Agronomic biofortification of crops with zinc and iron by Vajinder Pal Kalra

1. Agronomic biofortification of crops with zinc
and iron
Agronomic biofortification of crops with zinc
and iron
VAJINDER PAL KALRA
(L-2014-A-02-D)
Department of Agronomy
Punjab Agricultural University
Ludhiana

2. Biofortification
Increasing the bio-available concentrations of micronutrients in edible portions of plants
through crop management and genotype improvement
(WHO, 2002)
Agronomic biofortification is the application of
micronutrient-containing mineral fertilizer (blue
circles) to the soil and/or plant leaves (foliar), to
increase micronutrient contents of the edible part
of food crops
Valença et al (2017)
Glob Food Sec 12: 8–14

3. Need for
Biofortification

4. Dietary deficiency of essential micronutrients such as
zinc (Zn) and iron (Fe) affects more than two billion
people worldwide
(WHO, 2012)
 About 20% of deaths in children under five can be
attributed to vitamin A, Zn, Fe, and/or I deficiency
(Prentice et al, 2008)
It is estimated that over 60% of the world’s 6 billion
people are iron (Fe) deficient, over 30% are zinc (Zn)
deficient
(White and Broadley, 2009)

5. Cereals meet 60% of energy and protein needs of human
Up to 75% of the daily calorie intake of the developing world
people living in the rural areas comes only from cereal-based
foods with very low Zn concentrations (Cakmak, 2012)
Un-hulled rice -27-42 mg Zn kg-1
grain,
polished rice- 13-15 mg Zn kg-1
grain
wheat grains- 38-47 mg Zn kg-1
A diet of 300-400 g cereals day-1
will supply only 4-6 mg Zn
day-1
in the case of rice and11-18 mg Zn day-1
in the case of wheat
For a better ZnFor a better Zn nutrition ofnutrition of
human beingshuman beings,, cerealcereal grainsgrains
should contain aroundshould contain around
40-60 mg Zn kg-1
Current Situation:
10-40 mg kg-1

6. Micronutrients in whole cereal grains and legume seeds (pulses)
Plant food Fe Zn
(mg/ 100g)
Cereals Rice 4.31 1.09
Wheat 4.56 3.33
Oats 4.72 3.97
Maize 2.71 2.21
Pulses Mung bean 6.74 2.68
Chick pea 4.31 2.76
Cow pea 9.95 6.11
Soybean 15.70 4.89
Lentil 6.51 3.27
USDA Food Composition Databases
(2017) https://ndb.nal.usda.gov

7. Global mortality of children
under age of 5 years
Deficiency Deaths
Vitamin A 666,771
Zinc 453,207
Iron 20,854
Iodine 3,619
Risk factors Ratio (%)
Underweight 14.9
Unsafe sex 10.2
Unsafe water 5.5
Indoor smoke 3.7
Zinc deficiency 3.2
Iron deficiency 3.1
Vitamin A deficiency 3.0
Blood pressure 2.5
Tobacco 2.0
Cholesterol 1.9
Prasad et al (2013)
Agric. Res. 2: 111–118
Ten leading causes of illness and
disease in low income countries
Cakmak (2008)
Pl. Soil 302: 1–17

8. Cakmak (2012)
Better Crops 96: 17-19

9. Human Iron deficiency
Harvest Plus (2014)
http://www.harvestplus.org/content/iron

10. Micronutrient Deficiencies in Indian Soils
Number of
samples Zn Cu Fe Mn
East 54,061 47.3 1.4 0.4 4.9
North 64,906 51.2 1.3 12.8 3.1
South 68,863 59.9 5.1 21.6 9.6
West 63,717 34.7 19.4 7.6 2.4
All India 251,547 48.6 7.0 11.2 5.1
Percentage of soils deficient
Area
Singh, 2007
J Res Punjab Agric Univ 2 : 12-26
Nearly 50% of Indian soils are Zn-deficient

11. Temporal changes in Zn, Mn and Fe status of Punjab soils
Sadana et al (2010)
Better Crops-SA: 24-26

12. Possible solution to mineral deficiency in human population

13. Possible solution to mineral deficiency in human population
Intervention Scope Economics
Supplementation It is generally recommended during
pregnancy or in severe Zn deficiency for a
shorter period.
It is costly and only recommended when
a very quick response is required.
Fortification It is effective but limited to urban areas. It is very uneconomical if carried out for
longer period of times.
Food Diversification/
modification
It is applicable only where alternative food
products are available with high adoptability.
It is economically feasible and
sustainable intervention
Bio-fortification It is targeted and reachable It is cost effective and sustainable
approach.
It has added benefit of yield increase on
Zn deficient soils and seems permanent
solution to the problem
Hussain et al (2010)
Emir J Food Agric 22: 326-339

