Open Gerst.E.Novel.Submission.4.11

The Pennsylvania State University

The Graduate School

College of Engineering

A NOVEL APPROACH TO USE SECOND GENERATION BIOFUEL CROP PLANTS

(CAMELINA SATIVA, MISCANTHUS GIGANTEUS, AND PANICUM VIRGATUM) TO

REMEDIATE ABANDONDED MINE LANDS IN PENNSYLVANIA

A Thesis in

Environmental Pollution Control

Edward A. Gerst

Submitted in Partial Fulfillment of the Requirements for the Degree of:

Master of Science

May 2014

The thesis of Edward A. Gerst was reviewed and approved* by the following:

Sairam V. Rudrabhatla Associate Professor of Biology Director, Central Pennsylvania Laboratory for Biofuels Thesis Advisor

Shirley E. Clark Associate Professor of Science and Environmental Engineering

Shobha Devi Potlakayala Assistant Professor of Biology

Thomas Eberlein Associate Professor of Chemistry Program Chair

Alison Shuler Co-director of Environmental Training Center Special Signatory

Gregory Shuler, P.G. PA Department of Environmental Protection Bureau of Mining Programs Special Signatory

*Signatures are on file in the Graduate School.

ABSTRACT

We as humans should strive to develop ways for our impact on the environment to be minimized. In the past there has always been collateral damage to the environment during times when man was doing what they felt was necessary to harness available fuel resources. Barren and abandoned mine lands remind us of the coal waste left behind from mining operations. On these abandoned lands are the elemental remnants of what once took place. Dangerous levels of elements metallic elements include Silver (Ag), Arsenic (As), Barium (Ba), Cadmium (Cd), Chromium (Cr), Mercury (Hg), Lead (Pb), and Selenium (Se). These contaminants accompany Iron (Fe), Manganese (Mn), Sulfur (S), and Aluminum (AL).

Recently, scientists have explored the use of plants to naturally absorb toxic substances into their growth tissues through natural absorption of nutrients from the soil in which the plants live. This phenomenon is known as Phytoremediation. Our study focuses on phytoextraction, which is the use of plants to accumulate pollutants to remove metals or organics from the soil by concentrating them in the harvestable plant tissue parts. We chose to study three species of biofuel crops, Camelina sativa, Miscanthus giganteus, and Panicum virgatum. Each species has different characteristics that could prove useful in this research. Camelina sativa possesses the ability to fix nitrogen in there root zone. Miscanthus giganteus is very hearty and has a deep root structure. Panicum virgatum is also hearty and has long, fibrous and densely arranged roots.

The basis of our investigation is to observe the abilities of aforementioned biofuel crop species extract heavy metals to phytoremediate marginal soils affected by mining operations. To test this hypothesis we chose soil from three locations upon the land holdings of the Eastern Pennsylvania Coalition for Abandoned Mine Land Reclamation (EPCAMR) whose office is located in the Borough of Ashley, Luzerne County, PA. We believe that each of these species of biofuel crops will respectively show favorable removal of heavy metal contamination in each of three experimental soil types.

This experiment shall answer the following questions:

1. Which of the three (3) biofuel crops selected [Camelina sativa, Miscanthus giganteus, or Panicum virgatum] has the ability to survive in the marginal soils affected by mining operations? 2. Do any of the aforementioned species thrive in such conditions? 3. Do any of these biofuel crops have the ability to Phytoremediate soils with high concentrations of heavy metals? 4. Do any of the aforementioned plant species behave as a hyper-accumulator of any of the analytes studied? 5. What is the feasibility of using any of these plant species to phytoremediate the experimental soils and also as a source of energy following phytoextraction?

Our experiment involved growing the biofuel crops in contaminated soils under greenhouse conditions. We tested the initial and final contaminant concentrations in the sample soils after plant growth. We also tested plant tissues at the end of the experiment for contaminant concentration in the plant tissues. This would indicate the level of phytoextraction which occurred. Our experiment displayed results that indicated that the soils were indeed reclaimed through the utilization of biofuel crops to phytoextract some of the heavy metal contaminants. There were also instances where hyper-accumulation occurred. We are hopeful that these results could lead to further investigation to determine the feasibility of field stage application. Future experiments should also be conducted to determine how the environmental impact when these biofuel products are combusted.

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TABLE OF CONTENTS

List of Tables ...... vi List of Figures ...... vii Acknowledgements ...... viii

Chapter 1, INTRODUCTION ...... 1 Thesis Statement/Objective Questions ...... 3 Toxicity of Heavy Metal Contamination ...... 4 Phytoremediation ...... 5 Hyper-Accumulation ...... 10 Use of Biofuel Crops ...... 11

Chapter 2, METHODS AND MATERIALS ...... 13 Experimental Design ...... 14 Soil Collection and Preparation...... 15 Preparation of planting containers and plants ...... 17 Germination Records ...... 19 Growth Records ...... 19 Laboratory Analyses ...... 20 Methods for Calculations ...... 21 Variability Calculations...... 21 Percent Germination Calculations ...... 22 Mass-Volume Approximation Calculations ...... 22 Mass-Balance Calculations ...... 23 Chemical and Physical Properties ...... 24

Chapter 3, RESULTS ...... 26 Summary Results of Germination Records...... 27 Summary Results of Growth Records ...... 31 Summary Results of Laboratory Analyses ...... 37 Summary Results of Calculations ...... 40 Soil Variability Results ...... 41 Plant Tissue Variability Results...... 46 Percent Germination Results ...... 51 Mass-Balance Calculation Results ...... 52

Chapter 4, INTERPRETATION OF RESULT ...... 57 Discussion of Plant Species ...... 58 Discussion of Germination Results ...... 58 Discussion of Growth Record Results ...... 59 Discussion of Laboratory Results...... 60 Discussion of Variability Results ...... 62 Discussion of Mass-Balance Results ...... 63

Chapter 5, CONCLUSION ...... 65

Bibliography ...... 70 Appendix A, FORMS AND DEADLINES ...... 74 Master’s Approval Page...... 75

Appendix B, GROWTH RECORDS ...... 76 Spreadsheet of Daily Growth Records ...... 77 Field Data Sheets ...... 104

Appendix C, LABORATORY RESULTS ...... 124 Initial Soil Sample Results ...... 125 Initial Native Plant Tissue Sample Results ...... 132 Final Soil and Plant Tissue Sample Results for Trial 1 ...... 139 Final Soil and Plant Tissue Sample Results for Trials 2 – 4 ...... 166

Appendix D, CALCULATIONS ...... 188 Percent Germination Calculations ...... 189 Mass-Volume Calculation ...... 189 Approximate plant tissue mass calculations ...... 190 Mass-Balance Calculations ...... 192

LIST OF TABLES

EPA Reference Test Method: Table 1, EPA Reference Test Method ...... 21 Chemical and Physical Properties: Table 2, Chemical and Physical Properties...... 25 Summary Results of Germination Records: Table 3a, Results, Trial 1 ...... 27 Table 3b, Results, Trial 2 ...... 28 Table 3c, Results, Trial 3 ...... 29 Table 3d, Results, Trial 4 ...... 30 Summary Results of Chemical Analyses: Table 4a, Results, Initial Soil Analysis ...... 37 Table 4b, Results, Native Plant Tissue Analysis ...... 37 Table 5a, Results, Final Soil Analysis, Trial 1 ...... 38 Table 5b, Results, Final Soil Analysis, Trials 2 and 3 combined ...... 38 Table 6a, Results, Final Plant Tissue Analysis, Trial 1 ...... 39 Table 6b, Results, Final Plant Tissue Analysis, Trial 2 ...... 39 Table 6c, Results, Final Plant Tissue Analysis, Trial 3 ...... 40

Summary Results of Calculations:

Percent Germination: Table 7a, Results, Percent Germination, Trial 1 ...... 51 Table 7b, Results, Percent Germination, Trial 2 ...... 51 Table 7c, Results, Percent Germination, Trial 3 ...... 52 Table 7d, Results, Percent Germination, Trial 4 ...... 52

Mass-Balance:

Camelina sativa: Table 8a, Summary Results, Mass-Balance, Soil 1 ...... 52 Table 8b, Summary Results, Mass-Balance, Soil 2 ...... 53 Table 8c, Summary Results, Mass-Balance, Soil 3 ...... 53 Miscanthus giganteus: Table 9a, Summary Results, Mass-Balance, Soil 1 ...... 54 Table 9b, Summary Results, Mass-Balance, Soil 2 ...... 54 Table 9c, Summary Results, Mass-Balance, Soil 3 ...... 55 Panicum virgatum: Table 10a, Summary Results, Mass-Balance, Soil 1 ...... 55 Table 10b, Summary Results, Mass-Balance, Soil 2...... 56 Table 10c, Summary Results, Mass-Balance, Soil 3 ...... 56

LIST OF FIGURES

Figure 1, Coal Mining ...... 2 Figure 2, Effects of Acid Mine Drainage ...... 4 Figure 3, Remediation ...... 6 Figure 4, Types of Phytoremediation ...... 8 Figure 5, Phytoextraction ...... 9 Figure 6, Hyper-Accumulation ...... 11 Figure 7, Native Plant Species at EPCAMR ...... 14 Figure 8, Images from EPCAMR Soil excavation...... 15 Figure 9, Soil Preparation ...... 16 Figure 10, Plant Preparation ...... 17 Figure 11a, Monitoring of Plant Growth, Germination ...... 18 Figure 11b, Monitoring of Plant Growth, Vegetative ...... 19 Figure 12a, Results, Camelina sativa Growth Records, Trial 1 ...... 31 Figure 12b, Results, Camelina sativa Growth Records, Trial 2 ...... 31 Figure 12c, Results, Camelina sativa Growth Records, Trial 3 ...... 32 Figure 12d, Results, Camelina sativa Growth Records, Trial 4 ...... 32 Figure 13a, Results, Miscanthus giganteus Growth Records, Trial 1...... 33 Figure 13b, Results, Miscanthus giganteus Growth Records, Trial 2 ...... 33 Figure 13c, Results, Miscanthus giganteus Growth Records, Trial 3...... 34 Figure 13d, Results, Miscanthus giganteus Growth Records, Trial 4 ...... 34 Figure 14a, Results, Panicum virgatum Growth Records, Trial 1 ...... 35 Figure 14b, Results, Panicum virgatum Growth Records, Trial 2 ...... 35 Figure 14c, Results, Panicum virgatum Growth Records, Trial 3 ...... 36 Figure 14d, Results, Panicum virgatum Growth Records, Trial 4 ...... 36 Figure 15a, Aluminum Soil Variance ...... 41 Figure 15b, Arsenic Soil Variance ...... 41 Figure 15c, Barium Soil Variance ...... 42 Figure 15d, Cadmium Soil Variance ...... 42 Figure 15e, Chromium Soil Variance ...... 43 Figure 15f, Lead Soil Variance ...... 43 Figure 15g, Mercury Soil Variance...... 44 Figure 15h, Selenium Soil Variance ...... 44 Figure 15i, Silver Soil Variance ...... 45 Figure 15j, pH Soil Variance ...... 45 Figure 16a, Aluminum Plant Tissue Variance ...... 46 Figure 16b, Arsenic Plant Tissue Variance ...... 47 Figure 16c, Barium Plant Tissue Variance ...... 47 Figure 16d, Cadmium Plant Tissue Variance ...... 48 Figure 16e, Chromium Plant Tissue Variance ...... 48 Figure 16f, Lead Plant Tissue Variance ...... 49 Figure 16g, Mercury Plant Tissue Variance ...... 49 Figure 16h, Selenium Plant Tissue Variance...... 50 Figure 16i, Silver Plant Tissue Variance ...... 50 Figure 16j, Sulfur Plant Tissue Variance ...... 51

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ACKNOWLEDGEMENTS

This research would not have been possible without the support and guidance from a great number of amazing teams and individuals. No great deed is accomplished alone. The primary author would like to thank The Graduate School of Pennsylvania State University. Especially all the amazingly passionate instructors involved in the Science, Technology, Engineering and

Mathematics (STEM) disciplines. The Environmental and Civil Engineering department at Penn

State Harrisburg offered great insight to bring this research to fruition.

