Plant growth research for food production at Kennedy Space Center looks at how future residents of Mars and the Moon will enjoy the sight, smell, taste, and nutrition of plants. Overall, the goal is to provide a sustainable source of healthy food, on long-duration space flights, so astronauts can get the nutrition they need and produce food. The sustainable production of food will aid in the efforts of closed life support. Plants have a vital application for bio regenerative life support as demands for food and oxygen can be provided through photosynthesis, while the carbon dioxide from human respiration is removed. Transpiration is also used in life support processes as waste water that can be recycled through plant systems with the resultant humidity then condensed as clean water. Selected crops will provide the nutrient requirements needed for long duration space flight. Currently, projects in food production are investigating how plants grow in Martian regolith simulant, new crops testing with tomato and pepper cultivars, acquiring real-time photosynthetic data on crops, assessing plant growth in porous concrete media, and the use of plasma for surface decontamination of seeds

Massa, Gioia D.

Onsay, Daphne

Smith, Trent

NASA Technical Reports Server

NASA KSC – Internship Final Report 
Page 1  
NASA Kennedy Space Center   04/11/2019 
 
 
 
 
 
 
 
Researching Plant Growth in Amended Martian Regolith 
Simulant, Photosynthetic Rates of Plants, Seed Surface 
Decontamination by Plasma Methods, New Crop 
Development, and Porous Concrete Media 
Daphne Onsay  
NASA Kennedy Space Center 
Major: Environmental Biology/Plant Biology 
Exploration Research and Technology Programs (UB-A) Spring 
Session 
Date: April 12, 2019 
 
 
 
  
https://ntrs.nasa.gov/search.jsp?R=20190002674 2019-08-30T21:39:34+00:00Z
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Researching Plant Growth in Amended Martian Regolith 
Simulant, Photosynthetic Rates of plants, Seed Surface 
Decontamination by Plasma methods, New Crop 
Development, and Porous Concrete Media 
Daphne Onsay 1  
Michigan State University, East Lansing, MI, 48225 
Gioia Massa2 
NASA Kennedy Space Center, Florida, 32899 
Trent Smith3 
NASA Kennedy Space Center, Florida, 32899 
 
Nomenclature 
CFU=colony forming unit  
Ci = intercellular carbon dioxide 
HCL = hydrochloric acid 
JSC= Johnson Space Center 
MGS =Mars Global Simulant  
MSL= Mars Science Laboratory  
I. Abstract 
Plant growth research for food production at Kennedy Space Center looks at how future residents of Mars and the 
Moon will enjoy the sight, smell, taste, and nutrition of plants. Overall, the goal is to provide a sustainable source of 
healthy food, on long-duration space flights, so astronauts can get the nutrition they need and produce food. The 
sustainable production of food will aid in the efforts of closed life support.  Plants have a vital application for bio 
regenerative life support as demands for food and oxygen can be provided through photosynthesis, while the carbon 
dioxide from human respiration is removed. Transpiration is also used in life support processes as waste water that 
can be recycled through plant systems with the resultant humidity then condensed as clean water. Selected crops will 
provide the nutrient requirements needed for long duration space flight.  Currently, projects in food production are 
investigating how plants grow in Martian regolith simulant, new crops testing with tomato and pepper cultivars, 
acquiring real-time photosynthetic data on crops, assessing plant growth in porous concrete media, and the use of 
plasma for surface decontamination of seeds.  
II. Plant Growth in Amended Martian Regolith Simulant  
Future plant growth experiments in amended Martian regolith simulant will be completed after the safety has been 
reviewed in the processing and handling of the Martian regolith simulant. The goal of the Martian regolith research 
                                                             
1 NIFS Intern, Exploration Research and Technology Program (UB-A), Kennedy Space Center, and Michigan State 
University 
2 Project Scientist, Exploration Research and Technology Program (UB-A), Utilization and Life Sciences Office, 
Kennedy Space Center. 
3 Project Manager, Exploration Research and Technology Program (UB-A), Utilization and Life Sciences Office, 
Kennedy Space Center 
  