14. Hussain et al (2010)
Emir J Food Agric 22: 326-339
Approaches in Agronomic biofortification

15. Breeding approach
Genetic biofortification is a strategy that uses plant breeding
techniques to produce staple food crops with higher micronutrient
levels, reducing levels of anti-nutrients and increasing the levels of
substances that promote nutrient absorption
Advantage
Easily applicable
 Affordable in the target populations

16. Constraints to the breeding approach
Adverse soil chemical
properties
Release of high-yielding
zinc enriched cultivars
will face difficulties

17. Major approach for agronomic bio-fortification
Singh and Prasad (2014)
Biol Sci 84: 613–623
Sources of
fertilization
Quantity of
fertilization
Stage of
fertilization

18. Critical limits of micronutrients in soil and plant
Micronutrient Soil (ppm) Plant (ppm)
Zn Low - < 0.6 15
Fe
Low - <4.5
Medium - 4.5-18
High - >18
50
Source – Soil Practical Manual, Deptt. of Soil Science, PAU

19. Recommended daily allowance (RDA) for Indians
Group RDA (mg/day)
Zinc (Zn) Iron (Fe)
Adult man - 12 21
Adult Woman (NPNL) 10 17
Pregnant 12 35
Lactating 0-6m 12 25
Lactating 6-12m 12 5
Children 1-3 y 5 9
4-6 y 7 13
7-9 y 8 16
Adolescents Boys (10-18 y) 11-12 21-28
Girls (10-18 y) 9-12 26-27
ICMR (2010)
http://icmr.nic.in/final/rda-2010.pdf
Average daily requirement
Zn : 15-20 mg/day
Fe: 20 mg/day

20. Effects of micronutrients on crops
Dimpka and Bindraban (2016)
Agric Sustain Dev 36: 6-27

21. Grain Zn concentration of maize at different rates of Zn
application
Kanwal et al (2010)
Int J Agric Biol 12: 299–302
pH -7.8
OM- 0.72%
Extractable Zn- 0.72 mg kg-1

22. Effects of Zn application methods on the grain Zn concentration.
accumulation and grain zinc recovery (GZR) in wheat
Treatment Grain Zn concentration
(mg kg-1
)
Grain Zn accumulation
(g ha−1
)
Grain zinc recovery
GZR (%)
2008 2009 2008 2009 2008 2009
Control 18.79b 23.11d 94b 105c - -
S50 19.48b 29.11c 105b 143b 1.0b 1.7b
F4 24.40a 35.59b 118a 160a 26.4a 32.3a
F4 + S50 24.03a 43.61a 126a 193a 2.6b 3.6b
Wang et al (2012)
Field Crops Res 135:89–96
S50- Soil application of 50 kg ZnSO4·7H2O ha−1
F4- Foliar application of 4 kg ZnSO4·7H2O ha−1
(Stem elongation flowering Stage)
pH- 8.12
OM (g kg−1
)- 14.3
Extractable Zn (mg kg−1
)- 0.78

23. Effects of Zn application methods on the grain Zn
concentration and grain zinc recovery (GZR) in maize
Treatment Grain Zn concentration (mg kg−1
) Grain zinc recovery
GZR (%)
2008 2009 2008 2009
Control 14.36b 16.47c - -
S50 15.74b 19.22b 0.4b 1.6b
F4 18.50a 21.69ab 35.2a 42.9a
F4 + S50 18.28a 22.54a 2.9b 2.9b
Wang et al (2012)
Field Crops Res 135:89–96
S50- Soil application of 50 kg ZnSO4·7H2Oha−1
F4- Foliar application of 4 kg ZnSO4·7H2Oha−1
(Tasseling, 1 week after silking)
pH- 8.12
OM (g kg−1
)- 14.3
Extractable Zn (mg kg−1
)- 0.48

