Abstract
Conventional in vitro propagation may affect normal morphophysiological characteristics and survival during the acclimatization of commercial plants, such as Etlingera Elatior (Jack) R.M. Smith. CO2 enrichment, gas-permeable membranes, and reduced carbohydrate concentration in culture media can be used to mitigate these effects. Here, we aimed to determine the impact of photomixotrophic and photoautotrophic growth on morphophysiological responses of E. elatior and its survival during ex vitro acclimatization. In vitro-established plants were placed in flasks containing semisolid MS medium with 0.54 µM α-naphthalene acetic acid and sucrose (0 or 15 g L−1). The flasks were sealed with lids with or without orifices covered by gas-permeable membranes and kept for 45 days under forced-air ventilation (360 or 1000 µmol−1 CO2). Growth variables, anatomical and physiological characteristics, and survival rate were evaluated. Overall, 1000 µmol mol−1 CO2, gas-permeable membranes, and 15 g L−1 sucrose resulted in plants with higher fresh and dry mass, higher level of photosynthetic pigments, as well as more developed secondary ribs in the leaves, prominent midribs, highly organized and differentiated chlorenchyma cells (palisade and spongy parenchyma), more developed vascular bundles, and intercellular gaps were reduced, compared to conventionally in vitro-propagated plants. Moreover, these conditions promote morphophysiological responses and favored survival of E. elatior plants (75%) when transferred to ex vitro conditions.
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References
Abdelwahab SI, Zaman FQ, Mariod AA, Yaacob M, Abdelmageed AH, Khamis S (2010) Chemical composition, antioxidant and antibacterial properties of the essential oils of Etlingera elatior and Cinnamomum pubescens Kochummen. J Sci Food Agr 90:2682–2688. https://doi.org/10.1002/jsfa.4140
Albuquerque ESB, Neves LJ (2004) Anatomia foliar de Alpinia zerumbet (Pers.) Burtt & Smith (Zingiberaceae). Acta Bot Bras 18:109–121. https://doi.org/10.1590/S0102-33062004000100010
Alvarez C, Sáez P, Sáez K, Sánchez-Olate M, Ríos D (2012) Effects of light and ventilation on physiological par during in vitro acclimatization of Gevuina avellana mol. Plant Cell Tiss Organ Cult 110:93–101. https://doi.org/10.1007/s11240-012-0133-x
Arigita L, Cañal MJ, Tamés RS, González A (2010) CO2-enriched microenvironment affects sucrose and macronutrients absorption and promotes autotrophy in the in vitro culture of kiwi (Actinidia deliciosa Chev. Liang and Ferguson). In Vitro Cell Dev Biol-Plant 46:312–322. https://doi.org/10.1007/s11627-009-9267-x
Ayub RA, Santos JND, Zanlorensi Junior LA, Silva DMD, Carvalho TCD, Grimaldi F (2019) Sucrose concentration and volume of liquid medium on the in vitro growth and development of blackberry cv. Tupy in temporary immersion systems. Cienc Agrotec. 43:e007219. https://doi.org/10.1590/1413-7054201943007219
Badr A, Angers P, Desjardins Y (2011) Metabolic profiling of photoautotrophic and photomixotrophic potato plantlets (Solanum tuberosum) provides new insights into acclimatization. Plant Cell Tiss Organ Cult 107:13–24. https://doi.org/10.1007/s11240-011-9951-5
Baque MA, Elgirban A, Lee EJ, Paek KY (2012) Sucrose regulated enhanced induction of anthraquinone, phenolics, flavonoids biosynthesis and activities of antioxidant enzymes in adventitious root suspension cultures of Morinda citrifolia (L.). Acta Physiol Plant 34:405–415. https://doi.org/10.1007/s11738-011-0837-2
