Abstract
In conventional in vitro cultivation systems, the use of polyvinyl chloride plastic film to seal the culture flasks hinders gas exchange. Consequently, morphophysiological disturbances may occur, causing economic losses and, in some cases, failure to apply plant tissue culture techniques. Therefore, the use of commercial gas permeable membranes, or the development of alternative sealing systems, are necessary for improvements in physiological processes, anatomical aspects and growth of plants propagated in vitro. Here, we proposed a simple and alternative low-cost sealing system, and evaluate its effectiveness to promote the in vitro growth and improvement of the physio-anatomical traits of Croton lechleri. The treatments were arranged in a 2 × 2 factorial scheme, two tube sealing systems (triple fabric surgical mask and polyvinyl chloride film) and two explant types (nodal segment and apical bud). The Quoirin and Leproivre culture medium supplemented with indole-3-butyric acid (0.5 mg.L− 1), sucrose (30 g.L− 1) and gar (7 g.L− 1) was applied. The mixotrophic culture system, using the triple-layer surgical mask as a closure type, allowed more gas exchange and growth uniformity, and improvement of physio-anatomical traits of Croton lechleri. Nodal segments cultivated under alternative sealing system produced plantlets with more roots, higher chlorophyll a content, and increase in thickness of palisade parenchyma and leaf blade. Additionally, our sealing system has several advantages for micropropagation, such as efficiency, does not result in contamination, and is relatively inexpensive to implement when compared to imported membranes and other alternative systems developed by other authors.
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Data Availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- TFSM:
-
triple fabric surgical mask.
- PVC:
-
polyvinyl chloride.
- IBA:
-
indole-3-butyric acid.
- QL:
-
Quoirin and Leproivre medium.
- CO2 :
-
Carbon dioxide.
- KOH:
-
Potassium hydroxide.
- WVLR:
-
Water vapor loss rate.
- FAA:
-
Formaldehyde, acetic acid, and alcohol.
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Funding
This research was supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes/Embrapa 001-2011/Grant 39) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq 308731/2019-0).
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Oliveira Junior, J.B., Pessoa, C.M.P., Scherwinski-Pereira, J.E. et al. A simple, alternative and efficient sealing system to improve natural ventilation in culture vessels and the morphophysiological and anatomical quality of Croton lechleri (Muell. Arg.) grown in vitro. Biologia 77, 2945–2954 (2022). https://doi.org/10.1007/s11756-022-01140-5
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DOI: https://doi.org/10.1007/s11756-022-01140-5