24. Grain Zn concentration of wheat grown with Zn fertilizer treatment
Country Location pH and
DTPA-Zn
(mg kg−1
)
Year Grain Zn concentration (mg kg–1
) CD
(p=0.05)Nil Soil Zn Foliar Zn Soil +
foliar Zn
China Quzhou 7.8 & 0.32 2009 27.7 32.4 47.2 53.9 3.5
2010 29.5 40.9 44.3 52.0 6.8
Yongshou 8.0 & 0.31 2009 18.8 21.0 26.5 22.5 3.8
2010 19.5 22.1 31.4 34.0 3.5
India Varanasi 7.7 & 0.86 2008 29.0 32.0 44.0 47.0 12.0
Kapurthala 7.6 & 2.2 2010 49.0 52.0 64.8 65.3 7.4
2011 31.4 30.2 51.1 49.1 7.9
Ludhiana 7.6 & 6.5 2010 25.5 30.3 61.0 60.8 7.5
2011 27.3 36.7 58.3 57.0 6.4
Pakistan Faisalabad - & 1.3 2008 29.0 29.0 60.0 59.0 9.0
Mean 27.4 30.5 48.0 49.0
% increase over nil 12.3 83.5 89.7
Zou et al (2012)
Pl Soil 361:119–130
pH- 7.5-8.2
Extractable Zn (mg kg−1
)- 0.3-6.5
S50- Soil application of 50 kg ZnSO4·7H2Oha−1
F4- Foliar application of 0.5 % ZnSO4·7H2O

25. Effect of rate, source and method of Zn application on Zn concentration in oat grains and straw
Treatment (mg kg-1
grain) (mg kg-1
straw)
2009-10 2010-11 2009-10 2010-11
Control (no Zn) 22.0 22.5 31.3 32.2
2 kg Zn ha-1
as ZnSHHa, deep placed at sowing 25.1 (14.1) 25.6(13.8) 40.4 (29.1) 41.2 (27.9)
2 kg Zn ha-1
as ZnSHH, broadcast at final ploughing 23.8 (8.2) 24.3 (8.0) 38.5 (23.0) 39.3 (22.0)
2 kg Zn ha-1
as ZnSHH
(used for coating onto 100 kg seed required for sowing one hectare)
32.0 (45.5) 32.5 (44.4) 48.6 (55.3) 49.3 (53.1)
2 kg Zn ha-1
as ZnO deep placed at sowing 24.7 (12.3) 25.2 (12.0) 39.6 (26.5) 40.3 (25.2)
2 kg Zn ha-1
as ZnO broadcast at sowing 22.1 (0.5) 23.6 (4.9) 36.9 (17.9) 37.8 (17.4)
2 kg Zn ha-1
as ZnO
(used for coating onto seed required for sowing 1 ha)
31.6 (43.6) 32.3 (43.5) 47.5 (51.8) 48.2 (47.7)
5 kg Zn ha-1
as ZnSHH broadcast at sowing 29.5 (34.1) 30.3 (34.7) 45.9 (46.6) 46.8 (45.3)
5 kg Zn ha-1
as ZnSHH band placed before first irrigation (25 DAS) 29.0 (31.8) 29.5 (31.1) 42.5 (35.8) 42.5 (32.0)
5 kg Zn ha-1
as ZnSHH band placed after first irrigation (25 DAS) 27.3 (24.1) 27.8 (23.5) 40.4 (29.1) 41.3 (28.3)
5 kg Zn ha-1
as ZnO broadcast at sowing 28.6 (30.0) 29.1 (29.3) 42.7 (36.4) 43.5 (35.1)
5 kg Zn ha-1
as ZnO band placed before first irrigation (25 DAS) 28.0 (27.3) 28.4 (26.2) 41.8 (33.5) 42.3 (31.4)
5 kg Zn ha-1
as ZnO band placed after first irrigation (25 DAS) 26.8 (21.8) 27.3 (21.3) 40.6 (29.7) 41.2 (28.0)
LSD (P = 0.05) 1.90 1.87 1.61 1.47pH- 7.8, OM – 0.48% Extractable Zn (mg kg−1
)- 0.63 Shivay et al (2013) Agric Res 2:375–381

