Bio-fortification for high micronutrients in wheat breeding program in China
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Transcript of Bio-fortification for high micronutrients in wheat breeding program in China
Institute of Crop Science/National Wheat Improvement Centre, CAAS
Email: [email protected]
Bio-fortification for high micronutrients
in wheat breeding program in China
Yong Zhang and Zhonghu He
The 11th International Gluten Workshop. 12-15, 2012, Beijing
Outline
General information
Cultivar screening for high micronutrients
Fe/Zn consistency over locations
Nitrogen application on Fe/Zn transportation
Effect of milling on micronutrient contents
Fe bioavailability of wheat samples
Basic information for wheat in China
Largest producer and consumer in
the world
Main food for 50% of the
population, especially for northern
China’s less developed area
Area: 24 million ha; Yield: 4.8 t/ha
Production: 115 million tons in 2012,
with 78% for food
General information
Challenges for wheat production
Yield improvement, with wide adaptability
Resistance to bio- and abiotic stresses
Processing and nutrition qualities
Basic information for malnutrition HarvestPlus (IFPRI): Fe, Zn, Vitamin deficiency throughout
the world, with population over 2 billions, in undeveloped area
HEALTHGRAIN (EU): Phytochemicals, especially phenolics
for chronic diseases and metabolic syndrome, cardiovascular
incidence in urban China is 18%, doubled in the last 10 years
Immense economic and social costs, biofortification is a best way to solve the problem
Objectives of HarvestPlus China what program
To select high Fe/Zn varieties with good agronomic performance and broad adaptation - acceptable to farmers
To improve nutrient malnutrition, prevent or reduce Fe/Zn deficiency and relative diseases - beneficial to poor consumers
Strategy
Variety testing and screening, then breeding
Combination of various disciplines, i.e.,
breeding, plant nutrition, and human
nutrition
Link with HarvestPlus wheat
program through Dr. Ivan
Monasterio from CIMMYT
HarvestPlus Wheat Annual Meeting
Collaborative institutes
CAAS: germplasm screening, multi-location field
trial, effect of milling and processing on nutrients
CAU: Fe and Zn analysis, and their transportation
efficacy from root to shoot, and to grain
Sichuan University: bio-availability analysis
Eight lines with high Fe/Zn in Chinese wheat
In the 8 high Fe genotypes, 5 had high Zn
Euphytica, 2010, 174:303-313
Genotype Fe Genotype Zn
Jimai 26 60.2 Henong 326 58.2
Henong 326 58.1 Jimai 26 51.9
Han 3475 52.7 Nongda 3197 47.8
Jingdong 8 51.8 Jingdong 8 46.1
Zhoumai 3 49.4 Zhongmai 175 45.8
Jimai 3 49.4 Nongda 179 45.6
Nongda 3197 47.1 Lumai 23 45.5
Nongda 179 46.4 Weimai 8 42.4
265 genotypes were sown at Anyang in Henan province in two seasons
Cultivar screening
Cultivar screening for high micronutrients
Three cultivars Henong 326, Jimai 26 and Jingdong 8 have
high Fe/Zn, among which Jingdong 8 is the leading cultivar
as check in Northern China Plain before 2010
There is significant and positively correlations among Fe and
Zn, it is possible to combine high Fe/Zn in wheat breeding
Zhongmai 175 with high yield potential, released in Northern
Plain and Northern Yellow and Huai Valleys Drought and
Fertile Region, has high Zn, extend 0.2 million Ha in 2012
Six lines with high free and bound phenolic acids
Season
Cultivar
Free phenolic (mg/kg)
Cultivar
Bound phenolic (mg/kg)
2008-2009 Jishi 02-1 45 Kaimai 18 756
Taishan 23 39 Linyou 145 725
Xiaoyan 6 21 Zhongmai 895 684
Liangxing 66 20 Shaanyou 225 683
Xinong 979 20 Liangxing 66 674
Zhongmai 895 19 Zhangmai 349 666
2009-2010 Jimai 19 25 Kaimai 18 951
Liangxing 66 25 Jimai 19 923
Shaanyou 225 23 Xiaoyan 6 844
Zhongmai 895 19 Zhongmai 155 815
Zhongyu 5 16 Zhongmai 895 783
Zhongmai 875 16 Liangxing 66 777