Batista DS, de Castro KM, Koehler AD, Porto BN, da Silva AR, de Souza VC, Teixeira ML, Cardoso MG, Santos MO, Viccini LF, Otoni WC (2017a) Elevated CO2 improves growth, modifes anatomy, and modulates essential oil qualitative production and gene expression in Lippia alba (Verbenaceae). Plant Cell Tiss Organ Cult 128:357–368. https://doi.org/10.1007/s11240-016-1115-1
Batista DS, Dias LLC, Rêgo MMd, Saldanha CW, Otoni WC (2017b) Flask sealing on in vitro seed germination and morphogenesis of two types of ornamental pepper explants. Cienc Rural 47:e20150245. https://doi.org/10.1590/0103-8478cr20150245
Chan EWC, Ng VP, Tan VV, Low YY (2011) Antioxidant and antibacterial properties of Alpinia galanga, Curcuma longa, and Etlingera elatior (Zingiberaceae). Pharmacogn J 3:54–61. https://doi.org/10.5530/pj.2011.22.11
Chanemougasoundharam A, Sarkar D, Pandey SK, Al-Biski F, Helali O, Minhas JS (2004) Culture tube closure type affects potato plantlets growth and chlorophyll contents. Biol Plant 48:7–11
Cruz CD (2013) GENES - a software package for analysis in experimental statistics and quantitative genetics. Acta Sci-Agron 35:271–276. https://doi.org/10.4025/actasciagron.v35i3.21251
de la Viña G, Pliego-Alfaro F, Driscoll SP, Mitchell VJ, Parry MA, Lawlor DW (1999) Effects of CO2 and sugars on photosynthesis and composition of avocado leaves grown in vitro. Plant Physiol Biochem 37:587–595. https://doi.org/10.1016/S0981-9428(00)80111-4
Desjardins Y, Dubuc JF, Badr A (2009) In vitro culture of plants: a stressful activity. Acta Hortic 812:29–50. https://doi.org/10.17660/ActaHortic.2009.812.1
García-Ramírez Y, Barrera GP, Freire-Seijo M, Barbón R, Concepción-Hernández M, Mendoza-Rodríguez MF, Torres-García S (2019) Effect of sucrose on physiological and biochemical changes of proliferated shoots of Bambusa vulgaris Schrad. Ex Wendl in temporary immersion. Plant Cell Tiss Organ Cult 137:239–247. https://doi.org/10.1007/s11240-019-01564-z
Ghasemzadeh A, Jaafar HZ, Rahmat A, Ashkani S (2015) Secondary metabolites constituents and antioxidant, anticancer and antibacterial activities of Etlingera elatior (Jack) R.M.Sm grown in different locations of Malaysia. BMC Complement Altern Med. 15:335. https://doi.org/10.1186/s12906-015-0838-6
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 Tiss Organ Cult 110:227–238. https://doi.org/10.1007/s11240-012-0145-6
Ivanova M, Staden J (2010) Natural ventilation effectively reduces hyperhydricity in shoot cultures of Aloe polyphylla Schönland ex Pillans. Plant Growth Regul 60:143–150. https://doi.org/10.1007/s10725-009-9430-8
Karnovsky MJ (1965) A formaldehyde–glutaraldehyde fixative of high osmolarity for use in electron microscopy. J Cell Biol 27:127–128
Kozai T (2010) Photoautotrophic micropropagation—environmental control for promoting photosynthesis. Propag Ornam Plants 10:188–204
Kozai T, Kubota C (2001) Development a photoautotrophic micropropagation system for woody plants. J Plant Res 114:525–537. https://doi.org/10.1007/PL00014020
Lins SRO, Coelho RSB (2003) Antracnose em inflorescências de bastão do imperador (Etlingera elatior): Ocorrência e métodos de inoculação. Summa Phytop 29:355–358
Martins MBG, Caravante ALC, Appezzato-da-Glória B, Soares MKM, Moreira RRD, Santos LE (2010) Caracterização anatômica e fitoquímica de folhas e rizomas de Hedychium coronarium J. König (Zingiberaceae). Rev Bras Plant Med 12:179–187. https://doi.org/10.1590/S1516-05722010000200009
