This document summarizes breeding efforts to develop wheat germplasm with enhanced zinc and iron concentration. [1] The first international wheat yield trial identified lines with 50-100% of the zinc concentration target and some lines outyielding checks. [2] The lines showed high heritability for zinc and iron concentration across locations, indicating potential for widespread cultivation. [3] Positive correlations were found between zinc and iron concentration, grain yield, and protein content, demonstrating the lines possess both nutritional and agronomic advantages.

1. Breeding for enhanced Zinc and Iron
concentration in CIMMYT spring wheat
germplasm
Govindan Velu
Global Wheat Program, International Maize and
Wheat Improvement Center (CIMMYT)- Mexico
E-mail: velu@cgiar.org
MAZATLAN, JUNE 15, 2012

2. Achieving sustainable food and
nutritional security

3. HarvestPlus Impact & Product Pathway
Discovery
HPlus I
2004-2008
Product Development
and Delivery Plan
Development
HPlus II
2009-2013
Delivery
HPlus III
2014+

4. G8 summit supports for
biofortified crops

5. Breeding strategy

6. Potential donors for high Zn
HONG HUA MAI
HONG DUAN MANG

7. Successful shuttle breeding

8. Performance of 1st HarvestPlus International Wheat Yield
Trial in target environments for Zn, 2010-11
50
Mexico
45 50% breeding Pakistan
40 target
India
35
Pooled
30
100%
25 breeding
20 target
%
n
e
s
r
t
i
15
10
5
0
0 1-2 3-4 5-6 7-8
Zn concentration (ppm) above the checks
• >40% entries had atleast 50% target Zn (4 ppm) & 5-10% entries had
full target Zn (8 ppm).
• high heritability (0.7) and high genetic correlations between locations
suggests low G x E.

9. Grain yield potential of 1st HPYT
across locations
30
% of 3 checks mean
25 % of local check
20
15
%
n
e
E
s
r
t
i
10
5
0
<80 81-85 86-90 91-95 96-100 101-105 106-110
Grain yield (% over check)
• 2-3 best leads were identified in India & Pakistan with 75-100%
target Zn & better grain yield potential, rust resistance and
superior end-use quality.
• These leads are being evaluated in regional PVS/on-farm trials

10. Performance of 1st HPYT across
locationsAcross locations Zn (ppm)-Regional Global GY (% Agronomy Stem rust(Kenya) NJ End-use qualit
Cross
ppm % India Pak Asia Mex- Off-
Ent Zn Fe 9 Hea Matu GRNH GRNP
above target (5 (2 (7 2 PH seaso Main season
(ppm) (ppm) sites d r RD % RO %
checks level sites) sites sites) sites n
CROC_1/AE.SQUARROSA (210)//INQALAB
435 91*2/KUKUNA/3/PBW343*2/KUKUNA 38.7 8.7 109 7.7 9.5 9.3 6.9 44.4 86 78 119 107 5MSS 5 MS APR_R 45 14
OASIS/SKAUZ//4*BCN/3/2*PASTOR/4/T.SPELT
412 A 35.1 5.1 64 3.3 6.1 4.4 6.1 40.2 97 93 130 105 20MSS 30 MSS APR_MR 41 12
PI348449/5/BAV92/3/OASIS/SKAUZ//4*BCN/4/
FRET2/TUKURU//FRET2*2/3/T.SPELTA
422 PI348530 35.0 5.1 63 2.4 5.9 5.7 3.2 39.4 87 94 131 115 20MSS 10 M APR_R 44 12
REH/HARE//2*BCN/3/CROC_1/AE.SQUARROSA
(213)//PGO/4/HUITES/5/T.SPELTA
414 PI348599/6/REH/HARE//2*BCN/3/CROC_1/AE.S
35.0 5.0 63 3.6 3.2 5.7 2.4 40.7 91 96 136 105 20MSS 50 MSS MS 40 12
QUARROSA (213)//PGO/4/HUITES/7/QUAIU
WBLL1*2/TUKURU/3/T.DICOCCON
424 PI94624/AE.SQUARROSA 34.8 4.9 61 1.1 4.0 3.7 6.5 38.0 80 85 128 110 30MSS 20 M APR_R-MR 43 13
(409)//BCN/4/WBLL1*2/TUKURU
OASIS/SKAUZ//4*BCN/3/2*PASTOR/4/T.SPELT
413 A 34.7 4.8 60 3.0 7.4 4.2 4.1 39.7 96 93 130 109 20MSS 30 MSS APR_MR 36 12
PI348449/5/BAV92/3/OASIS/SKAUZ//4*BCN/4/
432 GTO95.1.20/KIRITATI//MUU 34.6 4.6 58 1.3 5.1 4.1 5.1 38.2 79 89 131 104 5MSS 1 MSS APR_NIR 39 13
CROC_1/AE.SQUARROSA
433 (210)//PBW343*2/KUKUNA/3/PBW343*2/KUK 34.3 4.3 54 4.9 4.9 4.6 2.5 42.0 86 86 128 119 5MSS 1 MS APR_NIR 40 14
UNA
CROC_1/AE.SQUARROSA
434 (210)//PBW343*2/KUKUNA/3/PBW343*2/KUK 34.2 4.2 52 3.0 6.9 4.0 4.1 41.4 91 88 129 119 40MSS 30 MSS APR_MR 41 13
UNA
Checks 30.0 36.0 100 94 132 109 30MSS 47 12
Mean 32.1 1.1 2.9 1.5 1.0 37.8 92 89 130 108 42.5 12.3
Min 28.3 -2.6 -1.9 -1.6 -2.2 32.6 76 78 119 96 35.5 10.9
Max 38.7 7.7 9.5 9.3 6.9 44.4 107 102 140 119 59.5 14.3
SD 2.17 1.9 2.6 2.5 2.2 2.5 7.7 5.7 4.3 5.4 5.30 0.81