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is to perform crop studies to assess the performance of full duration crop growth in an amended Martian regolith 
simulant. Mars is covered with regolith which is crushed volcanic rock that is composed of a high amount of 
perchlorate salts. Mars consists of basaltic rock that is mainly comprised of iron on the crust and a surface that is 
very similar to Earth’s thin crust in which we have iron, magnesium silicate igneous rocks  on the crust and 
surface.7Martian regolith simulants have been generated to further analyze plant growth and aim to replicate features 
of the reference sample. In Martian soil simulant, the following properties are simulated: mineralogy, bulk 
chemistry, and particle size. The simulant is generated in the terrestrial environment, so there are structural and 
chemical differences between the terrestrial simulant and extraterrestrial regolith samples.3 Last summer, Extra 
Dwarf Pac Choi was grown in Martian regolith simulant: JSC Mars1A. The plant growth experiment was completed 
with three treatments. The first treatment measured the overall performance of Extra Dwarf Pac Choi in JSC Mars 
1A autoclaved regolith simulant. The second treatment measured the growth in JSC Mars-1A in regolith that had 
been previously used in potato experiments. An image of the Extra Dwarf Pak Choi in the second treatment is shown 
in Figure 1. The third treatment was the control as it assessed plant growth in Arcillite, a calcined clay media that 
aids in water retention The MGS-1 resembles the soil analyzed at the Gale crater. It was developed based on the 
mineralogical data collected from the MSL Curiosity rover. This simulant has a chemical composition that is similar 
to other basaltic soils at different landing sites, so it was denoted with the “global” title. 3Red Romaine Lettuce will 
be grown in the amended MGS-1 simulant. MGS-1 will be amended with Mushroom Compost. Mushroom compost 
is known for being a nutrient reservoir with a high carbon to nitrogen ratio8 Red Romaine Lettuce will be grown for 
2 28-day growth periods. The following plant growth parameters will be measured: leaf area, shoot height, shoot 
diameter, chlorophyll quantity, anthocyanin quantity, fresh weight, and dry weight. 
 
 
 
 
Figure 1 : Extra Dwarf Pac Choi on DAP 21 in 400mL of Martian Regolith Simulant JSC Mars1A (100% treatment)  
 
 
III. New Crop Development 
 
 
For new crop testing, 6 cultivars of peppers were used: Espanola, Pompeii, Bulgarian Carrot, Mohawk, Baby Bell, 
and Big Jim. In addition to peppers, there were 5 cultivars of tomatoes used: Golden Heirloom Cherry, Tomato-97, 
Tomato-851, Sweet n Neat, and Red Robin. Pepper and tomato cultivars were grown under the following 
environmental conditions: 
Temperature-23 °C (constant) 
RH-50% (constant) 
CO2-3000ppm 
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Tomatoes provide carotene which the human body converts into vitamin A. In addition to containing carotene, 
tomatoes provide significant amounts of potassium to the human diet as potassium can decrease blood pressure, 
minimize the risk of kidney stones, and prevent bone loss. Individuals who consume diets that consist of tomatoes that 
are rich in carotenoid lycopene were found less likely to develop stomach and rectal cancers. 4 
Peppers contain vitamins C, K, carotenoids and flavonoids. Vitamins A and C help to prevent cell damage, cancer, 
and improve the functioning of the immune system. Vitamin K promotes proper blood clotting and provides rigidity 
in the bone structure. 4 
These crops will provide the vitamins necessary for long duration space travel.  
 
IV.  Radish and Lettuce Growth for Li-6800 Portable Photosynthesis System 
Radish and Lettuce were sown for the portable photosynthesis system Li-6800. The Li-6800 portable 
photosynthesis measures photosynthesis at the leaf-level in real-time measurements. 
Initially it measures the carbon dioxide uptake and transpiration of water by the leaf 
through infrared gas analyzers. In addition to these parameters, the instrument 
calculates the stomatal conductance and intercellular carbon dioxide concentration.  
Following the quantification of gas exchange and transpiration processes, the 
instrument quantifies the fluorescence yield. 6 Seedlings were sown on March 19th , 
2019 with four seeds in each pot. For each pot, a perched water table was designed 
and used arcillite for this method. The arcillite was placed 1.5-2.0 cm from the bottom 
of the pot. The cultivars used in this study were Radish, Dragoon Lettuce, Waldman’s 
Green Lettuce, and Outredgeous Lettuce. There were 5 pots consisting of radishes, 2 
pots of dragoon lettuce, 2 pots of the Waldmann’s Green Lettuce, and 1 pot of the 
Outredgeous Lettuce. The radish and lettuce seedlings were grown in a small Percival 
reach-in chamber with the following environmental settings: 23°C and 50% Relative 
Humidity.  
On DAP 15, light response and carbon dioxide response was measured for 5 radish 
plants. Fast light response data was acquired through the following set-points: 1000, 
750,500,350, 200, 100, 50, and 0 µmol m-2 s -1. The light response curves that are 
generated measure the plant’s response to varying light intensity. The light response 
curves can provide 
information on the 
quantum efficiency and 
the light saturated 
photosynthetic rate. The fast light response is used to 
analyze how the stomata remains open at lower light 
values and how the intercellular carbon dioxide levels 
rise throughout the measurement.  In this method, the 
leaf is exposed to a high light value such as 1000 µmol 
m-2 s -1 then the light values rapidly decrease after 1-2 
minutes at each light value. 10 The slow light response 
is used to allow time for the stomata to equilibrate at 
each light value through holding the measurement for 
15minutes. This allows for the intercellular to remain 
constant which is evidence for the stomata adjusting to 
the set light level. 10 
 
