Abstract
BackgroundDue 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.ResultsA 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.Conclusions3D 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.
Highlights
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
This study provides a detailed demonstration of the procedure to produce/test fused deposition modeling (FDM) 3D printed vessels and devices for developing new systems to grow in vitro plant cultures and demonstrates their utility in conducting properly replicated experiments to study the effects of light on plant growth and development
One of the major limitations of FDM 3D printing with respect to plant tissue culture is that most materials currently used have relatively low melting points and are not suited to heat sterilization or autoclaving
Summary
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. Plant tissue culture is the aseptic culture of cells, tissues, organs or whole plants under controlled nutritional and environmental conditions, allowing the growth and development of the cells or tissues to be manipulated for a variety of applications. These techniques provide powerful tools to study fundamental processes in plants and Shukla et al Plant Methods (2017) 13:6 multiplied to produce several thousand plants in a relatively short time period and little space on a year round basis. Though light quality is of key significance, experimentation with light qualities affecting in vitro growth of plants presents a number of challenges related to control over the light spectrum produced and difficulties in proper replication and experimental design
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