We would like bring special recognition

• National Science Foundation’s Research Experience for Undergraduate Programs Penn

State Harrisburg

• Mr. Robert Hughes, Executive Director of the Eastern Pennsylvania Coalition for

Abandoned Mine Reclamation (EPCAMR)

• *REU students Tyler Bowe and Matt Robinson

• Staff and students of the Central PA Laboratory for Biofuels at Penn State Harrisburg

• Analytical Laboratory Service (ALS), Middletown, PA

• Microbac Laboratories, Harrisburg, PA

• US Department of Agriculture

• Mr. Gregory Shuler, P.G., PA Department of Environmental Protection

• Cynthia Gerst and children (Son Kameron, Daughter Korynn and Son Korben)

• This research is dedicated in memory of Mrs. Diana Hudler and Mrs. Pamela Gerst

*NSF REU student interns

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Chapter 1: INTRODUCTION Thesis Statement and Questions

Toxicity of Heavy Metal Contamination

Phytoremediation

Hyper-Accumulation

Use of Biofuel Crops

Chapter 1: INTRODUCTION

In the world today there is a constant battle for a balance between wants and needs. For example, the need for energy to sustain man’s Figure 1 Coal Mining insatiable appetite for the comforts that we have become accustomed to is in constant conflict with the need to treat our environment with respect. The ability to view the overall picture and lead a balanced existence is imperative for a sustainable

Coal mining operations have caused many ecosystem. This master’s thesis will provide some environmental problems in Pennsylvania. ways to offset the anthropomorphic damage that Image from: http://quiet-environmentalist.com/wp- has occurred in the past and offer ways to develop a content/uploads/2011/02/Dirty-Coal.jpg more sustainable environmental stance for the future.

In Pennsylvania, vast amounts of land have been virtually disregarded (neglected?) due to the natural resources being stripped out of the land. What is left is a barren wasteland that is contaminated with a myriad of different chemicals and minerals. This study focuses on the effects of potentially toxic heavy metal contaminants left in the environment after previous mining ventures. These abandoned mine lands are left consisting of very marginal soil types.

The mine refuse formed from the coal waste lead to the formation of acid mine drainage (AMD).

This occurs when water passes over mining refuse and is exposed to the atmosphere. We have chosen to utilize Miscanthus giganteus, Camelina sativa, and Panicum virgatum biofuel crops to reclaim the compromised soil types and provide a viable locally-produced renewable energy source. Each of these plant species has characteristics that could benefit the aforementioned marginal soils. These crops can be used as a renewable energy source following the uptake of

potentially harmful concentrations of heavy metal waste products. Camelina sativa has the ability to fix nitrogen throughout its life cycle. Camelina also produces seeds that possess a high percentage of oil that can be refined into biodiesel. Miscanthus giganteus is extremely hearty with roots that penetrate deeply in the ground. It produces a large amount of biomass that can be used to form pellets that may be incinerated. The biomass may also be processed to form ethanol. Additionally, miscanthus may inhibit erosion if managed properly. Panicum virgatum is also hearty and possess long fibrous roots that are densely arranged. These roots may inhibit erosion and can act as a sponge to capture contaminants and/or nutrients that may be moving below the earth’s surface. The biomass from Panicum virgatum may be formed into pellets and/or used to produce ethanol.

An objective of this research is to determine answers for each of the following questions:

1. Which of the three (3) biofuel crops selected [Camelina sativa, Miscanthus giganteus, or

Panicum virgatum] has the ability to survive in the marginal soils affected by mining

operations?

2. Do any of the aforementioned species thrive in such conditions?

3. Do any of these biofuel crops have the ability to Phytoremediate soils with high

concentrations of heavy metals?

4. Do any of the aforementioned plant species behave as a hyper-accumulator of any of the

analytes studied?

5. What is the feasibility of using any of these plant species to phytoremediate the

experimental soils and also as a source of energy following phytoextraction?

Toxicity of Heavy Metal Contamination:

When minerals, or fossil fuels, were mined many years ago it caused byproducts to be released into the environment. Historically, precautions were not taken into consideration until the formation of the Resources Conservation and Recovery Act (RCRA) of 1976, which changed the way solid wastes were handled during mining enterprises. At this point many of the metal contaminants were identified. Some of these substances are beneficial and even needed for proper metabolism in animals and plants. “Some heavy metals namely, cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), nickel (Ni) and zinc (Zn) are considered to be essential for plants, whereas chromium (Cr), and antimony (Sb) are found to be essential for animals” (Misra and Mani 1991; Markert 1993). There are many more instances where these elements and compounds are considered to be toxic and harmful to plants and animals. The mechanism of the substances is varied and some can be related to metabolic pathways. For instance, “chromium toxicity in plants vary from the inhibition of enzymatic activity to mutagenesis” (Barcelo et al. 1993). The soil profile Figure 2 Effects of Acid Mine Drainage and make-up play an important role in the ability for species to survive in marginal soils.

Abandoned mine lands in Pennsylvania are a prime example of where this important balance went wrong. Amidst the industrial revolution and the years that followed, man’s way of life recklessly

took advantage of the vast supplies of coal. During Effects on environment from Acid Mine Drainage. (Gerst, 2012). these early years of mining, the techniques of mining operations did not consider the environment. Tracks of land, where coal waste materials

were discarded, left marginal soil devastated by low pH and high concentrations of heavy metals.

The mining waste products and coal refuse allows the production of AMD. “Generally, higher the clay and/or organic matter and soil pH, the metals will be firmly bound to soil with longer residence time and will be less bioavailable to plants” (Chang et al. 1987). Heavy metals are those elements that are dense and share particular physical properties with one another. “Heavy metals are defined as the elements having density greater than 5 grams per cm3” (Adriano 2001).

Plants and other organisms require small concentrations of these elements to perform essential metabolic mechanisms. “Although many metal elements are essential for the growth of plants in low concentrations, their excessive amounts in soil above threshold values can result in toxicity”

(Shah, Ahmad, Masood, Peralta-Videa & Ahmad, 2010).

The toxicity of each of these substances can vary significantly from substance to substance as does the molecular arrangement and structure. “Heavy metal toxicity in plants depends on the bioavailability of these elements in soil solution, which is a function of pH, organic matter and cation exchange capacity to the soil” (Shah, Ahmad, Masood, Peralta-Videa

& Ahmad, 2010). The ability for organisms to process the chemicals may also provide ill effects. “For example, the bioaccumulation of heavy metals in excessive concentrations may replace essential metals in pigments or enzymes disrupting their function and causing oxidative stress” (Shah, Ahmad, Masood, Peralta-Videa & Ahmad, 2010).

Phytoremediation:

Phytoremediation is the ability of certain plants to naturally remediate contaminated soils by the uptake of inorganic compounds from the soil into the plants tissues. Contamination stems from many sources, directly or indirectly caused by man. These anthropomorphic activities can be blamed for many of the world’s environmental perils that now have to be addressed. Many

different sites have been neglected due to the high cost of traditional remediation methods. In order to clean soils that are contaminated

Figure 3 with metals the cost is roughly “$250 per Remediation cubic yard, add explosive residues, and the cost jumps to about $1020 per cubic yard for incineration” (Wong, 2004).

Along with the heavy monetary costs come the production of greenhouse gases

(CO or CO2) and nitrous oxides (NOx) from the construction machinery and Traditional remediation methods include physical excavation working conditions. There are many and removal of contaminated soil. Image from: http://systemsbiology.usm.edu/BrachyWRKY/WRKY/IMG/ different ways that contaminants can Phytoremediation-01.jpg affect the environment, air, water, and soil being the most common forms of media. They each act hand in hand through daily interactions between phases that are dictated by partitioning coefficients (i.e. KOA, KOW, or KOC) and other chemical and physical properties (see Table #2). As these interactions occur contaminants are left in the environment. Traditionally, this pollution has been removed through physical remediation and restoration projects involving great economic, ecological, and energy investments. Luckily many ecosystems have a number of methods built in naturally to help with this issue.

For today’s stakeholders, the economic impact is a key determinant of which remediation technique to utilize. One such financially sound technique is phytoremediation, including methods such as bioremediation and phytoremediation. Bioremediation involves the

biodegradation of contaminants through the use of bacterial organisms. These microorganisms break down the organic pollutants to remove them from the environment. The amount of time it takes to break down is usually represented by half-lives and can be accounted for through mass- balance relationship equation calculations.

Phytoremediation is emerging as a preferred solution to soil contamination for a number of reasons. One reason is its potential economic impact. ”Phytoremediation is an attractive alternative to remediate contaminated soil naturally and cost effectively” (Wong, 2004).

Furthermore, phytoremediation has become a crucial part of the environmental market.

“Phytoremediation costs about $80 per cubic yard” (Wong, 2004). The lower cost of phytoremediation makes hazardous waste cleanup more feasible for many abandoned sites.

“This cost-effective plant-based approach to remediation takes advantage of the remarkable ability of plants to concentrate elements and compounds from the environment and metabolize various molecules in their tissues” (Salt, 2005). Another reason phytoremediation is beneficial to the environment is its ability to work instinctively by harnessing a naturally occurring process, which has had a great impact on the air quality for these areas. It positively affects atmosphere in a number of ways. One way is by significantly decreasing the necessity to use heavy construction equipment and eliminating their subsequent harmful emissions from the combustion of fossil fuels. The plants also sequester carbon dioxide (CO2) and replenish the atmosphere with oxygen (O2) through photosynthetic processes. This introduces the concept of CO2 abatement.

There are a number of ways that plants can act through Phytoremediation. Some of the types of phytoremediation are: phytodegradation, phytovolatilization, phytoextraction, phytostabilization, phytostimulation and rhizofiltration (See Figure 4). Each of the

aforementioned phytoremediation methods favor plants that grow rapidly, are hardy, have high Figure 4 Types of Phytoremediation biomass production, and are tolerant to harsh soil conditions.

Many different pollutants can be handled through phytoremediation. Metals are typically handled through phytoextraction. The degradation of organic pollutants may occur in the root zone of Image from: http://systemsbiology.usm.edu/BrachyWRKY/W

RKY/IMG/Phytoremediation-01.jpg plants or extracted, sequestered, or volatilized through the above ground plant tissues. The chemical properties determine the way the pollutant is treated. (Pilon-Smits, 2005)

Following is a breakdown of each of these methods of phytoremediation:

 Phytodegradation is when plants have the ability to breakdown harmful chemicals during

metabolism. Plants with large, tightly arranged roots systems and high enzymatic activity

are favored for this technique. (Pilon-Smits, 2005)

o This technique “involves the degradation of contaminants such as pesticides,

explosives, and organic solvents by the metabolic processes in plants. This

breakdown depends on the specific enzymes produced by the plant species”

(Wong, 2004).

 Phytovolatilization is the process when pollutants are taken into the plants vascular

system and emitted through the leaves in a volatile form. Some plants can absorb and

transpire pollutants after being carried through the phytosystem. (Pilon-smits, 2005).

This technique may take a soil or water problem and make it an air quality issue. The

chemical is degraded during the process through the process of phytodegradation.

 Phytoextraction is the ability of some plants to Figure 5 Phytoextraction take up pollutants and accumulate them in their

above ground leafy tissues. These can then be

harvested and disposed of or used properly.

This method is important for the removal of

inorganic or metal contaminants. (Pilon-Smits,

2005) Phytoextraction is an excellent way to remove heavy metal toxins from contaminated soils. o “Toxic heavy metals and organic Image from: Http://www.webapps.cee.vt.edu/ewr/environm pollutants are the major targets for ental/teach/gwprimer/group17/phyto/Extract.g if phytoremediation” (Salt, 2005). These

targets are best contacted through phytoextraction.

 Phytostabilization is the use of plants to prevent the migration of pollutants by preventing

erosion, leaching, or runoff. This can also involve the chemical conversion of pollutants

to less reactive forms. This could involve the precipitation of contaminant in the plants

root zone (aka: rhizosphere). (Pilon-Smits, 2005)

 Phytostimulation (or Rhizodegradation) is the ability of plants to form a sort of

mutualistic relationship with microorganism (bacteria) in the root zone of the plant. A

larger root area (Rhizosphere) favors this type of phytoremediation due to the promotion

of bacterial growth in the soil, a form of facilitated biodegradation. “For

phytostimulation of microbial degraders in the root zone, grasses such as fescue (Festuca

sp.), ryegrass (Lolium sp.), switchgrass (Pancium sp.), and prairie grasses (e.g., Buchloe

dactyloides, Bouteloua sp.) are very popular because they have dense and relatively deep

root systems and thus large root surface area” (Pilon-Smits, 2005).

 Rhizofiltration “is the absorption or adsorption of pollutants into the plants root zone”

(Wong, 2004). Some plants have the ability to isolate toxic substance in the soil

surrounding their roots. “To avoid the toxicity, plants have developed specific

mechanisms by which toxic elements are excluded, retained at root level, or transformed

into physiologically tolerant forms” (Shah, Ahmad, Masood, Peralta-Videa & Ahmad,

2010).