Liangxing 66 and Zhongmai 895 perform high and more consistent for
both free and bound phenolic acids
Journal of Cereal Science, 2012, in press
Consistency of Fe/Zn over locations and years
Source df Fe Zn P TKW Protein
Genotype (G) 23 9.7 2.8 7.7 50.6 37.1
Environment (E) 13 66.0 89.5 75.8 28.8 33.6
G×E 299 24.3 7.7 16.5 20.6 29.3
E/G 6.8 32.0 9.8 0.6 0.9
Euphytica, 2010, 174:303-313
Fe/Zn consistency
Fe of Zhongyou 9507 at 7 locations over 2 seasons
Zn of Zhongyou 9507 at 7 locations over 2 seasons
Cultivar performance over locations
Significant effects of genotype, environment, and G × E on Fe/Zn were observed
Significant variation for Fe/Zn in the same genotype was observed in 7 locations over years, however, the variation of Fe is much lower than Zn
QTL with Xwmc312 on chromosome 1A QTL with Xgwm251 on chromosome 4B
QTL Closest Marker Position AE LOD PVE (%) Total PVE (%)
QZinc.caas-1A Xwmc312 2.0 1.61 3.1 14.0 29.2
QZinc.caas-4B Xgwm251 6.5 -1.78 2.7 15.2
72 DH lines were sown at Anyang in Henan province in two seasons
QTL mapping of Zn
Effect of N on Fe in root and shoot
Optimized N application significantly increase Fe uptake of shoot and plant,
63%-82% of Fe is kept in root
Trait Check 174 kg/ha
Root
Fe concentration (mg/kg) 1654a 1431a
Fe uptake (mg/m2) 98a 137.0a
Shoot
Fe concentration (mg/kg) 65.3a 86.6a
Fe uptake (mg/m2) 21.3b 80.8a
Shoot/Root ratio of Fe concentration 0.04 0.06
Shoot/Root ratio of Fe uptake 21.77 58.99
Shoot/(Root+Shoot) ratio of Fe uptake 18% 37%
Total (Root+Shoot)
Fe concentration (mg/kg) 338.8a 309.1a
Fe uptake (mg/m2) 129.0b 217.8a
Liangxing 99 was sown at Quzhou in Hebei province in 2010-2011
Effect of nitrogen application
Effect of N on Zn in root and shoot
Optimized N application significantly increase Zn uptake in root, and Zn
concentration and uptake of shoot and plant. 16%-42% of Zn is kept in root
Trait Check 174 kg/ha
Root
Zn concentration (mg/kg) 44.8a 45.4a
Zn uptake (mg/m2) 2.7b 4.4a
Shoot
Zn concentration (mg/kg) 11.6b 24.3a
Zn uptake (mg/m2) 3.7b 22.6a
Shoot/Root ratio of Zn concentration 0.26 0.54
Shoot/Root ratio of Zn uptake 1.4 5.2
Shoot/(Root+Shoot) ratio of Zn uptake 58% 84%
Total (Root+Shoot)
Zn concentration (mg/kg) 17.0b 26.3a
Zn uptake (mg/m2) 6.4b 26.9a
Effect of N on Zn/Fe distribution in wheats
0
10
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0kg/ha 174kg/haZ
n D
istr
ibu
tio
n(%
)
Grain Stem+Leaves Husk Root
0
10
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30
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50
60
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0kg/ha 174kg/ha
Fe
Dis
trib
uti
on
(%)
Grain Stem+Leaves Husk Root
N application decrease root Fe and Zn by 9% and 6%, increase grain Fe
and Zn by 2% and 3% than control, significantly increase the efficacy of
Fe/Zn transportation from root to shoot and grain, but the efficiency for Zn
was much better than that for Fe
Source df Fe Zn P PAP Phytase
Milling 7 3324*** 2506*** 367306*** 67006*** 10134***
Genotype 42 20*** 28*** 1564** 126** 1157***
Error 294 62 65 6060 508 2956
There were significant effect of milling and genotype on all micronutrient
concentrations, with milling being the predominant
43 cultivars sown in Jinan, Shandong province in 2005-2006
ANOVA of milling on micronutrients
Effect of milling and processing
Fraction Fe Zn
Mean Range Mean Range
Break 1 6.9edf 4.6-10.9 7.3d 4.1-14.2
Break 2 7.9ed 3.6-12.1 7.9d 1.8-12.1
Break 3 15.7c 10.4-27.7 12.7c 9.4-17.7
Reduction 1 5.5f 3.2-13 7.8d 4.9-13.8
Reduction 2 6.5ef 2.5-10.4 8.9d 4.8-16
Reduction 3 8.9d 5-12.8 11.4c 6.9-17
Shorts 43.5b 30.8-61.3 49.1b 35.4-74.2
Bran 100.5a 77.4-124.4 86.3a 69.2-128.2
Fe/Zn contents of streams from Buhler mill
Fraction P PAP Phytase
Mean Range Mean Range Mean Range
Break 1 844de 555-1097 548d 445-649 233e 45-587