26. Treatment Grain
yield
(t ha-1
)
Zn concentration
in unhusked rice
(mg kg-1
)
Zn concentration
in polished rice
(mg kg-1
)
BREZn
(%)
Control 3.92 30.4 26.1 -
25 kg ZnSO4.7H2O ha-1
(5.3 kg Zn ha-1
) Soil application 5.20 47.5 40.3 1.42
0.2% ZnSO4.7H2O foliar application (1.2 kg Zn ha-1
) 4.99 52.6 28.8 2.42
Soil application of 1% ZnO-coated urea (2.6 kg Zn ha-1
) 4.48 38.2 32.4 1.16
Soil application of 2% ZnO-coated urea (5.2 kg Zn ha-1
) 5.13 44.7 37.9 1.24
Soil application of 1% ZnSO4.7H2O coated urea (2.6 kg Zn ha-1
) 4.69 40.3 34.1 1.55
Soil application of 2% ZnSO4.7H2O coated urea (5.2 kg Zn ha-1
) 5.27 49.7 42.1 1.61
CD (p=0.05) 0.45 4.5 - -
Effect of method, source, and rate of Zn application on grain yield, Zn content and bio-
fortification recovery efficiency of basmati rice
Prasad et al (2014)
Adv Agron 125: 55-91
pH- 8.2,
OC- 0.51%
DTPA extractable Zn- 0.36 mg kg-1
Urea-2 split
Foliar-1 spray

27. Grain Zn concentration in wheat due to degree of Zn enrichment of urea
Wheat
Prasad et al (2014)
Adv Agron 125: 55-91

28. Effect of source and method of Zn application on Zn concentration in grain
and stover in maize
Treatment (Zn ha-1
) Grain Zn concentration
(mg kg-1
grain)
Stover Zn concentration
(mg kg-1
DM)
Control 40.2 45.0
5 kg to soil 44.2 49.2
1 kg foliar 46.0 59.2
5 kg to soil + 1 kg foliar 49.2 64.5
2.83 kg through Zn-coated urea (to soil) 45.8 58.2
CD (p=0.05) 2.0 2.7
Shivay and Prasad (2014)
Egyptian J Biol 16: 72-78
pH- 8.3
OC- 0.37%
DTPA-extractable Zn- 0.36 mg kg-1
*Foliar Spray- Tasseling and Initiation of silking

29. Effect of various Zn fertilizer treatments on grain zinc (mg kg–1
) of rice during 2010-11
at four locations in Punjab, India
Location
(2011)
No Zn Soil Zn Soil Zn+
foliar Zn
Soil Zn +foliar Zn
+propiconazole
CD (p=0.05)
Patiala 20.1 22.1 28.6 28.5 3.3
Ferozepur 21.2 23.1 27.9 27.8 3.9
Bhagatpur 20.6 21.9 29.1 29.2 3.7
Mean 20.6 22.4 28.5 28.5 3.6
% Increase 8.7 38.3 38.4
Location
(2010)
No Zn Soil Zn+
foliar Zn
Soil Zn +foliar Zn
+propiconazole
CD (p=0.05)
Patiala 18.1 26.5 26.4 4.2
Ferozepur 22.2 29.5 29.6 5.1
Bhagatpur 18.2 25.9 26.1 5.3
Mean 19.5 27.3 27.4 4.9
% Increase 40.0 40.5
Ram et al (2015)
Current Sci 109
Soil Zinc-50 kg ZnSo4.7H2O/ha
Foliar- Two spray (0.5% ZnSo4.7H2O)

30. Effect of various Zn fertilizer treatments on grain zinc (mg kg–1
) of
wheat at four locations in Punjab, India
Location No Zn Soil Zn Soil Zn+
foliar Zn
Soil Zn +foliar Zn
+propiconazole
CD
(p=0.05)
Zn -deficient Bathinda 22.7 24.7 72.2 73.5 18.4
Gurdaspur 25.3 27.2 60.4 60.3 13.9
Mean 24.0 25.9 66.3 66.9 16.1
% Increase 7.9 175.8 178.5
Zn-sufficient Ferozepur 26.5 28.7 61.6 61.9 18.2
Patiala 30.2 31.2 61.2 59.2 16.6
Bhagatpur 22.4 25.3 64.2 63.2 13.0
Mean 26.3 28.4 62.4 61.4 15.9
% Increase 7.9 136.7 133.2
Overall mean 25.4 27.4 63.9 63.6 0.2
% Overall increase 7.9 151.5 150.3
Ram et al (2015)
Current Sci 109: 1-6
Soil Zinc-50 kg Znso4.7H2O/ha
Foliar- Ear formation, milking stage (0.5% Znso4.7H2O)

