Abstract
The term in vitro photomixotrophism refers to the ability of explants to obtain metabolic energy from the culture medium and as a product of photosynthesis. The objective of this research was to study the physiological and biochemical mechanisms of V. planifolia during in vitro multiplication using a photomixotrophic system with different sucrose contents (0, 15 and 30 g L–1) and CO2 supply levels (500, 800 and 1200 ppm) using a Temporary Immersion Modular System (SMIT®). After 45 days of multiplication, response percentage, number of shoots per explant, shoot length, number of leaves per shoot, stomatal index (%), percentage of closed stomata, and chlorophyll, β-carotene, Phosphoenolpyruvate (PEP) and Rubisco contents were evaluated. In addition, the survival rate during acclimatization at 60 days was evaluated. For the multiplication stage, the highest response percentage was obtained in the treatments with 15 g L–1 sucrose with 500 and 800 ppm CO2, and 30 g L–1 sucrose with 500 ppm CO2. In the latter treatment, the best development parameters were obtained, with 14.75 shoots per explant, a shoot length of 2.38 cm and 2.5 leaves per shoot. In general, the highest chlorophyll, β-carotene, PEP and Rubisco contents were observed with 30 g L–1 sucrose + CO2. No effects of the treatments on stomatal index (%) were observed, while the percentage of closed stomata showed differences among treatments. At the acclimatization stage, the highest survival percentages were obtained from the treatments of 30 g L–1 sucrose with 500 and 800 ppm CO2. In conclusion, this study demonstrates physiological and biochemical mechanisms for a better understanding of photomixotrophism during in vitro multiplication in vanilla and may be applied to other species.
Key message
In vitro photomixotrophism can be induced by injecting carbon dioxide to promote photosynthesis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Aragón CE, Escalona M, Rodriguez R et al (2010) Effect of sucrose, light, and carbon dioxide on plantain micropropagation in temporary immersion bioreactors. Invitro Cell Dev Biol Plant 46:89–94. https://doi.org/10.1007/s11627-009-9246-2
Arencibia AD, Vergara C, Quiroz K, Carrasco B, García-Gonzales R (2013) Establishment of photomixotrophic cultures for raspberry micropropagation in temporary immersion bioreactors (TIBs). Sci Hortic 160:49–53
Asayesh ZM, Vahdati K, Aliniaeifard S, Askari N (2017) Enhancement of ex vitro acclimation of walnut plantlets through modification of stomatal characteristics in vitro. Sci Hortic 220:114–121. https://doi.org/10.1016/j.scienta.2017.03.045
Bello-Bello JJ, Schettino-Salomón S, Ortega-Espinoza J et al (2021) A temporary immersion system for mass micropropagation of pitahaya (Hylocereus undatus). 3 Biotech 11:437. https://doi.org/10.1007/s13205-021-02984-5
Biehler E, Mayer F, Hoffmann L, Krause E, Bohn T (2010) Comparison of 3 spectrophotometric methods for carotenoid determination in frequently consumed fruits and vegetables. J Food Sci 75:55–61
Cha-um S, Chanseetis C, Chintakovid W et al (2011) Promoting root induction and growth of in vitro macadamia (Macadamia tetraphylla L. ‘Keaau’) plantlets using CO2-enriched photoautotrophic conditions. Plant Cell Tiss Organ Cult 106:435–444. https://doi.org/10.1007/s11240-011-9940-8
Cochard H, Coll L, Le Roux X, Améglio T (2002) Unraveling the effects of plant hydraulics on stomatal closure during water stress in walnut. Plant Physiol 128:282–350. https://doi.org/10.1104/pp.010400
de Paula Alves J, Pinheiro MVM, Corrêa TR et al (2023) Morphophysiology of Ananas comosus during in vitro photomixotrophic growth and ex vitro acclimatization. Vitro Cell Dev Biol-Plant 59:106–120. https://doi.org/10.1007/s11627-022-10321-5
