GMO plum trees are being evaluated at the Kennedy Space Center as a possible candidate for future space crops. Previously conducted horticultural testing compared the performance of several plum genotypes in controlled environment chambers, resulting in a down-selection to the NASA-11 genotype. Precursory studies determined the water use requirements to sustain the plants as well as the feasibility of grafting non-GMO plum scions onto GMO plum rootstocks of NASA-5, NASA-10, and NASA-11 genotypes. This study follows the growth and horticultural progress of plum trees and in-vitro cultures from August 2017 to November 2017, and provides supplemental support for future GMO plum studies. The presence of Hurricane Irma in early September 2017 resulted in the plants undergoing material deterioration from major changes to their overall horticultural progress

Mercado, Dianne

NASA Technical Reports Server

NASA KSC – Internship Final Report 
 
NASA KSC  1 17/11/2017 
 
 
 
 
 
 
 
 
 
 
 
Analysis of GMO Plum Plant Culture in System Operations 
Failure 
Dianne Mercado 
NASA Kennedy Space Center 
Major: Biotechnology 
NIFS Fall Session 
17/11/2017 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
https://ntrs.nasa.gov/search.jsp?R=20170011601 2019-08-31T01:20:08+00:00Z
NASA KSC – Internship Final Report 
 
NASA KSC  2 17/11/2017 
 
Analysis of GMO Plum Plant Culture in System Operations 
Failure 
 
Dianne Mercado1 and Lashelle Spencer2 
Kennedy Space Center, FL 32899 
 
GMO plum trees are being evaluated at the Kennedy Space Center as a possible candidate for 
future space crops. Previously conducted horticultural testing compared the performance of 
several plum genotypes in controlled environment chambers, resulting in a down-selection to 
the NASA-11 genotype. Precursory studies determined the water use requirements to sustain 
the plants as well as the feasibility of grafting non-GMO plum scions onto GMO plum 
rootstocks of NASA-5, NASA-10, and NASA-11 genotypes. This study follows the growth and 
horticultural progress of plum trees and in-vitro cultures from August 2017 to November 2017, 
and provides supplemental support for future GMO plum studies. The presence of Hurricane 
Irma in early September 2017 resulted in the plants undergoing material deterioration from 
major changes to their overall horticultural progress.  
 
Nomenclature 
BA =   6-Benzyladenine 
EtOH =   ethanol 
GMO =   genetically modified organism 
IBA =   indole 3-butyric acid    
KSC =   Kennedy Space Center 
L = liters 
NASA =   National Aeronautics and Space Administration 
NIFS =   NASA Interns, Fellows, and Scholars 
PtFT1 =   Flowering locus T1 
SGM =   standard growth media 
SRM =   standard rooting media 
USDA =   United States Department of Agriculture 
 
I. Introduction 
lum trees (Prunus domestica) genetically modified to overexpress the FT flowering gene are being evaluated 
as a candidate for future space crop selection. This overexpression removes seasonal dormancy requirements 
and allows the dwarf plants to mature and produce fruit from a young age (12 to 14 months), and exhibit compact 
continuous growth from seed to fruit. Prior studies have linked phytonutrients found in plums to prevent bone 
calcium loss in both animal and human models3,4. Successful fruit trees present a potentially sustainable and high-
impact addition to space food systems, especially those of long-term deep-space missions, in a two-fold form: 
providing a source of fresh food and different produce variety which contributes to stable psychological well-being, 
and enhancing nutritional benefits that help counteract one of the largest health-related obstacles of spaceflight 
missions3. Plum trees and in-vitro plum tissue cultures of the NASA-5, NASA-6, NASA-10, NASA-11, and 
Control-5 genotypes are currently growing and monitored at KSC under controlled environments. 
                                                          
1 NIFS Intern, Engineering Services Contract, Kennedy Space Center, University of Central Florida. 
2 Scientist and NIFS internship mentor, Engineering Services Contract, Kennedy Space Center. 
I. P 
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II. Methods and Materials 
A. Plum Trees  
Modified and unmodified varieties of plum trees (P. domestica, “Blue Byrd” cultivar) were kept and 
monitored in a controlled environment chamber at the Operations and Check Out (O&C) Building at KSC for the 
duration of the study. In addition to the plums, two tomato plants (Capsicum annum, “Red Robin” cultivar) were 
grown as well to provide a source of comparison to the plums. 
Grown specimens of NASA-5, NASA-6, NASA-10, NASA-11 (the down-selected line), and Control-5 were 
maintained in the chambers under a 16hr:8hr light:dark lighting schedule at temperatures of approximately 22 degrees 
Celsius and approximately 50% relative humidity. The plants were fertigated 6 times daily with Peter’s granulated 
nutrient mixture solution with an electric conductivity of 1200-1250 μL and pH of 6.00-7.00. An automatic pump 
irrigation system was developed and used to deliver nutrient solution to plants. 
 