31. Treatment Zn concentration in rice grain (mg kg-1
) Zn mobilization efficiency index (ZnMEI)
2010 2011 2010 2011
Absolute control 20.7 21.2 0.28 0.28
NPK (120 kg N+26.2 kg P+60 kg K) 23.1 23.6 0.29 0.29
NPK+5kg Zn ha-1
through ZnSHH as SA 26.4 26.9 0.30 0.30
NPK+ ZnSHH 0.2 % FSAT 24.8 25.3 0.29 0.29
NPK+ ZnSHH 0.2 % FSAT+B stages 26.3 26.8 0.30 0.30
NPK+ ZnSHH 0.2 % FSAT + B+ GF stages 26.8 27.3 0.30 0.30
NPK+ ZnSHH 0.5 % FSAT 25.4 25.9 0.30 0.30
NPK+ ZnSHH 0.5 % FSAT +B stages 26.6 27.1 0.29 0.30
NPK+ ZnSHH 0.5 % FSAT+ B+ GF stages 28.2 28.7 0.30 0.30
NPK+ 5kgZnha-1
through Zn–EDTA as SA 27.8 28.3 0.30 0.30
NPK+ Zn–EDTA 0.2 % FSAT 24.7 25.2 0.29 0.29
NPK + Zn–EDTA 0.2 % FSAT+B stages 26.6 27.1 0.30 0.30
NPK+ Zn–EDTA 0.2 % FSAT+ B+ GF stages 27.7 28.2 0.30 0.30
NPK+ Zn–EDTA 0.5 % FSAT 25.8 26.3 0.29 0.29
NPK+ Zn–EDTA 0.5 % FSAT+ B stages 28.2 28.7 0.30 0.30
NPK + Zn–EDTA 0.5 % FSAT +B +GF
stages
29.8 30.3 0.31 0.31
CD (p=0.05) 2.1 1.7 0.01 0.02
Effect of sources, time and method of zinc application on Zn concentration in grain of
basmati rice
Shivay et al (2015)
Proc Natl Acad Sci 86 (4): 973-984.
pH - 7.8, OC- 0.49 %
DTPA-extractable Zn- 0.43 mg kg-1

32. Effect of sources, time, and method of Zn application on Zn concentrations in grain
and straw of chickpea
Treatment Zn concentration (mg kg grain−1
) Zn concentration(mg kg straw−1
)
2011-12 2012-13 2011-12 2012-13
Check (no Zn) 42.6 41.4 18.3 17.1
ZnSHH soil at 5 kg Zn ha−1
51.9(21.8) 50.7(22.5) 22.6 21.3
ZnSHH one spray (V) 49.8(16.9) 48.5(17.1) 22.8 21.5
ZnSHH two sprays (V + F) 54.7(28.4) 53.4(29.0) 27.1 25.8
ZnSHH, three sprays (V + F + G) 58.4(37.1) 57.1(37.9) 32.5 31.2
Zn-EDTA soil at 2.5 kg Zn ha−1
52.6(23.5) 51.3(23.3) 24.6 23.4
Zn-EDTA one spray (V) 51.2(20.2) 50.1(21.0) 25.1 24.0
Zn-EDTA two sprays (V + F) 58.1(36.4) 56.7(39.4) 28.3 27.1
Zn-EDTA three sprays (V + F + G) 72.3(69.7) 63.5(53.4) 33.9 32.6
LSD (P = 0.05) 3.31 3.33 3.51 1.81
ZnSHH= Zn sulphate hepta hydrate V= active vegetative stage, F= flowering stage, G= grain filling stage a% increase over check (biofortification)
pH- 8.1, OM-0.47%
Available Zn (mg kg−1
)- 0.67
Shivay et al (2015)
Commun Soil Sci Plan 46:2191-2200

33. Effect of sources, time, and method of Zn application on Zn uptake in grain and
straw of chickpea
Treatment Zn uptake in grain (g ha−1
) Zn uptake in straw (g ha−1
)
2011-12 2012-13 2011-12 2012-13
Check (no Zn) 78.5 71.3 78.0 68.5
ZnSHH soil at 5 kg Zn ha−1
102.3 93.9 104.2 93.9
ZnSHH one spray (V) 96.3 87.9 103.3 92.8
ZnSHH two sprays (V + F) 112.3 103.2 128.6 116.2
ZnSHH, three sprays (V + F + G) 124.9 114.8 166.8 152.0
Zn-EDTA soil at 2.5 kg Zn ha−1
102.7 93.9 114.5 103.5
Zn-EDTA one spray (V) 98.8 90.9 117.0 106.0
Zn-EDTA two sprays (V + F) 125.4 115.8 139.2 126.6
Zn-EDTA three sprays (V + F + G) 162.8 135.4 181.0 148.9
LSD (P = 0.05) 14.93 15.52 10.45 20.25
ZnSHH= Zn sulfate hepta hydrate V= active vegetative stage, F= flowering stage, G= grain filling stage a% increase over check (biofortification)
pH- 8.1, OM-0.47%
Available Zn (mg kg−1
)- 0.67
Shivay et al (2015)
Commun Soil Sci Plan 46:2191-2200

