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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 CELL UPTAKE AND COMPARTIMENTATION HYSTOLOGICAL LOCALIZATION GOLGI TRICOMES NUCLEUS M M VACUOLE M M M M M PLASMA MEMBRANE M metalchaperonines xilem M M APOPLASTIC TRANSPORT M TRANSPORT XILEMATIC M Mn+ Mn+ M M SYMPLASTIC TRANSPORT MOBILIZATION DIFFUSION M n+ CELL WALL XILEM PARENCHIMATIC CELLS M floema M cell. transfer H+-ATPase APOPLASTIC + H LOADING ? MALIC ACID CITRIC ACID ROOT UPTAKE M PLASMA MEMBRANE CELL WALL SYMPLASTIC n+ MLOADING M H SOIL + M VACUOLE M n+ M M M M M Mn+ n+ M H+-ATPase H+ + H n+ M M M M CITOSOLIC CHELATORS 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