PLANTS USED FOR PHYTOREMEDIATION

1. PLANTS USED FOR PHYTOREMEDIATION Rashad Reed and Kenong Zhang BZ 572 Fall 2010 November 18, 2010

2. Plant Classification • Ornamental Plants • Pot marigold (Calendula officinalis) • Common hollyhock (Althaearosea) • Chinese brake fern (Pteris vittata ) • Aquatic Plants • Water Hyacinth (Eichhorniacrassipes ) • Eurasian Watermilfoil (Myriophyllumspicatum) • Fool's Watercress (Apiumnodiflorum) • Duckweed (Lemnatrisulca L.)

3. Pot Marigold and Common Hollyhock

4. Features of Ornamental Phytoremediation • Employs phytostabilization of pollutants • Found to phytoremediate heavy metals, i.e. Cd, Pb • Organic phyoremediation of petroleum (TPH) • Cultural background and influence (marketing, aesthetics) • Profitability to existing nursery, fertilizer, and gardening industries • Public acceptance • Advantages: Can be used in urban environments without changing existing landscape. • Relatively low costs • Non-food chain plants that can be periodically phytoextracted for pollutants. • Can be genetically modified to not produce pollen or set seed with minimal effect to ecology

5. Ornamental Phyto Case Study • Liu et. Al tested 3 ornamentals, Impatiens Balsamina, Althaearosea and Candulata officinalis for ability to tolerate and accumulate Cd and Pb. • Soil and hydro conditions tested • Cd levels ranged from 0, 10, 30, 50, 100 mg/Kg-1 • Pb was added to Cd in the following concentrations: 0 + 0 (control), 1 + 50, 3 + 50, 5 + 50, 10 + 50, 1 + 100, 3 + 100, 5 + 100 and 10 + 100 mg L-1 • Pb(NO3)2 form of bioavailable lead • CdCl2·2.5H2O form of bioavailable cadmium

6. Ornamental Phyto Case Study Soil • Impatiens balsamina had leaves turn brown under >50 mg/Kg • C. officinalis height increased as Cd levels increased • A. roseaheightslightly decreased for levels above 50 mg/Kg (insignificant) • C. officinalis and I. balsamina accumulates more Cd in roots than the shoot at every Cd level • A. rosea accumulates more Cd in shoots for 10, 30, and 50 mg/Kg, but more Cd in the roots under 100 mg/Kg CdCl2·2.5H2O

7. Ornamental Phyto Case Study

8. Ornamental Phyto Case Study • A. roseahad the highest ability to accumulate Pb, the maximal Pb concentration in the shoots and roots was 24 and 640 mg kg−1 • Phytotoxicity shown (Cd and Pb = 10 + 50 and 10 + 100 mg L−1) • C. officinalis had the highest ability to accumulate Cd, the Cd concentration in the shoots and roots reached 825 and 1585 mg kg−1 in TP4 treatment, 700 and 1492 mg kg−1 in TP8 treatment, respectively • All three plants had a lower general affinity for Pb

9. Ornamental Phyto Case Study • Case study conclusions • C. offinalisis tolerant to Cd levels, but is not a hyperaccumulator since more Cd was found in the roots than in the shoots. Phytostabilization • A. rosea, tolerant to heavy metals and is a hyperaccumulater of Cd when conc. < 100 mg kg-1 • Efficacy of ornamentals was examined and has potential for use in urban environments with moderate to high Cd/Pb contamination levels

10. Fern Phytoremediation • Classification: Marattiales, Ophioglossales, and leptosporangiate ferns • Best used in areas of high humidity • Ability to uptake high levels of arsenic • Industrial applications in phytoremediation • Humidity requirements reduces water necessity • Fast growing species and varieties available (increased biomass) • Ornamental use: Japanese painted fern

