Final Report: Woody Biomass Crops for Bioremediating Hydrocarbons and Metals

EPA Grant Number: R829479C010
Subproject: this is subproject number 010 , established and managed by the Center Director under grant R829479
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).

Center: The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program
Center Director: Schumacher, Dorin
Title: Woody Biomass Crops for Bioremediating Hydrocarbons and Metals
Investigators: Rockwood, Donald L.
Institution: University of Florida
EPA Project Officer: Lasat, Mitch
Project Period: August 12, 2002 through December 31, 2003
RFA: The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program (2001) RFA Text |  Recipients Lists
Research Category: Targeted Research , Hazardous Waste/Remediation

Objective:

The overall objective of this research project was to evaluate and characterize fast-growing tree-based systems for maximizing phytoremediation of hydrocarbon and heavy metal-contaminated soil and groundwater through a series of field and greenhouse studies that started with six objectives. The specific objectives of this research project were to:

  1. determine long-term nitrogen and water use efficiency for cottonwood Populus deltoides (PD), Eucalyptus amplifolia (EA), and E. grandis (EG) under four silvicultural options (no compost or mulch [W]; compost only [WC]; mulch only [WM]; and compost plus mulch [WCM]) in a 2.8 ha sewage effluent study (Water Conserv II) west of Orlando, Florida, initiated in March 1998;
  2. assess ongoing remediation of copper, chromium, and arsenic (As) contaminated soil by PDclones at a former wood preservative treatment facility in Archer, Florida;
  3. compare phytoremediation potential of PD clones and EA progenies planted in July 2000 at a trichloroethylene (TCE)-contaminated site near St. Augustine, Florida;
  4. evaluate early effectiveness of poplar and willow clones in remediating TCE- and perchloroethylene (PCE)-contaminated sites at LaSalle, Illinois;
  5. quantify variation between and within PD, EA, EG, willow, and cypress and assess mechanisms for tolerance to and remediation of varying TCE levels in a greenhouse study;
  6. and select PDclones in collaboration with Florida A&M University (FAMU), promising for southwide phytoremediation from a clone test near Quincy, Florida.

Summary/Accomplishments (Outputs/Outcomes):

Objective 1: Determine Nitrogen and Water Use Efficiency for PD, EA, and EG Under Four Silvicultural Options in Water Conserv II

Preharvest species and culture differences included EG being much larger than PD, the WCM culture considerably enhancing tree growth, and EG being much more effective for water and nutrient uptake (Rockwood et al., 2001). PD, EA, and EG representing various coppice rotation management strategies (1-year-old first coppice, 2-year-old first coppice, 1-year-old second coppice) were felled in May 2003. Tree size and dry weight data suggest that first and second coppice rotation yields can equal or exceed first-rotation yields, peak coppice productivity will not be achieved in the first year of coppice growth, and root biomass does not significantly increase with coppicing. Nutrient analyses to assess differences among coppice rotations and ages remain in progress.

Objective 2: Assess Remediation of As Contaminated Soil by PD Clones at Archer, Florida

A tendency for fast-growing clones to have lower As concentrations suggests that total As uptake by PD will be a balance between biomass production and As concentration. Certain clones, however, such as ST-229, that combine fast growth with high concentration would maximize uptake.

In reconstructed portions of the Archer site, PD, EA, and EG genotypes were planted with compost and no-compost amended soils in April 2003. Because of the high PD mortality, Chinese brake fern (Pteris vittata [PV], an As hyperaccumulator) and selected PD, EA, and EG genotypes were established in a fern-tree mixing study with and without compost amendment in August 2003, to determine if fern-tree mixes took up more As than each species alone. To date, compost amendment has increased As concentration in the soil by about two-fold, but lowered As concentration in PV by nearly 40 percent (see Table 1). PV still has demonstrated its capability to hyperaccumulate As, with PVconcentrations of as much as 80 times soil levels. Compost has not caused more rapid tree growth for 3 months, but PD has thus far been more vigorous than EA and EG. As concentrations in the trees are yet to be determined.