34. Zinc concentrations in brown rice from plants grown without Zn treatment and with foliar Zn
treatment alone or in combination with pesticide in 3 countries
Country Location
(pH, OM (%) and
Extractable Zn
(mg kg-1
)
Year Brown rice Zn (mg kg-1
) LSD (p=0.05)
No Zn Foliar Zn Foliar Zn+ Pesticide
India Ludhiana
(7.6, 0.25 and 0.58)
2012 19.8b 25.1a 26.5a 3.1
2013 19.1b 23.5a 23.0a 1.5
Gurdaspur
(7.5, 0.29 and 0.55)
2012 18.7b 23.5a 23.4a 2.0
2013 17.8b 21.8a 22.1a 2.2
China Jiangsu-Rudong
(8.4, 0.82 and 0.33)
2012 17.3b 22.7a 20.1a 2.3
2013 19.8b 22.0a 23.2a 2.2
Anhui-Changfeng
(6.4, 0.46 and 0.37)
2012 19.8b 22.9a 21.1ab 1.9
2013 23.0b 31.9a 31.7a 3.4
Thailand CMU
(7.7, 1.50 and 0.90)
2011 21.2c 30.2a 25.4b 3.1
2012 26.0a 28.2a 28.1a NS
Takli
(6.2, 3.70 and 0.50)
2011 13.9b 22.5a 21.0a 2.8
2012 12.5a 14.9a 17.3a NS
Mean 19.1b 24.1a 23.6a 1.3
Ram et al (2016)
Pl Soil 403: 389-401

35. Foliar application Grain Yield (g plant-1
) Fe concentration in Grain (mg kg-1
)
Control 2.71 36
Control + Urea 3.34 36
FeSO4 2.73 38
FeSO4+ Urea 2.69 43
FeEDTA 3.07 38
FeEDTA + Urea 3.38 42
FeEDDHA 3.11 35
FeEDDHA + Urea 2.61 39
Fe Citrate 2.54 36
Fe Citrate + Urea 2.97 37
CD (p=0.05) N.S 5
Changes in grain yield and Fe concentrations in wheat treated by various
foliar Fe fertilizers with and without urea
Aciksoz et al (2011)
Pl Soil 349:215–225
pH- 8
DTPA-Extractable Fe- 2.1 mg kg-1
DTPA-extractable Zn- 0.1 mg kg-1
Fe-0.25% (w/v)
Urea -1 % (w/v)

36. Effect of Ferti-fortification with Fe on grain yield, grain Fe concentration and
uptake in different maize cultivars
Maize
cultivars
Grain yield
(q ha-1
)
Grain Fe Conc.
(mg kg-1
)
Grain uptake
(g ha-1
)
Control Fe spray Mean Control Fe spray Mean Control Fe spray Mean
PMH 1 42.82 45.60 44.21 23.53 38.23 30.88 1007.56 1743.29 1375.42
JH 3459 43.11 44.12 43.62 32.57 39.90 36.24 1404.0 1760.39 1582.24
30V92 46.33 49.00 47.67 31.23 38.53 34.88 1446.89 3814.47 2630.68
Prabhat 38.71 40.33 39.52 25.80 36.23 31.02 998.72 1461.16 1229.94
Navjot 41.89 43.56 42.73 28.37 39.57 33.97 1188.42 1723.67 1456.04
Mean 42.57 44.52 48.55 28.30 38.49 33.40 1209.13 2100.59 1654.87
CD (0.5%) 6.55 4.86 - 3.65 3.84 - 45.66 55.67 -
Dhaliwal et al 2013
Ind J Fert 9(8): 52-57.Fe spray-1% (3 sprays)
Knee high stage, Pre tasseling, Post tasseling

37. Effect of foliar spray of FeSO4.7H2O on Fe concentration in different rice cultivars
Treatment Rice cultivars
PR 113 PR 116 PR 118 PR 120 PAU 201
Fe concentration in rice (mg kg-1
)
Control 15.2 14.8 13.0 17.8 12.5
0.5 % FeSO4 18.8 20.5 19.7 20.2 19.8
% increase over control 23.6 38.5 51.5 13.4 58.4
1 % FeSO4 26.4 25.8 26.5 28.2 28.8
% increase over control 73.6 74.3 103.8 58.4 130.4
CD (p=0.05) NS 3.1 1.1 6.2 5.7
Singh et al (2013)
J Res Punjab Agric Univ 50: 32-38
pH- 7.9
OC- 0.22%
Extractable Fe (mg kg-1
) - 5.28
Spray-Maximum tillering, Pre-anthesis and Post-anthesis stages

