This document summarizes research on biofortified maize varieties in southern China. It discusses issues like iron, zinc, and vitamin A deficiencies in the region. It then describes breeding efforts to develop high-oil, quality protein, and provitamin A (ProVA) biofortified maize varieties. Two successful varieties developed are Yunrui 8, a high-oil variety, and Yunrui 1, a quality protein variety. Both varieties have good yields, disease resistance, and nutritional quality. They have been widely adopted by farmers, increasing production and improving nutrition. The document also discusses molecular breeding efforts to develop ProVA maize through marker-assisted selection of genes controlling carotenoid levels. This has

1. Impact and Use of Biofortified Maize
in Southern China
Prof. Xingming Fan, Yaqi Bi, Daniel Jeffers,
Yudong Zhang, Li Liu
Institute of Food Crops (IFC)
Yunnan Academy of Agricultural Sciences (YAAS)

2. Background
Iron Deficiency
Vitamin A Deficiency
Zinc Deficiency
From Boitshepo Bibi Giyose(FAO)
More than 1/3
of the world’s
population suffered
iron, vitamin A, and
zinc deficiency

3. Background
From HarvestPlus
In sub-Saharan Africa, over 40 varieties of biofortified maize have
been released onto the market
the first-ever varieties of biofortified zinc maize were released by
the Colombian and Guatemalan governments.

4. Background
RNI of Zinc
12.5mg/d
RNI of Iron
12mg/d
Poor VillageRural AreaTownCity
Daily intake of Zinc and Iron in China
Iron (mg)
Zinc (mg)
National Nutrition Survey 2010-2012 ， RNI ：
16.29
7.56
Yunnan

5. Background
Daily intake of Retinol in China
National Nutrition Survey 2010-2012 ， RNI ：
Retinol (μg)
Poor VillageRural AreaTownCity
381.2

6. Background
 Iron deficiency:
heart disease, cancer, anemia
 Zinc deficiency:
cognitive disorder
 VA deficiency:
birth defect, blindness
Deficiency Zinc Iron VA
Urban 34.8% 12.7% 3.0%
Rural 36.7% 20.8% 11.1%
Nutrition Deficiency for kids under 6 years old
National Nutrition Survey 2010-2012

7. Background
 Micronutrient supplements: Vintamin A and Zinc
 Micronutrient fortification: Iron and salt iodization
 Expanded immunization coverage for children
 Biofortification

8. Background
Biofortified
maize
High oil
maize
Quality protein
maize
Provitamin A
Zinc
&
Iron
Hopkins et al. 1903; Smith 1908;
Miller et al. 1981; Song et al. 1998;
Laurie et al. 2004; Li et al. 2013; etc.
Mertz et al. 1964; Paez et al. 1969; Vasal
et al. 1980; Prasanna et al. 2001; etc.
Bouis and Welch, 2010; Pandey
et al. 2015; Chomba et al. 2015;
Hindu et al. 2018; etc.
Maziya-Dixon et al. 2000; Harjes et
al. 2008; Yan et al. 2010; Babu et
al. 2013; Li et al. 2015; etc.

9. HarvestPlus Program
Coordinated by the International Center for
Tropical Agriculture (CIAT) and the International
Food Policy Research Institute (IFPRI), 2003
Country program:
HarvestPlus-Brazil
HarvestPlus-India
HarvestPlus-China (HPC)
http:www.harvestplus-china.org
HPC: aims to reduce “hidden hunger” and to improve overall
nutrition in China through germplasm screening, breeding,
evaluation and delivery of the biofortified crops.

10. Background
Yunnan Province,
Southwest China

11. Background
Maize is a staple food for minority people in Yunnan

12.  Maize is the most important food
crop in Yunnan province.
 area of 1.4 million ha.
 accounts for 32% of total arable area.
 Maize is a staple food of people
living in mountain areas and is an
important feed source.
Maize production in Yunnan

13.  Maize is vulnerable to many devastating diseases
due to the humid subtropical climate.
Turcicum leaf blight, gray leaf spot, leaf rusts,
Fusarium and Gibberella ear rots
 Most of the cultivated varieties have poor quality.
Maize production in Yunnan

14. The germplasm improvement
High-oil Maize
 Germplasm innovation the original
tropical High-oil
maize inbreds
has been created.

