6th INTERNATIONAL AGROSPACE
WORKSHOP
Sperlonga, 22-23 May 2014
The growth of higher plants in space
greenhouses: cultivation strategies to minimise
plant sensitivity to space stressors and maximise
resource use efficiency
De Pascale S., Paradiso R., De Micco V., Aronne G.
University of Naples Federico II
Department of Agricultural and Food Sciences
Context
The sustainability of long duration
manned missions in space relies on
plant-based Bioregenerative Life
Support Systems (BLSSs).
Resources
Psychology
Multiple stressors
Plants on Earth
Plants in Space
Temperature
Humidity
Altered gravity
Light
Radiations
Space factors may modify plant biology and availability of
resources for plant growth
Interaction between factors
Whenever acting at suboptimal levels, such factors may operate
synergistically to low-gravity and radiation, thus enhancing the effects of
space factors.
T = 18 °C
1g
T = 21 °C and 24 °C
Cl
1g
Cl
(De Micco et al. 2006. Acta Astronautica 58: 139-148)
The current research efforts in agrotechnology aim to realise space
greenhouses where environmental factors can be precisely modulated
and optimised in order to maximise plant growth efficiency and reduce
plant sensitivity to space stressors.
Ground-based Projects
Past projects
Effect of simulated microgravity on plant growth
Project
Sprouts production
Phaseolus vulgaris
Glycine max
Project
Species selection
Gametophyte of
various crops
Project
Agro-Technology
SpaceGreenHouse
Space Projects
Effect of real microgravity on plant growth
Project
Sprouts production
Glycine max
Plant adaptation on Earth and in Space
!
1. Identification of Critical Phases
2. Identification of tresholds per each CP
3. Evaluation of the effects on crop productivity
4. Definition of countermeasures (agro-technologies)
!
Analysed critical stages
1) Seedling development in microgravity
1g
Organogenesis and biometry
Development of the vascular system
Cell wall formation and lignification
Nutritional value as fresh food (starch and phenolics)
Early stress signaling
Space
Project
Sprouts production
Phaseolus vulgaris
Glycine max
Project
Sprouts production
Glycine max
2) Pollen germination and tube development
1g
Space
Evaluation of the reproductive success
Application of the gametophytic selection
Project
Species selection
Various crops
Clinostat rotation
Microgravity: FOTON M2
Simulated microgravity affects the development of pollen tubes: this
can constrain plant reproduction
Tomato
Eggplant
Broad bean
Broccoli
Apple
Apricot
Cherry
Plum
Almond
Peach
Pear
Perturbations were dependent on the species
• De Micco et al. 2006. Protoplasma 228: 121-126
• De Micco et al 2006. Acta Astronautica 58: 464-470
Morpho-functional response
of soysprouts to microgravity
1g-seedlings
Space-seedlings
The process of proper
cell-wall building
(i.e. cellulose deposition),
although not prevented, is
perturbed at the early
stage of development
of xylem.
Several growth
phenomena are slowed
in microgravity
De Micco V, Aronne G, Joseleau JP, Ruel K. 2008. Xylem development and cell wall changes of soybean
seedlings grown in Space. Annals of Botany. 101: 661-669
Ground-based Projects
Past projects
MELISSA
Micro-Ecological Life Support System Alternative
Food Characterisation Phase 1
Aims of MELiSSA FCP1
I. to develop an objective theoretical procedure for a
preliminary identification of four candidate cultivars of
soybean for seed production
II. to design and realize an hydroponic system for the
optimization of soybean cultivation
III. to evaluate the plant behaviour of the four selected
cultivars in hydroponic system through cultivation trials
Why soybean?
Nutritional value of seeds
Hoff et al., 1982
NASA Contract Report 166324
Osteoporosis risk
Khalil et al., 2002
J. Nutr. 132: 2605-2608
Peroxidation products in brain
Chowdhury and Soulsby, 2002
Ann. Cl. Lab. Sci. 32(2):188-192
Proteolysis of myofibrillar proteins
Tada and Yokogoshi, 2002
J. Nutr. Sci. Vit. 48(2): 115-119
Main issues:
Processing
Hydroponics
(Cultivar selection)
Cultivar selection
Aim
 to develop an objective and repeatable procedure to select the best European
cultivars (cvs) for cultivation in BLSSs. This purpose was pursued with a two-steps
approach:
a) the development of a theoretical procedure for a preliminary identification of cvs
b) the evaluation of cvs behaviour in hydroponics, in controlled environment
Results
 A preliminary screening, based on a scientific and technical literature survey and the
criteria suggested by ESA, allowed to select 93 cvs among the 297 admitted in EU.
