Food Security and Nu00on
The 19th Triennial Conference Brussels, 6 – 11 July 2014
Exploring biodiversity to introduce nutri0onal quality criteria in potato breeding program Piñeros C., Peña C.B., Del Castillo S., Rodríguez E., Restrepo P., Kubow S., Sabally, K., Kushalappa A., Mosquera-Vásquez T.
Project undertaken with the financial support of the Interna7onal Development Research Centre (IDRC), www.idrc.ca, and the Government of Canada, provided through the Department of Foreign Affairs, Trade and Development Canada (DFATD), Canada, www.interna7onal.gc.ca
Content 1. 2. 3. 4.
Introduction Objective General methodology Results 4.1. Nutritional status of the population 4.2. Proximal analysis 4.3. Functional food analysis 4.4. Potato breeding program 5. Conclusions and perspectives
1. Introduction
Where are we working?
Working with Andean communities
Pasto
Why are we working in nutritional quality? v Nariño is the second undernourished province in Colombia. v High percentage of food insecurity. v 21.5% of the population younger than five years suffers malnutrition. v There is not accurate data to design public policies about food security and nutrition. v Breeding programs have not considered nutritional quality criteria for selecting new cultivars. v Nariño is a potato biodiversity center.
Food security is built on three pillars We have worked on:
1. Food use: appropriate use based on knowledge of basic nutrition and care, as well as adequate water and sanitation.
v Nutritional status of the population. v Analysis of potato intake.
2. Food availability: sufficient quantities of food available on a consistent basis.
v Analysis of nutritional quality for potato tubers. v Producing tuber seeds of good quality for potato production.
3. Food access: having sufficient resources to obtain appropriate foods for a nutritious diet.
v Delivering new potato cultivars, with high yield and reducing costs.
2. Objective
Improve food security in indigenous communities by selection of potato cultivars with high yield and nutritional qualities to improve their daily diet.
Photo: Ernesto Rodríguez
3. General methodology Social study Nutritional analysis for population
Advanced technologies Nutritional quality of potatoes
Food consumption pattern
Biochemical analysis
Nutritional status of population
• Micronutrientes • Macronutrients • Polyphenols
Functional analysis Potato breeding program New potato cultivars with better nutritional quality
• Metabolomics: metabolites • Biotransformation and bioaccessibility
4. Results
4.1. Nutritional status of the population Percentage of boys and girls under nutritional standards
Nutritional situation by anthropometry Stunting Overweight Obesity Underweight
Boys
Girls
41.6 23.4 18.0 3.1
30.8 44.4 7.0 1.8
Percentage of the population with deficient intake Age groups Under 5 From 5 to 12 years Adults Total
Calories
Proteins
56.6 73.8 88 72.9
15.8 33.2 85.1 41.9
Fe 21.8 26.1 42.7 29.0
Minerals Zn 81.6 91.0 99.8 91.1
Ca 50.8 92.1 91.9 82.7
Vitamins A C 45.7 5.9 72.5 17.3 87.9 42.7 70.4 20.8
Samples preparation for the analysis Planting and harvesting
1 Grind and pulverization
6
Cooking according to the tuber size
Clasification and washing
3
2
Cutting in slides
Lyophilization
4 5
4.2. Proximal analysis: Macronutrients content in Solanum phureja genotypes 25"
Clara%Peña% 20"
CONTENT"mg/100g"
Phureja collec7on Phu$collec)on$ Advanced clones Advanced$ clones$ Commercial Commercial$ clones$ cul7vars
15"
10"
5"
0" Dry"Ma+er""
Protein"
Ash"
Fat"
4.2. Proximal analysis micronutrients: mineral content in Solanum phureja genotypes 25
a
Content mg/kg DW
20
a
b
Phureja collec7on Advanced clones Advanced clones Commercial Commercial clones cul7vars
15
Phu collec7on
a
10
a
5
0
Fe
Mn
Cu
Zn
4.3. Phenolic compounds in Solanum tuberosum group Phureja Clara Piñeros
Chromatograms at 320 nm of two potato extracts ChA
Cripto-‐ChA Neo-‐ChA
CaA
Chlorogenic acid (ChA) Crypto-‐ChA Neo-‐ChA Caffeic acid (CaA)
Col 23 Col 112
Dionex Ultimate 3000 UHPLC system (Thermo Scientific corp) coupled to diode array detector
Chlorogenic acid content in Solanum tuberosum group Phureja
Genotypes Â
4.3. Functional food analysis Materials and methods Metabolite extraction & LC-MS analysis
ü Lyophilized potato flesh ü Eight advanced clones
ground in liquid N
Liyao-‐Ji Kushalappa A.