15. Inbred Original Oil content (%) Ecology type
Y46 Suwan1 --- Tropical
CML161 Pool25QPM --- Tropical
CML171 Pool25QPM 7.72 Tropical
CMl166 Pop66QPM --- Tropical
GY276 BHO --- Temperate
GY717 BHO --- Temperate
GY923 ALEXHO 10.66 Temperate
GY220 ALEXHO 10.13 Temperate
GY237 ALEXHO 13.21 Temperate
GY798 ALEXHO 9.13 Temperate
Classification of heterosis group
——Diallel mating design
High-oil Maize

16. Classification of heterotic groups
 There were significant genetic differences between
temperate and tropical High-oil maize germplasm.
 There was a higher chance to develop High-oil
maize hybrids by using temperate x tropical
inbreds.
High-oil Maize
Analysis on combining ability between temperate high-oil maize inbred lines and tropical miaze
germplasms. J. Maize Sciences, 2010, 5-10.

17. Yunrui 8Yunrui 8
Yunrui 21Yunrui 21
Yunyou 19Yunyou 19
The Successful Experience in
High-oil Maize Breeding
Yunrui 21Yunrui 8

18. The Development of Yunrui 8
Inbred (RR)
Resistant to diseases
Inbred (rr)
Susceptible to Disease×
F1(Rr)
×F4 ( Rr, with resistant segment)
F4 ( RR or Rr or rr )
Selfing for 4 times
Phenotyping+Foreground
selection
BC1F4 ( Rr)
Phenotyping+Foreground+Background selection
BC1F5 ( Rr)
BC2F5 ( Rr, with highest recovery rate)
BC2F6 ( RR)
Improved inbreds
Resistance to diseases
×
Phenotyping+Foreground selection
Phenotyping+Foreground+Background selection
Phenotyping+Foreground selection
Susceptible inbred (rr)
Susceptible inbred(rr)
Improve the resistance of temperate High-oil inbred to gray leaf spot

19. CML171-1-1-2-1-1 × YML107 Yunrui 8
The Development of Yunrui 8

20.  Yield potential.
The Development of Yunrui 8
Variety comparative
testing trial, 2001
Regional testing trials,
2002-2003
Average yield
(kg/ha.)
9848 8435
Yield increase over
the check
10.4% 0.4%
Rank 1 3

21. The Development of Yunrui 8
 National disease resistance evaluation, Chinese
Academy of Agricultural Sciences (CAAS).
 high level of resistance to ear rot and head smut.
 resistant to northern leaf blight and southern leaf blight.

22. The Development of Yunrui 8
 Quality testing by the Inspection and Testing Center
for Quality of Cereals and Their Products, MOA.
Reached the first level of national High-oil maize standard.
Protein content 10.99%
Lysine 0.39
Oil content 9.16%
Protein content 10.99%

23.  Released in Yunnan in 2005.
 Received “variety intellectual
rights” in China in 2010.
 Recommended as the leading
hybrid by MOA in 2010.

24. Demonstration and Application
 Yunrui 8 serves a very
important role in the
development of animal
husbandry and food safety for
farmers living in the less-
developed mountainous areas.

25. Yunrui 8 has widely
recognized at the
national and
international level.

26.  Reported in CIMMYT Informa due to the good
performance, good quality, and resistance to diseases.
Demonstration and Application

27.  From 2005 to 2018, Yunrui 8 has occupied a
cumulative area of 1 million ha. in the region.
 Yield increase of 0.7 million tons and an increased
value of 165 million US dollars.
Demonstration and Application

28.  Quality protein maize (QPM) is a valuable crop for
resource-poor families who depend on maize as their
staple.
The advantages of QPM maize
QPM Maize
 2X lysine and tryptophan in maize
endosperm.
 doubles wild-type level in nitrogen
balance index.
 90% quality value of milk.