As with all methods of remediation these techniques have pros and cons. Since plants are inhibited in movement they have to be near the pollutant. They also have to be able to react with the chemicals thus the soil properties, toxicity level, and climate should be suitable for the particular plant species growth. This is linked to the selection of the plant species type. It is crucial that the plant can grow properly under the adversity of the contaminated ecosystem. The effectiveness of phytoremediation also depends on the dispersion of the pollutant. Where the pollutant is in relation to the plant is key. This is related to the root zone distribution. Another limitation can be linked to the speed of the process. It can be very slow depending upon the biological processes that are applied. More traditional remediation techniques are much faster

(such as excavation, incineration, or pump-and-treat systems.) (Pilon-Smits, 2005)

Hyper-Accumulation

Some plants have the uncanny ability to make very good use of their process when accumulating one or more inorganic elements. When a plant can absorb 100-fold higher amounts of contaminants it is called a hyper accumulator. “Hyper accumulators have been reported for Arsenic (As), Cobalt (Co), Copper (Cu), Manganese (Mn), Nickel (Ni), Lead (Pb),

Selenium (Se), and Zinc (Zn)” (citation needed). Although these plants have amazing abilities, they are not commonly selected due to their Figure 6 slow growth and low biomass generation. Hyper-Accumulation

These types of plants could be very useful in attaining the conditions of EPA’s Resource

Conservation and Recovery Act (RCRA). An example of a hyper-accumulator can be seen in

Pteris vittata which is a variety of a fern. It has Hyper accumulators have the ability to absorb up to 100 times as much contaminants as compared to other been identified to be an Arsenic (As) hyper- plants grown under similar conditions. accumulator. (Pilon-Smits, 2005) Image from: http://t3.gstatic.com/images?q=tbn:ANd9GcSp1GZz Use of Biofuel crops: AyP0XZnr2cppn_BVhypBMJ5nfIzDgy1BvRmHSUs 77En8

Using biofuel crops for phytoremediation purposes is a relatively new topic. It focuses on the importance of the potential for increased sustainability by using biofuel crops for phytoremediation. We hope to offer some further research into this area. Using biofuel plant species for phytoremediation also expands on the topic of carbon dioxide (CO2) abatement. The use of biofuel crops for phytoremediation would offset CO2 production by decreasing the use of heavy equipment used in conventional forms of remediation. Furthermore, biofuel crops could compound this positive impact by also supplying a viable source of fuel. These fuel products could potentially burn much more cleanly than conventional fossil fuel based products. They could also be produced locally, which would allow for more energy independence on the local level. Ethanol is already being used as an additive for fuels and is gaining commercial acceptance. The emissions from combusted biofuels will align with the use of conventional fuel types.

Another concept to consider is the importance of replacing current biofuel crops.

Currently the majority of biofuel is derived from food based crop such as sugar cane or corn.

There is a distinct benefit from by using non-food crops, such as Camelina sativa, Miscanthus giganteus, Panicum virgatum, along with a number of other possibilities. Each of these species has different benefits and challenges. Camelina sativa has great success once established, but typically has difficulty thriving in adverse conditions (i.e. low pH). There are many varieties (at least 50) Camelina sativa which each may have different survival capabilities. Camelina sativa also has the ability to fix nitrogen throughout its life cycle. Miscanthus giganteus is a very hearty plant with a deep root system with rhizomes. Panicum virgatum have long roots that may be very densely arranged in the root zone of each plant. It is also important to not take away from the worlds food supply while attempting to solve energy or pollution issues. In the future it would be great to see a push to use phytoremediation as a tool for environmental pollution control. The practical application of government based financial initiatives may aid in bringing this to fruition.

The thesis will progress in the following manner: Chapter 2, Methods and Materials, will go into great detail upon how biofuel crops can be used for the phytoremediation of heavy metal contaminants from acid mine drainage affected soils in Pennsylvania; Chapter 3, Results, will present the findings of this experiment; Chapter 4, Interpretation of results, will elaborate on the meaning of these findings; Chapter 5, Conclusion, will explain the final findings of the project, offer answers to the theoretical questions earlier, and provide future avenues for research.

Chapter 2: METHODS AND MATERIALS

Experimental Design

Soil Collection and Preparation

Preparation of planting containers and plants

Germination records

Growth records

Laboratory Analysis

Calculation Methods

Chemical and Physical Properties

Chapter 2: METHODS AND MATERIALS

Experimental design:

We selected three species of Biofuel crops for this experiment. They were Camelina sativa, Miscanthus giganteus, and Panicum virgatum. Each of the selected biofuel crops interact with the soil in slightly different ways. The root system of the Miscanthus giganteus has different properties than those of the Camelina sativa Figure 7 which also differ from those of the Panicum virgatum. Native plant species at EPCAMR:

Miscanthus plants have very deep root systems while

Panicum have long tightly arranged roots. Camelina roots are very thin and fragile. These differences (and other physical attributes) from species to species play an enormous role in how well each will survive, thrive, and ultimately phytoremediate the contaminated soil samples that we are studying.

One area of interest to be addressed in this experiment is the ability of each species to survive in different contaminated soil types. This will play an enormous role in how well each species will first germinate and establish a viable root system. Next they can gain the ability to grow well through the vegetative growth stages and produce biomass in plant tissue. After each specimen was established the plant (Gerst, 2012) tissues grew and had the ability to remove

contaminants (through phytoremediation) from the experimental soils and translocate the heavy metals into the plant leaf tissues. Figure 8 Soil Collection and Preparation: Images from EPCAMR Soil Excavation: The first order of business at hand was to design Location 1: The Office the experiment in a way to account for each stage of growth. To accomplish this task we first travelled to the

Eastern Pennsylvania Coalition for Abandoned Mine land Reclamation (EPCAMR) located in Ashley Location 2: Loomis Bank Borough, located in Luzerne County, PA. This area in

Northeastern Pennsylvania was a hotbed for both surface and deep mining operations of Anthracite Coal throughout the early 1900’s.

Soil was collected for the experiments after we

were given permission. Native plant tissue samples were Location 3: Honey Pot also taken to serve as additional experimental control samples. The initial soil sample sizes were two five gallon buckets (placed in black trash bags) from each location.

We selected three different locations throughout

EPCAMR’s properties holdings throughout Luzerne (Gerst, 2012)

County, PA to diversify the soil survey throughout the site and provide different soil types. The first region (Location 1) from which we sampled was located near the EPCAMR office in Ashley

Borough, Luzerne County, PA. This was right next to the old coal elevator and breaker plant.

This soil was very sandy and had a large amount of small gravel sized rocks present. There were

a number of native plant species growing very sparsely throughout this area. The second region

(Location 2) from which we sampled was located on what was called the “Loomis Bank”. The

“Loomis Bank” is located in Hanover Township, Luzerne Figure 9 Soil Preparation: County, PA. This area was where a great deal of waste coal Soil Blending: was piled up creating a hill side. There was a small layer (about

3 inches) of topsoil on top of the waste coal.

Surprisingly, there was a great deal of vegetation growing in this topsoil (including poison ivy). This sample was Potting Preparation: obtained from a hill side with a steep slope. This soil had a lot of boney coal and rocks. Digging and collecting these samples was difficult due to the very thin soil layer. The third and last region (Location 3) from which we sampled was called the

“Honey Pot” and was located at a bottom of a valley. The

“Honey Pot” is located in Newport Township, Luzerne County,

PA. A nearby stream showed signs of AMD. This soil appeared much darker (black in color) and a high amount of Initial Soil Sample preparation: (For analysis at lab) shimmery silt. The native vegetation was limited to mostly larger evergreens (pines and hemlocks) and some sparsely located bunch grasses. Larger amounts of soil were required to repeat the experiment for three (3) more trials.

After the soil was collected we headed to the greenhouse (Gerst, 2012) to prepare the soil. The two bags from each site were combined in large (approximately 25 - 30 gallon) planters and mixed thoroughly. Grab samples (about ½ quart sized sandwich bag) were

taken from of each soil type to be sent off for laboratory analysis. The samples were analyzed for the concentrations of Sulfur (S), Aluminum (Al), Arsenic (As), Barium (Ba), Cadmium (Cd),

Chromium (Cr), Lead (Pb), Mercury (Hg), Selenium (Se), and Silver (Ag). Each of these concentrations was used respectively as the initial soil concentrations throughout this experiment. Each sample Figure 10 Plant Preparation: was also analyzed for pH. Sowing of seeds (Both Seeded species only): Preparation of planting containers and plants:

The soil from each location was then divided equally between (approximately 3 gallon) sample planters for each of the plant species. This planter pot size was selected to allow for the mature plants to be able to thrive throughout the experiment. This used up the initial soil harvested rather Separation and Cutting (Rhizome species only): quickly. There was only enough growth media to provide for one trial. This trial consisted of twelve (12) total potted samples, with nine (9) experimental potted samples and three (3) control potted samples. The control soil planter pots were prepared with virgin potting soil. (Robinson, 2012)

To solidify our results we prepared subsequent samples for three (3) more trials (for Trials 2-4). The planting containers were then placed on trays to collect any possible run-through caused by watering. A great deal of run-through could affect the soil chemistry. This leachate was not considered for the chemical transport and transfer of the inorganic heavy metal contaminants. The leachate was unfortunately discarded and not analyzed.

After the planting containers were prepared for each of the three experimental and control soil types for each of the three Figure 11a Monitoring of plant growth: plant species the pots were readied for planting. The Miscanthus Germination phase: (Both Seeded species only) giganteus required preparation of split root portions of parent plants due their rhizosomal reproductive properties. This technique was used due to the long sexual reproductive cycle typical of this species of plant. Four (4) Miscanthus giganteus Measurement of growth: Miscanthus giganteus specimens separated by splitting a parent plant were prepared for each trial. These sample specimens (trimmed to 7.2 cm for trial 1 and 10 cm for trials 2-4) were planted each of the experimental and control planters. A small amount (approximately 50 ml) of Camelina sativa water was added to the soil to assist with acclimation to the soil.

Approximately one-hundred (100) seeds of the other two plant species (Sunson variety for Camelina sativa and Blackwell Panicum virgatum variety for Panicum virgatum) were counted and prepared to be sown into the experimental and control soil planters. The sowing of the seeds included distributing the seeds and mixing them evenly and covering them with approximately an inch of soil.

Then the seeds were sown into each of the planters and were (Gerst and Robinson, 2012) watered in with a small amount (approximately 50 ml) of water.

Germination Records:

The two plant species that were grown from seed Figure 11b Monitoring of plant growth: were Camelina sativa and Panicum virgatum. Each of Initial these species was monitored for the first few weeks for percent germination. This was accomplished by simply making periodic observations to count seed emergence. ≈Two Weeks Growth Growth Records:

Now that the samples have been prepared a rigorous watering schedule was established to provide the

≈Ten Weeks Growth necessary moisture required successful plant germination and ultimately growth. Initial watering volumes added to each sample were kept very small (200 ml) to discourage run-through. The initial goal was to eliminate any loss of water through run-through or leaching. Each Miscanthus giganteus was observed through measuring the growth from the beginning. The Camelina sativa and Panicum (Gerst and Robinson, 2012) virgatum were initially observed for percent germination.

Percent germination was determined by observing (counting) the number of seeds that germinated over time. This was repeated until the germination period was completed and observations could be made by measuring growth. Growth was measured every few days depending on availability. All measurements were conducted with the same laboratory ruler and subsequent meter stick in the same manner.

Laboratory analyses:

In order to test the hypothesis, and answer the postulated questions for our research, each of the initial soil and final soil and plant tissue samples were analyzed for inorganic (heavy metal) contamination. Each sample was analyzed in accordance with the EPA Reference Test

Methods listed in Table #1. The following laboratory analyses were conducted:

1. Laboratory testing for the initial soil samples was performed by ALS Environmental in

Middletown, PA.

2. Laboratory testing for the initial plant tissue samples from native plant species was

performed also by ALS Environmental in Middletown, PA.

3. Testing for the final soil samples and plant tissue samples for Trial 1 was performed by

ALS Environmental in Middletown, PA.

4. Testing for the final soil samples and plant tissue samples for Trials 2 and 3 was

performed by Microbac Laboratories, Inc. in Harrisburg, PA.

Table #1, EPA Reference Test Methods Parameter Epa Method Laboratory Corrosivity, pH SW846-9045D ALS Environmental and Microbac Laboratories, Inc. Moisture SM20-2540G ALS Environmental Total Solids SM20-2540G ALS Environmental and Microbac Laboratories, Inc. Aluminum, Al SW846-6010C ALS Environmental Aluminum, Al SW846-6010B Microbac Laboratories, Inc. Arsenic, As SW846-6020A ALS Environmental Arsenic, As SW846-6010B Microbac Laboratories, Inc. Barium, Ba SW846-6020A ALS Environmental Barium, Ba SW846-6010B Microbac Laboratories, Inc. Cadmium, Cd SW846-6020A ALS Environmental Cadmium, Cd SW846-6010B Microbac Laboratories, Inc. Chromium, Cr SW846-6020A ALS Environmental Chromium, Cr SW846-6010B Microbac Laboratories, Inc. Lead, Pb SW846-6020A ALS Environmental Lead, Pb SW846-6010B Microbac Laboratories, Inc. Mercury, Hg SW846-7471B ALS Environmental Mercury, Hg SW846-7471A Microbac Laboratories, Inc. Selenium, Se SW846-6020A ALS Environmental Selenium, Se SW846-6010B Microbac Laboratories, Inc. Silver, Ag SW846-6020A ALS Environmental Silver, Ag SW846-6010B Microbac Laboratories, Inc. Sulfur, S SW846-6010C ALS Environmental Sulfur, S SW846-6010B Microbac Laboratories, Inc. All EPA Methods are accessible at the technology transfer network. The website is: www.EPA.gov/ttn. The equation that is used to calculate the standard deviation is: Methods for Calculations:

Variability Calculations:

https://encrypted- The variability among the experimental conditions were tbn0.gstatic.com/images?q=tbn:ANd9GcT1I nGUZe7alrtIYUOiuyp18zgYC4rbrdPZ1fp approximated by calculating the standard deviation Mk2ZFKJvYyO5b between each soil type. The standard deviation is a statistical function that assists with data analysis. It indicates the variation between data points and the average values in the experimental data set. The standard deviation calculations were perfomed by a function in Excel.

