Tomato and mini-cucumber tolerance to photoperiodic injury involves photorespiration and the engagement of nighttime cyclic electron flow from dynamic LEDs.

Controlled environment agriculture (CEA) is critical for achieving year-round food security in many regions of the world. CEA is a resource-intensive endeavor, with lighting consuming a large fraction of the energy. To lessen the burden on the grid and save costs, an extended photoperiod strategy can take advantage of off-peak time-of-day options from utility suppliers. However, extending the photoperiod limits crop production morphologically and physiologically if pushed too long. Here, we present a continuous-light dynamic light-emitting diode (LED) strategy (involving changes in spectra, intensity, and timing), that overcomes these limitations. We focused on tomato, a well described photoperiodic injury-sensitive species, and mini-cucumber, a photoperiodic injury-tolerant species to first assess morphological responses under control (16-h photoperiod, unchanging spectrum), constant (24-h photoperiod, unchanging spectrum), and two variations of a dynamic LED strategy, dynamic 1 (16-h "day", 3-h "peak", 8-h "night" spectra) and dynamic 2 (20-h "day", 5-h "peak", 4-h "night" spectra). Next, we tested the hypothesis of photorespiration's involvement in photoperiodic injury by using a leaf gas exchange coupled with chlorophyll fluorescence protocol. We further explored Adenosine triphosphate (ATP): Nicotinamide adenine dinucleotide phosphate (NADPH) ratio supply/demand responses by probing photosynthetic electron flow and proton flow with the MultispeQ instrument. We found canopy architecture can be tuned by minor variations of the same dynamic LED strategy, and we highlight dynamic 1 as the optimal choice for both tomato and mini-cucumber as it improved biomass/architecture and first-yield, respectively. A central discovery was that dynamic 1 had a significantly higher level of photorespiration than control, for both species. Unexpectedly, photorespiration was comparable between species under the same treatments, except under constant. However, preliminary data on a fully tolerant tomato genotype grown under constant treatment upregulated photorespiration similar to mini-cucumber. These results suggest that photoperiodic injury tolerance involves a sustained higher level of photorespiration under extended photoperiods. Interestingly, diurnal MultispeQ measurements point to the importance of cyclic electron flow at subjective nighttime that may also partially explain why dynamic LED strategies mitigate photoperiodic injury. We propose an ontology of photoperiodic injury involving photorespiration, triose phosphate utilization, peroxisomal H2O2-catalase balance, and a circadian external coincidence model of sensitivity that initiates programmed cell death.

Tissue culture coupled with a gas exchange system offers new perspectives on phenotyping the developmental biology of Solanum lycopersicum L. cv. 'MicroTom'.

Solanum lycopersicum L. cv. 'Microtom' (MicroTom) is a model organism with a relatively rapid life cycle, and wide library of genetic mutants available to study different aspects of plant development. Despite its small stature, conventional MicroTom research often requires expensive growth cabinets and/or expansive greenhouse space, limiting the number of experimental and control replications needed for experiments, and can render plants susceptible to pests and disease. Thus, alternative experimental approaches must be devised to reduce the footprint of experimental units and limit the occurrence problematic confounding variables. Here, tissue culture is presented as a powerful option for MicroTom research that can quell the complications associated with conventional MicroTom research methods. A previously established, non-invasive, analytical tissue culture system is used to compare in vitro and conventionally produced MicroTom by assessing photosynthesis, respiration, diurnal carbon gain, and fruit pigments. To our knowledge, this is the first publication that measures in vitro MicroTom fruit pigments and compares diurnal photosynthetic/respiration responses to abiotic factors between in vitro and ex vitro MicroTom. Comparable trends would validate tissue culture as a new benchmark method in MicroTom research, as it is like Arabidopsis, allowing replicable, statistically valid, high throughput genotyping and selective phenotyping experiments. Combining the model plant MicroTom with advanced tissue culture methods makes it possible to study bonsai-style MicroTom responses to light, temperature, and atmospheric stimuli in the absence of confounding abiotic stress factors that would otherwise be unachievable using conventional methods.

A Noninvasive Gas Exchange Method to Test and Model Photosynthetic Proficiency and Growth Rates of In Vitro Plant Cultures: Preliminary Implication for Cannabis sativa L.

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.