29. —Digestibility ：
• Normal maize ： 71.45%
• QPM ： 95.00 %
—The biological valence:
• Normal maize ： 19.45%
• QPM ： 78.25%
—Net protein utilization:
• Normal maize ： 13.7%,
• QPM ： 73.8%
The nutritional value of QPM
QPM Maize

30. Dietary quality
QPM has played very important role in health improvement and
prevent Pellagra for lysine and tryptophan deficiency
QPM Maize

31. Feed quality
 QPM is efficient and economical in increasing animal weight and
sell rate.
 QPM increases 30% of live pigs daily weight gain, and 15% of
chicken egg production.
QPM Maize

32. Classification of heterotic groups: 18 QPM maize inbreds
QPM Maize
Inbred Pedigree Resource Ecology
CML149 G24QMH159-2-2-2-B-2-B-B-B-#-B Pool 24QPM Tropical
CML147 Pob63c2HC53-1-1-B-B-B-9-B-B-# Pop63 Tropical
CML140 Pob62c3HC87-2-1-#-#-1-B-#-B Pop62 Tropical
CML154 [EV8762-SR]-17-1-B-B-# Pop62 Tropical
YML23 Pob66c1HC216-4-1-2-B-B-2-B-B-B Pob66 Tropical
YML29 G25QS4B-MH13-2-B-1-2-B-1-B-B-B Pool 25QPM Tropical
YML12 G25Qc18MH520-1-1#-1-2#-5-3-B-1-B-B-B-B-# Pool 25QPM Tropical
YML102 Pob65c2HC193-2-7-2#-1-B-1-2-1-B-3-B-1-B-B Pop 65 Tropical
CML166 Introgression between tropical and temperate germplasm S. Africa & temperate germplasm Subtropical
QCML171 Introgression between tropical and temperate germplasm S. Africa & temperate germplasm Subtropical
CML161 Pool 31 QPM Subtropical
CML164 Pool 31 QPM Subtropical
CML194 UYO11-3-2-1-B S. Africa Subtropical
CA339 Derived from improved Pool 33QPM (Zhongqun 13) Pool 33 Temperate
Chang631/O2 - Temperate
Zhongxi096/O2 - Temperate
Qi205 (Wei141*Zhongxi017) *Pop70 (Zhongqun 14) Pool 34 Temperate
Xin9101/O2 - Temperate

33.  Grouping of 18 QPM inbreds and four normal maize inbreds.
QPM Maize
Classification of heterotic groups
Heterotic grouping of quality protein maize inbreds divided by SSR markers.
Acta Agro. Sin. 2003, 105-100

34. The Successful Experience in
QPM Maize Breeding

35. The Development of Yunrui 1
 National disease resistance evaluation, CAAS.
 high level of resistance to gray leaf spot.
Zhongdan
9409
Soft
endosperm
Yunrui 1
Hard
endosperm

36. The Development of Yunrui 1
 Quality testing by the Inspection and Testing Center
for Quality of Cereals and Their Products, MOA.
Reached the first level of national QPM maize standard.
Lysine 0.40
Bulk density 776.5 g/L
Protein content 9.35%

37.  Yunrui 1 has occupied a cumulative area of about
0.6 million ha.
 Yunnan
 Guangxi
 Guizhou
 Yield increase of about 0.8 million tons and an
increased value of 164 million US dollars.
Leading technology and famous product in maize production.
Demonstration and Application

38. Yunrui 1 serves a very important role in the development
of animal husbandry and nutritional improvement for farmers
living in the less-developed mountainous areas.

39. Minutes
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
mAU
0
20
40
60
80
mAU
0
20
40
60
80
Lutein
Zeaxanthin
beta-cryptoxanthin
alpha-carotene
beta-carotene
2: 450 nm, 4 nm
774
774.data
Name
High ProVA Maize
 In developing countries, nearly one quarter of all pre-
school children suffer from Vitamin A deficiency (VAD).
 A preliminary target level of 15 μg g-1 for ProVA in
maize has been set for the international agricultural
research HarvestPlus project.