Table 1. Three-Month PD, EA, and EG Tree Heights (m) and As Concentrations (mg/kg) in PV and Soil With and Without Compost for Various Fern-Tree Mixes at Archer, Florida, in November 2003
Mix
No Compost
Compost
Height PV Soil Height PV Soil

PD

1.66 - 39.0 1.58 - 91.6

PD-PV

1.59 2355 43.9 1.29 1457 79.2

EA

0.65 - 33.4 0.74 - 69.6

EA-PV

0.60 2917 36.5 0.60 2096 73.8

EG

1.02 - - 1.39 - -

EG-PV

0.70 3139 41.4 0.73 1810 56.1

PV

- 3010 45.5 - 1923 71.5

Objective 3: Compare Phytoremediation Potential of PD Clones and EA Progenies at a TCE-Contaminated Site Near St. Augustine, Florida

After 29 and especially 40 months, 15 cm diameter plastic “training” tubes inhibited above-ground growth of PD and EA (see Table 2), and presumably root growth and access to groundwater. Trees planted in 2-, 3-, and 4-ft tubes had up to 16 percent less survival at 29 months and, after 40 months, were 1.2 m shorter and as much as 1.1 cm less in diameter at breast height (DBH). PD and EA were equally vigorous statistically at both ages, but EA progeny 4938 was the most productive genotype by 1 m in height at 40 months. Tree survival was not correlated with toluene concentrations. Toluene was detected in leaf and branch samples of EA but not PD. Toluene detected in air samples could not be traced to transpiration.

Table 2. Number of Trees (in parentheses), Means, 95 Percent Significance for Species (capital letters) and Tube Depths (lowercase letters), and Range Among Genotype Means Within PD and EA for Height (H in m), DBH (D in cm), and/or Survival (S in %) at 29 and 40 Months at St. Augustine, Florida
Trait
PD
EA
Tube Depth
Mean Range Mean Range 0 2 3 4
H29 2.11A (241)

1.47 — 2.61

2.12A (272)

1.40 — 3.88

2.45a (311) 1.62b (70) 1.61b (63) 1.59b (69)
D29 1.27A (166)

0.53 — 1.67

1.81A (178)

0.88 — 3.05

1.73a (243) 1.33ab (31) 1.06b (32) 0.94b (38)
S29 76.5A (315)

35.7 — 100.0

86.3A (315)

61.9 — 100.0

86.4a (360) 77.8ab (90) 70.0b (90) 76.7ab (90)
H40 2.70

1.51 — 4.19

2.88

1.72 — 5.18

3.29 2.14 2.12 2.08
D40 1.55

0.88 — 2.39

2.54

1.43 — 4.62

2.50 1.44 1.36 1.37

Objective 4: Evaluate Effectiveness of Poplar and Willow Clones in Remediating TCE- and PCE-Contaminated Sites at LaSalle, Illinois

Unrooted willow cuttings averaged only 10.9 percent survival 4 months after planting in the PCE-contaminated northwest unit because of exceptionally late and severe spring freezes; the only willow clone with more than 50 percent survival grew poorly (see Table 3). Although rooted poplar cuttings had only 47.8 percent survival at 4 months, several poplar clones all survived and grew rapidly in the PCE-contaminated clay soil, averaging 2 m in height, with individual ramets reaching 3.3 m, a reflection of improved soil texture and organic matter content as well as the advantage of planting mini-barbatelles. Mini-barbatelles interplanted in the summer, however, typically lagged far behind the spring planting. Of 19 baldcypress (Taxodium distichum var. distichum) trees, 63.2 percent of surviving trees that were 0.59 m tall after 4 months were only 1.2 m tall after 20 months.

In the northwest unit, most trees surviving from the spring 2002 planting grew well for 20 months. The poplars were the most vigorous, averaging more than 4 m tall, with some clones exceeding 5 m in height and 4 cm in DBH (see Table 3). Willow clones, in contrast, all were under 2 m in height. Three vigorous poplar clones in the area of highest PCE concentration contained PCE in basal stem cores and various tree parts (see Table 4). This coring procedure, developed in conjunction with our sponsor Ecology & Environment, offers a quick, inexpensive means for assessing hydrocarbon uptake by trees.