Mills D, Yanqing Z, Benzioni A (2004) Improvement of jojoba shoot multiplication in vitro by ventilation. Vitro Cell Dev Biol-Plant 40:386–402. https://doi.org/10.1079/IVP2004537
Mohamed MAH, Alsadon AA (2010) Influence of ventilation and sucrose on growth and leaf anatomy of micropropagated potato plantlets. Sci Hortic 123:295–300. 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 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Nguyen QT, Kozai T (2005) Photoautotrophic micropropagation of woody species. In: Kozai T, Afreen F, Zobayed SMA (eds) Photoautotrophic (sugar-free medium) micropropagation as a new micropropagation and transplant production system. Springer, Dordrecht, pp 123–146. doi:https://doi.org/10.1007/1-4020-3126-2_8
Nguyen QT, Kozai T, Niu G, Nguyen UV (1999) Photosynthetic characteristics of coffee (Coffea arabusta) plantlets in vitro in response to different CO2 concentrations and light intensities. Plant Cell Tiss Organ Cult 55:133–139. https://doi.org/10.1023/A:1006224026639
O’Brien TP, McCully ME (1981) The study of plant structure principles and select methods. Termarcarphi Pty Ltd, Melbourne. https://doi.org/10.1111/1365-3040.ep11572627
Pinheiro MVM, Martins FB, xavier A, Otoni WC, (2013) Trocas gasosas influenciam na morfogênese in vitro de duas cultivares de oliveira (Olea europaea L.). Rev Arvore 37:19–29. https://doi.org/10.1590/S0100-67622013000100003
Quinaya DCP, d’Almeida JRM (2019) Possibility of exploring and applying wastes from some ornamental plants (Elatior etlingera; Costus comosus; Heliconia bihai) as sources of natural cellulosic fibers. J Nat Fibers. https://doi.org/10.1080/15440478.2019.1581118
Ribeiro APO, Picoli EAT, Lani ERG, Vendrame WA, Otoni WC (2009) The influence of flask sealing on in vitro morphogenesis of eggplant (Solanum melongena L.). In Vitro Cell Dev Biol - Plant 45:421–428
Ríos-Ríos AM, da Silva JVS, Fernandes JVM, Batista DS, Silva TD, Chagas K, Pinheiro MVM, Faria DV, Otoni WC, Fernandes AS (2019) Micropropagation of Piper crassinervium: an improved protocol for faster growth and augmented production of phenolic compounds. Plant Cell Tiss Organ Cult 137:495–509. https://doi.org/10.1007/s11240-019-01585-8
Rodrigues M, Costa THF, Festucci-Buselli RA, Silva LC, Otoni WC (2012) Effects of flask sealing and growth regulators on in vitro propagation of neem (Azadirachta indica A. Juss.). In Vitro Cell Dev Biol-Plant 48:67–72. https://doi.org/10.1007/s11627-011-9398-8
Rodrigues M, Rocha DI, Mendonça AMDC, Silva LCD, Festucci-Buselli RA, Otoni WC (2020) Leaf anatomy micromorphometry plasticity and histochemistry of Azadirachta indica during acclimatization. Rodriguésia 71:e01902018. https://doi.org/10.1590/2175-7860202071019
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 Tiss Organ Cult 110:413–422. https://doi.org/10.1007/s11240-012-0162-5
Saldanha CW, Otoni CG, Notini MM, Kuki KN, Cruz ACF, Rubio Neto A, Dias LLC, Otoni WC (2013) A CO2-enriched atmosphere improves in vitro growth of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen]. In Vitro Cell Dev Biol-Plant 49:433–444. https://doi.org/10.1007/s11627-013-9529-5
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 Tiss Organ Cult 118:87–99. https://doi.org/10.1007/s11240-014-0464-x
Santos RP, Cruz ACF, Iarema L, Kuki KN, Otoni WC (2008) Protocolo para extração de pigmentos foliares em porta-enxertos de videira micropropagados. Rev Ceres 55:356–364