11. Chinese brake fern and Japanese painted fern

12. Arsenic accumulation in ferns • Ma et al. (2001) reported the first known arsenic hyperaccumulator Chinese brake fern (Pteris vittata L.). • Tu et al. 2002 found that P. vittata grown in an As-contaminated soil accumulated total dry biomass of 18 g plant-1 after 18 weeks of growth • Chinese brake fern tolerant of high concentrations of arsenic, up to 1,500 mg As kg-1 soil • Ferns operate by phytoextracting Arsenic, As(V) or As(III), storing As in the fronds • Ferns hyperaccumulate As through phosphate transporters and do not have phosphate deficiency symptoms at high levels of As

13. Fern phytoremediation (mechanism) • Contains arsenate reductase (AR) genes to reduce arsenate to arsenite • AR was not detectable in the fronds, suggests that arsenate reduction, occurs in roots • Arsenite is then transported to shoots, where it may be stored in the vacuoles (Lombi et al., 2002) • 75-95% As in the fronds is present in the form of arsenite (Ma et al. 2001, Zhang et al. 2002).

14. Fern Phytoremediation in Practice • 10 μg As/L-1is the limit set by US EPA • Over 29 million people in Bangladesh may be exposed to over 50 μg/L As. • Bangladesh, India, and Pakistan have problems with ground water contamination from As • Up to 7500 mg As/kg on a contaminated site without showing toxicity symptoms. (Ma et al, 2001) • Contamination comes from industrial and chemical plants, dumping effluent into groundwaters (lack of regulation)

15. Scope of Arsenic contaminaton

16. Practical Urban Phytoremediation • Hyde Park neighborhood: Augusta, GA

17. Phyoremediation in practice • Low-lying geographic area containing approximately 200 houses and surrounded by industry, abandoned industry, railroad lines and large highways. • Problems of high poverty levels (>70%) • Abandoned houses, overgrowth of weeds, snakes, etc. • Illegal dumping and drain contamination • Lack of community cohesiveness and/or solid leadership • Phytoremedation can be used as a part of a larger objective in urban environmental renewal

18. Hyde Park Pictures

19. Bob Safay. ATSDR Regional Representative. U.S. EPA, Region IV released a report in 1994. • The most serious contamination found near the Goldberg site. • From ditch sediments: Highest lead detected was 1800 mg/kg, and the highest level of PCBs was 13.2 mg/kg • From soil samples: lead (1100 mg/kg), arsenic (59 mg/kg), and dioxin/furans (0.0001 mg/kg Agency for Toxic Substances and Disease Registry report, APPENDIX 3 - MARCH 1994 HEALTH CONSULTATION

20. Phytoremediation plan Perennials • Alpine Pennygrass, Thlaspicaerulescens • Hairy Goldenrod, Solidagohispida • Yellow Tuft, Alyssum lesbiacum • Bladder Campion, Silene vulgaris • Horse bean, Viciafaba Trees and Shrubs • Aspen, Populustremula • Shrub violet, Hybanthusfloribundus Grasses • Indian grass, Sorghastrumnutans • Kleingrass, Panicumcoloratum • Little bluestem, Schizachyriumscoparius • Bent grass, Agrostiscastellana

21. Aquatic plants Aquatic plants are those plants living in and adapted to aquatic environments, which can only grow in water or permanently saturated soil. Bodies of aquatic plants can be either floating or submerged. Aquatic plants are often viewed as indicators of aquatic environment pollution.

22. Living environment of aquatic plants Rivers Lakes Where do they live? Hydroponic systems Constructed Wetlands

23. Mechanisms of aquatic plants phytoremediation Inorganic pollutants Rhizofiltration Phytostabilization Phytoextraction Organic pollutants Phytodegradation Phytovolatilization Rhizodegradation

24. Constructed Wetlands High substrate heterogeneity Wide application for pollutants removal and phytoremediation Act as a cover above contaminated areas Rhizofiltration techniques are the most commonly used Pollutant removal efficiencies are significantly related to plant species present

25. Pollutant Removal Efficiencies of Constructed Wetlands (Otte and Jacob, 2006)

26. Benefits and drawback of constructed wetland Benefits: Aesthetical functions Provide habitat for wildlife animal species Educational resources Little maintenance required Increased Cost-efficiency Drawbacks: Limited by plant tolerance and pollutant bioavailability Limited plant life expectancy Susceptible to climate change, pollution, and disease