The 2 m tall poplar and willow whips for the GTU unit rooted well in fiber pots, and many exceeded 4 m in height when planted as deep as 2 m in 0.61 m wide plastic pipes in September and October 2002. The one poplar clone in the GTU, I 45\51, a below average performer in the northwest, was as tall after 12 months as the 2.4 m average of the 21 willow clones, some of which surpassed 3 m in height. While the GTU trees, typically with root systems fully occupying their “containers,” have effectively lowered the water table in the area and contained the TCE plume onsite, no TCE was detected in basal stem cores or other tree tissues.

Table 3. Number of Trees, Species Means for Height (H in m), DBH (D in cm), and/or Survival (S in %), and Range Among Means of 18 Poplar and 24 Willow Clones at 4, 8, and 20 Months in the Northwest Unit and Among Whips of 1 Poplar and 21 Willow Clones at 4 and 12 Months in the GTU Unit of the LaSalle Study
Trait
Poplar
Willow
No Mean Range No Mean Range
NW

H4

149 0.59 0.43 — 0.70 34 0.34 0.15 — 0.58

H8

- 1.46 0.73 — 2.00 - - -

H20

- 4.1 2.3 — 5.1 - 1.2 1.0 — 1.5

D20

- 3.3 2.0 — 4.5 - 0.8 0.5 — 1.1

S4

312 47.8 25.0 — 100.0 312 10.9 0.0 — 87.5

S8

312 82.1 50.0 — 100.0 - - -
GTU

H4

24 1.9 - 126 1.4 0.9 – 1.9

H12

24 2.4 - 126 2.4 2.1 – 3.2
Table 4. PCE Concentrations (μg/g) in Stem and Branch Tissues (wood + bark) at Upper, Middle, and Lower Stem Positions of Three Poplar Clones in the Northwest “Hot” Zone at LaSalle Previously Determined to Contain PCE in Basal Stem Cores
Clone (Position)
Upper
Middle
Lower
Stem Branch Stem Branch Stem Branch

Eugenii (3-15)

0.14 - <.04 0.14 <.04 0.08

ISU 25-R5 (4-24)

<.04 - <.04 0.08 <.04 0.07

I 45/51 (5-21)

0.13 - <.04 0.18 <.04 0.13

Objective 5: Quantify Variation Between and Within PD, EA, EG, Willow, and Cypress and Assess Mechanisms for Tolerance to and Remediation of Varying TCE Levels in a Greenhouse Study

A study, involving saturating PD, EA, EG, and cypress root masses with 0, 50, and 100 ppm TCE concentrations from mid-May to early August 2002, detected differences among and within species for tree vigor assessments such as height, stem diameter, and leaf number. Determination of tree component dry weights is in progress.

Objective 6: In Collaboration With FAMU, Select PD Clones Promising for Southwide Phytoremediation From a Clone Test Near Quincy, Florida

To evaluate propagation container and media options, unrooted cuttings of PD clones, selected to represent the range of As uptake, and new clones were placed at FAMU in April 2003. Because of disease and related problems, the study was repeated in September 2003 using greenwood cuttings of many of the same clones from the Quincy test. Because this effort also failed, the study will be initiated again in February 2004.

Conclusions:

EGis much more effective for wastewater and nutrient uptake in central Florida. PD, EA, and EG coppice yields may equal or exceed first-rotation yields, but peak coppice productivity will not be achieved in the first year of coppice growth, and root biomass does not increase with coppicing.

PD clones combining fast growth with high concentration would maximize As uptake. Compost appears to increase As concentration in the soil, but lowers As concentration in PV. PVhyperaccumulates As as much as 80 times soil levels.

PD and EA were equally vigorous through 40 months, but EA progeny 4938 was the most productive genotype at a toluene-contaminated site near St. Augustine, Florida. Toluene was detected in leaf and branch samples of EA but not PD. Root trainers restrict above, and presumably below, ground growth.

Several poplar clones planted as mini-barbatelles in the PCE-contaminated site at LaSalle, Illinois exceeded 5 m in height and 4 cm in DBH in 20 months. Three contained PCE in basal stem cores and various tree parts. Poplar and willow whips planted in 0.61 m wide plastic pipes in the TCE-contaminated site averaged 2.4 m tall after 12 months, with some willow clones surpassing 3 m, and effectively lowered the water table and contained the TCE plume onsite. Basal stem cores can quickly and inexpensively indicate hydrocarbon uptake by trees. A greenhouse study suggests variations between and within PD, EA, EG, willow, and cypress for TCE tolerance.