They are displayed on the variance graphs in the form of error bars. If the standard deviation value is larger, then the greater the variance and vice versa.

Percent Germination Calculation:

The percent germination was calculated in order to determine the ability for the seeded species

(Camelina sativa and Panicum virgatum) to survive in the experimental soils. Each of the trials was evaluated for percent germination. We selected a seed sowing value of approximately one hundred (100) to allow for a simple percent germination calculation. Essentially the maximum number of germinated seeds was equal to the maximum germination for the seeded plant species.

This was accomplished by the following equation:

(Number of germinated seeds)] / (Total number seeds sewn) * 100 = germination rate

[(Nsgerminated) / (Nssewn)] * 100 % Germination

Mass-Volume Approximation Calculations:

The mass and volume values to be considered for each of the media involved in this experiment (soil and plant tissue) were approximated using standard unit conversions. Each of the pots used for the experiments were considered to be three (3) gallons. The pots were assumed to be ninety percent (90%) full. Values for the plant tissues were estimated based on yield values found in other studies.

The following equations were used to approximate the volume of soil:

Msoil = Vsoil * ρsoil

3 3 Vsoil = 3 gallons * (3.785 liters/gallon) * (1 m / 1000 L) * (0.90) = 0.0102 m

The following values were used to approximate the expected biomass yield for each plant species:

Camelina sativa:

Mplants-tissue = 2134.5 kg/ha/year (averaged from 1638, 3106, 1987, 3320, 1096, 1660

kg/ha) (Vakulabharanam, 2010)

Miscanthus giganteus:

Mplant-tissue = 8.2 dry ton/acre/year (averaged from 6.6 and 9.8 dry ton/acre/year)

(Caslin et. al., 2010)

Panicum virgatum:

Mplants-tissue = 6 Metric tons/acre/year (Jensen et al., 2005)

These values were converted to reflect the estimated yield output from our experiment. The resulting calculated yield value was then used in order to approximate mass balance.

Mass-Balance Calculations:

The Mass-Balance approach was applied to the soil and plant tissues to determine the amount of phytoextraction that occurred. This principle is based on the Law of Conservation of

Mass (principle of mass conservation) and the First Law of Thermodynamics, which states that energy is conserved in a closed system. For this research we did not consider the potential loss of mass to the leachate. The experiment was designed in a manner to limit the amount of water added to the system. This was an attempt to minimize the leaching effect. Each of the trials was evaluated for the mass balance transfer and calculations were performed for each of the contaminants that demonstrated phytoextraction. The approximate values for volume and density for the soils and plant tissues was used for this purpose. Average plant tissue volume was approximated by growth portion of this study along with average yields provided and supported through other studies. Lab analyses with non-detect (ND) were calculated with the

reporting detection limit (RDL). Lab analyses with no-analysis (NA) were calculated with a zero value to represent the lack of sample analysis.

The following equations were used to help interpret the results of this experiment:

Total Massin = Total Massout

Total [(Concin)*(Volumein )*(Densityin) ] = Total [(Concout)*(Volumeout)* (Densityout]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

3 Approximate density of the soils: Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Chemical and Physical Properties:

The chemical and physical properties offer the ability to determine how chemicals behave in the environment. These properties are crucial in modeling the chemical fate and transport through the environment and they offer insight to the mobility of chemicals.

Table # 2, Chemical and Physical Properties Partitioning Coefficients Molar Dens Vapor Prop- Melting Point, Log Henry’s law Mass, -ity, Water solubility pressure, erty Tm S KOW constant, H M ρ P L KOW KOC

Para- g/100 ml Other g/mol °C K g/cm3 Pa na L/kg Pa m3/mol meter H2O solubility

1.27E- Ag 107.87 964.78 1238 10.5 70,480 na 0.23 1.698 Calc. 07

As 77.95 817 1090 5.75 34,710 na 0.68 4.786 14 0.0245

7.97E- Ba 139.36 727 1000 3.62 54,760 na 0.23 1.698 14 Calc. 06

2.80E- Cd 112.4 321.07 594 8.69 122,800 React in acid -0.07 0.851 14 0.0308 04

React in 2.45E- Cr 52 1907 2180 7.15 88,670 0.23 1.698 14 Calc. dilute acid 08

Hg 200.59 -38.83 234 13.54 0.06 na 140 0.62 4.169 14 0.008622

5.54E- Pb 209.21 327.46 601 11.3 9581 Conc. Acid 0.73 5.370 14 0.0245 07

Se 78.96 220.8 494 4.81 2,063 CS2 0.24 1.738 14 0.00974

Sl in EtOH, S 32.06 115.21 388 2.07 i in Bz, Eth, s CS2 Acid or 3.06E- Al 26.98 660.32 933 2.70 59,400 0.33 2.138 14 0.0245 Alkaline 10

Chapter 3: RESULTS

Summary results of germination records

Summary results of growth records

Summary results of chemical analysis

Summary results of calculations

Chapter 3: RESULTS

Summary results of germination records:

Table 3a: Summary results of Germination records, Trial 1 Plant Species Date Camelina sativa Panicum virgatum Soil 1 10 0 Soil 2 44 0 June 13, 2012 Soil 3 18 0 Control 17 0 Soil 1 28 0 Soil 2 73 0 June 14, 2012 Soil 3 43 0 Control 95 0 Soil 1 * 100 Soil 2 100 June 15, 2012 Soil 3 100 Control 100 Notes: Seeds were sown on June 10, 2012. *Indicates only growth was measured after 3 cm was reached

Table 3b: Summary results of Germination records, Trial 2 Plant Species Date Camelina sativa Panicum virgatum Soil 1 37 0 Soil 2 4 0 September 18, 2012 Soil 3 53 0 Control 0 0 Soil 1 59 0 Soil 2 12 0 September 21, 2012 Soil 3 83 0 Control 0 11 Soil 1 14 45 Soil 2 73 78 October 1, 2012 Soil 3 8 31 Control 34 * Soil 1 28 20 Soil 2 71 48 October 3, 2012 Soil 3 14 39 Control 31 38 Soil 1 20 25 Soil 2 * 52 October 8, 2012 Soil 3 0 27 Control * *Growing well Soil 1 10 Soil 2 * October 11, 2012 Soil 3 5 Control * Soil 1 12 Soil 2 * October 15, 2012 Soil 3 0 Control * Soil 1 8 Soil 2 * October 18, 2012 Soil 3 0 Control * Soil 1 7 Soil 2 * October 22, 2012 Soil 3 0 Control * Notes: Seeds were sown on September 14, 2012 *Indicates only growth was measured (after 3 cm was reached)

Table 3c: Summary results of Germination records, Trial 3 Plant Species Date Camelina sativa Panicum virgatum Soil 1 52 0 Soil 2 2 0 September 18, 2012 Soil 3 38 0 Control 0 0 Soil 1 68 0 Soil 2 21 0 September 21, 2012 Soil 3 45 0 Control 0 13 Soil 1 15 15 Soil 2 62 53 October 1, 2012 Soil 3 15 52 Control 90 * Soil 1 39 42 Soil 2 82 32 October 3, 2012 Soil 3 15 27 Control 95 48 Soil 1 25 45 Soil 2 * 40 October 8, 2012 Soil 3 0 31 Control * *Growing well Soil 1 18 Soil 2 * October 11, 2012 Soil 3 0 Control * Soil 1 16 Soil 2 * October 15, 2012 Soil 3 0 Control * Soil 1 13 Soil 2 * October 18, 2012 Soil 3 0 Control * Soil 1 12* Soil 2 * October 22, 2012 Soil 3 3 Control * Notes: Seeds were sown on September 14, 2012 *Indicates only growth was measured (after 3 cm was reached)

Table 3d: Summary results of Germination records, Trial 4 Plant Species Date Camelina sativa Panicum virgatum Soil 1 55 0 Soil 2 10 0 September 18, 2012 Soil 3 82 0 Control 0 0 Soil 1 4 0 Soil 2 73 0 September 21, 2012 Soil 3 36 0 Control 3 3 Soil 1 4 10 Soil 2 58 82 October 1, 2012 Soil 3 17 43 Control 95 * Soil 1 19 51 Soil 2 93 51 October 3, 2012 Soil 3 12 42 Control 100 51 Soil 1 15 62 Soil 2 * 61 October 8, 2012 Soil 3 4 39 Control * * Growing well Soil 1 12 Soil 2 * October 11, 2012 Soil 3 0 Control * Soil 1 15 Soil 2 * October 15, 2012 Soil 3 3 Control * Soil 1 13 Soil 2 * October 18, 2012 Soil 3 3 Control * Soil 1 11 Soil 2 * October 22, 2012 Soil 3 3 Control * Notes: Seeds were sown on September 14, 2012 *Indicates only growth was measured (after 3 cm was reached)

Summary Results of Growth Records:

Figure 12a: Camelina sativa Growth Records, Trial 1 90

50 Soil 1

40 Soil 2 Height , cm Height 30 Soil 3 Control 20

0 1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 Time, Days

Figure 12b: Camelina sativa Growth Records, Trial 2 70

40 Soil 2 30

Height, cm Height, Soil 3 Control 20

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Figure 12c: Camelina sativa Growth Records, Trial 3 70

40 Soil 1 Soil 2 30 Height, cm Height, Soil 3 20 Control

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Figure 12d: Camelina sativa Growth Records, Trial 4 70

40 Soil 1 Soil 2 30 Height, cm Height, Soil 3 20 Control

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Figure 13a: Miscanthus giganteus Growth Records, Trial 1 180

160

140

120

100 Soil 1

80 Soil 2 Height, cm Height, Soil 3 60 Control 40

0 1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 Time, Days

Figure 13b: Miscanthus giganteus Growth Records, Trial 2 90

50 Soil 1

40 Soil 2 Height, cm Height, Soil 3 30 Control 20

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Figure 13c: Miscanthus giganteus Growth Records, Trial 3 120

100

Soil 1 60 Soil 2 Height, cm Height, Soil 3 40 Control

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Figure 13d: Miscanthus giganteus Growth Records, Trial 4 90

50 Soil 1

40 Soil 2 Height, cm Height, Soil 3 30 Control 20

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Figure 14a: Panicum virgatum Growth Records, Trial 1 90

50 Soil 1

40 Soil 2 Height, cm Height, Soil 3 30 Control 20

0 1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 Time, Days

Figure 14b: Panicum virgatum Growth Records, Trial 2 90

50 Soil 1

40 Soil 2 Height, cm Height, Soil 3 30 Control 20

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Figure 14c: Panicum virgatum Growth Records, Trial 3 80

50 Soil 1 40 Soil 2 Height, cm Height, 30 Soil 3 Control 20

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Figure 14d: Panicum virgatum Growth Records, Trial 4 70

40 Soil 1 Soil 2 30 Height, cm Height, Soil 3

20 Control

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Time, Days

Summary results of Laboratory analyses: The results from each of the laboratory analyses demonstrate the chemical concentrations of each analyte under each experimental condition respectively.

Table 4a: Results, Initial Soil Analysis, [ALS Environmental] Parameter Result Units Soil 1 Soil 2 Soil 3 mg/kg dry Aluminum, Al 12800 8200 12800 mg/kg dry Arsenic, As 14.3 21.9 29.6 mg/kg dry Barium, Ba 103 81.9 82.5 mg/kg dry Cadmium, Cd <0.52 <0.48 <0.58 mg/kg dry Chromium, Cr 13.7 11.0 9.1 mg/kg dry Lead, Pb 85.7 16.7 46.0 mg/kg dry Mercury, Hg 0.38 0.16 0.21 mg/kg dry Selenium, Se <2.6 <2.4 6.8 mg/kg dry Silver, Ag <1.0 <0.96 <1.2 mg/kg dry Sulfur, S 1010 411 2440 mg/kg dry pH 4.24 5.50 4.15 mg/kg dry Note: The initial soil results indicate the concentrations of each analyte in the soil at the beginning of the experiment. These results serve as a starting point for the experiment.