27. Heavy Metal Heavy metal ions of Cd2+, Hg2+, and Pb2+ are nonessential and toxic to plants Cu2+, Zn2+, Mn2+, Fe2+, Ni2+, and Co2+ are essential micronutrients for plants, but toxic when present in high concentration Hyperaccumulators: plant species tolerate, uptake, and translocate high levels of certain heavy metals that is toxic to other species. Contain >100 mg/kg of Cd, >1000 mg/kg of Cu, or >10,000 mg/kg of Zn and Mn(dry weight in leaves)

28. Heavy Metal Toxicity Mercury toxicity symptoms concentration deficits Impaired motor function Lead toxicity symptoms Learning disability Mental retardation Chromium toxicity symptoms Damage DNA Damage kidney Hg Pb

29. Typical Species for Heavy Metal Removal Water Hyacinth (Eichhorniacrassipes ) Fool's Watercress (Apiumnodiflorum) Eurasian Watermilfoil (Myriophyllumspicatum) Duckweed (Lemnatrisulca L.)

30. Water Hyacinth Floating plant with broad ,thick, and glossy leaves that the plant body can grow as much as 1m high. Able to phytoaccumulate metal pollutants contain Ag, Pb, Cd and Zn in municipal and agricultural wastewater. Known as one of the plants with fastest growth rate that can double population in 2 weeks. High invasive potential.

31. Case Study: Removal of Cadmium and Zinc by Water Hyacinth The stock solution was prepared in distilled water with analytical grade CdCl2. 2½ H2O and ZnSO4.7H2O which was later diluted as required. The plants were maintained in tap water with concentrations of 0.5, 1, 2, 4 mg/L of Cd and 5, 10, 20, 40 mg/L of Zn. The test durations were 0 (two hours), 4, 8 and 12 days. Relative growth, metal accumulation, and bioconcentration factor (BCF) are evaluated. Relative growth (above) and BCF (below)

32. Relative plant growth Metal Accumulation BCF Cd Cd Cd Zn Zn Zn (Lu et al., 2004)

33. Eurasian watermilfoil Submerged aquatic perennial plant which grows in still or slow-moving water. Slender stems up to 3 m long with numerous leaflets thread-like, 4-13 mm long. Introduced to North America between the 1950s and 1980s where it has become invasive species. Able to uptake and remove lead, zinc, and copper from wastewater.

34. Plant tissue was washed with 3% HCl solution previously. Metal source were provided by CuSO4, ZnSO4, and Pb(NO3)2 to prepare the stock solutions with concentration of 2, 4, 8, 16, 32 and 64 mg/L (doubling). Standard curves were made. Absorption tests were conducted in 250 ml conical flasks placed on orbital shaker and contact for 2 hours. Filtrate was analyzed by atomic absorption spectrophotometer (AAS) to determine sample metal concentrations. Case Study: Removal of Lead, Zinc, and Copper by Eurasian watermilfoil (Keskinkan et al., 2003)

35. According to figure 1 (left), equilibrium were reached after about 20 mins after beginning; after that the data was adhered well to the Langmuir equation (see below) which means that absorption was as a monolayer. The value of metal concentration of solution on time t over the beginning concentration Ct/C0 is shown below: Compare of the metal uptake capacities (qmax, mg/g) of Myriophyllumspicatumto other plant species: (Keskinkan et al., 2003)

36. Fool's Watercress Hollow stems rooting at base and finely grooved. About 0.3-1m high. Simply pinnate leaves in shiny bright green in color with 2-4 pairs of  lobes. Usually find in grown ditches, shallow ponds or very damp places. Capable to uptake and remove various heavy metals such as Hg, Cr, Pb, Cu, and Zn. (Vlyssides et al., 2005)