 
  
  
 
 Daily checks twice a day, one in the morning (timestamps 
ranging from 10:00am-11:00am) and the other in the afternoon 
(timestamps ranging from 1:00pm-2:00pm), were carried out to 
observe and maintain horticultural progress of the plants, with 
the exception of the duration of Hurricane Irma in September 
2017. 
 
B. Plum Tissue Cultures 
 Fresh plant material from the four selected GMO plum lines 
(excluding NASA-6) and an unmodified plum line called 
Control-5 were transferred aseptically into in-vitro Murashige 
and Skoog-based IBA/BA SGM and had been exhibiting leafy 
plantlet and callus growth prior to the start of this study. 6-8 node 
bud tissues for each genotype were transferred into Magenta 
Plant Culture boxes with IBA/BA SGM and monitored daily 
under ambient lab temperature (~23°C) and 50 µmol of light. 
 
   
 
 
Figure 1. Controlled environment 
chambers for plum plants.  
Figure 2. Automatic pump irrigation system for nutrient 
solution delivery. Main pumps pulled up intake from manually 
filled reservoir and auxiliary tubes were attached to growing pots 
of plums. 
Figure 3. Magenta boxes containing 
transferred plant material in IBA/BA 
SGM. Continuous lightning from AIBC LED 
Grow Light fixtures and ambient environment 
temperature were maintained throughout this 
study.  
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III. Results and Discussion 
A. Plum Plants 
 Specimens of NASA-5, NASA-6, NASA-10, the down-
selected line NASA-11, and Control-5 were kept and maintained in 
controlled environment chambers at KSC under the specified 
parameters. Prior to Hurricane Irma in early September 2017, all 
genotypes exhibit healthy nodal tissues, production of chlorophyll-
rich leaves, and growth of secondary and tertiary branches from the 
main plant. Plants were watered manually while the automatic 
irrigation system was under development.  
 Before Hurricane Irma reached main-land Florida and 
initiated statewide environmental emergency conditions, immediate 
steps were taken to increase the optimal viability of the plants 
through the hurricane. Due to the automatic irrigation system 
undergoing technical complications, as well as facing the possibility 
of losing access to power and water for up to several weeks, the 
potted plants were placed in growth trays and submerged in 5 L of 
nutrient solution for sub-irrigation. The chambers were set to be 
active throughout the hurricane.  
 After Hurricane Irma, assessments were carried out to 
analyze the health of the plants. The chambers had lost power during 
the hurricane and increased the humidity of the growing 
environment, resulting in mold and fungal growth on all the plants. 
In addition, it is estimated that the plants drained their nutrient 
solution three to four days into the hurricane duration and two to 
three days prior to any possible initial assessments being made. In 
the days following Hurricane Irma, the combination of 
dehydration and fungal growth resulted in all genotypes losing 
leaves and developing infected tissue.  
  
 
Figure 4. Magenta box containing three plant 
material samples from genotype NASA-10 in 
SGM. While node tissues were initially used for 
tissue culture study, calluses (example: sample to 
the right of the box) were also separated from the 
main tissue and grown in the same conditions.  
Figure 5. Tissue cultures growing under 24/7 
LED light in ambient room temperature. 
Cultures were separated on trays by genotype, 
clonal propagation date, and transfer date based on 
aseptic transfers into new media every 4 to 6 weeks.  
Figure 6. NASA-11 and Control-5 three days 
after Hurricane Irma. While automatic 
irrigation system was established soon after the 
hurricane, the plants still suffered from fungal 
growth which reached the xylem and phloem of 
the inner stem and resulted in plant 
deterioration.  
 