Table 4b: Results, Native plant species, [ALS Environmental] Parameter Result Units Location 1 Location Location 3 mg/kg dry 2 Aluminum, Al 43.5 420 <37.9 mg/kg dry Arsenic, As <4.1 <36.2 <5.6 mg/kg dry Barium, Ba 22.6 38.4 9.4 mg/kg dry Cadmium, Cd 2.2 <1.8 <1.9 mg/kg dry Chromium, Cr <2.7 <3.6 <3.7 mg/kg dry Lead, Pb <2.7 <3.6 <3.7 mg/kg dry Mercury, Hg <0.15 <0.18 <0.17 mg/kg dry Selenium, Se <6.8 <8.9 <9.3 mg/kg dry Silver, Ag <2.7 <3.6 <3.7 mg/kg dry Sulfur, S NA NA NA mg/kg dry pH 5.63 4.10 4.00 Note: The native plant tissue results indicate the concentrations of each analyte in the leaf tissues of native plants at the beginning of the experiment. These results serve an additional control and indicate the phytoextraction abilities of the native plant species.

Table 5a: Results Final Soil Analysis, Trial 1 [ALS Environmental] Parameter Camelina sativa Miscanthus giganteus Panicum virgatum Soil 1 Soil 2 Soil 3 Soil 1 Soil 2 Soil 3 Soil 1 Soil 2 Soil 3 Aluminum, Al 5840 7110 7020 6390 6700 4380 8650 5840 4120 Arsenic, As 27.5 13.0 33.9 16.7 11.8 43.2 17.0 11.8 36.2 Barium, Ba 139 113 120 98.7 153 148 93.7 67.8 98.9 Cadmium, Cd <0.55 <0.55 <0.57 <0.57 <0.46 <0.61 <0.53 <0.54 <0.63 Chromium, Cr 83.6 14.5 15.8 11.8 11.4 19.5 12.2 7.8 12.1 Lead, Pb 189 22.5 48.4 96.7 22.2 60.3 86.9 18.4 71.3 Mercury, Hg 0.36 0.072 0.22 0.46 0.12 0.30 0.39 0.098 0.14 Selenium, Se 4.0 <2.7 6.6 3.4 <2.3 9.5 <2.7 <2.7 11.0 Silver, Ag <1.1 <1.1 <1.1 <1.1 <0.92 <1.2 <1.1 <1.1 <1.3 Sulfur, S 793 545 4580 782 441 2540 754 523 2720 pH 4.48 6.22 4.82 4.61 5.84 4.65 5.19 6.01 4.76 Notes: All concentrations are in mg/kg on a dry basis. The initial soil results indicate the concentrations of each analyte under the original conditions. These results serve as a starting point for the experiment.

Table 5b: Results Final Soil Analysis, Trials 2 & 3 combined, [Microbac Laboratories, Inc.] Parameter Result Units Soil 1 Soil 2 Soil 3 mg/kg dry Aluminum, Al 8360 5190 3830 mg/kg dry Arsenic, As 21.4 7.13 14.0 mg/kg dry Barium, Ba 101 56.5 65.2 mg/kg dry Cadmium, Cd <1.78 <1.80 <2.00 mg/kg dry Chromium, Cr 12.1 <8.98 <9.95 mg/kg dry Lead, Pb 79.3 21.2 35.7 mg/kg dry Mercury, Hg 0.302 <0.0478 0.0616 mg/kg dry Selenium, Se <8.87 <8.98 <9.95 mg/kg dry Silver, Ag <4.44 <4.49 <4.98 mg/kg dry Sulfur, S 1040 735 2600 mg/kg dry pH 4.75 6.11 4.97 Note: The final soil results indicate the concentrations of each analyte under the final conditions. These results serve as an ending point for the experiment.

Table 6a: Results, Final Plant Tissue Analysis, Trial 1 [ALS Environmental]

Parameter Camelina sativa Miscanthus giganteus Panicum virgatum Soil 2 Soil C Soil 1 Soil 2 Soil 3 Soil C Soil 1 Soil 2 Soil 3 Soil C Al 146 181 33.7 <33.1 34.1 61.2 1220 173 807 106 As <10.3 <10.0 <5.0 <4.9 <4.7 <5.6 <6.9 <6.0 <7.5 <8.6 Ba <17.2 49.3 50.8 14.6 <7.9 <9.3 26.9 13.7 16.3 <14.3 Cd <3.4 <3.3 <1.7 <1.6 <1.6 <1.9 <2.3 <2.0 <2.5 <2.9 Cr <6.9 <6.7 <3.4 <3.2 <3.2 <3.7 <4.6 <4.0 19.7 6.4 Pb <6.9 <6.7 <3.4 <3.2 <3.2 <3.7 <4.6 <4.0 <5.0 <5.7 Hg <0.35 <0.35 <0.17 <0.17 <0.16 <0.15 <0.19 <0.20 <0.25 <0.26 Se <17.2 <16.7 <8.4 <8.1 <7.9 <9.3 <11.5 <10 <12.6 <14.3 Ag <6.9 <6.7 <3.4 <3.2 <3.2 <3.7 <4.6 <4.0 <5.0 <5.7 S 6150 11400 2290 1770 2240 2180 1880 2540 3550 4270 pH 7.07 6.28 6.48 6.55 6.64 6.97 6.02 6.11 6.35 6.20 Notes: All concentrations are in mg/kg on a dry basis. Incidence of phytoextraction highlighted in bold. The final plant tissue results indicate the concentrations of each analyte in the plant tissues at the end of the experiment.

Table 6b: Results, Final Plant Tissue Analysis, Trial 2 [Microbac Laboratories, Inc.] Para- Camelina sativa Miscanthus giganteus Panicum virgatum meter Soil 1 Soil 2 Soil C Soil 1 Soil 2 Soil 3 Soil C Soil 1 Soil 2 Soil 3 Soil C Al 2800 3790 24.0 <15.7 <21.7 <27.4 <16.5 <16.0 27.7 741 <15.6 As <22.6 <11.4 <4.10 <3.91 <5.40 <6.84 <4.10 <3.98 <3.91 <3.65 <3.89 Ba 87.8 81.9 <8.21 36.9 12.1 18.7 <8.21 10.7 <7.84 17.4 <7.78 Cd <9.08 <4.56 <1.65 <1.57 <2.17 <2.74 <1.65 <1.60 <1.57 <1.47 <1.56 Cr <45.3 <22.7 <8.21 <7.84 <10.8 <13.7 <8.21 <7.98 <7.84 <7.32 <7.78 Pb <45.3 <22.7 <8.21 <7.84 <10.8 <13.7 <8.21 <7.98 <7.84 8.28 <7.78 Hg <45.3 NA <0.0524 <0.0501 <0.0537 <0.299 <0.0344 <0.0661 <0.0344 0.0489 <0.0394 Se <45.3 <22.7 <8.21 <7.84 <10.8 <13.7 <8.21 <7.98 <7.84 <7.32 <7.78 Ag <22.7 <11.4 <4.11 <3.92 <5.41 <6.85 <4.11 <3.99 <3.92 <3.66 <3.90 S 6900 10100 10800 1640 1800 1060 1220 1780 1880 2050 1360 Notes: All concentrations are in mg/kg on a dry basis. Incidence of phytoextraction highlighted in bold. The final plant tissue results indicate the concentrations of each analyte in the plant tissues at the end of the experiment.

Table 6c: Results from plant tissue analysis, Trial 3 [Microbac Laboratories, Inc.] Para- Camelina sativa Miscanthus giganteus Panicum virgatum meter Soil 1 Soil 2 Soil 3 Soil 1 Soil 2 Soil 3 Soil 1 Soil 2 Soil 3 Soil C Al 1860 NA NA <26.1 <20.5 <27.1 57.7 <16.2 293 <15.6 As <21.8 NA NA <6.50 <5.10 <6.72 <4.11 <4.05 <6.01 <3.89 Ba 43.8 NA NA 25.5 20.0 14.1 12.9 <8.10 <12.0 <7.78 Cd <8.77 NA NA <2.61 <2.05 <2.70 <1.65 <1.62 <2.41 <1.56 Cr <43.7 NA NA <13.0 <10.2 <13.5 <8.22 <8.10 <12.0 <7.78 Pb <43.7 NA NA <13.0 <10.2 <13.5 <8.22 <8.10 <12.0 <7.78 Hg NA NA NA <0.109 <0.0785 <0.0472 <0.0368 <0.0366 <0.247 <0.0394 Se <43.7 NA NA <13.0 <10.2 <13.5 <8.22 <8.10 <12.0 <7.78 Ag <21.9 NA NA <6.52 <5.11 <6.74 <4.12 <4.06 <6.03 <3.90 S 10500 NA NA 1810 1500 1160 2460 1830 2190 1360 Notes: All concentrations are in mg/kg on a dry basis. Incidence of phytoextraction highlighted in bold. The final plant tissue results indicate the concentrations of each analyte in the plant tissues at the end of the experiment.

Summary of Calculation Results

Summary of Soil Variability Results:

The Experimental Conditions are represented in each of the following graphs as follows:

1. Initial soil laboratory results, (Initial). 2. Final soil laboratory results for Camelina sativa after Trial 1, (FCamT1). 3. Final soil laboratory results for Miscanthus giganteus after Trial 1, (FMisT1). 4. Final soil laboratory results for Panicum virgatum after Trial 1, (FPanT1). 5. Final combined soil laboratory results after Trial 2 and 3, (FT2/3).

The error bars on each of the graphs represent the standard deviation for each experimental condition, respectively.

Figure 15a: Aluminum Soil Variance

Aluminum Soil Variance

15000

10000

Soil 1

Soil 2 5000

Concentration, mg/Kg Concentration, Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Figure 15b: Arsenic Soil Variance

Arsenic Soil Variance

Soil 1

20 Soil 2

Concentration, mg/Kg Concentration, Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Figure 15c: Barium Soil Variance

Barium Soil Variance

200

150

100 Soil 1

Soil 2 Concentration, mg/Kg Concentration,

50 Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Figure 15d: Cadmium Soil Variance

Cadmium Soil Variance

Soil 1 1 Soil 2

Concentration, mg/Kg Concentration, Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Figure 15e: Chromium Soil Variance

Chromium Soil Variance

100

80 60

Soil 1 40 Soil 2

Concentration, mg/Kg Concentration, Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Figure 15f: Lead Soil Variance

Lead Soil Variance

200

150

100 Soil 1 Soil 2

Concentration, mg/Kg Concentration, Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Figure 15g: Mercury Soil Variance

Mercury Soil Variance

0.5000

0.4000 0.3000 Soil 1

0.2000 Soil 2

Soil 3

Concentration, mg/Kg Concentration,

0.1000

0.0000

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condtion

Figure 15h: Selenium Soil Variance

Selenium Soil Variance

8 6 Soil 1

Soil 2 4

Concentration, mg/Kg Concentration, Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Figure 15i: Silver Soil Variance

Silver Soil Variance

Soil 1

2 Soil 2

Concentration, mg/Kg Concentration, Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Figure 15j: pH Soil Variance

pH Soil Variance

4 log [H+] log - Soil 1

pH, pH, Soil 2

2 Soil 3

Initial

FT2/3

FMisT1

FPanT1 FCamT1 Experimental Condition

Summary of Plant Tissue Variability Results:

The Experimental Conditions are represented in each of the following graphs as follows:

1. Native plant tissue laboratory results, (NPTl). 2. Final soil laboratory results for Camelina sativa after Trial 1, (FCamT1). 3. Final soil laboratory results for Miscanthus giganteus after Trial 1, (FMisT1). 4. Final soil laboratory results for Panicum virgatum after Trial 1, (FPanT1). 5. Final soil laboratory results for Camelina sativa after Trial 1, (FCamT2). 6. Final soil laboratory results for Miscanthus giganteus after Trial 1, (FMisT2). 7. Final soil laboratory results for Panicum virgatum after Trial 1, (FPanT2). 8. Final soil laboratory results for Camelina sativa after Trial 1, (FCamT3). 9. Final soil laboratory results for Miscanthus giganteus after Trial 1, (FMisT3). 10. Final soil laboratory results for Panicum virgatum after Trial 1, (FPanT3).

The error bars on each of the graphs represent the standard deviation.