60% methanol, 0.1% formic acid
Supernatant filtered and injected
LC-ESI-LTQ Orbitrap, Thermo Fisher, Waltham, MA, USA High resolution hybrid mass spectrometer system
Functional food analysis Materials and methods
v
Identification and classification of metabolites ü Accurate mass error <5 ppm ü MS/MS fragmentation pattern ü Classified based on their chemical groups
v Statistical analysis ü ANOVA will be done to identify significant metabolites.
v Identification of functional foods ü Metabolites were searched for functional foods properties in the literature.
Preliminary results v 302 metabolites were detected in cooked potatoes in eight genotypes. v 99 were found to have beneficial roles in human health: ü Anti-cancer ü Anti-HIV ü Anti-hypertension ü Anti-inflammatory ü Anti-malarial ü Antimicrobial ü Anti-diabetic
Metabolites chemical groups Important chemical groups Glycerolipids 1% Pesticides 1% Sulfur Acids 2%
Terpenoids 11%
Sterol Lipids 1% Prenol Lipids 3%
Alkaloids Amino acid 6% 4%
Polyketides 5%
Carbohydrates 3% Carboxylic Acids 2% Fatty acids and fatty acyls 20%
Phenylpropanoids 4% Nucleic acids 4% Hydrocarbons 4% Heterocyclic Compounds 9%
Glycerophospholipids 8%
Flavonoids 10%
Relative intensity of some compounds in S. tuberosum group Phureja Relative intensity (* 100,000)
Phenylpropanoids: chlorogenate
Health benefits: antioxiant, anti-diabetic and anti-lipidemic effects. (Bouayed et al., 2007; Shafi and Tabassum, 2013)
Relative intensity (* 100,000)
Flavonoids: Dihydrokaempferol 1. 4 1. 2 1
Health benefits: antimicrobial
0. 8 0. 6
(Malterud et al., 1985)
0. 4 0. 2 0
CA04 CA09 CA50 CA51 CA52 CA59 CA63 CA64
Limitations in identifying the health benefits of polyphenolics Stan Kubow
1. Most of the studies are in vitro studies. Pure polyphenolic compound as found in foods.
Variation in polyphenolic structure and absorption in the human gut.
2.â&#x20AC;Ż Lack of fundamental knowledge regarding bioactive polyphenols formed in the human gut.
Saura-Calixto et al., 2007
Pancreatic solution
Base solution
Acid solution
Pump
pH probe
Control panel Water bath
1 Computer
Food
Stomach
2 Small Intestine
3
4
Ascending Transverse colon colon
5 Descending colon
Nitrogen cylinder
Effluent
• 1 g/L arabinolactan • 2 g/L pectin • 1 g/L xylan GI • 3 g/L starch food • 0.4 g/L glucose • 3 g/L yeast extracts • 1 g/L peptone • 4 g/L mucin • 0.5 g/L cystein powders
Step 1: Inoculation Fecal slurry from the fecal samples of five volunteers Step 2: Stabilization period (two weeks)
Pancreatic solution
Base solution
Acid solution
Pump
pH probe
Control panel Water bath
1 Computer
Food
Stomach
GI food + Treatment /Potato
Feeding protocol: 3 times/24 hours
2 Small Intestine
3
4
Ascending Transverse colon colon
Sampling T= 0 hours T= 8 hours T= 24 hours
5 Descending colon
Nitrogen cylinder
Effluent
Analyzed by LC-‐MS TOF Liquid chromatography coupled to time-of-flight mass spectrometry.