Silva AR, de Melo NF, Yano-Melo AM (2017) Acclimatization of micropropagated plants of Etlingera elatior (Jack) R. M. Sm. inoculated with arbuscular mycorrhizal fungi. S Afr J Bot 113:164–169. https://doi.org/10.1016/j.sajb.2017.08.014
Soni V, Keswani K, Bhatt U, Kumar D, Singh H (2021) In vitro propagation and analysis of mixotrophic potential to improve survival rate of Dolichandra unguis-cati under ex vitro conditions. Heliyon 7:e06101. https://doi.org/10.1016/j.heliyon.2021.e06101
Suthar RK, Rathore P, Purohit SD (2009) In vitro growth and shoot multiplication in Terminalia bellerica Roxb under controlled carbon dioxide environment. Indian J Exp Biol 47:204–209
Victorio CP, Arruda RCO, Riehl CAS, Lage CLS (2011) Leaf volatiles and secretory cells of Alpinia zerumbet (Pers.) Burtt et Smith (Zingiberaceae). Nat Prod Res 25:939–948. https://doi.org/10.1080/14786419.2010.514575
Wellburn AR (1994) The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol 144:307–313. https://doi.org/10.1016/S0176-1617(11)81192-2
Xiao Y, Niu G, Kozai T (2011) Development and application of photoautotrophic micropropagation plant system. Plant Cell Tiss Organ Cult 105:149–158. https://doi.org/10.1007/s11240-010-9863-9
Yoon Y-J, Mobin M, Hahn E-J, Paek K-Y (2009) Impact of in vitro CO2 enrichment and sugar deprivation on acclimatory responses of Phalaenopsis plantlets to ex vitro conditions. Environ Exp Bot 65:183–188. https://doi.org/10.1016/j.envexpbot.2008.08.001
Yunus MF, Aziz MA, Kadir MA, Rashid AA (2012) In vitro propagation of Etlingera elatior (Jack) (torch ginger). Sci Hortic 135:145–150. https://doi.org/10.1016/j.scienta.2011.12.016
Zhang B, Song L, Bekele LD, Shi J, Jia Q, Zhang B, Jin L, Duns GJ, Chen J (2018) Optimizing factors affecting development and propagation of Bletilla striata in a temporary immersion bioreactor system. Sci Hortic 232:121–126. https://doi.org/10.1016/j.scienta.2018.01.007
Zobayed S (2006) Aeration in plant tissue culture. In: Dutta Gupta S, Ibaraki Y (eds) Plant tissue culture engineering, vol 6. Focus on Biotechnology. Springer, Dordrecht, pp 313–327. doi:https://doi.org/10.1007/1-4020-3694-9_16
Zobayed SMA, Afreen F, Kozai T (2001a) Physiology of Eucalyptus plantlets grown photoautotrophically in a scaled-up vessel. In Vitro Cell Dev Biol-Plant 37:807–813. https://doi.org/10.1007/s11627-001-0134-7
Zobayed SMA, Armstrong J, Armstrong W (2001b) Leaf anatomy of in vitro tobacco and cauliflower plantlets as affected by different types of ventilation. Plant Sci 161:537–548. https://doi.org/10.1016/S0168-9452(01)00438-1
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The authors thank the Brazilian Federal Agency for Support and Evaluation of Graduate Education (CAPES) for granting a scholarship, and the Minas Gerais Research Foundation (FAPEMIG) for providing financial support.
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MVMP, ACFC, MYO, AMRR, and CWS performed the experiments. MVMP and ACFC raised the in vitro plants for the experiments. MVMP, ACPPC, DSB, DIR, and WCO contributed to the design and interpretation of this study and the writing of the paper.
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Pinheiro, M.V.M., Ríos-Ríos, A.M., da Cruz, A.C.F. et al. CO2 enrichment alters morphophysiology and improves growth and acclimatization in Etlingera Elatior (Jack) R.M. Smith micropropagated plants. Braz. J. Bot 44, 799–809 (2021). https://doi.org/10.1007/s40415-021-00741-9
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DOI: https://doi.org/10.1007/s40415-021-00741-9