The carbon dioxide response allows for an analysis 
into the intercellular carbon dioxide concentration. 
Light is kept constant throughout the carbon dioxide measurements. In starting the measurements, the carbon 
dioxide is set to a low concentration and then measured at increasing concentrations. At the low concentrations, 
Rubisco is deactivated whereas at high concentrations, s tomata will close. 10  
The fan light response allows for an analysis into the relationship between fan speed and carbon dioxide assimilation. 
Light is kept constant through the fan-light response measurements. When the fan speed is increased, the boundary 
layer decreases and this increases gas exchange. This results in a higher concentration of carbon dioxide being 
assimilated. 10 The plants were harvested on DAP 21 and the following growth parameters were observed: plant height, 
plant width, chlorophyll content, fresh weight, dry weight, stem length, hypocotyl length, and leaf area.  
Figure 2 : The leaf is clamped to allow 
for carbon dioxide, light response and 
light-fan response measurements to 
take place. 
Figure 3: Licor 6800 Portable Photosynthesis System. 
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As carbon dioxide increased, the stomatal conductance decreased as shown in Figure 5 .  When the carbon dioxide 
levels rose, the leaf temperature increased as shown in  Figure 6. In Figure 7, the carbon dioxide levels showed an 
inverse relationship with transpiration such that when carbon dioxide levels increased, transpiration decreased. At 
warmer temperatures, plants will open their stomata and release more water vapor thus resulting in higher transpiration 
rates. In regard to humid environments, the transpiration rates will decrease. When there is a higher concentration of 
carbon dioxide present, this increases the humidity and results in decreased transpiration rates. In the environment, 
plants in high carbon dioxide environments have adapted to have fewer stomata to further conserve water. 8 
  
 
Figure 4: Radish on DAP 21.  
 
 
 
 
 
 
Figure 5 : Stomatal conductance to water vapor as a function of carbon dioxide set points.   
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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Figure 6: Average leaf temperature as a function of carbon dioxide set points.  
 
 
 
 
Figure 7: Transpiration rate as a function of carbon dioxide set points.  
 
V.  Porous Concrete Media for Assessing Plant Growth 
 
Porous concrete cubes provide an environment that stimulates plant growth as air, water, and plant roots can 
navigate through the pores. 6 Porous concrete provides for a robust structure for root architecture in plants and aids in 
root development through the available pores 5 If there is a high percentage of connected pores, then moisture and 
nutrient solution can be retained and utilized as needed by plants 2 
Porous concrete cubes of 0.5m x 0.5m x 0.05m will be used to assess the growth of Outredgeous Lettuce for a 28-day 
growth period. The concrete cubes will be soaked in deionized water for 24 hours to allow for improved germination 
rates as water will be retained through the small pores present. 15 seeds will be sowed in each cube. A Hoagland’s 
nutrient solution will be added to the growing tray where the cubes will be placed. On April 4th, Barese Swiss Chard, 
Babyleaf Outredgeous Lettuce, and Shungiku were germinated on petri dishes as this allows for more control on the 
germination environment and time-course of the germination process. The results of this experiment are in progress.  
 
 
 
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Figure 8 : Outredgeous seedlings that are DAP 7 in the porous concrete media. Prior to placement on the porous 
concrete cube, the seedlings were germinated on petri dishes. 
 
 
 
Figure 9 : Prior to sowing the seeds on the porous concrete media, the seeds were germinated in petri dishes.  
 
