1. Environmental control to increase the nutritional value of vegetables in BLSS Walter Stefanoni 1, Simona Proietti 1, Stefano Moscatello 1, Guglielmo Santi 1, Giuseppe Colla 2, Alberto Battistelli 1 1 CNR Istituto di Biologia Agroambientale e Forestale, via Guglielmo Marconi, 05010 Porano (TR) Italy. Tel. +39 0763- 374910. Email: alberto.battistelli@ibaf.cnr.it 2 Dipartimento di scienze e tecnologie per l'Agricoltura, le Foreste, la Natura e l'Energia, Università della Tuscia, via San Camillo De Lellis snc, 01100 Viterbo (VT) Italy. Tel: +39 0761-357536. Part of this work was funded by the Italian Ministry of Foreign Affairs under the Italy -USA scientific and technological cooperation agreement. Project: Sviluppo di una serra gonfiabile per la crescita delle piante, la produzione di cibo e il supporto alla vita nello spazio. ID 00071 Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

2. Gas exchange Photosynthesis Mineral nutrition Water relations Secondary metabolism (Estimated >200000 compounds defense and interaction with the environment) Flavonoids Phenolic acids Lignans Carotenoids Tocopherols and tocotrienols Quinones Sterols Alkaloids Glucosinolates Primary metabolism (Photosynthesis. respiration, growth ….) Synthesis of carbohydrates Synthesis of amino acids Synthesis of organic acids Synthesis of fatty acids Mineral ions Source tissues Sink tissues Fruit and vegetables contain most if not all essential components of human nutrition They can also contain metabolites that can positively or negatively affect our nutritional and health status. Metabolism Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

3. Carbohydrates Β-carotene Proteins Ascorbic acid Lycopene Anthocyanins Fiber-fructans Polyphenols Tocopherols Polyalcohols Workshop on Bioregenerative Life Support Turin, 18-19 May 2015 Oxalic acidNitrate

4. Plant metabolism is strongly dependent on environmental factors (optimal and stress conditions), including those related to cultivation techniques a.Light (quantity, quality and photoperiod) b.Temperature (all enzymatic processes, water relations and gas exchange, cell cycle and respiration) c.Atmosphere composition (CO 2, O 2, H 2 O, O 3, VOCs, pollutants) d.Composition of the growing medium (soil, water, mineral nutrients, osmotic potential, pollutants) e.Biotic environment (other organisms in the same environment, other cultivated species, symbiotic microorganisms, pathogens) The aim of our work is to study how environmental factors that we have to control in BLSS, can be used to increase the positive characteristics of plant food while decreasing the negative ones. Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

5. At the lower light intensity the dry and fresh weight and the leaf area were reduced. Leaves were thinner under low light as showed by the specific leaf dry weight (SLDW) and they showed a higher shoot/root ratio. Low light High light 200 ± 50 PFD 800 ± 50 PFD LIGHT Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

6. High light increased carbohydrate content of spinach leaves. Low light High light 200 ± 50 PFD 800 ± 50 PFD LIGHT

7. At the lower temperature dry weight and specific leaf dry weight (SLDW) increased, while the chlorophyll content is higher Carbohydrate content increased in spinach leaves growth under low temperature. 10 °C 25 °C TEMPERATURE Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

8. What about positive VS negative metabolites composition in relation to the environment ?

9. High light increased the ascorbate content while decreasing the content of oxalic acid and nitrate ion. Workshop on Bioregenerative Life Support Turin, 18-19 May 2015 Low light High light 200 ± 50 PFD 800 ± 50 PFD LIGHT

10. Workshop on Bioregenerative Life Support Turin, 18-19 May 2015 10 °C 25 °C TEMPERATURE Acclimation of spinach plants to low temperature (5 days) caused an increase of the ascorbic acid content but no changes in the oxalic acid content.

11. What about the interactive effects of different environmental parameters on plant growth and food quality? Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

12. Interaction of light intensity and CO 2 partial pressure on spinach No interaction Interaction Additive effects on productivity (strong by both light and CO 2 ) Interaction only on chlorophyll. Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

13. Additive effects on ascorbic acid (stronger by CO 2 ) Workshop on Bioregenerative Life Support Turin, 18-19 May 2015 Interaction of light intensity and CO 2 partial pressure on spinach

14. Workshop on Bioregenerative Life Support Turin, 18-19 May 2015 Light intensity and CO 2 partial pressure in spinach No interaction between light intensity and CO 2 partial pressure on the leaf content of carbohydrates

15. Workshop on Bioregenerative Life Support Turin, 18-19 May 2015 Interactive effects on the contentof Nitrate and oxalic acid

16. Current activities Investigation of environmental control of growth conditions on valuable metabolites accumulation Light quality Metabolites with nutraceutical value (vitamins, antioxydants, fibres…..) Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

17. CONCLUSIONS The control of growing environmental parameter can be used for the modulation of the desired food outputs from a fully controlled plant growing unit, not only in quantitative but also in qualitative AND PARTICULARLY NUTRITIONAL terms. This is also true for the control of growing condition on hearth. Positive and negative nutritional traits must be considered, as shown here for the effect of light intensity temperature, CO 2 partial pressure and the interaction among light and CO 2 partial pressure, on the content of SUGARS, ASCORBIC ACID, NITRATE and OXALIC ACID. Environmental control has a very high potential for future application in greenhouse agriculture, functional food production, nutraceuticals and plant derived drugs production. Workshop on Bioregenerative Life Support Turin, 18-19 May 2015

18. …to be continued in EDEN –ISS…… EDEN ISS Project Reference: 636501 Funded under: H2020-COMPET-2014 - Competitiveness of the European Space Sector: Technology and Science Title: Ground Demonstration of Plant Cultivation Technologies and Operation in Space for Safe Food Production on-board ISS and Future Human Space Exploration Vehicles and Planetary Outposts