Cell Tissue Organ Cult 129(3):501–510. https://doi.org/10.1007/s11240-017-1195-6
Snow MD, Bard RR, Olszyk DM, Minster LM, Hager AN, Tingey DT (2003) Monoterpene levels in needles of Douglas fir exposed to elevated CO2 and temperature. Physiol Plant 117(3):352–358. https://doi.org/10.1034/j.1399-3054.2003.00035.x
Souza ADS, Albuquerque UP, Nascimento ALBD, Santoro FR, Torres-Avilez WM, Lucena RFPD, Monteiro JM (2017) Temporal evaluation of the conservation priority Index for medicinal plants. Acta Bot Bras 31(2):169–179. https://doi.org/10.1590/0102-33062017abb0027
Trevisan MT, Marques RA, Silva MG, Scherer D, Haubner R, Ulrich CM, Owen RW (2016) Composition of essential oils and ethanol extracts of the leaves of Lippia species: identification, quantitation and antioxidant capacity. Rec Nat Prod 10(4):485
Us-Camas R, Rivera-Solís G, Duarte-Aké F, De-la-Peña C (2014) In vitro culture: an epigenetic challenge for plants. Plant Cell Tissue Organ Cult 118(2):187–201. https://doi.org/10.1007/s11240-014-0482-8
Vahdati K, Hassankhah A (2014) Developing a photomixotrophic system for micropropagation of persian walnut. Acta Hortic 1050:181–187. https://doi.org/10.17660/ActaHortic.2014.1050.23
Van den Dool H, Kratz PD (1963) A generalization of the retention index system including linear temperature programmed gas–liquid partition chromatography. J Chromatogr A 11:463–471
Wilkinson MJ, Monson RK, Trahan N, Lee S, Brown E, Jackson RB, Polley HW, Fay PA, Fall R (2009) Leaf isoprene emission rate as a function of atmospheric CO2 concentration. Glob Chang Biol 15(5):1189–1200. https://doi.org/10.1111/j.1365-2486.2008.01803.x
Xiao Y, Niu G, Kozai T (2011) Development and application of photoautotrophic micropropagation plant system. Plant Cell Tissue Organ Cult 105(2):149–158. https://doi.org/10.1007/s11240-010-9863-9
Ziska LH, Panicker S, Wojno HL (2008) Recent and projected increases in atmospheric carbon dioxide and the potential impacts on growth and alkaloid production in wild poppy (Papaver setigerum DC.). Clim Change 91(3–4):395. https://doi.org/10.1007/s10584-008-9418-9
Zobayed S, Armstrong J, Armstrong W (2002) Multiple shoot induction and leaf and flower bud abscission of Annonacultures as affected by types of ventilation. Plant Cell Tissue Organ Cult 69(2):155–165. https://doi.org/10.1023/A:1015275718908
Acknowledgements
This study was financed in part by the National Council for Scientific and Technological Development (CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico), the Minas Gerais State Research Foundation (FAPEMIG - Fundação de Pesquisa do Estado de Minas Gerais), and the Coordination for the Improvement of Higher Education Personnel (CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil) (CAPES – Finance Code 001).
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LESL, ACS and FVP performed the laboratory work and the experimental work. SKVB performed the chemical analyses. AAC participated in the statistical analysis. BY participated in the correction of the manuscript. JEBPP wrote and corrected the manuscript. The authors accepted the final version of our manuscript.
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Key message This research reports on the influence of explant type and natural ventilation systems on growth and content of volatile compounds of Lippia gracilis, a threatened medicinal plant. Growth results were higher for explants with leaves and four membranes. The natural ventilation system with four membranes promoted greater accumulation of carvacrol and thymol.
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Lazzarini, L.E.S., Bertolucci, S.K.V., de Carvalho, A.A. et al. Explant type and natural ventilation systems influence growth and content of carvacrol and thymol of Lippia gracilis Schauer. Plant Cell Tiss Organ Cult 137, 33–43 (2019). https://doi.org/10.1007/s11240-018-01548-5
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DOI: https://doi.org/10.1007/s11240-018-01548-5