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Application of 3D Printing to Prototype and Develop Novel Plant Tissue Culture Systems

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Application of 3D Printing to Prototype and Develop Novel Plant Tissue Culture Systems

Mukund R Shukla et al. Plant Methods.

Abstract

Background: Due to the complex process of designing and manufacturing new plant tissue culture vessels through conventional means there have been limited efforts to innovate improved designs. Further, development and availability of low cost, energy efficient LEDs of various spectra has made it a promising light source for plant growth in controlled environments. However, direct replacement of conventional lighting sources with LEDs does not address problems with uniformity, spectral control, or the challenges in conducting statistically valid experiments to assess the effects of light. Prototyping using 3D printing and LED based light sources could help overcome these limitations and lead to improved culture systems.

Results: A modular culture vessel design in which the fluence rate and spectrum of light are independently controlled was designed, prototyped using 3D printing, and evaluated for plant growth. This design is compatible with semi-solid and liquid based culture systems. Observations on morphology, chlorophyll content, and chlorophyll fluorescence based stress parameters from in vitro plants cultured under different light spectra with similar overall fluence rate indicated different responses in Nicotiana tabacum and Artemisia annua plantlets. This experiment validates the utility of 3D printing to design and test functional vessels and demonstrated that optimal light spectra for in vitro plant growth is species-specific.

Conclusions: 3D printing was successfully used to prototype novel culture vessels with independently controlled variable fluence rate/spectra LED lighting. This system addresses several limitations associated with current lighting systems, providing more uniform lighting and allowing proper replication/randomization for experimental plant biology while increasing energy efficiency. A complete procedure including the design and prototyping of a culture vessel using 3D printing, commercial scale injection molding of the prototype, and conducting a properly replicated experiment are discussed. This open source design has the scope for further improvement and adaptation and demonstrates the power of 3D printing to improve the design of culture systems.

Keywords: 3D printing; Culture vessel design; LED lighting system; Light quality; Micropropagation; Plant tissue culture; Prototyping.

Figures

Fig. 1
Fig. 1
A typical tissue culture room shelf (120 × 60 × 40 cm) with two florescent bulbs on a ballast at the center of the shelf (a), Heat map of light fluence rate (b). Each square represents a 10 cm2 area measured from the center with a light meter 31 cm from the light
Fig. 2
Fig. 2
Dimensional drawing and design of culture vessels with lid and its 3D view before printing (a), and injection molded (left) and 3D printed culture vessels (b). Injection molded vessels was based on 3D printed design and produced following initial experiments
Fig. 3
Fig. 3
3D printed lid housing which is used to hold LED RBG strips and connector for power supply with provision for exhaust fan (a), whole assembly on 3D printed culture vessel (b). Several 3D printed units set at different light spectra stacked in a completely randomized design (c) and injection moulded culture vessels set at different light spectra stacked in completely randomized design (d)
Fig. 4
Fig. 4
Tobacco and Artemisia plants cultured under in vitro condition with lid having various light spectra: a red:blue 3:1, b red:blue 1:1, c red:blue 1:3, d white with their respective graphs and fluence rate data and showing growth after 3 weeks period
Fig. 5
Fig. 5
Differences in plant height, no. of shoots, no. of nodes and fresh weight measured after 3 weeks of growth of tobacco and artemisia growing under white and red/blue combination with the fluence rate 35 μmol m−2 s−1. Data presented as mean ± SE and different letters in the figures indicate significant differences at α = 0.05 using Tukey’s test (lower and upper case letters are used for artemisia and tobacco, respectively)
Fig. 6
Fig. 6
Average chlorophyll content of artemisia and tobacco plant 3 weeks of growth under different red/blue light combination and white with the fluence rate 35 μmol m−2 s−1. Data presented as mean ± SE and different letters in the figures indicate significant differences at α = 0.05 using Tukey’s test (lower and upper case letters are used for artemisia and tobacco, respectively)

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