Biofortification of Edible Crops with Zinc · Biofortification of Edible Crops with Zinc Philip J....

Biofortification of Edible Crops with Zinc

Philip J. White

Plant Nutritional Genomics

5th June 2012

COST FA0905, ETH Zurich

Martin R. Broadley

University of Nottingham

Mineral Malnutrition – The Hidden Hunger

INVESTMENT 1 Bundled Micronutrient Interventions, to fight hunger and improve education 2 Expanding the Subsidy for Malaria Combination Treatment 3 Expanded Childhood Immunization Coverage 4 Deworming of Schoolchildren, to improve educational and health outcomes 5 Expanding Tuberculosis Treatment 6

R&D to Increase Yield Enhancements, to decrease hunger, fight biodiversity destruction, and lessen the effects of climate change

7 Investing in Effective Early Warning Systems to protect against natural disaster 8 Strengthening Surgical Capacity 9 Hepatitis B Immunization

10 Using Low‐Cost Drugs in the case of Acute Heart Attacks in poorer nations

http://www.copenhagenconsensus.com/Projects/CC12.aspx

“If we had an extra $75 billion to put to good use, which problems would we solve first?”

Mineral Malnutrition – The Hidden Hunger

“The greatest concern lies with deficiencies in Vitamin A, iron, iodine and zinc.”

Of the 7 billion people in the world, 60-80% are Fe deficient

30% are I deficient >30% are Zn deficient

Of the agricultural soils in the world, 25-30% are alkaline with

low Fe, Zn, Cu and Mn availability

White & Broadley (2005) Trends in Plant Science 10, 586-593 White & Broadley (2009) New Phytologist 182, 49-84

Increasing Mineral Concentrations In Edible Crops

• If mineral elements are absent from the soil they must be applied to crops as soil or foliar fertilisers

• If mineral elements are present in the soil, either agronomic or genetic strategies can be developed to increase their acquisition, or mineral elements can be added as soil or foliar fertilisers


Increasing Zinc Concentrations In Edible Crops

Through Agronomy

the application of zinc fertilisers Biofortification of cereals

brassicas & potatoes

Screening potato genotypes Screening brassica genotypes

Through Genetics

select or breed varieties that accumulate zinc


Mineral Concentrations of Edible Portions Physiological Constraints


White & Broadley (2011) Frontiers in Plant Science 2:80

Movement of mineral elements to edible tissues

Mineral Concentrations of Edible Portions Physiological Constraints

White & Broadley (2011) Frontiers in Plant Science 2:80

Genotypic Variation USDA Food Composition

HarvestPlus Target

Brassica oleracea – a model crop brassica

Borecole - kale

Cauliflower and Broccoli

Cabbages

Kohlrabi

Brussels sprout

Screening Brassica oleracea (2002-2009)

10 standard genotypes at many P-fertilisation rates screened in glasshouse and field

Core collection 376 genotypes at two P-fertilisation rates in the glasshouse

90 informative genotypes AGDH genetic mapping population at two P-fertilisation rates in the glasshouse and field

74 commercial genotypes at two P-fertilisation rates in the glasshouse and field

Genetic Variation in Zinc Concentrations In Shoots of Brassica oleracea

Broadley et al. (2010) J. Hort. Sci. Biotech. 85, 375-380

h=18.5% h=0.7% h=12.4% h=13.4%

Genetic Loci Impacting Zinc Concentrations In Shoots of Brassica oleracea


Experiment Chromosome LOD Additive Effect (A12DHa allele)

GE2 (Zn both P) 2 (82.2 cM) 3.2 -5.92

9 (69.2 cM) 2.5 -7.16

GE2 (Zn high P) 5 (54.7 cM) 2.6 -4.20

FE2 (Zn all P) 1 (91.2 cM) 3.3 -1.40

3 (21.1 cM) 4.1 -1.48

7 (38.5 cM) 3.3 +1.64

FE2 (298 kg ha-1 P) 1 (89.2 cM) 2.6 -1.76

3 (21.1 cM) 3.9 -2.41

3 (43.4 cM) 4.2 -3.64

7 (52.4 cM) 3.0 +1.79

FE2 (1125 kg ha-1 P) 7 (58.4 cM) 2.8 +1.68

Effects of Phosphorus Fertilisation on Shoot Zinc Concentrations of B. oleracea

DFS, Glasshouse (GE1)


Increasing Zinc Concentrations in Shoots of B. oleracea with Zn-fertilisers

DFS, Glasshouse (GE4)


MacNicol & Beckett (1985) Plant & Soil 85, 107-129 White & Broadley (2011) Frontiers in Plant Science 2:80

Phytotoxicity Limits Zinc Biofortification of Shoots

0

200

400

600

800

1000C

ritic

al S

hoot

Zin

c (m

g / k

g)

bras

sica

lettu

ce

spin

ach

legu

mes

cere

als

sola

num

0102030405060708090

100

0 5 10 15 20

female

male

LRNI female

LRNI Male

Zinc in the UK Diet

Henderson et al. (2003) The National Diet & Nutrition Survey: Adults Aged 19 to 64 Years. London, HMSO.