38. Effects of different treatment of foliar application of iron sulfate on iron
contents (mg kg-1
) in leaves, stems and grains of mungbean
Treatment Iron content (mg kg-1
)
Leaves Stems Grains
T1 (Control) 511.37h 380.07g 78.50g
T2 (0.5% FeSO4 at branching) 601.73g 470.42f 90.43f
T3 (0.5% FeSO4 at flowering) 623.70f 488.17e 96.10e
T4 (0.5% FeSO4 at branching + 0.5% FeSO4 at flowering) 675.43d 520.24d 101.50e
T5 (1.0% FeSO4 at branching) 654.07e 515.22d 96.83e
T6 (1.0% FeSO4 at flowering) 668.37de 505.16de 99.60e
T7 (1.0% FeSO4 at branching + 1.0% FeSO4 at flowering) 717.17b 585.54b 127.80b
T8 (1.5% FeSO4 at branching) 672.60c 550.33c 115.73d
T9 (1.5% FeSO4 at flowering) 698.70c 559.51c 121.43c
T10 (1.5% FeSO4 at branching + 1.5% FeSO4 at flowering) 794.90a 634.27a 146.43a
Ali et al (2014)
Pak J Life Soc Sci 12(1): 20-25.
pH- 7.5
OC- 0.96%
DTP A- extractable Fe (mg kg-1
) - 3.0

39. Application of PGPR alone and in combination with iron (Fe) on iron
uptake indifferent plants parts of chickpea
Fe Concentration (mg 100 g-1
)
Grains Shoot Root
Absolute control 1.20g 0.66g 0.14g
Fe @ 5.6 kg ha-1
2.40f 1.80f 0.86f
S1 3.26de 2.23e 1.40ce
S2 3.30ce 2.50cd 1.30e
S3 3.36be 2.26e 1.33de
S4 3.20e 2.36de 1.36ce
S5 3.40be 2.40b 1.30e
S1+Fe @ 5.6 kg ha-1
3.60bc 2.73b 1.70a
S2+Fe @ 5.6 kg ha-1
4.36a 3.16a 1.56ab
S3+Fe @ 5.6 kg ha-1
3.50be 2.80b 1.50bc
S4+Fe @ 5.6 kg ha-1
3.53bd 2.70b 1.50bc
S5+Fe @ 5.6 kg ha-1
3.63b 2.63bc 1.46bd
Khalid et al (2015)
Pak J Bot 47: 1191-1194
pH- 6.5
OC- 0.6%
DTP A– extractable Fe (mg kg-1
) - 4.5

40. Effects of Zn application methods on the Fe concentrations (mg kg−1
)of
spring maize and winter wheat grains
Treatment Spring maize Wheat
2008 2009 2008 2009
Control 17.7b 18.5b 31.4a 34.4a
S50 18.7b 18.4b 31.3a 35.7a
F4 21.6a 20.9a 32.3a 35.8a
F4 + S50 20.9a 20.5a 29.6a 36.4a
S50- Soil application of 50 kg ZnSO4·7H2O ha−1
F4- Foliar application of 4 kg ZnSO4·7H2Oha−1
Wang et al (2012)
Field Crops Res 135:89-96
Parameters Wheat Maize
2008 & 2009 2008 2009
pH 8.12 8.20 8.27
OM (%) 1.43 1.17 1.60
Extractable Zn (mg kg-1
) 0.78 0.63 0.48
Extractable Fe (mg kg-1
) 0.94 5.84 4.99
Tasseling and 1 week after silking for maize,
Stem elongation and flowering stage for wheat

41. Effects of Zn application methods on the Fe concentrations (mg kg−1
)of
spring maize and winter wheat grains
Maize grain Wheat grain

