UPTAKE AND MINIMIZE

CADMIUM IN RICE
Maulana Malik Nashrulloh

BIT 691

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 OUTLINES
• Introduction
o Essential and non-essential elements in plant.
o Role of Transition Metals in Plant
o Brief introduction of Cadmium contamination in rice
o Brief introduction about methods in reducing cadmium uptake by
rice

• Uptake of Cadmium (Cd) in rice

• Selected methods for minimizing Cd uptake in rice

o Soil Amendments
o Bioremediation 2
 ESSENTIAL AND NON-ESSENTIAL
ELEMENTS IN PLANT
ESSENTIAL ELEMENTS

C, O, H
Structural Elements
BENEFICIAL, NON-BENEFICIAL,
NON-ESSENTIAL NON-ESSENTIAL
ELEMENTS N, P, K ELEMENTS
Primary Macronutrients
Na, Al, Si, Se, V Cd, Pb, Hg, Cr
Ca, Mg, S
Secondary Macronutrients

Some of them are

Fe, Mn, Transition Metals
Co, Zn,
Ni, Mo, B,
Cl
Micronutrients 3

Fageria, 2013; Marschner, 2017; White and Pongrac, 2017

 ROLE OF TRANSITION METALS IN PLANT
Critical leaf concentration (mg/g DM)
Element Biological Functions
Sufficiency Toxicity
Fe Photosynthesis; mitochondrial respiration; C and N metabolism; 50–150 × 10–3 >0.5
production and scavenging of ROS; regulation of transcription and
translation; hormone biosynthesis
Mn Photosystem II; enzyme activation in photosynthesis, C and N 10–20 × 10–3 0.2–5.3
metabolism, RNA polymerase
Cu Photosynthesis; mitochondrial respiration; C and N metabolism; 1–5 × 10–3 15–30 × 10–3
protection against oxidative stress
Zn Structural stability of proteins; regulation of transcription and 15–30 × 10–3 100–700 × 10–3
translation; oxidoreductases and hydrolytic enzymes
Ni Constituent of Urease 0.01 × 10–3 20–30 × 10–3
Mo Catalytic site of nitrate reductase, aldehyde oxidase, xanthine 0.1–1.0 × 10–3 1
dehydrogenase and sulfite oxidase
Co Nitrogen fixation (Trace)4 10–20 × 10–3
White and Pongrac, 2017
 ROLE OF TRANSITION METALS IN PLANT
Critical leaf concentration (mg/g DM)
Element Biological Functions
Sufficiency Toxicity
Pb - - 10–20 × 10–3
Cd - - 5–10 × 10–3
Hg - - 2–5 × 10–3
Cr - - 1–2 × 10–3

White and Pongrac, 2017

 RICE
Global Staple Food Demand
(% Population)

Rice
23%
Wheat

Corn
50%
17% Other sources

10%

Khush, 2003
 BIOGEOCHEMICAL CYCLE OF
CADMIUM AND RICE CONTAMINATION

Sebastian et al., 2013

 WATER REGIME AFFECT Cd UPTAKE

Slamet-Loedin et al., 2015

 CADMIUM TRANSLOCATION TO RICE

Clemens et al., 2013

 CADMIUM EFFECT TO RICE PLANT
Observable Phenotypic
Performance:
1. Height
2. Number of Tillers
3. Root Length
4. Root Number
Normal Cd
Normal Cd

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Normal
Zhang et al., 2018 Cd Normal
Cd
 CADMIUM CONCENTRATION
PARTITIONING BY ORGAN IN RICE

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Zhang et al., 2018

 CADMIUM CONTAMINATED RICE
COMPROMISE HUMAN HEALTH

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Khaokaew et al., 2011
Ruppel, 2011
REGULATIONS OF MAXIMUM TRESHOLD
OF CADMIUM IN SOIL
Organizations Maximum Limit (mg/kg) Regulation
China 0.3 – 0.6 GB15168-1995
Country-specific Indonesia 2 MoA Regulation 70/2011
Finland 1 Gov. Regulation 214/2007