Break 2 878de 80-1224 558d 479-690 259e 107-762
Break 3 1314c 985-1715 684c 578-832 617c 221-936
Reduction 1 777e 321-978 524d 437-606 209e 8-437
Reduction 2 1033de 458-8199 574d 484-697 337de 74-452
Reduction 3 1072dc 461-1382 579d 510-667 403d 112-594
Shorts 3860b 886-4999 1262b 967-1684 1449b 580-3061
Bran 10819a 8541-12960 4839a 4166-5579 1683a 630-3726
Phosphorus contents of streams from Buhler mill
Effect of milling on micronutrients
In the same flour stream, at least 2-3 times difference on Fe/Zn among cultivars was observed
Very good (B1, B2, R1 and R2) and fine quality (B1, B2, B3, R1, R2 and R3) flour has low Fe/Zn
Micronutrients concentrate in short and bran fraction
Relationship between nutrient contents and flour yield
0
20
40
60
80
100
120
55 60 65 70 75 80 85 90 95 100
Fe
Zn
Total P
PAP
Phytase
Flour yield
% o
f to
tal
in g
rain
Journal of Cereal Science, 2008, 48:821-828
Total phosphorus decreased the most with decreasing flour yield
Phytic acid (PAP) and phytase activity decreased the least
Relationship of phenolic acids and flour yield
Flour yield
mg
/kg
Journal of Cereal Science, 2012, in review
Ferulic was the predominant, and Caffeic and P-coumaric
was important in phenolic acids
Phenolic acids concentrate in short and bran fraction
0
100
200
300
400
500
600
700
60% 70% 80% 90% 100%
Ferulic
Caffeic
Chlorogenic
Syringic
Gentisic
P-coumaric
Phenolics
Effect of processing method on micronutrients
Process Fe Zn P PAP
Flour 6.7c 8.4c 857a 565 a
Bread 8.4b 9.2b 860a 1.5 b
Steamed bread 8.2b 9.6a 867a 1.6 b
Noodle 9.5a 9.7a 855a 1.7 b
Processing method significantly increase concentration of
Fe/Zn, but decrease phytic acid P very deeply at the same
Sample information for in vitro digestion/Caco-2 cell model
Biomedical and Environmental Sciences, 2012, in press
Sample Flour yield (%) Fe (mg/kg) PAP
Yumai 18 78 15.8 0.78
Jingdong 8 78 20.8 0.82
Zhongmai 9 78 22.7 0.87
Zhongmai 175 78 20.9 0.79
Yumai 2 78 16.0 0.68
Zhongyou 9507 78 29.2 0.91
Fe bioavailability
Bioavailability in In-Vitro Digestion/Caco-2 cell model
d
b
c
cd
d
a
c
a
bc c
d
ab
0
50
100
150
200
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300
Yumai 18 Jingdong 8 Zhongmai 9 Zhongmai 175 Yumai 2 Zhongyou 9507
BV
& B
VP
G (
% o
f co
ntr
ol)
Fe BVPG (% of control) Fe BV (% of control)
Zhongyou 9507 and Jingdong 8 have high Fe bioavailability
Phytate is an important Fe BV inhibitors, as reductions in phytate
content in cereals lead to strong increases in Fe BV
Conclusions
Jingdong 8 and Zhongyou 9507 have high Fe/Zn. Liangxing
66 and Zhongmai 895 have high and consistent phenolic acids
Fe, Zn and PAP are significantly influenced by G, E, milling,
and processing
Two QTLs on 1A and 4B contribute 29.2% of the PVE of Zn
Nitrogen can significantly increase Fe/Zn transportation
efficacy from soil to shoot and grain, but the efficiency for Zn
is much better than that for Fe
Zhongyou 9507 and Jingdong 8 have high Fe bioavailability
based on in-Vitro digestion/Caco-2 cell model
Perspective
More genotypes need to be screened and their stability across
environments need to be investigated
QTL s on Zn should be confirmed, molecular marker will be
validated for breeding program. How about Fe?
Effects of other fertilizer and their interaction on Fe/Zn
transportation efficacy will be done next
Effects of milling and processing on bioavailability will be
done next, and how about animal and human trial?
Need to meet efforts to combine high yield potential and
major disease resistance with nutrition quality, farmer’s need
Acknowledgements
Prof. Yan Zhang, Mr. Jianwei
Tang, and Xiaoyong Shen, Lan
Wang, CAAS
Drs. Rongli Shi and Qichao Song,
CAU
Drs. Lin Bai and Dr. Ji Lei, SCU
Dr. Raymond P. Glahn, Cornell
University
Funding from HPC and Chinese Ministry of Agriculture