Lemnaceae; from green nuisance ……… ………to green technology Dr. Marcel Jansen Lemnaceae Wolffia globosa Wolffiella gladiata Lemna minor Spirodela polyrhiza http://waynesword.palomar.edu Smallest flowering plants http://world-flowers-plants.blogspot.com/2011/04/smallestcrop-in-world.html http://waynesword.palomar.edu Small plant – fast grower! Doubling time at UCC Lemna minor Lemna gibba Landoltia puntata Wolffia brasiliensis 2.27 day 2.18 day 2.11 day 3.35 day Doubling time optimal conditions <20 hours http://www.mobot.org/jwcross/duckweed/duckweed.htm Fast growth; problem or opportunity ? Pennsylvania Lake Maracaibo - Venezuela http://www.voltairenet.org/The-Promise-of-Restitution-of Cork http://imageshack.us/ Lemnaceae; problem or opportunity ? Lemnaceae in traditional /sustainable agriculture (FAO 2011) http://www.fao.org/ag/againfo/resources/documents/DW/Dw2.htm Lemnaceae; problem or opportunity? Protein (43% DW) feed aquaculture Essential amino acids (Lys, Leu, Ile, Phe, Thr, Val) Bio-crude (renewable fuel) Technology for growth, harvesting and processing Yield up to 4 ton FW / hectare / day PetroAlgae Inc Florida Lemna production http://www.petroalgae.com/ Lemnaceae; problem or opportunity ? Devils Lake – North Dakota – 10.000 inhab. Tertiary wastewater treatment (N/P removal) Plant nutrient uptake Anaerobic microbial processes Marriage between biology & engineering http://www.ci.devils-lake.nd.us/departments/sewer-department.html Phytoremediation–I - Zinc http://www.imdb.com/ Metals and heavy metals Lemnaceae as bioaccumulators – – Phytoremediation Feed fortification Zinc uptake by Lemna minor zinc Time in days Linear model with first order kinetics Based on 0.2 mg/L BCF=1000 kg/ha/day Zinc bioaccumulation by Lemna minor Literature; accumulation factors Zn > 35000 Zinc in the food chain Available for trophic transfer From phytoremediation to …. ……….food fortification Zinc in Gammarus pulex fed on Lemna minor and after 24h depuration Phytoremediation-II - TCP Chlorophenols Priority pollutant Genotoxic, mutagenic, carcinogenic Ubiquitous in aqueous environment – – – Sawmills (wood impregnation agents) Precursors / degradation products chemicals Landfill sites 2,4,6, trichlorophenol TCP is phytotoxic EC50 (µM) Specific growth rate (day-1) L.punctata L.minor L.gibba POD activity (nmol ABTS min-1 mg-1) Peroxidase activity in medium L. punctata 100 90 80 70 60 50 40 TCP 30 20 TCP TCP 10 0 Landoltia punctata Lemna minor Lemna gibba Response to TCP concentration that gives ca. 15% growth inhibition UV/VIS spectrophotometry 0.15 SpEx 0.1 Enzyme + 2,4,6, TCP 0.05 0 -0.05 0.5 HRP-C 0.4 Oxidation to 2,6 dichloro1,4 benzoquinone 0.3 0.2 0.1 0 240 280 320 Wavelength (nm) 360 400 m/z 195 TCP (+2, +4, +6 for 37Cl) TCP - no enzyme LC-MS TCP - peroxidase Isomer dichloro benzoquinone Substrates for class III peroxidases Three components of Lemnaceae response system Lemnaceae frond (1) Recognition & Induction (2) Enzyme secretion Trichlorophenol (3) Oxidative detoxification by Mono and Dichloro benzoquinones Class III peroxidase Lemnaceae bioreactor? Range of potential substrates? TCP breakdown products? Capacity Lemnaceae reactor? ------------------------ Secrete other desirable proteins http://www.lemnagene.com/ ………..Lemnaceae and the Green Economy…..? Molecular transformation technologies Understanding Physiological potential Engineering solutions DOE-JGI genome sequence Green technology Protein/biofuel Phytoremediation Food fortification Pharmaceuticals Acknowledgements TCP (JIC, Norwich, UK) • Roger Thorneley • Lionel Hill TCP (UCC) • Dilip Biswas (BEES) • Gillian Scannell (BEES) • Nurlan Akhmetov (Chemistry) • Dara Fitzpatrick (Chemistry) Zinc (UCC) • Elma Lahive (BEES) • John O’Halloran (BEES) Funding EPA and IRCSET