Journal Articles:

No journal articles submitted with this report: View all 7 publications for this subproject

Supplemental Keywords:

phytoremediation, dendroremediation, compost, arsenic, PCE, TCE, toluene, cottonwood, willow, Eucalyptus amplifolia, Eucalyptus grandis, Florida, Illinois, FL, IL, bioremediation, bioaccumulation, plant biotechnology,, Scientific Discipline, Waste, TREATMENT/CONTROL, POLLUTANTS/TOXICS, Sustainable Industry/Business, Treatment Technologies, Geochemistry, Chemicals, Technology, Bioremediation, New/Innovative technologies, Environmental Engineering, Agricultural Engineering, bioengineering, transgenic plants, biodegradation, woody biomass crops, plant biotechnology, photoremediation, biotechnology, remediation, hydrocarbons, bioacummulation, phytoremediation, bioenergy, heavy metals

Relevant Websites:

http://www.floridacenter.org Exit
http://www.cpbr.org Exit

Progress and Final Reports:

Original Abstract
  • 2003 Progress Report

  • Main Center Abstract and Reports:

    R829479    The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program

    Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
    R829479C001 Plant Genes and Agrobacterium T-DNA Integration
    R829479C002 Designing Promoters for Precision Targeting of Gene Expression
    R829479C003 aka R829479C011 Biological Effects of Epoxy Fatty Acids
    R829479C004 Negative Sense Viral Vectors for Improved Expression of Foreign Genes in Insects and Plants
    R829479C005 Development of Novel Plastics From Agricultural Oils
    R829479C006 Conversion of Paper Sludge to Ethanol
    R829479C007 Enhanced Production of Biodegradable Plastics in Plants
    R829479C008 Engineering Design of Stable Immobilized Enzymes for the Hydrolysis and Transesterification of Triglycerides
    R829479C009 Discovery and Evaluation of SNP Variation in Resistance-Gene Analogs and Other Candidate Genes in Cotton
    R829479C010 Woody Biomass Crops for Bioremediating Hydrocarbons and Metals
    R829479C011 Biological Effects of Epoxy Fatty Acids
    R829479C012 High Strength Degradable Plastics From Starch and Poly(lactic acid)
    R829479C013 Development of Herbicide-Tolerant Energy and Biomass Crops
    R829479C014 Identification of Receptors of Bacillus Thuringiensis Toxins in Midguts of the European Corn Borer
    R829479C015 Coordinated Expression of Multiple Anti-Pest Proteins
    R829479C016 A Novel Fermentation Process for Butyric Acid and Butanol Production from Plant Biomass
    R829479C017 Molecular Improvement of an Environmentally Friendly Turfgrass
    R829479C018 Woody Biomass Crops for Bioremediating Hydrocarbons and Metals. II.
    R829479C019 Transgenic Plants for Bioremediation of Atrazine and Related Herbicides
    R829479C020 Root Exudate Biostimulation for Polyaromatic Hydrocarbon Phytoremediation
    R829479C021 Phytoremediation of Heavy Metal Contamination by Metallohistins, a New Class of Plant Metal-Binding Proteins
    R829479C022 Development of Herbicide-Tolerant Energy and Biomass Crops
    R829479C023 A Novel Fermentation Process for Butyric Acid and Butanol Production from Plant Biomass
    R829479C024 Development of Vectors for the Stoichiometric Accumulation of Multiple Proteins in Transgenic Crops
    R829479C025 Chemical Induction of Disease Resistance in Trees
    R829479C026 Development of Herbicide-Tolerant Hardwoods
    R829479C027 Environmentally Superior Soybean Genome Development
    R829479C028 Development of Efficient Methods for the Genetic Transformation of Willow and Cottonwood for Increased Remediation of Pollutants
    R829479C029 Development of Tightly Regulated Ecdysone Receptor-Based Gene Switches for Use in Agriculture
    R829479C030 Engineered Plant Virus Proteins for Biotechnology