 An algorithm, based on the relevance of the considered features, was created to
attribute a score to each cv: the first 4 cvs were selected for subsequent cultivation
trials
PR91m10 – Regir – Atlantic - Cresir
De Micco et al., 2012 - Adv. in Space Res. 49: 1415-21
Criteria for the
cultivar selection
Cultivar selection: validation of
theoretical procedure with
experimental data
Xi
Algorithm
S1
Relevance of criteria
for Space cultivation
Pi
Preliminary
list of
cultivars
and data
from
available
resources
4 candidate
cultivars
S2
Experimental data
per each cultivar
and criterion
Cultivation trials
Aim
Algorithm
re-running
S3
Candidate cultivars:
final score
On the 4 candidate cvs:
 to validate the algorithm with data from hydroponic cultivation in controlled conditions
 to compare the experimental scores with the theoretical scores from data in open-field
(literature)
Cultivar
Results
 the relative ranking was the same in
the two cases
 the 4 cvs reached higher scores in
hydroponics than in soil in open-field
Theoretical
Score
Experimental
Score
PR91m10
30.75
34.92
Regir
23.42
33.22
Atlantic
23.05
32.62
Cresir
22.37
31.5
Agronomical and nutritional characterization of soybean for BLSS:
lessons learned from the MELiSSA project – FC I
De Micco V., Paradiso R., Aronne G., Fogliano V., De Pascale S.
Proceedings 63rd International Astronautical Congress (IAF), Naples, Italy, 1-5 October 2012, 7 pp.
Closed Hydroponic Systems
WUE = Y/W
Evapotranspiration (E)
Growth
Water
use (W)
Water
source
W  I Irrigation (I)
Yield (Y)
Mixing
tank
WU
Soil/substrate
Recycling
Leaching (drainage) (L)
• Soybean (field crop,
Rhizobium?)
• Salinity build up due
Advantages:
to the accumulation of
• Faster plant growth rate
non-essential salts
• Higher yield and quality
• Root pathogens
• Higher water and nutrient use efficiency
• Phytoxicity
• Automatization
Leaching fraction = L/W
Runoff

 Drawbacks of water
recycling:
Hydroponic cultivation
Aim
 To evaluate the plant performance of the 4 selected cvs in
terms of seed germination, growth and yield in an on-purpose
designed NFT system, in controlled environment
Results
 the NFT system and the nutrient solution management were
efficient in growing healthy soybean plants
 the 4 cvs chosen in the theoretical selection phase showed
good performances in hydroponics
 ‘Pr91m10’ was the best cv, coupling short size and high
resource use efficiency to good yield and quality of seeds
University of
Naples growth
chamber
Nitrogen nutrition
and root bacterial symbiosis
in cooperation with
School of Environmental Sciences
(University of Guelph)
Soybean cultivation at the
University of Guelph
Aim
 to evaluate the effect of urea, as alternative N-source to
nitrate, and of root inoculation with Bradirhizobium
japonicum on seeds yield and quality in NFT
Results
 Urea as sole N-source reduced plant growth and seed yield
compared to nitrate
 Root inoculation with B. japonicum did not influence plant
performance, regardless of the N-source
 The positive effect of urea on symbiosis did not improve the
performance of inoculated plants
Nitrogen nutrition
and hydroponic system
in cooperation with
School of Environmental Sciences
(University of Guelph)
Aim
 to evaluate the effects of 2 hydroponic systems, NFT
and cultivation on rockwool, and of urea as alternative
N-source to nitrate, on root nodulation and seeds yield
and quality of plants inoculated with B. japonicum.
Results
 cultivation on rockwool positively influenced root
nodulation and plant growth and yield, without
affecting the composition of seeds, compared to NFT
 urea improved root symbiosis but it reduced plant
growth and yield while affected positively the protein
content of seeds, compared to nitrate.
Soybean cultivation at the
University of Guelph
Seed-to-Food
Hydroponic cultivation improves the nutritional quality of
soybean and its products…
Seed nutritional quality
Aim
 To determine the nutritional composition (protein, fat, dietary
fiber, phytic acid and isoflavones) of seeds, soy milk and okara,
in the 4 cvs grown hydroponically
 To compare the nutrient quality of these products with those
obtained from the same cvs cultivated in soil in open field
Results
 ‘Pr91m10’ showed the highest quality of seeds ( protein content)
 Hydroponic cultivation improved nutritional quality (fats and dietary fiber content) in
seeds and okara, while did not influence soy milk composition compared to soil.