Zn Intake (dietary sources, mg d-1)

Per

cent

age

of P

opul

atio

n

4% males < LRNI 4% females < LRNI

vegetables 6% potatoes 5% fruit & nuts 2%

Agronomic Biofortification of Potato Tubers with Zinc

Scottish Government Programme 7 (2011-2016)

2012

2011

Agronomic Biofortification of Potato Tubers with Zinc

White et al. (2012) J. Hort. Sci. Biotech. 87, 123-129

Biofortification of Rice Grain with Zinc

Gra

in Z

inc

(mg

kg-1

DM

)

Solution Zinc (µM)

Handao297

K150

Jiang, Struik, van Keulen, Zhao, Jin, Stomph (2008) Ann. Appl. Biol. 153, 135-147

Relationship Between Tuber Zinc and Nitrogen Concentrations

0.0 0.5 1.0 1.5

Tuber N concentration (% DM)

0.0 0.5 1.0 1.5

Tube

r Zn

conc

entr

atio

n (m

g kg

-1 D

M)

0

5

10

15

0

5

10

15

202006

y = 6.8x + 3.3R2 = 0.28

2007

2008

2009

y = 6.7x + 2.1R2 = 0.66

y = 6.3x + 3.0R2 = 0.62

y = 6.0x + 2.7R2 = 0.59

White et al. (2012) J. Hort. Sci. Biotech. 87, 123-129

Agronomic Biofortification of Cereal Grain with Zinc

Kutman et al. (2010) Cereal Chemistry 87, 1-9

Zinc biofortification of durum wheat

through soil and foliar

applications of nitrogen-fertilisers

Strategies for the Biofortification of Potato Tubers with Zinc


zinc transport in the phloem is the process limiting Zn biofortification of potato tubers

zinc is transported as Zn-nicotianamide

Option A foliar N fertilisation

Option B biosynthesis of nicotianamide

Cereals overexpressing nicotianamine synthase (NAS)

often have greater grain Zn concentrations

Overexpression of OsNAS Genes Increases Zinc Concentrations in Unpolished Rice

Johnson, Kyriacou, Callahan, Carruthers, Stangoulis, Lombi, Tester (2011) PLoS ONE 6(9): e24476

Johnson et al. (2011) PLoS ONE 6(9): e24476

Overexpression of OsNAS Genes Increases Zinc Concentrations in Unpolished Rice

Biofortification of Rice Grain with Zinc

Gra

in Z

inc

(mg

kg-1

DM

)

Solution Zinc (µM)

Handao297

K150

Jiang et al. (2008) Ann. Appl. Biol. 153, 135-147

The Genetics of Biofortification Potatoes, Brassicas & Cereals

Potatoes

Commonwealth Potato Collection Neotuberosum Collection

Commercial Core Collection Mapping Populations

Brassicas (B. oleracea)

WHRI Core Collection Mapping Populations

Cereals (barley)

Commercial Collection Mapping Populations

Bowman Mutant Collection Induced Mutant Collections

‘Phureja’ Potatoes

• Diploid

• Popular in parts of the Andes

• Selected at SCRI for UK conditions

• Excellent flavour

• Reduced cooking time

• Commercial Varieties: Mayan Gold & Inca Sun (2001), Inca Dawn (2003), Mayan Queen, Mayan Star & Mayan Twilight (2008)

Tuber Zinc Concentrations in S. tuberosum groups Phureja & Tuberosum

Subramanian (2012) PhD Thesis. University of Nottingham

Solanum tuberosum Group

Tube

r Zin

c (m

g g-

1 DM

)

H P T

16

14

12

10

8

Zinc in Tubers of Tuberosum Potato Varieties (effects of increased yield)

White et al. (2009) HortScience 44, 6 -11 Subramanian et al., data from four field trials of 23 genotypes

Tube

r Zin

c (m

g kg

-1 D

M)

Tuber Yield (kg / plot)

Genetic Loci Impacting Tuber Mineral Concentrations

Tetraploid mapping population (12601 ab1 x Stirling )

- Bradshaw et al. (2008) Theor Appl Genetics 116,193-211

Software (http://www.bioss.ac.uk/knowledge/tetraploidmap/)

QTLs impacting tuber mineral concentrations

- LG V of Stirling (QTLs for minerals, yield and maturity)

- QTLs for several mineral elements

Stirling LG V contains QTL for both maturity and several mineral elements

Zn (1

3% v

ar)

S (2

7% v

ar)