37. A mathematical model was established to evaluate the inherent capacity of watercress to uptake heavy metals. Plant uptake rate follows the first order kinetic model depending on the heavy metal concentration in the plant biomass. This model allowed to evaluate the specific uptake rate and the maximum content within plant biomass. The relationship between metal concentration in solution Es (mg/L) and plant biomass Ep (mg/g) is: Case Study: Removal of Copper, Lead etc. by Fool’s Watercress (Vlyssides et al., 2005)

38. Change of heavy metal concentrations in solution and plant biomass (Vlyssides et al., 2005)

39. E∞ = the saturation heavy metal concentrations in the plant biomass km = maximum uptake rate of metal KS = saturation constant (Vlyssides et al., 2005) The estimated absorption kinetic parameters for various heavy metals, by ApiumNodiflorum using the data acquired

40. Duckweed Has a very simple structure that lacks obvious stems or leaves, with small plate-shaped structure floating on water surface. Reproduction is mainly rely on asexual budding. High pollutant removal potential due to small size, fast growth, and easy to cluture. (Kara and Kara, 2004)

41. The duckweed obtained from natural lake was acclimatized to laboratory conditions for one week before starting research. Solution of Cadmium was prepared using Cd(NO3)2 and contact with plant sample for different length. After absorption, water samples were analyzed by AAS at 228.8nm. Case Study: Removal of Cadmium by Duckweed Cd removal efficiencies (Kara and Kara, 2004)

42. Aquatic Plants VS. Terrestrial Plants in Phytoremediation • Advantages of aquatic plants • Faster growth and larger biomass production rate • Relative higher capability of pollutant uptake • Better water purification effects due to direct contact GO AQUATIC!

43. Aquatic Plants VS. Terrestrial Plants in Phytoremediation Advantages of terrestrial plants More plant-soil microbe interactions to enhance pollutant uptake Higher tolerance against severe weather and temperature change More sophisticated root system GO TERRESTRIAL!

44. Conclusions • Ornamental phytoremediation is beautiful, profitable and effective: utilizing phytoextraction and phytostabilization • Ferns hyperaccumulate As which can be useful in contaminated groundwater regions, such as Bangladesh • Hyde Park, Augusta GA has a current and ongoing contamination problem that ornamentals may help to alleviate without affecting food chain or food supply

45. Conclusions • Most of the aquatic plants showed high heavy metal phytoremediation potential are usually considered as invasive species, which indicates that there are numerous positive aspects of those species that is able to take advantage of • To achieve better remediate effects, more effort is required to accelerate the pace of phytoremediation techniques from laboratory experiment to practical use.

46. References: 1. Otte, ML and Jacob, DL (2006) Constructed Wetlands for Phytoremediation. Phytoremediation Rhizoremediation57-67. 2. Kamal, M, Ghaly, AE, Mahmoud, N, Côté, R (2004) Phytoaccumulation of heavy metals by aquatic plants. Environment International 29: 1029-1039. 3. Skinner, K, Wright, N, Porter-Goff, E (2007) Mercury uptake and accumulation by four species of aquatic plants. Environmental Pollution 145: 234-237. 4. Titus JE and Urban RA (2009) Aquatic Plants: A General Introduction. Encyclopedia of Inland Waters 43-51. 5. Keskinkan O, Goksu MZL, Yuceer A, Basibuyuk M, Forster CF (2003) Heavy metal adsorption characteristics of a submerged aquatic plant (Myriophyllumspicatum). Process Biochemistry 39(2): 179-183. 6. Vlyssides A, Barampouti EM, Mai S (2005) Heavy Communications in Soil Science and Plant Analysis metal removal from water resources using the aquatic plant Apiumnodiflorum. 36: 1075-1081. 7. Lu X, Kruatrachue M, Pokethitiyook P, Homyok K (2004) Removal of Cadmium and Zinc by Water Hyacinth, Eichhorniacrassipes. Science Asia 30: 93-103. 8. Kara Y, Kara I (2004) Removal of Cadmium from water using Duckweed (Lemnatrisulca L.). International Journal of Agriculture & Biology 7: 660-662. 9.Duan G, Zhu Y, Tong Y, Cai C, Kneer R (2005) Characterization of arsenate reductase in the extract of roots and fronds of Chinese brake fern, an arsenic hyperaccumulator. Plant Physiology 138, 461-469 10.Ma, L.Q., K.M. Komar, C. Tu, W. Zhang,and Y Cai. 2001. A fern that hyperaccumulates arsenic. Nature. 409:579. 11. Internet source, Banglopedia.org http://www.banglapedia.org/httpdocs/HT/A_0308.HTM 12.Internet source, Slide 15. http://www.hamitekllc.com/sites/mountainmovers.org/files/img/arsenic-poisoning.gif

