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POTATO BIOFORTIFICATION INTRODUCTION OF GENETIC GAINS FOR IRON&ZINC CONCENTRATIONfrom a Diploid population to advanced disease
resistant tetraploid potatoes
Elisa Salas Murrugarra International Potato Center (CIP). Germplasm Enhancement and Crop Improvement-Crop Management DivisionAv. La Molina 1895. La Molina. Lima 12. PeruCorresponding author’s email: [email protected]
WPC | ALAP | CUSCO, PERU | 2018
WPC | ALAP | CUSCO, PERU | 2018
Women of reproductive age
Rwanda: 59 %Kenya: 24 %
Ethiopia: 19 %Peru: >50 %
Children under 5 years old
Rwanda: 38 %Kenya: 46 %
Ethiopia: 50 %Peru: 43 %
• Maternal mortality
• Mental development and learning
• Physical labor
• Stunting
• Susceptibility to infections
Micronutrient Deficiencies ”Hidden Hunger”
Introduction
WPC | ALAP | CUSCO, PERU | 2018
WHO 2008
Anaemia as a public health problem by country
Preschool-age children
WPC | ALAP | CUSCO, PERU | 2018
Genetic Resources and Improved Populations
Lowland Tropics
Virus Resistant
(LTVR)
Population
Highland
Tropics
Late Blight
Resistant (B3)
Population
Biofortified
Population:
High Micronutrient
Concentrations
Promoters
and Inhibitors
of Mineral
Absorption
Iron (Fe)
Zinc (Zn)
Vitamin C
Carotenoids
Phenolics
Genetic diversity assessment
WPC | ALAP | CUSCO, PERU | 2018
FEDW
Gro
up
s
30272421181512
LB
LTVR
LTVR_LB
Var. World
IRON_DW
Biofortification: Base Population
FEDW
Sp
p
30272421181512
Adg
Gon
Phu
Stn
Each symbol represents up to 2 observations.
IRON_DW
X=18.03
n=64
n=81
n=195
n=242
n=140
n=65
n=41
n=30
IRON (mg/k,DW)
Lan
dra
ceA
dva
nce
d c
lon
es
WPC | ALAP | CUSCO, PERU | 2018
Breeding for Iron and Zinc in Diploid Potato- 3 cycles of recurrent selection -
Base Population and
Crosses (2x-2x)
Cycle I: 16 families
NCD I (4 male groups)
Heritability, selection
Cycle II: 32 families
NCD II (2 sets 4 x 4)
Parental value, selection
Cycle III: Crosses
Selected clones (n=20)
Cycle 0: Selected diploid cultivarsStn, Gon, Phu
Cycle III: Select diploid clones38 - 43 ppm Fe
30 – 40 ppm Fe
Cycle II: Selected diploid clones
Timeline
2005
2010
2008
2006
2013
Breeding at diploid level
WPC | ALAP | CUSCO, PERU | 2018
30383
2
30382
6
30384
1
30384
2
30384
5
30384
630382
8
30383
5
703421
Stn
702815
Stn
703291
Phu
703825
Gon
704393
Gon
X
Group I
♂
♀s
703168
Gon
703831
Gon
703352
Gon
701165
Phu
703825
Gon
X
Group
II♂
♀s
Development of Base Population at Diploid Level: NCD I
WPC | ALAP | CUSCO, PERU | 2018
GCA effects for Fe and Zn concentration
Genetic evaluation of Micronutrient Traits in Diploid base
population
Parental value:
WPC | ALAP | CUSCO, PERU | 2018
Progress: Fe and Zn contents of potato increased
through
3 Cycles of Recurrent Selection at Diploid Level
Year 2011 : 26 Diploid Clones Selected Cycle II
Year 2015: 34 Diploid Clones Selected Cycle III
Fe: 19-32 ppmZn: 17-27 ppmYield:8-23 t ha-1
Fe: 25-35 ppmZn=18-32 ppmYield:9-25 t ha-1
Fe mg/kg DW
Fre
qu
en
cy
403530252015
7
6
5
4
3
2
1
0
21.81 3.348 17
25.97 3.571 21
29.15 2.791 22
Mean StDev N
Cycle 0
Cycle2
Cycle3
Treatment
Zn mg/kg DW
Fre
qu
en
cy
353025201510
6
5
4
3
2
1
0
18.67 3.502 17
21.59 3.236 21
24.45 3.290 22
Mean StDev N
Cycle 0
Cycle2
Cycle3
Treatment
25 % Genetic Gain for ZINC
27 % Genetic Gain for IRON
WPC | ALAP | CUSCO, PERU | 2018
Estimates of Gains (%) per Cycles
for Iron, Zinc and Yield Components
12.92 10.84 11.41-2.84
-23.73-2.22
-24.28
9.83 10.65
35.93
4.80
-20.73
-3.32
27.40
27.10 24.86
56.67
1.89
-33.57
-5.35
2.41
-100
-50
0
50
100
150
Iron Zinc Number oftubers/plant
Tuberweight/
plant
Averagetuber weight
Dry matter Vitamin C
Cycle 2 (M2-Base Population) Cycle 3 (M3-M2)
Total (M3-Base Population)
WPC | ALAP | CUSCO, PERU | 2018