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REGULATIONS OF MAXIMUM THRESHOLD
OF CADMIUM IN POLISHED RICE
Organizations Maximum Limit (mg/kg) Regulation
Codex Alimentarius 0.4 CXS 193-1995
Supranational Bodies Comission (FAO/WHO)
European Commission (EU) 0.2 EU 488/2014
China 0.2 GB2762-2012
Indonesia 0.4 SNI 7387:2009
Australia & New Zealand 0.1 Australia New Zealand
Food Standards Code,
Country-specific Schedule 19, 2016
Hongkong 0.1 Cap. 132v, Section 55(1),
1983
Japan 0.4 Japan Food Standards,
2011
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 APPROACHES IN MINIMIZING Cd
UPTAKE IN RICE

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Sebastian et al., 2013

Cd UPTAKE BY RICE

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Ishikawa et al., 2011

 RICE SEEDLINGS PREPARATION
Rice Seed Moved to indoor
Indica cultivars Japonica cultivars
(High Cadmium (Low Cadmium
accumulation) accumulation) Exposed to a short-day treatment with
an 8-hour photoperiod, day/night
temperatures of 30°C/25°C, relative
Seeded in Kimura B solution-dipped humidity of 70%, and light
mesh intensity of 400 μmol m -2 s -1

Grown in a greenhouse using

natural sunlight (2 – 3 w)

Rice Seedling

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Ishikawa et al., 2011

 POSITRON-EMITTING TRACER
IMAGING SYSTEM (PETIS) IMAGING
Rice Seedling γ-ray emission from 107Cd decay
recorded using PETIS apparatus
Indica cultivars Japonica cultivars
(4 min interval, until 36 h)
(High Cadmium (Low Cadmium
accumulation) accumulation)
Image analysis using NIH Image J
Moved to 0.5 mM CaCl2 solution, 24 h
prior application of 107Cd

Fed by 7.6 - 60.2 MBq 107Cd carried by

0.1 mM non-irradiated Cd

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Ishikawa et al., 2011

 PETIS SETUP
PETIS
Target
Pump Region
Siphon
Tube

Secondary Primary
Reservoir Reservoir

Culture PETIS
Solution Sensor

Automatic
Balance
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Figure taken from Fujimaki et al., 2010, same setup used in experiment
 Japonica Indica Japonica Indica

Root

Leaf

Stem

Panicle and
Flag Leaf

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Ishikawa et al., 2011

 SUMMARY
• Cd is really mobile element that can be traveled from lower part to the upper
part of the rice plant.
• Cd accumulation and translocation depend on the variety and cultivars used.
Japonica cultivars absorb less Cd than Indica cultivars.
• Cd accumulation in rice organs accumulated in different part. Upon uptake,
Cd mainly accumulated in root, stem nodes, grains.

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 EFFECT OF SOIL AMENDMENTS IN
LIMITING UPTAKE OF Cd IN RICE BY
IMMOBILIZATION

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Mori et al. 2016

FERMENTED BARK AMENDMENT
(FBA) PREPARATION
Cedar Wood Bark

Fermented with White Rod Fungus and

Bean Curd (1:1:1)
Fermented until reach Moisture Content
15%

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 RICE CULTIVATION AND Cd
DETERMINATION
FBA
(0.1 – 2% Rice Sampled for soil and plant tissues
w/w)
Rice Bran
5 kg Soil Analysis using ICP-OES
(1% w/w) Planted separately
Bark
Compost
(1% w/w) Fertilized by 0.5 g N:P:K
(14:14:14)/pot

Divided into watered (3), watered

with midterm-drying (3), and
unwatered (3)

Grown for 120 d

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 FBA SUPPLEMENTATION EFFECT TO
CADMIUM UPTAKE INTO BROWN RICE

FBA = Fermented Bark Amendment (% w/w)

RB = Rice Bran (1% w/w)
BC = Bark Compost (1% w/w) 27

Mori et al. 2016

 FBA AFFECT Cd TRANSLOCATION IN
RICE PLANT

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Mori et al. 2016

 FRACTIONATION OF Cd BY
SEQUENTIAL EXTRACTION
Sample (Soil)