Delucia EH, Sasek TW, Strain BR (1985) Photosynthetic inhibition after long-term exposure to elevated levels of atmospheric carbon dioxide. Photosynth Res 7:175–184
Escalona M, Samson G, Borroto C et al (2003) Physiology of effects of temporary immersion bioreactors on micropropagated pineapple plantlets. In Vitro Cell Dev Biol-Plant 39:651–656. https://doi.org/10.1079/IVP2003473
Fortini EA, Batista DS, Mamedes-Rodrigues TC et al (2021) Gas exchange rates and sucrose concentrations affect plant growth and production of flavonoids in Vernonia condensata grown in vitro. Plant Cell Tiss Organ Cult 144:593–605. https://doi.org/10.1007/s11240-020-01981-5
Gago D, Vilavert S, Bernal M, Sánchez C, Aldrey A, Vidal N (2021) The effect of sucrose supplementation on the micropropagation of Salix viminalis L. shoots in semisolid medium and temporary immersion bioreactors. Forests 12:1408. https://doi.org/10.3390/f12101408
Gago D, Sánchez C, Aldrey A, Christie CB, Bernal M, Vidal N (2022) Micropropagation of plum (Prunus domestica L.) in bioreactors using photomixotrophic and photoautotrophic conditions. Horticulturae 8:286. https://doi.org/10.3390/horticulturae8040286
Gago D, Bernal M, Sánchez C, Aldrey A, Cuenca B, Christie CB, Vidal N (2022) Effect of sucrose on growth and stress status of Castanea sativa x C. crenata shoots cultured in liquid medium. Plants 11:965. https://doi.org/10.3390/plants11070965
Giuliani R, Karki S, Covshoff S et al (2019) Transgenic maize phosphoenolpyruvate carboxylase alters leaf–atmosphere CO2 and 13CO2 exchanges in Oryza sativa. Photosynth Res 142:153–167. https://doi.org/10.1007/s11120-019-00655-4
Harborne JB (1973) Compuestos fenólicos en Métodos fitoquímicos. Springer, Dordrecht, Holanda, pp 33–88
Hazarika BN (2006) Morpho-physiological disorders in in vitro culture of plants. Sci Hortic 108:105–120. https://doi.org/10.1016/j.scienta.2006.01.038
Hirschberg J (2001) Carotenoid biosynthesis in flowering plants. Curr Opin Plant Biol 4:210–218
Hurtado-Gaitán E, Sellés-Marchart S, Hartwell J, Martínez-Esteso MJ, Bru-Martínez R (2021) Down-regulation of phosphoenolpyruvate carboxylase kinase in grapevine cell cultures and leaves is linked to enhanced resveratrol biosynthesis. Biomolecules 11:1641. https://doi.org/10.3390/biom11111641
Kume A, Akitsu T, Nasahara KN (2018) Why is chlorophyll b only used in light-harvesting systems? J Plant Res 131:961–972
Langlé-Argüello LA, Martínez-Gutiérrez GA, Santiago-García PA, Escamirosa-Tinoco C, Morales I, Enríquez-del-Valle JR (2019) Nutrient solutions and drought in plant growth and fructans content of Agave potatorum Zucc. HortScience horts 54:1581–1584. https://doi.org/10.21273/HORTSCI14129-19
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
Netto AT, Campostrini E, De Oliveira JG, Bressan-Smith RE (2005) Photosynthetic pigments, nitrogen, chlorophyll a fluorescence and SPAD-502 readings in coffee leaves. Sci Hortic 104:199–209.https://doi.org/10.1016/j.scienta.2004.08.013
Pinheiro MVM, Ríos-Ríos AM, da Cruz ACF et al (2021) CO2 enrichment alters morphophysiology and improves growth and acclimatization in Etlingera Elatior (Jack) R.M. Smith micropropagated plants. Braz J Bot 44:799–809. https://doi.org/10.1007/s40415-021-00741-9
Raffi A, Abdullah NAP, Noor-Syaheera MY, Go R (2019) Preliminary foliar anatomical assessment of Four Vanilla Species (Orchidaceae) from Perak, Malaysia. Pertanika J Trop Agric 42(2):807–816
Ramírez-Mosqueda MA, Bello-Bello JJ (2021) SETIS™ bioreactor increases in vitro multiplication and shoot length in vanilla (Vanilla planifolia Jacks. Ex Andrews) Acta Physiol Plant 43:1–8. https://doi.org/10.1007/s11738-021-03227-z
Ramírez-Mosqueda MA, Iglesias-Andreu LG, Ramírez-Madero G, Hernández-Rincón EU (2016) Micropropagation of Stevia rebaudiana Bert. in temporary immersion systems and evaluation of genetic fidelity. S Afr J Bot 106:238–243. https://doi.org/10.1016/j.sajb.2016.07.015