a) Inductively Coupled Plasma Spectroscopy
(ICP)
High-Throughput Methods for Mineral Analysis
Inductively coupled plasma mass spectrometry
Burgos et al 2007. The Science of Food
and Agriculture.87(4):668-675
b) X-ray fluorescence Spectroscopy (XRF)Zinc (Zn)
Iron (Fe)
o 50 Samples/dayo $ 25 / Sample
o 300 Samples/dayo $ 15 / Samples
Fluorescence Spectroscopy (XRF)
T. zum Felde et al (Por publicar)
WPC | ALAP | CUSCO, PERU | 2018
Taking advantage of Agroecological Diversity in Peru
Selection of clones with resistance to LB under field conditions
Soil texture Sandy Loam
Soil EC (dS/m) 0.68
Soil PH 4.6
Zinc ppm 2.1
Iron ppm 169.05
Soil texture Sandy Clay Loam
Soil EC (dS/m) 11.58
Soil PH 7.6
Zinc ppm 13.2
Iron ppm 2.36
Soil texture Clay Loam
Soil EC (dS/m) 1.49
Soil PH 7.59
Zinc ppm 32.34
Iron ppm 7.72
WPC | ALAP | CUSCO, PERU | 2018
Bioaccessibility of Iron
WPC | ALAP | CUSCO, PERU | 2018
(17) Selected 2x Male
Parents from Cycle II(33) Advanced 4x
Female Parents
♂♀
• Disease
Resistance
• Stress tolerant
• High stable yield
• Processing quality
• Tuber appearance
• High level / GCA of
Fe & Zn
concentration
• High frequency 2n
pollen
230 families 4x -2x (11 829 genotypes)
Seed
Incorporation of Micronutrients from Diploid into Tetraploid populations
WPC | ALAP | CUSCO, PERU | 2018
Ploidy level identification by chloroplast counting in guard cells
Sowing seeds families Transplanting seedlingsTransplanting to pots Samples of 3 folioles
13 284 genotypes
Peel off the surfacecell layer from theundersurface of the leaf.
One or two drops of iodine solution (iodine-potassium iodide) were placed in the center of a slide
Chloroplast Number in Stomatal Guard Cells
12-14 9 – 11 6 – 8
WPC | ALAP | CUSCO, PERU | 2018
TETRAPLOIDIZATION
Crosses 4x - 2x
• Ploidy identification
(11 829 seedlings)
Tuber families
Clean copy
Loc 1-LM230 masl
2012
2013
2014 Micronutrient Screening &
Field trials
Biofortification at Tetraploid Level- Cycle 1
6700 genotypes
Loc 2-PAUC3300 masl
2015
2016
Clean copy
Clean copy
Loc 32600 masl
2017
4500genotypes
100-200genotypes
Disease screening
Loc 32600 masl
Loc 42800 masl
2018
Loc 5-OXALate blight
Loc 6-SR Potato Virus Y
Clean copy
100-200genotypes Loc 4
2800 masl
Invitrointroduction
International Distribution
for Variety SelectionIntercrossing
Start cycle 2
Loc 1- LM (GH)Late blight & Citometry
Analysis
Loc 5-OXALate blight
Loc 6-SR Potato Virus Y
MET’s of 30 Biofortified
clones in 8 localities of
the highland in Peru
WPC | ALAP | CUSCO, PERU | 2018More than 50 new generation of biofortified tetraploid potato clones
adapted to tropical highland and mid-elevation ecologies were selected for international distribution:
Buthan, Nepal, Kenya, Malawi, India & PeruHigh Fe contents (30 - 45 mg/kg)
High yield (from 20 – 50 ton/ha)
Cusco 2018
Resistance to late blight and/or PVY
Cusco 2018
Zn contents (26 - 35 mg/kg)
WPC | ALAP | CUSCO, PERU | 2018
0
5
10
15
20
25
30
35
40
45
CIP
31
27
28
CIP
31
26
42
CIP
31
27
21
CIP
31
27
51
CIP
31
26
37
CIP
31
27
64
CIP
31
27
63
CIP
31
26
82
CIP
31
26
61
CIP
31
28
15
CIP
31
28
26
CIP
31
27
99
CIP
31
24
79
CIP
31
25
14
CIP
31
25
35
CIP
31
25
47
CIP
31
25
86
CIP
31
26
11
CIP
31
26
26
CIP
31
26
40
CIP
31
26
71
CIP
31
27
00
CIP
31
27
20
CIP
31
27
57
CIP
31
27
70
CIP
31
28
02
CIP
31
28
21
CIP
31
28
39
CIP
31
28
49
CIP
31
28
63
% S
ele
cti
on
Parents 4x
CIP393083.2
CIP393536.13
CIP393613.2
CIP394908.13
CIP395017.229
CIP395017.242
CIP395112.32
CIP395446.1
Parents 2x
CIP306018.4
CIP306085.114
CIP306087.11
CIP306087.82
CIP306140.14
CIP306143.62
CIP306416.68
CIP306418.53
CIP306418.69
CIP306514.64
Top families and parentsTraits: Iron & Zinc, yield, tuber appearance
Families (4x-2X)