VI. Seed Sanitization by Plasma Methods 
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Researchers at Kennedy Space Center are investigating the surface decontamination properties 
of plasma on seeds to compare to the current seed sanitization procedure. Plasma consists of 
bactericide properties that decrease microbial concentrations on surfaces as well as removing 
biofilm. Biofilm is a mechanism by which bacteria mobilize and colonize their host through the 
deposition of an EPS (extracellular polymeric substance).13 The proposed mechanism of 
inactivation of the microbial organisms through plasma is a result in damaged microbial DNA 
and quorum sensing capabilities. In damaging the quorum sensing capabilities, microbial 
organisms are unable to increase their population as the plasma inhibits this mechanism of 
communication 1 In this study, Radish Cherry Bomb II Hybrid, Mizuna Mustard, Red Romaine 
Lettuce, and Barese Swiss Chard are being investigated for surface decontamination upon plasma 
exposure. Currently, the sanitization procedure for Red Romaine Lettuce and Mizuna Mustard 
consists of using hydrochloric acid (HCl) and a bleach solution to generate an oxidizing vapor 
that inhibits microbial colonization of the seed coat. 11 Seeds are placed in a jar and 30mL of 
bleach is added to each jar in addition to the 0.5mL of concentrated HCl. The dish containing the 
seeds is placed on top of the inverted beaker and the lid is sealed. The seeds remain in the 
sealed jar for an hour to allow for the reaction between the bleach and HCl to take place. 12 
 
Upon completion of the treatment, the seeds are transferred to a laminar flow hood to 
outgas overnight. Barese Swiss Chard and Radish Cherry Bomb II Hybrid seedlings have not 
been sanitized by the bleach and HCl method.8 The objective of this study is to observe reduced 
microbial populations on the seeds treated with plasma. The seedlings were prepped in glass petri dishes prior to entry 
into the plasma systems. There were 45 seeds of each crop treated for 2 minutes in each treatment. The treatments 
consisted of an atmospheric plasma system, sub-atmospheric plasma system, and control system. Upon treatment of 
the plasma system, 10 seeds were sectioned off for germination tests. 35 seeds were analyzed for microbial content 
and to determine CFU levels on each of the seeds.  
 
 
Figure 11: Sub-atmospheric plasma system  
 
 
 
 
 
References 
 
 
 
1. Bafoil, Maxime, et al. "Effects of low temperature plasmas and plasma activated waters on Arabidopsis thaliana 
germination and growth." PloS one. 2018. 
2. Bao, Xiaohua et al. “Development of Vegetation -Pervious Concrete in Grid Beam System for Soil Slope 
Protection.” Materials (Basel, Switzerland). 2017.  
 
3. Cannon, Kevin M., et al. "Mars global simulant MGS-1: A Rocknest-based open standard for basaltic martian regolith 
simulants." Icarus .2019. 
 
4. Dias, João Silva. "Nutritional quality and health benefits of vegetables: a review."  Food and Nutrition Sciences .2012. 
Figure 10 : Radish seedlings 
in a sub-atmospheric plasma 
treatment.  
NASA KSC – Internship Final Report 
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NASA Kennedy Space Center   04/11/2019 
 
5. Hitti, Yasmeen. Bio-receptive Ground Granulated Blast-furnace Slag Porous Concrete Substrate. Diss. McGill 
University Libraries, 2018. 
6. Kim, Hwang-Hee, and Chan-Gi Park. "Plant growth and water purification of porous vegetation concrete formed of blast 
furnace slag, natural jute fiber and styrene butadiene latex."  Sustainability.2016.  
 
7. Kounaves, S., Chaniotakis,N., Chevrier, V., et al., “Identification of the perchlorate parent salts at the Phoenix Mars 
landing site and possible implications”Icarus.  2014.  
 
8. Körner, Christian. "Does global increase of CO2 alter stomatal density?." Flora.1988.  
 
9. Lau, K. Lꎬ, Y. Yꎬ Tsang, and S. W. Chiu. "Use of spent mushroom compost to bioremediate PAH-contaminated 
samples." Chemosphere.2003. 
 
 
10. LI-COR Biosciences. Using the LI-6800 Portable Photosynthesis System. LI-COR Biosciences, Nebraska, 2017.  
 
11. Lindsey III, Benson E., et al. "Standardized method for high-throughput sterilization of Arabidopsis seeds." Journal of 
visualized experiments: JoVE.2017.  
 
12. Massa GD, Newsham G, Hummerick ME, Morrow RC, Wheeler RM Plant Pillow Preparation for the Veggie Plant 
Growth System on the International Space Station. Gravitational and Space Research.2017.  
 
 
13. Ziuzina, Dana, et al. "Cold plasma inactivation of bacterial biofilms and reduction of quorum sensing regulated 
virulence factors." PLoS One.2015.  
 


Researching Plant Growth in Amended Martian Regolith Simulant, Photosynthetic Rates of Plants, Seed Surface Decontamination by Plasma Methods, New Crop Development, and Porous Concrete Media

Researching Plant Growth in Amended Martian Regolith Simulant, Photosynthetic Rates of Plants, Seed Surface Decontamination by Plasma Methods, New Crop Development, and Porous Concrete Media

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