Transformation of ingested polyphenols Sample taken from the stomach analyzed for ferulic acid- (trace)
Samples from the small intestine analyze for 3-(3-hydroxyphenyl)-propionic acid â&#x20AC;&#x201C; (x)
Sample from stomach analyzed for chlorogenic acid- (xx)
Transformation of ingested polyphenols Chlorogenic acid conversion to 3-‐phenylpriopionic acid Colombian Cultivar - 24hr
Small
Compound
m/z (-ve)
Stomach
Intestine
Colon
Colon
Colon
Chlorogenic Acid
353.088
xx
xx
x
TRACE
TRACE
CONVERTED
Neochlorogenic acid
353.088
x
x
xx
x
0
CONVERTED
Cryptochlorogenic acid
353.088
x
x
xx
TRACE
TRACE
CONVERTED
Caffeic acid 179.033 3-Hydroxyphenylpropionic acid (3-HPP) 165.056
x
xx
xx
xx
x
CONVERTED
x
xx
xx
xx
xx
FINAL METABOLITE
Ascending Transverse Descending Possible metabolic fate
3-Hydroxyphenylacetic acid
151.04
TRACE
TRACE
x
xx
xx
FINAL METABOLITE
3-Phenylpropionic acid
149.061
-
-
-
xx
xxx
FINAL METABOLITE
Chlorogenic acids
Pathway of ingested polyphenols
Quinic acid
Caffeic acid
Dihydrocaffeic acid
Syringic acid
Ferulic acid
Vanilic acid
Dihydroferulic acid
Benzoic acid
3-‐Hydroxyphenylpropionic acid (3-‐HPP) 3-‐hydroxy-‐benzoic acid
Ru7n 3-‐Phenylpropionic acid Querce7n
3,4-‐dihydroxyphenylace7c acid
3-‐hydroxyphenylace7c acid
Protocatechuic acid
4.4. Moving results to field Ernesto Rodríguez
Participative research for selecting new potato cultivars Evaluation was carried out in eight localities during four crop cycles.
Some traits evaluated in advanced potato clones 41.000
Yield Kg/ha
39.000 37.000 35.000 33.000 31.000
Col.
Gal.
4
9
50
51
52
59
63
64
Code of genotypes 4"
Sem."A"
Frying quality
3,5"
Sem."2"
3" 2,5" 2"
Scale for the evalua7on
1,5" 1" 0,5" 0"
Col.
Gal.
4
9
50 51 52 Code of genotypes
59
63
64
1
2
3
4
5
Cultivar code 59
New cultivars for linking agriculture to nutrition Cultivar code 64 Vegetative period: 120 das; Yield: 37.3 ton/ha, Dry matter: 23.8; Frying quality: 2.7
Cultivar code 4
Vegetative period: 120 das; Yield: 32.9 ton/ha, Dry matter 25; Frying quality: 1.7
Vegetative period: 120 das; yield: 37.1 ton/ha, dry matter: 24.7; frying quality: 2.5
das: days after sowing
5. Conclusions and perspectives ü Variability in the content of macronutrients, micronutrients and functional foods were found in potato. This variability allows making pre-selection and selection of advanced breeding clones for new cultivars. ü Nutritional quality criteria were introduced in the potato breeding program, in order to strengthen the link agriculture-nutrition-health. ü To measure the incidence of these new cultivars in iron and zinc assimilation for children under five. ü To find genetic associations using data from GBS and 2bRAD.
Acknowledges
Contract to access gene7c resources 53
Thanks for your attention