A Noninvasive Gas Exchange Method to Test and Model Photosynthetic Proficiency and Growth Rates of In Vitro Plant...

Supplemental sugar additives for plant tissue culture cause mixotrophic growth, complicating carbohydrate metabolism and photosynthetic relationships. A unique platform to test and model the photosynthetic proficiency and biomass accumulation of micropropagated plantlets was introduced and applied to Cannabis sativa L. (cannabis), an emerging crop with high economic interest. Conventional in vitro systems can hinder the photoautotrophic ability of plantlets due to low light intensity, low vapor pressure deficit, and limited CO₂ availability. Though exogenous sucrose is routinely added to improve in vitro growth despite reduced photosynthetic capacity, reliance on sugar as a carbon source can also trigger negative responses that are species-dependent. By increasing photosynthetic activity in vitro, these negative consequences can likely be mitigated, facilitating the production of superior specimens with enhanced survivability. The presented methods use an open-flow/force-ventilated gas exchange system and infrared gas analysis to measure the impact of [CO₂], light, and additional factors on in vitro photosynthesis. This system can be used to answer previously overlooked questions regarding the nature of in vitro plant physiology to enhance plant tissue culture and the overall understanding of in vitro processes, facilitating new research methods and idealized protocols for commercial tissue culture.