Figure 16a: Aluminum Plant Tissue Variance

Aluminum Plant Tissue Variance

4001

3001

Soil 1 2001 Soil 2 Soil 3

Control

1001

Concentration, mg/Kg on a ln scale ln a on mg/Kg Concentration,

NPT

FMisT1 FMisT2 FMisT3

FPanT1 FPanT2 FPanT3

FCamT1 FCamT2 FCamT3 Experimental Condition

Figure 16b: Arsenic Plant Tissue Variance

Arsenic Plant Tissue Variance

20 Soil 1 Soil 2

Soil 3 Concentration, mg/Kg Concentration,

10 Control

NPT

FMisT3 FMisT1 FMisT2

FPanT1 FPanT2 FPanT3

FCamT2 FCamT1 FCamT3 Experimental Condition

Figure 16c: Barium Plant Tissue Variance

Barium Plant Tissue Variance

100

Soil 1

Soil 2 40

Soil 3 Concentration, mg/Kg Concentration,

Control

NPT

FMisT1 FMisT2 FMisT3

FPanT2 FPanT1 FPanT3

FCamT1 FCamT2 FCamT3 Experimental Condition

Figure 16d: Cadmium Plant Tissue Variance

Cadmium Plant Tissue Variance

Soil 1

4 Soil 2

Soil 3 Concentrations, mg/Kg Concentrations,

Control

NPT

FMisT2 FMisT1 FMisT3

FPanT3 FPanT1 FPanT2

FCamT1 FCamT2 FCamT3 Experimental Condition

Figure 16e: Chromium Plant Tissue Variance

Chromium Plant Tissue Variance

Soil 1

Soil 2 20

Soil 3 Concentration, mg/Kg Concentration,

Control

NPT

FMisT1 FMisT2 FMisT3

FPanT1 FPanT2 FPanT3

FCamT1 FCamT2 FCamT3 Experimental Condition

Figure 16f: Lead Plant Tissue Variance

Lead Plant Tissue Variance

Soil 1

Soil 2 20

Soil 3 Concentration, mg/Kg Concentration,

Control

NPT

FMisT1 FMisT2 FMisT3

FPanT1 FPanT2 FPanT3

FCamT3 FCamT1 FCamT2 Experimental Condition

Figure 16g: Mercury Plant Tissue Variance

Mercury Plant Tissue Variance

0.4000

0.3000

Soil 1

0.2000 Soil 2 Soil 3

Concentration, mg/Kg Concentration, Control

0.1000

0.0000

NPT

FMisT3 FMisT1 FMisT2

FPanT1 FPanT2 FPanT3

FCamT1 FCamT2 FCamT3 Experimental Condition

Figure 16h: Selenium Plant Tissue Variance

Selenium Plant Tissue Variance

Soil 1

20 Soil 2

Soil 3 Concentration, mg/Kg Concentration,

Control

NPT

FMisT1 FMisT2 FMisT3

FPanT1 FPanT2 FPanT3

FCamT3 FCamT1 FCamT2 Experimental Condition

Figure 16i: Silver Plant Tissue Variance

Silver Plant Tissue Variance

15 Soil 1

10 Soil 2

Soil 3 Concentration, mg/Kg Concentration,

5 Control

NPT

FMisT1 FMisT2 FMisT3

FPanT1 FPanT2 FPanT3

FCamT1 FCamT2 FCamT3 Experimental Condition

Figure 16j: Sulfur Plant Tissue Variance

Sulfur Plant Tissue Variance

4096 512

Soil 1

64 Soil 2 Soil 3

8 Control

Concentration, mg/Kg on a ln scale ln a on mg/Kg Concentration,

NPT

FMisT1 FMisT2 FMisT3

FPanT1 FPanT2 FPanT3

FCamT1 FCamT2 FCamT3 Experimental Condition

Percent germination results:

Table 7a: Results, Percent Germination, Trial 1 Percent Time Percent Time Plant Camelina Panicum germination, elapsed, germination, elapsed, Species sativa virgatum % days % days Soil 1 28 28 4 100 100 5 Soil 2 73 73 4 100 100 5 Soil 3 43 43 4 100 100 5 Control 95 95 4 100 100 5

Table 7b: Results, Percent germination, Trial 2 Percent Time Percent Time Plant Camelina Panicum germination, elapsed, germination, elapsed, Species sativa virgatum % days % days Soil 1 59 59 7 45 45 17 Soil 2 73 73 17 78 78 17 Soil 3 83 83 7 39 39 19 Control 31 31 19 39 38 19

Table 7c: Results, Percent germination, Trial 3 Percent Time Percent Time Plant Camelina Panicum germination, elapsed, germination, elapsed, Species sativa virgatum % days % days Soil 1 68 68 7 45 45 24 Soil 2 82 82 19 53 53 17 Soil 3 45 45 7 52 52 17 Control 95 95 19 100 100 7

Table 7d: Results, Percent germination, Trial 4 Percent Time Percent Time Plant Camelina Panicum germination, elapsed, germination, elapsed, Species sativa virgatum % days % days Soil 1 19 19 19 62 62 24 Soil 2 93 93 19 82 82 17 Soil 3 36 36 7 43 43 17 Control 100 100 19 100 100 17

Mass-Balance Calculation Results:

Camelina sativa:

Table 8a: Summary Results, Camelina sativa, Mass-Balance, Soil 1

Trial 1 Trial 2 Trial 3 Average

Parameter Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked * marked * marked * Al 128.0 * 54.38 82.0 * 34.68 82.0 * 34.68 97.0 * 41.25 As -242.4 -92.31 -130.4 -49.68 -130.4 -49.68 -167.7 -63.89 Ba -661.0 -34.95 36.4 1.93 35.3 1.87 -196.4 -10.38 Cd -23.1 -242.31 -23.2 -242.6 -23.2 -242.6 -23.2 -242.50 Cr -1283.4 -510.22 29.2 11.62 29.2 11.62 -408.3 -162.33 Pb -1896.6 -120.54 117.4 7.46 117.4 7.46 -553.9 -35.21 Hg 0.4 5.26 1.3 18.45 1.4 20.53 1.03 14.75 Se -25.7 -53.85 -115.3 -241.46 -115.3 -241.45 -85.4 -178.92 Ag -1.8 -10.00 -63.2 -344.40 -63.3 -344.38 -42.8 -232.93 S 4.0 * 21.49 -0.6 * -3.09 -0.6 * -3.15 0.9 * 5.08 *Al and S mass units are in grams.

Table 8b: Summary Results, Camelina Sativa Mass-Balance, Soil 2

Trial 1 Trial 2 Trial 3 Average

Parameter Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked * marked * marked * Al 10.0 * 5.98 61.0 * 37.70 61.0 * 36.71 44.0 * 26.46 As 181.5 40.64 301.3 67.43 301.3 67.44 261.3 58.50 Ba -634.5 -38.0 517.9 31.0 518.2 31.0 133.9 8.0 Cd -26.9 -275.04 -26.9 -275.15 -26.9 -275.00 -26.9 275.06 Cr -71.4 -31.82 41.1 18.33 41.2 18.36 3.6 1.62 Pb -118.3 -34.73 -91.9 -27.0 -89.8 -26.4 -100.0 -29.38 Hg 1.8 55.00 2.3 70.13 2.3 70.13 2.1 65.09 Se -6.1 -12.54 -134.3 -274.32 -134.2 -274.17 -91.6 -187.01 Ag -2.9 -14.62 -71.4 -364.40 -71.4 -364.58 -48.6 -247.87 S -2.7 * -32.68 -6.6 * -79.22 -6.6 * -78.8 -5.3 * -63.6 *Al and S mass units are in grams.

Table 8c: Summary Results, Camelina sativa, Mass-Balance, Soil 3

Trial 1 Trial 2 Trial 3 Average

Parameter Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked * marked * marked * Al 124.0 * 45.16 82.0 * 34.68 82.0 * 34.68 96.0 * 38.17 As -92.1 -14.53 334.2 52.70 334.2 52.70 192.1 30.29 Ba -803.3 -45.45 370.6 21.0 370.6 21.0 -20.7 -1.15 Cd -30.4 -244.83 -30.4 -245.04 -30.4 -244.83 -30.4 -244.90 Cr -143.5 -73.63 -18.2 -9.35 -18.2 -9.34 -60.0 -30.77 Pb -51.4 -5.22 220.6 22.39 220.6 22.39 129.9 13.19 Hg -0.2 -4.76 3.2 70.66 3.2 70.67 2.1 45.52 Se -14.8 -30.96 -67.5 -46.32 -67.5 -46.32 -49.9 -41.20 Ag 2.1 8.33 -81.0 -315.00 -81.0 -315.00 -53.3 -207.22 S -45.8 * -87.70 -3.4 * -6.56 46.7 * 89.34 -0.8 * -1.64 *Al and S mass units are in grams.

Miscanthus giganteus:

Table 9a: Summary Results, Miscanthus giganteus, Mass-Balance, Soil 1

Trial 1 Trial 2 Trial 3 Average

Parameter Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked * marked * marked * Al 0.118* 50.08 0.082* 34.69 0.082* 34.69 0.094 * 39.82 As -44.2 -16.80 -130.5 -49.70 -130.6 -49.70 -101.8 -38.73 Ba 78.4 4.15 35.6 1.88 35.95 1.90 49.9 2.64 Cd -23.1 -242.50 -23.2 -242.80 -23.2 -243.13 -23.2 -242.81 Cr 34.8 13.85 29.1 11.59 29.0 11.52 31.0 12.32 Pb -202.0 -12.84 117.3 7.45 117.1 7.44 10.8 0.68 Hg -1.5 -21.08 1.4 20.50 1.4 20.48 0.4 6.63 Se 2.0 3.98 -115.4 -241.65 -115.5 -242.00 -76.3 -159.89 Ag -1.9 -10.20 -63.3 -344.64 -63.4 -345.07 -42.9 -233.30 S 4.2 * 22.44 -0.6 * -3.24 -0.6 * -3.26 1.0 * 5.3 *Al and S mass units are in grams.

Table 9b: Summary Results, Miscanthus giganteus Mass-Balance, Soil 2

Trial 1 Trial 2 Trial 3 Average

Parameter Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked * marked * marked * Al 0.031 * 18.29 0.061 * 36.71 0.061 * 36.71 0.051 * 30.57 As 206.0 46.11 301.2 67.41 301.2 67.41 269.5 60.31 Ba -1450.6 -86.82 517.8 31.0 517.6 31.0 -138.4 -8.27 Cd -27.0 -275.18 -27.0 -275.66 -27.0 -275.63 -27.0 -275.49 Cr -8.2 -3.65 40.9 18.22 40.9 18.23 24.5 10.93 Pb -112.2 -32.94 -92.1 -27.0 -90.1 -26.44 -98.1 -28.79 Hg 0.8 24.94 2.3 70.08 2.3 70.05 1.8 55.03 Se -57.9 -39.77 -134.6 -274.83 -134.5 -274.79 -109.0 196.46 Ag 0.8 3.99 -71.6 -365.41 -71.6 -365.37 -47.5 -242.26 S -0.6 * -7.53 -6.7 * -79.48 -6.7 * -79.37 -4.7 * -55.46 *Al and S mass units are in grams.

Table 9c: Summary Results, Miscanthus giganteus, Mass-Balance, Soil 3

Trial 1 Trial 2 Trial 3 Average

Parameter Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked * marked * marked * Al 0.180* 65.78 0.192* 70.08 0.192* 70.08 0.188* 68.65 As -291.4 -45.95 333.9 52.67 334.0 52.67 125.5 19.80 Ba -1403.1 -79.4 370.0 20.9 370.1 20.95 -221.0 -12.52 Cd -30.4 -245.00 -30.5 -245.49 -30.5 -245.48 -30.5 -245.32 Cr -222.8 -114.30 -18.6 -9.55 -18.6 -9.55 -86.7 -44.5 Pb -306.3 -31.09 220.2 22.35 220.2 22.35 44.7 4.54 Hg -1.9 -42.90 3.2 70.47 3.2 70.64 1.5 32.74 Se -57.9 -39.77 -67.9 -46.61 -67.9 -46.60 -64.6 -44.34 Ag -0.04 -0.14 -81.2 -315.80 -81.2 -315.79 -54.1 -210.58 S -2.2 * -4.15 46.7 * 89.28 46.7 * 89.28 30.4 * 58.14 *Al and S mass units are in grams.

Panicum virgatum:

Table 10a: Summary Results, Panicum virgatum, Mass-Balance, Soil 1

Parameter Trial 1 Trial 2 Trial 3 Average

Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked * marked * marked * Al 0.076* 32.42 0.082* 34.69 0.082* 34.69 0.08* 34.6 As -49.6 -18.90 -130.4 -49.68 -130.4 -49.68 -103.5 -39.42 Ba 170.5 9.02 36.5 1.93 36.4 1.93 81.1 4.29 Cd -23.2 -242.50 -23.2 -242.68 -23.2 -242.69 -23.2 242.62 Cr 27.5 10.93 29.2 11.61 29.2 11.61 28.6 11.38 Pb -22.1 -1.40 117.3 7.46 117.3 7.46 70.8 4.51 Hg -0.2 -2.65 1.4 20.51 1.4 20.51 0.9 12.79 Se -1.9 -4.04 -115.3 -241.52 -115.3 -241.53 187.5 162.36 Ag -1.9 -10.20 -63.2 -344.48 -63.2 -344.49 42.7 -233.06 S 4.7 * 25.27 -0.6 * -3.18 -0.6 * -3.26 1.2 * 6.28 *Al and S mass units are in grams.