Sequential Extraction Chloride Extraction Chloride Fraction

AAS (λ = 228.8 nm) Carbonate Extraction Carbonate Fraction

Bound with Fe/Mn-Oxides Bound with Fe/Mn-Oxides

Extraction Fraction

Bound with Organic Matter Bound with Organic Matter

and Sulfides Extraction and Sulfides Fraction

Residual Residual Fraction

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 FBA SUPPLEMENTATION EFFECT IN Cd
FRACTION IN SOIL

F1 = Chloride (Exchangeable)
F2 = Bound with Carbonate
F3 = Bound with Fe/Mn Oxide
F4 = Bound with organic matter and sulphide 30
F5 = Residual
Mori et al. 2016
 SUMMARY
• FBA supplementation help immobilization of Cd uptake by rice.
• Best result achieved by applying FBA 1% with watering.
• Cd concentration in rice in treated rice lower (0.039 mg/kg), compared to
untreated rice (0.39 mg/kg).
• FBA supplementation increase Cd bound with organic matter and sulphide.

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 EFFECT OF MICROBIAL-ASSISTED
PHYTOREMEDIATION IN REDUCING
Cd IN SOIL

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Li et al., 2017
 INFORMATION REGARDING USED
STRAINS
Bacillus megaterium H3 Neorhizobium huatlense T1-17
IAA Production Yes Yes
Siderophore Production Yes Yes
Arginine Decarboxylase Yes N/A
ACC Deaminase No Yes

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Li et al., 2016
Chen et al., 2016
 HEAVY METAL IMMOBILIZATION
MECHANISMS BY BACTERIA

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Ahemad, 2014
Cd IMMOBILIZATION BY BACTERIA
ASSAY
B. megaterium H3 and N. huautlense T1-
17

Grown in 50 ml Luria Broth (LB) +

10 µM CdSO4

Cultured in 30°C, 180 rpm

Sampled in 0, 7, 14, and 21 d

Centrifuged 12000 rpm, 5 min

Analysis using by ICP-MS

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 Cd IMMOBILIZATION TEST OF B. megaterium
H3 AND N. huautlense T1-17

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Li et al., 2017
 RICE CULTIVATION AND Cd
DETERMINATION
0 mg/kg
CdSO4 Rice Sampled for soil and plant tissues

2.5 mg/kg
9 kg Soil Analysis using ICP-MS
CdSO4 Planted separately

5 mg/kg
CdSO4
Added with 108 CFU/ml
B. megaterium H3 and N. huautlense
TI-17 (separately)

Grown for 110 d

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 EFFECT OF BACTERIA SUPPLEMENTATION
TO Cd CONTENT IN ROOT

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Li et al., 2017
 EFFECT OF BACTERIA SUPPLEMENTATION TO
Cd CONTENT IN ABOVE-GROUND TISSUE

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Li et al., 2017
 EFFECT OF BACTERIA SUPPLEMENTATION TO
Cd CONTENT IN POLISHED RICE

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Li et al., 2017
 FRACTIONATION OF Cd BY
SEQUENTIAL EXTRACTION
Sample (Soil)

Sequential Extraction Chloride Extraction Chloride Fraction

AAS (λ = 228.8 nm) Carbonate Extraction Carbonate Fraction

Bound with Fe/Mn-Oxides Bound with Fe/Mn-Oxides

Extraction Fraction

Bound with Organic Matter Bound with Organic Matter

and Sulfides Extraction and Sulfides Fraction

Residual Residual Fraction

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 Cd FRACTION IN SOIL AFTER
SUPPLEMENTATION
2.5 mg/kg CdSO4 5 mg/kg CdSO4

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Li et al., 2017
 SUMMARY
• Heavy-metal resistant bacteria B. megaterium H3 and N. huautlense T1-17 can
help immobilization of Cd by uptake to their cell.
• Heavy-Metal Bacteria supplementation give effect in reducing Cd uptake by
Rice.
• Cd concentration in rice in treated rice lower (0.07 – 0.16 mg/kg), compared
to untreated rice (≥ 0.25 mg/kg).

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 CONCLUSIONS
• Cd is the mobile element and can be uptake by rice. It also can be
translocated into the grains.
• Cd in rice can be reduced by:
o By soil amendments
o By microbial-assisted remediation
• FBA supplementation is better compared with bioremediation method, as
can reduce Cd uptake to the grain to very low concentration.

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THANK YOU
For your attention

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