Copper localization in Cannabis

sativa grown in a copper-rich

solution
Piera Bonatti, Laura Arru, Sara Rognoni,
Micaela Baroncini, Pierdomenico Perata

University of Modena & Reggio Emilia

PHYTOREMEDIATION
The use of plants to remove heavy metals and
other pollutants from soils and waters

C E L L U P T A K E A N D C O M P A R T IM E N T A T IO N

H Y S T O L O G IC A L L O C A L IZ A T IO N

G O LG I
T R IC O M E S

NUCLEUS
M
M

VA CU O LE
M

PLASM A
M EM BRANE

m e ta lc h a p e r o n in e s

x ile m

X I L E MM A T I C T R A N S P O R T
M

A P O P L A S T IC
TRANSPOR T

S Y M P L A S T IC
TRANSPO RT

M O B IL IZ A T IO N
D IF F U S IO N
M n+

flo e m a
M
M

S O IL

CELL
W ALL
M

VA C U O LE
M

n+

n+

PLASM A
M EM BRANE

S Y M P L A S T IC
n+
M L O A MD I N G
H

n+

R O O T U P TA K E

CELL
W ALL
X IL E M
P A R E N C H IM A T IC
CELLS

c e ll.
tr a n s fe r
H +-A T P a s e
A P O P L A S T I +C
L O A D I N GH
?
M A L IC A C ID
C IT R IC A C ID

n+

H +- A T P a s e
H

n+

M
M

C IT O S O L IC
CH ELATO RS

n+

PHYTOEXTRACTION OF HEAVY METALS

The most common heavy metals are:

Cd, Co, Cr, Cu, Hg, Mn, Mo, Ni, Pb, Sn, Zn
Are often very toxic to living organisms over a certain
concentration threshold

WHICH PLANT SPECIES FOR PHYTOEXTRACTION?

HYPERACCUMULATOR SPECIES
Represent <0,2% of all angiosperms
~400 species are hyperaccumulators
HYPERACCUMULATION THRESHOLDS:
Zn, Mn:
10 000 mG/KG
Co, Cu, Ni, Se:
1 000 mG/KG
Cd:
100 mG/KG
These thresholds are 2-3 orders of magnitude higher than in normal plant species

Alyssum serpyllifolium

Brassica juncea

HYPERACCUMULATOR
SPECIES &
PHYTOREMEDIATION
PLANTS

Thlaspi
caerulescens
Liriodendron tulipifera

Pteris vittata

WHICH PLANT SPECIES FOR PHYTOEXTRACTION?

Hyperaccumulator species must be hypertolerant

to heavy metals

Pteris vittata accumulates up to

22 000 mG/KG of arsenic, with no
phytotoxicity simptoms up to 10 000
mG/KG As
(Ma et al, Nature, 2001; Wang et al., Plant Physiol., 2002)

WHICH PLANT SPECIES FOR PHYTOEXTRACTION?

Hyperaccumulator species = low biomass?

Thlaspi caerulescens, a Zn/Cd
hyperaccumulator, produces little biomass
(2-5 t/ha)
(McGrath et al. Adv. Agronom. 2002)

Alyssum bertolonii and Berkheya coddii, Ni

hyperaccumulators, produce up to 9-22 t/ha
biomass (Robinson, 1997)

WHICH PLANT SPECIES FOR PHYTOEXTRACTION?

Can high biomass compensate for lower accumulation?
Thlaspi caerulescens
can extract 2000 g Cd / ha / year

Cannabis sativa
Can extract 126 g
Cd / ha / year

WHICH PLANT SPECIES FOR PHYTOEXTRACTION?

Hemp as a high biomass plant for phytoextraction?
Phytoxicity?

Use of fibers after

phytoextraction?
Zn toxicity on hemp plants

WHICH PLANT SPECIES FOR PHYTOEXTRACTION?

Hemp as a high biomass plant for phytoextraction?

Not hyperaccumulator

High biomass

Could be used for phytoremediation if fibers can be

commercialized

COPPER SOIL CONTAMINATION

Copper is required for the physiology of the plant
An EXCESS of copper in the soil results in phytotoxic effects
Copper is an essential cofactor for many enzymatic activities in
animals too
An EXCESS of copper is excreted by animals (liver, bile)
Genetic diseases result in altered Cu homeostasis (Wilson
disease)
Exposure to the metal copper may increase the risk of
Alzheimer's disease

COPPER SOIL CONTAMINATION

Copper sulfate is used in
agricuture, including organic
farming
Possible copper build-up in the soil
which at high levels can be
harmful to earth worm populations.
In response to such environmental
concerns copper should be
phased out of usage as soon as
possible

The aim of this work is the

localization of Cu in hemp plants
grown in a copper-enriched solution

CuSO4

Energy Dispersive X-ray Spectra (X-EDS)

STEM

Cu

LEAF EPIDERMIS

Cu

HYSTOLOGICAL LOCALIZATION OF Cu

LEAF EPIDERMIS

40 m

HYSTOLOGICAL LOCALIZATION OF Cu

TRICOMES

80 m

CITOLOGICAL LOCALIZATION OF Cu

CELL WALL

VACUOLE

PLASTID

CONCLUSIONS
Hemp grown in hydoponics accumulates cooper in the leaves (tricomes)

Copper was not detected in fibers

Further work is needed to evaluate the fibers quality from hemp grown
in open field in the presence of copper sulfate