Table 10b: Summary Results, Panicum virgatum Mass-Balance, Soil 2

Trial 1 Trial 2 Trial 3 Average

Parameter Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked * marked * marked * Al 0.048* 28.78 0.061* 36.71 0.061* 36.71 0.057* 34.07 As 206.0 46.11 301.2 67.42 301.2 67.41 269.5 60.31 Ba 287.5 17.21 518.0 31.00 518.0 31.00 441.2 26.40 Cd -26.9 -275.16 -27.0 -275.35 -27.0 -275.36 -27.0 -275.29 Cr 65.2 29.08 41.0 18.29 41.0 18.28 49.1 21.88 Pb -34.7 -10.19 -89.9 -26.40 -89.9 -26.40 -71.5 -21.00 Hg 1.3 38.70 2.3 70.10 2.3 70.10 2.0 59.63 Se -6.2 -12.66 -134.4 -274.52 -134.4 -274.53 -91.7 -187.23 Ag -2.9 -14.75 -71.6 -365.02 -71.5 -365.04 -48.7 -248.27 S -2.3 * -27.49 -6.7 * -79.33 -6.6 * -79.31 -5.2 * -62.04 *Al and S mass units are in grams.

Table 10c: Summary Results, Panicum virgatum, Mass-Balance, Soil 3

Parameter Trial 1 Trial 2 Trial 3 Average

Mass Mass Mass Mass Percent Percent Percent Percent Balance, Balance, Balance, Balance, of input, of input, of input, of input, mg unless mg unless mg unless mg unless % % % % marked * marked marked * marked * Al 0.186* 67.81 0.192* 70.08 0.192* 70.08 0.190* 69.32 As -141.43 -22.31 334.1 52.69 334.1 52.69 175.6 27.69 Ba -351.4 -19.89 370.2 20.95 370.3 20.95 129.7 7.34 Cd -30.4 -245.00 -30.5 -245.09 -30.5 -245.25 -30.5 -245.11 Cr -64.4 -33.05 -18.4 -9.42 -18.5 -9.48 -27.7 -17.32 Pb -542.0 -55.0 220.4 22.37 220.4 22.36 -33.7 -3.42 Hg 1.5 33.29 3.2 70.64 3.2 70.55 2.6 58.16 Se -90.1 -61.83 -67.6 -46.43 -67.7 -46.50 -75.1 -51.59 Ag -2.2 -8.49 -81.05 -315.31 -81.1 -315.52 -54.8 -213.11 S -0.6 * -11.53 46.7 * 89.26 -3.5 * -6.65 14.2 * 23.69 *Al and S mass units are in grams.

CHAPTER 4, INTERPRETATION OF RESULTS

Discussion of Plant Species

Discussion of Germination Results

Discussion of Growth Results

Discussion of Laboratory Results

Discussion of Mass-Balance Results

Discussion of Plant Species:

The plants that we selected to study were Camelina sativa, Miscanthus giganteus and

Panicum virgatum. Each of these species offers different characteristics that were useful in reclaiming the marginal soils produced by mining operations. Camelina sativa displayed the ability to germinate easily in the experimental conditions but had difficulty adapting to the harsh soil environments. This could possibly be related to the low pH rather than the heavy metals present. Miscanthus giganteus displayed the ability to grow well in each of the experimental soils. This may be related to the deep root structures which could also inhibit erosion and/or the chemical transport within the soil. Panicum virgatum displayed the ability to grow well in each of the experimental soils. After the germination lag these plants grew very well. The delayed germination could have been related to lower initial watering amounts. The consistent growth patterns may be related to the dense root structures which would also inhibit erosion and/or the chemical transport within the soil.

Discussion of Germination Results:

The results from the germination phase of this experiment offered some useful information for the ability of each of the seeded species (Camelina sativa and Panicum virgatum) to survive in the harsh experimental environments. The extremely low pH and high concentration of heavy metals could be the main parameter causing inhibited germination. In each of the trials there were instances where the percent germination for both species appeared to be rather successful. In fact, the Panicum virgatum was extremely successful (at or near 100%) after a sustained germination period under optimal conditions. The Camelina sativa specimens had successful instances where there appeared to be very successful at first, only to taper out

after a few days. Camelina sativa appears to be very sensitive to adverse environmental conditions.

Discussion of Growth Results:

The results from the growth phase of this experiment offered very good data to aid in the determination of sustained survivability of each of the studied plant species. Each of the trials offered similar results for the ability for each species to survive under experimental conditions.

It was recognized that the extremely low pH in the initial soil samples could impact successful growth. The Miscanthus giganteus had the greatest success to the production of biomass over time. In each of the first three experimental trials they showed consistently increasing growth curve throughout the experimental time frames. The fourth such trial had instances where each of the experimental specimens died off and the control grew at a steadily increasing curve, as expected. These failures were most likely linked to the rhizome splitting process. This species had the ability to adapt to each of the experimental and control soil types. One drawback with using Miscanthus was the high demand for water. As the individual plants’ grew, larger amounts of water were required to sustain the increased biomass. The Camelina sativa grew well in each of the trials under experimental soil type #2 and the control. They did not do well in experimental soil type #1 or soil type #3, where the young plants died off shortly after depleting the post-emergent seed energy supply. This variety of Camelina sativa appeared to be intolerant the harsh environments of each of these soil types. The Panicum virgatum specimens grew very well under each of the soil types during each other experimental trials showing steady growth throughout. These growth patterns were also continuously increasing growth curve, although to a lesser degree than that of the Miscanthus giganteus specimens.

Discussion of Laboratory Results:

The results from the laboratory analysis portion of this experiment offered a great deal of useful information. They offer the ability of each of the studied species to phytoextract heavy metals from each of the experimental soil types. On the surface, the lab results offer a glance as to what is present under each of the analytical parameters. The initial soil lab results offer a starting point to show the initial concentrations for each of the parameters. Cadmium and silver were not detected in any of the experimental soil types. Selenium was not detected in soils 1 or

2, but was detected in soil type 3. Aluminum, arsenic, barium, chromium, lead, mercury, and sulfur offered responses above the detection limits of the inductively coupled plasma mass spectrometry (ICP-MS) analyzer.

The analysis of the native plant tissue samples offered an additional experimental control.

Each of the parameters was below the detection limit of the analyzer, with the exception of aluminum at locations 1 and 2, barium at each of the locations, and cadmium at location 1.

These concentrations were lower than the experimental analysis results, as expected.

The final soil lab analysis results give insight into the potential for phytoextraction.

Lower concentrations for each of the parameters would be expected after the experiment. This was not always the case. The “grab” samples taken for analysis from each of the experimental plant pots may have been taken from a different batch of soil than that of the initial soil samples.

There were many instances where the output was actually higher than the input. These instances are as follows:

1. Aluminum: none

2. Arsenic: Camelina trial 1, soils 1, 2 and 3 and trials 2 and 3, soils 2 and 3; Miscanthus

trial 1, soil 3 and trials 2 and 3, soils 2 and 3; Panicum, trial 1, soils 1 and 3 and trials 2

and 3, soils 2 and 3.

3. Barium: Camelina trial 1, soils 1, 2 and 3; Miscanthus trial 1, soils 2 and 3; Panicum,

trial 1, soils 3.

4. Cadmium: All readings were non-detects (ND)

5. Chromium: Camelina trial 1, soils 1, 2 and 3; Miscanthus trial 1, soils 1, 2 and 3;

Panicum, trial 1, soils 1 and 3.

6. Lead: Camelina trial 1, soils 1, 2 and 3 and trial 2, soil 2; Miscanthus trial 1, soil 3 and

trial 2, soil 2; Panicum, trial 1, soils 1, 2 and 3 and trial 2, soil 2.

7. Mercury: Camelina trial 1, soil 3; Miscanthus trial 1, soils 1 and 3; Panicum, trial 1, soil

8. Selenium: Camelina trial 1, soil 1; Miscanthus trial 1, soil 1; Panicum, trial 1, soil 3.

9. Silver: All readings were non-detects (ND)

10. Sulfur: Camelina trial 1, soils 2 and 3 and trials 2 and 3, soils 1, 2 and 3; Miscanthus trial

1, soil 2 and 3 and trials 2 and 3, soils 1, 2 and 3; Panicum, trial 1, soils 2 and 3 and trials

2 and 3, soils 1, 2 and 3.

At first glance, the presence of any of the chemicals in the plant tissues appears to indicate phytoextraction. Throughout the three trials study there were instances where aluminum, barium, chromium, lead, mercury and sulfur were detected in the plant tissues. Here are the instances where these parameters were detected by the analyzer:

1. Aluminum: Camelina trial 1, soils 2 and control, trial 2, soils 1, 2 (2 magnitudes

higher) and control, and trial 3, soil 1; Miscanthus trial 1, soils 1, 2 and control;

Panicum trial 1, soils 1, 2, 3 (magnitude higher) and control, trial 2, soils 2 and 3, and

trial 2, soils 1 and 3. There were results that indicated a full magnitude higher than

controls.

2. Barium: Camelina trial 1, control, trial 2, soils 1 and 2 (magnitude higher), and trial

3, soil 1; Miscanthus trial 1, soils 1 and 2, trials 2 and 3, soils 1, 2 and 3; Panicum

trial 1, soils 1, 2 and 3, trials 2 and 3, soil 1. There were results that indicated a full

magnitude higher than controls.

3. Chromium: Panicum trial 1, soils 3 and control.

4. Lead: Panicum trial 2, soil 3.

5. Mercury: Panicum trial 2, soil 3.

6. Sulfur: All of the samples that were analyzed. Although the Camelina samples

indicated up to a full magnitude higher than the rest of the samples.

Taking these at face value offers an indication as to whether phytoextraction has taken place. The lab results also offer the raw data necessary to extrapolate and interpret the environmental impact through further calculation. They also allow for the identification of instances of hyper-accumulation.

Discussion of Variability Discussion:

The results from the analyses for the variability between the experimental conditions, soils and plant tissues offered great insight into the reaching conclusions for objective questions.

The statistical analysis of each data set is crucial in determining the significance of the laboratory results for each parameter that was analyzed. The large standard deviations for most of the results indicated that there is a great amount of variance between the laboratory results from soil

types and plant tissues and the mean values from each data set, respectively. Smaller standard deviations indicate lower variability.

Discussion of Mass-Balance Results:

The results from the mass-balance calculations for each of the experiment conditions offered an extrapolated view into scientific merit of this research. Assumptions were made in order to realize the desired result outputs. These calculations offer a great amount of feedback for future applications in this type of research. The goal is reach a net zero for mass (or energy) input versus mass (or energy) output. There were some key missing details (leachate volume and concentrations) and plant tissue masses, which were not measured during the experiment.

Approximations were made based on assumed values derived from literature yield numbers.

This gave us a rough estimate on the mass-volume throughput for each sample. The output concentrations for a lot of the final soil lab analysis were higher than the input. This resulted in negative (larger output than input) mass balance numbers. This could be related to the samples being taken from a slightly different area for each geographical site at the EPCAMR property holdings. The discrepancy may also be related to different calibrations when the samples were analyzed with the ICP-MS instrument. Having results from two different laboratories could have played a role.

There were some meaningful results though. The parameters that showed the most promising results (closest to net zero) were those that had valid inputs for each of the mass balance equation components. As a general rule they should be within close proximity to the net zero value while having an acceptable percentage (± 10 %) of input value. This would include the following trials for each heavy metal parameter:

1. Aluminum: Camelina trial 1, soil 2.

2. Arsenic: none

3. Cadmium: none

4. Barium: Camelina trial 2 and 3, soil 1; Miscanthus trial 1, 2 and 3, soil 1; and Panicum

trial 1, 2 and 3, soil 1.

5. Chromium: Camelina trials 2 and 3, soil 3; Miscanthus trial 1, soil 2 and trial 2 and 3,

soil 3; and Panicum Trials 2 and 3, soil 3.

6. Lead: Camelina trial 1, soil 3, and trials 2 and 3, soil 1; Miscanthus trials 2 and 3, soil 1;

and Panicum Trials 1, 2 and 3, soil 1.

7. Mercury: Camelina trial 1, soil 1 and 3 and Panicum Trial 1, soil 1.

8. Selenium: Camelina trial 1, soil 3; Miscanthus trial 1, soil 2; and Panicum trial 1, soil 1.

9. Silver: Camelina trial 1, soil 3; Miscanthus trial 1, soils 2 and 3; and Panicum trial 1, soil

10. Sulfur: Camelina trial 2, soils 1 and 3 and trial 3, soil 1; Miscanthus trial 1, soils 2 and 3

and trials 2 and 3, soil 1; and Panicum trials 2 and 3, soil 1 and trial 3, soil 3.