Okara (Soy Pulp)
Ongoing Projects
FARO Project: Effect of ionizing radiation on tomato growth:
food countermeasures to support life in Space
Ground-based Projects FARO Project: Effect of ionizing radiation on tomato
growth: food countermeasures to support life in Space
Analyse the responses of tomato microtom plants to
increasing doses of X-rays
Plant growth
Stem elongation , internodes,
leaf area, leaf anatomy
Photosynthesis
Photochemistry, chlorophylls,
Rubisco expression and activity
Oxidative stress
and DNA damage
PARP activity, antioxidants
The effects of X-rays on tomato growth and metabolism depend on the
phenological stage at the time of irradiation
PHENOLOGICAL
PHASE AT THE
TIME OF
IRRADIATION
Fruits are
produced
Dry seed
Vegetative
phase
During
flowering
At high doses
plant growth is
blocked.
Possible recovery
At high doses
plant growth is
blocked.
Recovery not
possible
De Micco et al 2014. Scientific World Journal http://dx.doi.org/10.1155/2014/428141
Ongoing Projects
MELISSA
Micro-Ecological Life Support System Alternative
Food Characterisation Phase 2
MELiSSA FC2
CulSel Project official start on
the 14th of April 2013 (Kick off Meeting)
Duration 20 months
Participant Country Capacity
UGent
BE
Prime Contractor, in charge of the overall
management and covering tasks regarding to growth
parameter monitoring of potato cultivation, and
microbial rhizosphere analysis
UGuelph
CND Subcontractor, concentrating on durum wheat
cultivation and processing by the Cereal Research
Centre - Agriculture and Agri-Food (Canada)
UNapoli
I
Subcontractor, responsible for soybean cultivation
and processing
UBern
CH
Subcontractor, responsible for bread wheat
cultivation and processing
IPL
BE
Subcontractor, responsible for nutritional requirement
establishment, coordinates the nutrient-related
activities for all crop growth and processing activities
Scientist in charge
Danny Geelen
Mike Dixon
Stefania De Pascale
Urs Feller
Serge Pieters
UNapoli Research Activity
Potential beneficial interactions
between soybean plants and microorganisms
General Aims
 to analyse potential beneficial interaction between selected soybean cvs and specific
groups of microorganisms
 to evaluate the impact of microorganisms on plant growth, seed yield and nutritional
quality.
Specific Aims
 Literature survey for the interaction between soybean and symbiotic
microorganisms
 Set-up of the NFT system for cultivation of soybean in interaction with
microorganisms
 Set-up of the inoculation
 Bench tests to evaluate the interaction between cultivars and microorganisms
strains in comparison with cultivation without microorganisms
 Characterization of plants with specific focus on root/microorganism interaction
GROWTH CHAMBER &
ENVIRONMENTAL CONTROL
Integrated HPS – LEDs system
Wheeler et al., 1991. Soybean stem growth under high pressure
sodium with supplemental blue lighting. Agronomy J., 83, 903-906
NFT recirculating system
Modified Hoagland & Arnon ½ strength
(Wheeler et al., 2008)
N
K
P
Ca
Mg
S
Fe
Mn
Zn
Cu
B
Mo
mM
7.5
3.0
0.25 - 0.5
2.5
1.0
1.0
M
60.0
7.40
0.96
1.04
7.13
0.01
Fertigation management
EC target 1.2 dS m-1
pH target 5.8
Optimized cultivation protocol
- ↑ PPFD: 350 to 500 mmol m-2s-1
- + LEDs
- ↓ EC: 2.0 to 1.2 dS m-1
- ↓ P: 0.50 to 0.25 mM
•
Recirculating solution
controlled manually and
adjusted every day:
+ deionized water
and/or fresh solution
+ nitric acid
Seed inoculation protocol
SOYBEAN STERILIZED SEEDS
Myco Madness mix preparation
Cell counting (5x105 cells/ml)
Seeds inoculation (12 h)
Washing in sterilized distilled water
Sowing
(22 °C, darkness, 8 days)
(Bashan, 1986)
Projects in progress
EPO – HIP
EPO – Higher Plants
The actions proposed by this education project are:
1) Promotion of healthy eating on Earth and in Space
2) Experience seed germination in simulated microgravity and in 1-g
High School students will follow a cycle of lessons on the Space issues and
will perform an easy-experiment of plant growth in simulated Space
conditions
Projects in progress
EPO – HIP
EPO – Higher Plants
Experiment in the schools : growing sprouts under simulated space
condition with the support of Italian Universities and comparison with
sprouts produced in 1g
University of Naples
University of Milan
University of Sassari
Acknowledgements
This work is dedicated to the memory of Claude Chipaux
(1935-2010), father of the MELiSSA project.
Funding was provided by the European Space Agency and by
The Italian Space Agency
Carmen Arena
Roberta Buonomo
Vincenzo Fogliano
Maria Giordano
Maria Antonella Palermo