40.  Molecular breeding for high ProVA concentration.
High ProVA Maize
 increase the ratio of β- to α-
carotenoids in grain.
 reduce the conversion of β-
carotene.
Yuan et al., 2010, Nature geneticsHarjes et al., Science, 2008

41. LCYE
crtRB1
crtRB1 5’TE crtRB1 D4 crtRB1 3’TE
 PCR based markers are available.
Yuan et al., 2010, Nature genetics
High ProVA Maize

42. M: Marker; -: Negative control, H2O; 1-22: Maize inbred lines.
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 - M
2000bp
1000bp
750bp
500bp
250bp
100bp
2000bp
1000bp
750bp
500bp
250bp
100bp
 Line 14 has the favorable variation.
 Lines such as 2, 4, 5, etc. have the unfavorable variation.
FavorableUnfavorable
 PCR detection of favorable lcyE 5’TE allelic variation.
High ProVA Maize
Marker-assisted selection

43.  The effects of crt-RB1 3’TE allelic variation on proVA.
Favorable allelic
variation: 543bp
Unfavorable allelic
variation: 296+1221bp
Medium variation:
296+875bp
High ProVA Maize
Marker-assisted selection
Favorable Unfavorable
Pro VA
(μg/g)

44.  The effects of lcyE 5’TE allelic variation on proVA.
Favorable allelic
variation: 150+280bp
Unfavorable allelic
variation: 250+380bp
Medium variation:
250bp
High ProVA Maize
Marker-assisted selection
Favorable Unfavorable

45.  The increase of ProVA concentrations in the QPM maize though
molecular marker assisted foreground and background selections.
High ProVA Maize
Marker-assisted selection
BMRS YMRS BMRS YMRS
A B
A: CML161 population; B: CML171 population.
Introgression of the crtRB1 gene into quality protein maiaze inbred liens using molecular markers.
Molecular Breeding, 2015, 35: 154

46.  The increasements of ProVA concentrations do NOT affect the
concentrations of proteins in the QPM maize
High ProVA Maize
Marker-assisted selection
A: CML161 population; B: CML171 population.
Introgression of the crtRB1 gene into quality protein maiaze inbred liens using molecular markers.
Molecular Breeding, 2015, 35: 154
Lysine Tryptophan TryptophanLysine
A B

47. A619
(GY807/Y32)/GY807/
GY807/GY807
(GY220/Y32)/GY220/
GY220/GY220
GY807 backcross
(A619/Y32)/Y32/Y32 (GY923/Y32)/GY923/
GY923/GY923
High ProVA Maize

48. Favorable allelic
lcyE -5’TE-2
crtRB1-3’TE-1
Recurrent Parents
BC1F1
2010 summer, Kunming
Elite Maize Inbred lines
High ProVA Maize
Hp321-1×
F1
×
Foreground selection
Foreground+Background selection
Improved inbreds
Resistance to diseases
Foreground selection
Foreground+Background selection
Selfing for several generations
High ProVA Maize
×
Recurrent Parents
2010 winter, Jinghong
2011 summer, Kunming
BC2F1
BC2F2
BC2F3
2011 winter, Jinghong
2012 summer, Kunming and Yanshan
The Successful Experience in
High ProVA Maize Breeding

49. YR 506
(Y46 × A619)
 High proVA maize based on
HPLC analysis in CAU.
 ProVA concentration: >15 ug/g.
 Varietal testing in 2010: 10% yield
increase compared to check.
 Multi-locational trials in 2011-
2015 found the hybrid performed
well.
The Successful Experience in
High ProVA Maize Breeding

50.  The yield performance of YR 506 in multi-
locational testing in 2013.
Mean yield (kg/ha.) 10,032
Yield increase
Over the check (%)
8.14
Rank 10 (total 29)
The Development of YR 506

52. Qujing
YR 506
Multi-locational trials

53. Zhaotong
YR 506
Multi-locational trials

54. Conclusions
 A combination of nutritional traits are now being formed to
allow maize to provide more of the nutritional needs for use in
human and animal consumption.
 Exploiting heterosis between tropical and temperate germplasm
led to the development of competitive commercial hybrids with
improved adaptation in Southern China.

55. Acknowledgements

56. Thank you for your attention!