11. Comparison of grain yield of BWIR advanced lines with H+
improved lines
30
H+ lines yield potential
Irrigated BW lines yield
25
potential
20
Wild x Elite (N =>600)
% entries
15
10
Elite x Elite (N =>4900)
5
0
5
0
0
5
0
5
0
5
5
5
0
1
<7
0
-7
-8
-8
-9
-9
10
11
11
>1
-1
80
85
90
70
75
0-
5-
0-
95
10
10
11
Grain yield (% checks mean)

12. 60
Landraces derivative
50 Synthetic derivative
T Spelta derivative
40
% entries from HPYT
30
20
10
0
<20 21-25 26-30 31-35 36-40 <45
Zn concentration (mg/kg)

13. High H2 across and individual
environment
Trait Sites
Entry Residual Grand
LSD CV Heritability
Variance Variance Mean
E1 2.5 1.7 26.8 2.7 5.0 0.74
E2 6.7 4.2 34.0 4.3 6.2 0.76
E3 5.5 9.4 34.2 6.4 9.2 0.54
Grain E4 5.5 8.8 29.3 6.3 10.6 0.56
Zn E5 8.7 14.5 28.5 8.2 14.1 0.54
(mg/kg) E6 20.5 18.5 32.1 9.2 14.1 0.69
E7 0.8 19.7 31.3 9.2 14.4 0.07
E8 1.6 11.0 32.3 6.7 10.2 0.22
E9 18.1 10.6 44.1 6.5 7.2 0.77
Across
3.70 11.50 32.5 2.90 4.60 0.78
locations
E1 9.6 3.6 32.6 3.8 5.8 0.84
E2 17.3 7.7 42.7 6.1 7.0 0.82
Grain E3 7.2 10.1 34.8 7.0 9.8 0.59
Fe E4 16.8 8.7 37.8 6.4 8.3 0.79
(mg/kg) E5 2.1 2.5 32.4 3.3 5.0 0.62
E6 11.3 3.7 37.3 4.0 5.3 0.86
E7 1.6 4.5 35.5 4.3 6.0 0.42
E8 0.1 13.8 41.3 7.9 9.4 0.01
E9 7.1 2.5 38.9 3.3 4.1 0.85
Across
5.10 6.60 37.0 2.40 3.30 0.88
locations

14. 50
Associations y = 1.1761x
R² = 0.59
40
30
m
G
p
o
F
n
c
e
a
r
)
(
t
i
20
20 25 30 35 40
Grain Zn concentration (ppm)
40
Zn y = 1.3384x + 15.704
concentration R² = 0.2491
35 (ppm)
30
25
20
10.0 11.0 12.0 13.0 14.0 15.0
Protein %