47. References: 13. Tu, C., L.Q. Ma, A.O. Fayiga and EJ Zillioux. (2004) Phytoremediation of Arsenic-Contaminated Groundwater by the Arsenic Hyperaccumulating Fern Pteris vittata L . International Journal of Phytoremediation, 6(1):35–47. 14. Internet source, Slide 3(b). http://www.types-of-flowers.org/pictures/alcea_rosea.jpg 15. Internet source, Slide 3(a). http://pics.davesgarden.com/pics/2009/03/02/purplesun/ed602b.jpg 16. Hyde Park Charettte Report. October 2008. The University of Georgia College of Environment and Design Center for Community Design and Preservation. www.ced.uga.edu/charrettes.html 17. Rathinasabapathi, Bali, L.Q. Ma and M Srivastava. (2006) Floriculture, Ornamental and Plant Biotechnology Volume III. Global Science Books, UK. 18. Tu C, Ma LQ (2005) Effects of arsenic on concentration and distribution of nutrients in the fronds of the arsenic hyperaccumulatorPteris vittata L. Environmental Pollution. 135, 333-340. 19. Internet source, slide 11(b). http://www.shadesofgreenusa.com/Priclist_files/painted_fern1.jpg 20. J. Liu, Q. Zhou, T. Sun, L.Q. Ma and S. Wang. (2008) Identification and Chemical Enhancement of Two Ornamental Plants for Phytoremediation. Bull Environ Contam Toxicol80:260–265. 21. J. Liu, Q. Zhou, T. Sun, L.Q. Ma and S. Wang. (2008) Journal of Hazardous Materials 151:261–267. 22. Fiegl, J., Bryan P. McDonnell, Jill A. Kostel, Mary E. Finster, and Dr. Kimberly Gray "A Resource Guide: The Phytoremediation of Lead to Urban, Residential Soils". http://www.civil.northwestern.edu/EHE/HTML_KAG/Kimweb/MEOP/INDEX.HTM

48. References: 23. Lombi E., F. Zhao, M. Furhrmann, S.Q. Ma and S. McGrath. (2002) Arsenic distribution and speciation in the fronds of the hyperaccumulatorPteris vittata. New Phytologist. 156: 195–203. 24. Zhang WH, Cai Y, Tu C, Ma LQ. (2002) Arsenic speciation and distribution in an arsenic hyper accumulating plant. Sci Total Environ.; 300(1-3):167–177. 25. Zhao FJ, Dunham SJ, McGrath SP. Arsenic hyper accumulation by different fern species. New Phytologist. 2002;156(1):27–31. 26. I. Alkorta, J. Hernández-Allica, and C. Garbisu (2004) Environment International. 30: 7, 949-951. 27. Safey, Bob. Agency for Toxic Substances and Disease Registry. (1994)Appendix 3 – March 1994 Health Consultation. http://www.atsdr.cdc.gov/HAC/pha/pha.asp?docid=1029&pg=6 Suggested reading material Checker, Mellissa. (2005) From Friend to Foe and Back Again: Industry and environmental action in the urban south http://www.augustaneeds.com/files/ASU_HydePark_MelissaChecker_2005.pdf Checker, Mesllisa (2005) Polluted Promises: Environmental Racism and the Search for Justice in a Southern Town. NYU Press, NewYork, NY.