 
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 Out of all genotypes, NASA-6 remained the most 
unaffected by the effects of Hurricane Irma, maintaining 
70% of its healthy leaves and nodes. This is most likely due 
to its heavily modified genetic growth and may be a point of 
interest to explore in the future. NASA-5 also showed less 
plant deterioration than NASA-10, NASA-11, and Control-
5, with one plant maintaining 60% of its healthy leaves and 
nodes. Interestingly, the NASA-5 plants were observed to 
recover faster from the hurricane before the other plum lines, 
with two NASA-5 plants exhibiting healthy nodes and leaf 
growth approximately one and a half week after the 
hurricane. As of the writing of this paper, NASA-10 and 
NASA-11, the down-selected genotype, have not yet fully 
recovered. 
  
B. Plum Tissue Cultures 
 The plum tissue cultures did not undergo any adverse 
effect from Hurricane Irma. However, due to the plants in 
the chambers suffering from fungal growth and dehydration 
which led to wilting and further deterioration, there was a 
need to preserve and maintain all tissue cultures for future 
GMO plum studies. Cultures both node-based and callus-
based that had outgrown their boxes were cut in half (or in 
some cases of particularly large growths, three times) and 
aseptically allocated to additional boxes with SGM. The 
number of samples were doubled within two to three weeks.  
 In addition, healthy node tissues were extracted from 
the NASA-6 plants and processed into petri dishes with 
SGM for comparison to the other genotypes due to its 
horticultural performance despite the contextual effects of 
Hurricane Irma. Stem nodes containing buds were put 
through a tissue sterilization process: soaking in 70% EtOH 
for 30 seconds followed by 10% bleach for 10 minutes. 
Node tissues were rinsed in sterile nanopure water between 
treatments to decrease oxidative tissue damage as well as 
chances of microbial contamination, then aseptically 
transferred to petri dishes with IBA/BA SGM. 
Approximately 20% of all individual NASA-6 tissue 
cultures developed microbial contamination, most likely 
due to the fungal infection remaining in the plant tissue even 
after sterilization. 
 Since Hurricane Irma, all tissue cultures have been 
growing and monitored daily in IBA/BA. At the end of 
October 2017, 2 to 3 tissue cultures of all genotypes 
(excluding NASA-6) were processed into SRM. As of 
November 6, these cultures remained healthy. 
 
 
 
 
 
Figure 7. NASA-6 plant three days after 
Hurricane Irma. Despite fungal growth and lack 
of sufficient water supply, NASA-6 remained the 
healthiest of all tested genotypes and has to date 
produced flowers. 
 
Figure 8. NASA-6 plant tissue cultures in petri 
dish wrapped with parafilm to maintain 
moisture content. Daily observations indicated 
fast (one to two week) nodal growth and 
development of initial leafy tissue. 
 
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IV. Conclusion 
 Studies have shown GMO dwarf plums to be a potentially high-yield space crop candidate in terms of 
biomass production and nutrition content. This study points out the protocols used to maintain plum plants in 
controlled environments, as well as those used to preserve and propagate in-vitro tissue cultures, which require less 
space, materials, and labor to maintain and may be applied to a space food production setting. Developing a more 
precise and efficient start-to-finish protocol for the in-vitro process (taking healthy plant material from the main plant 
to processing healthy nodal tissues into SRM to develop roots) may help in furthering GMO plum studies. 
Additionally, there is room for future research on the long-term viability of not only GMO plums but other space crop 
candidates on the occasion that controlled system operations fail to operate. While Hurricane Irma resulted in major 
plant material loss and deterioration, it indicates a need for research regarding plant life viability and how that 
transitions to long-term deep-space food production systems. 
Acknowledgments 
 The author would like to thank Lashelle Spencer for her mentorship and unfailing guidance throughout this 
project. Additional thanks go to Mary Hummerick, Lawrence Koss, Dr. Christina Khodadad, and the members of the 
Kennedy Space Center Veggie team for their assistance and input. The author would also like to thank faculty and 
academic mentors at the University of Central Florida for their professional support and preparation for this 
opportunity. Funding for this internship was provided by the NIFS program. 
References 
 3 Spencer, L., Graham, T., Stutte, G., Massa, G., Mickens, M., and Raymond W., “Space Pomology: Dwarf Plums for 
Fresh Food Production,” Oral/Visual Presentation, 2017. 
 4 Globus, R. “Dried Plum Protects from Radiation-Induced Bone Loss by Attenuating Pro-Osteoclatic and Oxidative 
Stress Responses.,” Conference Paper, 2015. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Analysis of GMO Plum Plant Culture in System Operations Failure

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