15. 170
y = 31.918x
160 R² = 0.06
150
-1)
140 Positive association of
130 Zn/Fe yield with grain
120 yield
Z
h
k
d
n
a
g
y
e
(
l
i
110
100
3.0 3.5 4.0 4.5 5.0
Grain yield (tons ha-1)
200
y = 37.645x
190
R² = 0.33
180
170
-1)
160
150
140
130
F
h
k
d
a
g
y
e
(
l
i
120
110
100
3.0 3.5 4.0 4.5 5.0
Grain yield (tons ha-1)

16. Zn ppm Fe ppm GY PH
XRF- ICP- XRF- ICP- % Zn and Fe
t/ha cm
CIM WAS CIM WAS checks concentration of
Mean 33.9 41.1 31.0 31.7 6.8 101 102 Candidate lines
Min 22.9 29.3 23.5 27.5 6.3 96 79
(C2HPYT) Y10-11
Max 65.4 58.7 41.2 37.6 7.5 114 116
SE 0.37 0.4 0.14 0.12 0.27 0.19 0.37
60
N =420 Zn ppm
50
Fe ppm
40
30
%
n
e
s
r
t
20
i
10
0
25 30 35 40 45 50 >55
Zn and Fe concentration, ppm

17. Special emphasize on rust resistance
• Luckily some of the 1st HPYT
best bets possess Ug99
resistance
• 2nd HPYT entries were selected
considering Ug 99 data
Singh et al.,
2011

19. R² = 0.7367
60
50
40
30
M
20
R
X
n
C
Y
T
Z
F
)
(
I
10
0
0 10 20 30 40 50 60 70
ICP Zn (WAS)
50
45 R² = 0.684 N >200 samples
40 Correlation b/n ICP
Sets No of samples
& XRF (r2)
35 Set 1 570 0.9
30 Set 2 800 0.8
M
R
X
C
Y
e
T
F
Set 3 1204 0.7
)
(
I
25
Set 4 180 0.7
20 Set 5 140 0.6
15 Total 2894
15 20 25 30 35 40 45
ICP Fe (WAS)

20. New opportunities within CIMMYT initiatives
(SeeD, “WHEAT CRP”)

21. PVS: one of the ways for faster seed delivery

22. Product Testing & Release Schedule
Positive correlation between Fe and Zn - range for
crops: r = 0.3 to 0.8 - allows simultaneous

23. Molecular approach
Association mapping
14
Diverse origin of spelts,
dicoccon, synthetic and 12
landraces derived
10
% observations
COP confirms good 8
enough diversity in the
panel 6
 4
Phenotyping: 3 reps
randomized trial (Alpha 2
lattice-incomplete blocks) 0
in Cd. Obregon 0.0 0.1 0.2 0.3 0.4 0.5
COP - value
0.6 0.7 0.8 0.9 1.0
Genotyping: SNP 90K illumina platform – Largest consortium
covering diverse material
Mapping/map position of SNP will be available in mid 2012
Also SSR genotyping with Dr Klaus, SARDI, Australia

24. Challenges
● Soil Zn heterogeneity
● Applied common ZnSO4
100 kg/ha
● Soil Sampling in each check
plots (Waxwing)
● Spatial experimental design
& grid check approach

25. Product Attributes - Breeding trait in future
Goal: micronutrient density as core Objectives
breeding
will lead to Biofortification of all future varieties
micronutrients are not subject to genetic erosion
Wolfgang H PFEIFFER

26. Conclusion
 Multi-location HPYT results were encouraging –
Moderately high H2, high genetic correlations
 Leads were identified with high Zn and desirable yield
potential
 Time to promote biofortified products in target countries
and additional G x E testing in other countries
 Increasing trend for yield potential (and other desirable
traits) over the years from new set of materials (with
high Zn)
 Expanding breeding operations (↑ no. of crosses, double
the size of YT)
 Second cycle of breeding enhance possibility of
obtaining high Zn improved lines with good agronomy
 XRF allows fast-track screening of Zn and Fe