42. Effect of Zn and Fe sprays on their respective concentration in grains of
different wheat cultivars
Treatment PBW 343 PBW 550 PBW 17 PDW 233 PDW 274 PDW 291 Average
Concentration of Zn (mg kg-1
) in wheat grains with foliar Zn
-Zn 21.42 20.56 21.38 20.35 23.89 23.36 21.91
+Zn (F) 24.18 26.14 26.39 21.60 25.56 24.56 24.74
% increase 12.62 27.15 20.81 6.16 7.07 5.15 13.16
Concentration of Fe (mg kg-1
) in wheat grains with foliar Fe
-Fe 37.42 39.14 40.47 38.90 39.14 41.99 39.51
+Fe 47.70 45.27 48.90 44.27 46.65 45.89 46.45
% increase 28.00 15.66 20.99 13.76 19.17 9.27 17.81
Dhaliwal et al (2014)
VEGETOS 27: 139-145
pH- 7.6
Extractable Zn (mg kg-1
)- 0.74
Ectractabe Fe (mg kg-1
)- 4.76
4 sprays- Tillering, flower initiation, milk, dough

43. The iron content in the seed of cowpea bean
cultivar in response to different application rates
of ferrous sulfate and ferrous chelate
Márquez-Quiroz et al (2015)
J Soil Sci Plant Nutr 15 : 839-847
Percentage increase over control
100 μM L-1
ferrous sulphate = 29.4
50 μM L-1
ferrous chelate = 32.0
The effects of Zinc, iron, and manganese fertilizer
applications on concentration of these elements in
soybean seed
Kobraee et al (2013)
Annu Rev Bio 3: 83-91
pH- 7.6, OM-2.3%
Extractable Zn (mg kg−1
)- 0.71
Extractable Fe (mg kg−1
)- 2.6
Extractable Mn (mg kg−1
)- 6.3
Ferrous sulphate
Ferrous chelate

44. Effect of Fe, B and Zn on nutrient concentration in cowpea
Treatment Fe B Zn
mg kg-1
Control 40.00 16.00 8.00
Fe 1 ppm 90.00 31.00 13.00
Fe 2 ppm 154.00 47.00 17.00
B 1 ppm 51.00 31.00 18.00
B 2 ppm 58.00 40.00 24.00
Zn 1 ppm 47.00 26.00 25.00
Zn 2 ppm 50.00 37.00 42.00
LSD (P = 0.05) 1.28 1.35 1.35
Salih (2013)
J Agric Vet Sci 6(3): 42-46.
pH- 7.9 and OM (%)- 0.8
Extractable Zn (mg kg-1
)- 20
Extractable Fe (mg kg-1
)- 30
Extractable B (mg kg-1
)- 5

45. Parameters N levels (kg/ha)
N-0 N-low N-opt N-over
Grain ZnC (mg kg-1
) 15.2b 15.2b 16.6ab 17.3a
SZnC (mg kg-1
) 18.2ab 17.6b 20.9ab 21.8a
Grain FeC (mg kg-1
) 13.4c 15.5b 16.0ab 17.2a
SFeC (mg kg-1
) 63.3a 69.6a 58.3a 60.6a
Effect of nitrogen levels on Zn and Fe content in grain and stover of
maize
N-low= 75% of N-opt
N-opt= 240 kg N ha-1
N-over= 150 % N-opt
Xue et al (2014)
PLOS ONE 9: 1-12
pH 8.3
OM- 1.4%

46. Shoot and root iron contents, and iron translocation index of 14 day old plants of wheat cultivars
raised under different zinc and iron treatment combinations
Hanjagi and Singh (2016)
Ind J Pl Physio (Online) Dec 2016
T1= Fe 0 µM and Zn 0 µM
T2= Zn 1 µM and Fe 0 µM
T3= Zn 1 µM and Fe 10 µM
T4= Zn 1 µM and Fe 50 µM
T5= Fe 10 µM and Zn 0.5 µM
T6= Fe 100 µM and Zn 0 µM
T7= Fe 100 µM and Zn 0.5 µM
T8= Fe 100 µM and Zn 1.0 µM
T9= Fe 100 µM and Zn 10 µM
In nutrient solution culture
pH: 5.6-5.8
( mg kg-1
dry wt. )

47. Conclusion
Biofortification offer sustainable solutions to the escalating micronutrient-related
malnutrition problems
Two-three foliar sprays of Zn and Fe (0.5% ZnSO4 and FeSO4) on later growth stages offers a
practical and useful means for bio-fortification with Zn and Fe
Concentration of micronutrients increases 60-80% in cereal grains and 50-65% in pulses
over control
Foliar application of micronutrients results significantly higher micronutrient recovery
percent over soil application

48. Focus onFocus on betterbetter food,food,
not onlynot only moremore foodfood