Mg

(40%

var

) M

atur

ity (5

3% v

ar)

Yie

ld (

7% v

ar)

Mn

(20%

var

)

K (2

2% v

ar)

Cu

(10%

var

)

Ca

(10%

var

)

http://www.bioss.ac.uk/knowledge/tetraploidmap/�

Zinc in Tubers of Tetraploid Potato Varieties (effects of maturity)

Subramanian et al., unpublished. (mean data from field trials in 2007 and 2008)

0

4

8

12

16

0 2 4 6 8

Maturity Score

Tube

r Zin

c (m

g kg

-1 D

M)

Potatoes (crop longevity increases tuber yield)

Intercepted radiation (MJ m-2)

0 500 1000 1500 2000

Tube

r dry

mat

ter (

g m

-2)

0

500

1000

1500

2000

Wilja

Cara

Harris (1992) The Potato Crop. Chapman & Hall, London

Genetic Loci Impacting Tuber Zinc Concentrations

Subramanian et al., unpublished.

LG B (XII) LG XIa

Zn-2

007

Zn-2

007

Zn-2

007

Zn- 2

008

LG VI LG II LG III LG XIc

Zn

-200

7

Zn

-200

9

Zn-2

009

Zn-2

007

Zn-2

007

Zn

-200

8

Zn-2

008

Zn-2

008

Zn-2

009

Stirling 12601 ab1

AtOPT3

LeNRAMP1

LeNRAMP3, HMA, YSL

LeIRT1 & LeIRT2

YSL

YSL

Combining Agronomy and Genetics To Increase Zinc Concentrations in Potatoes


0

10

20

30

40

50

60

UK Food Tables

Vales E

verest

VE-ZC-D2X

4

VE-ZC-D2X

4+U

VE-ZS-D2X

4

VE-ZS-D2X

4+U

1260

1ab1

126-Z

C-D2X

4

126-Z

C-D2X

4+U

126-Z

S-D2X

4

126-Z

S-D2X

4+U

Golden Mille

nium

GM-ZC-D2X

4

GM-ZC-D2X

4+U

GM-ZS-D2X4

GM-ZS-D2X4+

USax

on

SX-ZC-D2X

4

SX-ZC-D2X

4+U

SX-ZS-D2X

4

SX-ZS-D2X

4+U

Tube

r Zn

(μg

g-1 D

M)



0

10

20

30

40

50

Flesh Skin Flesh Skin Flesh Skincontrol D2X4 D2X4+U

Zin

c (μ

g g-

1 DM

)

Saxon (30-45 mm)

peeled potatoes = 10 mg kg-1 DM peeled biofortified Saxon potatoes = 50 mg kg-1 DM increase UK dietary Zn intake 20%



0102030405060708090

100

0 5 10 15 20

conventional

biofortified

LRNI Male

Zn Intake (dietary sources, mg d-1)

Per

cent

age

of P

opul

atio

n 4.0% to 2.2% males < LRNI

Summary

Many people’s diets lack sufficient zinc

Zinc concentrations in edible crops can be increased by agronomic or genetic strategies

Leaves generally have greater Zn concentrations than seeds or tubers

Leaf Zn concentration limited by phytotoxicity

Seed and tuber Zn concentrations limited by phloem mobility Seed and tuber Zn concentrations correlate with N concentrations

Significant natural genetic variation in Zn concentrations of brassicas

but huge environmental effects (e.g. P or Zn supply)

Zn concentration of potato tubers is associated with maturity class Minor QTL perhaps linked to genes for transport proteins


Zinc in Potato Nithya Subramanian, Gavin Ramsay, John Bradshaw, Finlay Dale, Glenn Bryan, Ralph Wilson, Jackie Thompson, Gladys Wright (JHI); Christine Hackett (BioSS); Martin Broadley, John Hammond (Nottingham).

Acknowledgements

Zinc in Brassica Martin Broadley, John Hammond, Joe Ó Lochlainn (Nottingham); Helen Bowen (WHRI); Graham King (Southern Cross); Ismail Cakmak (Sabanci); Selim Eker, Halil Erdem (Cukurova).

Summary

Many people’s diets lack sufficient zinc

Zinc concentrations in edible crops can be increased by agronomic or genetic strategies

Leaves generally have greater Zn concentrations than seeds or tubers

Leaf Zn concentration limited by phytotoxicity

Seed and tuber Zn concentrations limited by phloem mobility Seed and tuber Zn concentrations correlate with N concentrations

Significant natural genetic variation in Zn concentrations of brassicas

but huge environmental effects (e.g. P or Zn supply)

Zn concentration of potato tubers is associated with maturity class Minor QTL perhaps linked to genes for transport proteins


Biofortification of Edible Crops with Zinc · Biofortification of Edible Crops with Zinc Philip J....

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