Ramos-Castellá A, Iglesias-Andreu LG, Bello-Bello J, Lee-Espinosa H (2014) Improved propagation of vanilla (Vanilla planifolia jacks. Ex Andrews) using a temporary immersion system. Invitro Cell Dev Biol-Plant 50:576–581. https://doi.org/10.1007/s11627-014-9602-8
Regueira M, Rial E, Blanco B, Bogo B, Aldrey A, Correa B, Varas E, Sánchez C, Vidal N (2018) Micropropagation of axillary shoots of Salix viminalis using a temporary immersion system. Trees 32:61–71. https://doi.org/10.1007/s00468-017-1611-x
Schulze ED, Beck E, Buchmann N, Clemens S, Müller-Hohenstein K, Scherer Lorenzen M (2019) Water deficiency (drought). Plant Ecol 165–202. https://doi.org/10.1007/978-3-662-56233-8_6
Ševčíková H, Lhotáková Z, Hamet J, Lipavská H (2019) Mixotrophic in vitro cultivations: the way to go astray in plant physiology. Physiol Plant 167:365–377
Shi J, Yi K, Liu Y, Xie L, Zhou Z, Chen Y, Hu Z, Zheng T, Liu R, Chen Y, Chen J (2015) Phosphoenolpyruvate carboxylase in Arabidopsis leaves plays a crucial role in carbon and nitrogen metabolism. Plant Physiol 167:671–681. https://doi.org/10.1104/pp.114.254474
Silvera K, Neubig KM, Whitten WM, Williams NH, Winter K, Cushman JC (2010) Evolution along the crassulacean acid metabolism continuum. Funct Plant Biol 37:995–1010. https://doi.org/10.1071/FP10084
Spinoso-Castillo JL, Chávez-Santoscoy RA, Bogdanchikova N, Pérez-Sato JA, Morales-Ramos V, Bello-Bello JJ (2017) Antimicrobial and hormetic effects of silver nanoparticles on in vitro regeneration of vanilla (Vanilla planifolia jacks. Ex Andrews) using a temporary immersion system. Plant Cell Tiss Organ Cult 129:195–207. https://doi.org/10.1007/s11240-017-1169-8
Trauger M, Hile A, Sreenivas K et al (2022) CO2 supplementation eliminates sugar-rich media requirement for plant propagation using a simple inexpensive temporary immersion photobioreactor. Plant Cell Tiss Organ Cult 150:57–71. https://doi.org/10.1007/s11240-021-02210-3
Vitlin-Gruber A, Feiz L (2018) Rubisco assembly in the chloroplast. Front Mol Biosci 5:24. https://doi.org/10.3389/fmolb.2018.00024
Wilkinson H (1979) The plant surface (mainly leaf). In: Metcalfe CR, Chalk L (eds) Anatomy of dicotyledons. Claredon Press, Oxford, pp 97–165
Wu HC, Lin CC (2013) Carbon dioxide enrichment during photoautotrophic micropropagation of Protea cynaroides L. plantlets improves in vitro growth, net photosynthetic rate, and acclimatization. HortScience 48:1293–1297
Xu Z, Zhou G (2008) Responses of leaf stomatal density to water status and its relationship with photosynthesis in a grass. J Exp Bot 59:3317–3325. https://doi.org/10.1093/jxb/ern185
Zhao X, Li WF, Wang Y et al (2019) Elevated CO2 concentration promotes photosynthesis of grape (Vitis vinifera L. cv. ‘Pinot noir’) plantlet in vitro by regulating RbcS and Rca revealed by proteomic and transcriptomic profiles. BMC Plant Biol 19:42. https://doi.org/10.1186/s12870-019-1644-y
Funding
The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.
Author information
Authors and Affiliations
Contributions
All authors listed have made substantial, direct and intellectual contribution to the work, and approved it for publication.
Corresponding author
Ethics declarations
Competing interests
The authors have no relevant financial or non-financial interests to disclose.
Additional information
Communicated by Ming-Tsair Chan.
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Luis, S.J., Jabín, B.J. CO2-enriched air in a temporary immersion system induces photomixotrophism during in vitro multiplication in vanilla. Plant Cell Tiss Organ Cult 155, 29–39 (2023). https://doi.org/10.1007/s11240-023-02546-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-023-02546-y