Here are few lessons to be applied to the mass balance segment of this type of research:

1. The collection, measurement of volume and analysis of the leachate is one area that

would offer more accurate mass-balance approximations.

2. The planter pots should have been weighed at the beginning and end of the experiment to

offer experiment mass values.

3. The experimental soil quantities should be sufficient for the duration of the experiment.

This may offer more consistency for concentrations between trials.

Chapter 5, CONCLUSION

In conclusion, this experiment provided an excellent opportunity to evaluate the phytoremediation properties for each of the three biofuel crop plant species that were studied.

There were instances where each species offered pros and cons. It was recognized that the ability for each of the species to survive in the experimental soils may have been related to the low pH coupled with the elevated heavy metal concentrations. In order to scientifically develop useful interpretations of the aforementioned results we hoped to answer five (5) questions throughout this experiment.

Here are the answers to each of these questions:

1. Which of the three (3) biofuel crops we selected [Camelina sativa, Miscanthus

giganteus, or Panicum virgatum] has the ability to survive in the marginal soils

affected by mining operations?

Each of the selected plant species have some ability to adapt to some of the harsh

environmental present in the experimental soils we studied. The Camelina sativa

specimens had the most difficulty adjusting to the other than optimal growth conditions.

These specimens only thrived in the control and experimental soil from location # 2.

The Miscanthus giganteus was rather hardy and showed the most potential to adapt to

adverse soil conditions. In all of the trials, with the exception of the extra trial #4, these

specimens thrived and showed consistent growth patterns. They did require the most

water to sustain the massive biomass creation. The Panicum virgatum showed

consistent growth throughout the experiment. There was a lag time at the beginning

while the seeds were in germination phase, but following that lag they were rather

consistent over each of the trials.

2. Do any of the aforementioned species thrive in such conditions?

The Miscanthus giganteus showed the greatest ability to thrive in the

experimental soil types. One drawback for using this species is the high water demand.

The specimens even produced seeds over the longer experimental time in trials 2 – 4.

The seeding did not occur until after the growth period for this study was complete. The

Camelina sativa had the most difficulty, but did grow well in soil from location #2. The

specimens that survived did produce flowers and seeds very well over times. The seeds

even self-propagated to extend the survival in the pots from trial #1 that were kept alive

through the growth periods for trials 2 – 4. The Panicum virgatum offered great

stability throughout the study without a large water requirement.

3. Do any of these biofuel crops have the ability to Phytoremediate soils with high

concentrations of heavy metals?

There were instances of phytoextraction by each of the biofuel crops that were

selected for this research. Specific species showed differing levels of abilities to

phytoextract heavy metals from the soil. Certain specimens displayed a greater affinity

to remove specific parameters. The Camelina sativa displayed the ability to absorb each

of the following chemical parameters very well: aluminum (trial 1, soils 2; trial 2, soils

1 and 2; and trial 3, soil 1); barium (trial 2, soils 1 and 2 and trial 3, soil 1); and sulfur.

Miscanthus giganteus showed a lesser affinity to extract barium and sulfur in trials 2

and 3. Panicum virgatum displayed the ability to extract aluminum very well (trial 1,

soils 1, 2 and 3; trial 2, soils 2 and 3; and trial 3, soil 3), and barium, lead (trial 2, soil 3),

mercury (trial 2, soil 3) and sulfur to a lesser degree.

These behaviors could be attributed to the chemical and physical properties of

each parameter. Perhaps this could also be related to each chemical interaction at the

root zone, through the plant into the leaf tissues. There also appear to be a great deal of

variability in how well each plant species responded to the analytes individually.

Instances that appeared to be statistically significant were recognized in experimental

conditions. An example of such an interaction is the relationship where Barium

interacted with Camelina within the variance range of one standard deviation during

Trial 2. The graphs indicate numerous relationships such as this.

4. Do any of the aforementioned plant species behave as a hyper-accumulator of any

of the pollutants studied?

Yes, there appeared to be a number of situations that could be considered

instances of hyper-accumulation. For this study anything above one magnitude higher

than control is considered hyper-accumulation. This was determined strictly by

comparing concentration found in the experimental plant tissues versus those seen in the

control plant tissues. Camelina sativa appeared to possibly be a hyper-accumulator of

aluminum, barium in trial 2 and 3, soil 1, trial 2, soil 2, and sulfur in trial 3, soil 1.

Sulfur results for trials 1 and 2 are confusing due to the large amount found in the

control sample. Miscanthus giganteus appeared to possibly be a hyper-accumulator of

barium in trials 2 and 3 for each experimental soil type. Panicum virgatum appeared to

possibly be a hyper-accumulator of: aluminum in trial 2, soils 2 and 3, trial 3, soils 1 and

3 and barium in trials 2 and 3, soil 1; and lead for trial 2, soil 3. These considerations

are based on non-detects with zero concentrations.

There would be less incidence of hyper-accumulation recognized if the reported

detection limits (RDL) are considered. Sulfur was present at high concentration for the

majority of the samples. Only the very large sulfur concentration in Camelina sativa for

trial 2 soil 1 is being considered an instance of hyper-accumulation.

5. What is the feasibility of using any of these plant species to phytoremediate

contaminated soils and also as a source of energy following phytoextraction?

The feasibility for applying these crops to in-situ process would be very

challenging. The lands that would be selected for such applications would have to be of

sufficient grade to operate the agricultural machinery required to plant, maintain, and

harvest such crops. A lot of the lands that would be candidates for such research are

traditionally being treated with the passive limestone alkaline treatment and wetland

filtration systems with great success. In order to support the capital investment required

to farm these lands properly, an in-depth cost/benefit analysis would have to be

conducted prior to considering each specific situation.

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Appendix A, FORMS AND DEADLINES

Master’s Approval Page

MASTER’S APPROVAL PAGE

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We accept and approve the thesis of the student named above and agree to distribution as indicated.

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Appendix B, GROWTH RECORDS

Spreadsheet of Daily Growth Records

Field Data Sheets

Spreadsheet of Daily Growth Records:

Additional Observations No recognized changes from the initial planting. The Miscanthus shoots are green.

Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 13- Shoots MR/EG Jun 10.8 15.2 11.8 14.2 N/A N/A N/A N/A N/A N/A N/A N/A recorded Lowest Shoots with 6.8 6.8 7.3 7.5 N/A N/A N/A N/A N/A N/A N/A N/A growth # of buds 9a, 7a, (a = alive & 5d 5d 5a 11 N/A N/A N/A N/A N/A N/A N/A N/A d =dead ) Watering Volumes in N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A ml. Additional Observations Pictures taken by MR. (To be taken approximately every week or as needed.) Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 14- Shoots MR/EG Jun 13.1 17.0 13.5 17.3 N/A N/A N/A N/A N/A N/A N/A N/A recorded Lowest Shoots with MR/EG 6.3 6.8 7.8 12.4 0.7 1.8 1.0 1.6 N/A N/A N/A N/A growth # of buds 9a, 7a, (a = alive & MR/EG 5d 5d 5a 11 28 73 43 95 0 0 1 0 d = dead ) MR/EG 0 0 0 0 0 0 0 300 250 250 250 250 Additional Observations 18/

Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C 15- Highest Shoots MR/TB Jun 16.0 20.2 16.2 21.0 1.1 3.1 1.6 3.2 0.0 0.0 0.0 0.0 recorded Lowest Shoots 10.0 10.6 N/A 11.3 N/A N/A N/A N/A N/A N/A N/A N/A with growth # of Not recorded buds/Germination Watering Volumes N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A in ml. Additional Observations The change in germination rate was not recorded, to be recorded weekly.(eg) Daily Record Keeping 16- Watering Volumes MR/TB Jun 500 500 500 500 500 500 500 500 500 500 500 500 in ml.

Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C 18- Highest Shoots TB/MR/EG Jun 20.3 27.5 21.8 25.8 1.4 4.2 2.2 5.3 2.0 2.0 2.1 1.1 recorded Lowest Shoots 11.0 10.5 16.1 11.8 N/A N/A N/A N/A N/A N/A N/A N/A with growth # of germinated N/A N/A N/A N/A N/A N/A N/A N/A 100+ 100+ 100+ 100+ seeds Watering Volumes 500 500 500 500 500 500 500 500 500 500 500 500 in ml. • When watering each of the species in soil type 2 the water ran out of into the overflow tray. Perhaps this is due to the soil composition. • Miscanthus giganteus o All soil types appear to be doing well. • Camelina Sativa o Soil type 1 is struggling to survive post germination. o Soil type 2 and control are doing well. o Soil type 3 is struggling to survive post germination. Additional • Switchgrass Observations EG o All soil types appear to being doing well. Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 20- Shoots MR/TB/EG Jun 28.8 35.8 28.2 32.1 1.4 5.3 2.0 7.0 3.3 3.5 2.9 3.9 recorded Lowest Shoots with 9.1 13.0 15.7 9.0 N/A N/A N/A N/A N/A N/A N/A N/A growth Watering Volumes in 250 250 250 250 250 250 250 250 250 250 250 250 ml. Additional Water ran through soil type 2 for Camelina and Switchgrass. All species and soil types are doing well Observations EG except for Camelina Sativa soil type 1 and 3.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 22- Shoots MR/TB Jun 37.0 50.2 35.4 43.2 1.4 4.9 2.0 6.7 5.1 5.9 5.0 5.8 recorded Lowest Shoots with 7.8 8.5 12.3 8.2 N/A N/A N/A N/A N/A N/A N/A N/A growth Watering Volumes 250 250 250 250 250 250 250 250 250 250 250 250 in ml. Additional Observations Nothing Recorded.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 25- Shoots MR/TB Jun 48.9 59.0 45.6 50.3 1.4 7.0 1.9 9.3 6.8 6.0 7.1 10.0 recorded Lowest Shoots with 7.0 8.3 8.8 8.8 N/A N/A N/A N/A N/A N/A N/A N/A growth Watering Volumes 250 250 250 250 250 250 250 250 250 250 250 250 in ml. Additional Observations Nothing recorded.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 27- Shoots EG Jun 51.0 64.5 51.8 56.5 1.4 7.6 3.0 12.8 8.8 10.8 9.8 14.3 recorded Lowest Shoots with 7.0 6.8 7.3 7.8 N/A N/A N/A N/A N/A N/A N/A N/A growth Watering Volumes 250 250 250 250 250 250 250 250 250 250 250 250 in ml. Additional The species Camelina Sativa in soil types 1 and 3 have very low survival rate. They are struggling very Observations EG badly. All other samples appear to be thriving with excellent growth.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 29- Shoots MR/TB Jun 52.7 74.1 58.0 67.0 1.0 9.0 3.1 13.4 11.8 13.6 11.0 16.0 recorded Lowest Shoots with 5.2 5.5 9.2 8.5 N/A N/A N/A N/A N/A N/A N/A N/A growth Watering Volumes 250 250 250 250 250 250 250 250 250 250 250 250 in ml. Additional observations Nothing recorded.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 64.3 85.2 67.1 80.4 1.3 11.0 3.1 18.0 17.0 17.6 14.9 20.4 Shoots MR/TB 2-Jul recorded Lowest Shoots 7.5 6.1 12.5 9.5 N/A N/A N/A N/A N/A N/A N/A N/A with growth Watering Volumes 250 250 250 250 250 250 250 250 250 250 250 250 in ml. Additional observations Nothing recorded.

Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest Shoots MR/TB 3-Jul recorded Lowest Shoots with Nothing recorded. growth Watering 250 250 250 250 250 250 250 250 250 250 250 250 Volumes in ml. Additional observations Nothing recorded.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 67.0 91.5 68.9 83.3 1.1 14.6 3.1 23.2 18.0 20.7 18.1 22.5 Shoots MR/TB 4-Jul recorded Lowest Shoots 7.0 6.5 9.3 8.6 N/A N/A N/A N/A N/A N/A N/A N/A with growth Watering Volumes 250 250 250 250 250 250 250 250 250 250 250 250 in ml. Additional observations Nothing recorded.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 81.7 94.3 71.1 94.4 1.2 16.0 2.0 25.1 27.5 22.8 20.6 27.0 Shoots MR/TB 7-Jul recorded Lowest Shoots 7.4 10.8 12.8 9.2 N/A N/A N/A N/A N/A N/A N/A N/A with growth Watering Volumes Not watered. Growing well. in ml. Additional observations Nothing recorded.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 21.1 Shoots MR/TB 9-Jul 85.0 98.0 76.0 99.7 1.0 23.1 ? 29.4 29.8 17.1 22.5 29.2 recorded Lowest Shoots with 8.0 12.0 12.6 9.6 N/A N/A N/A N/A N/A N/A N/A N/A growth Watering Volumes Not watered. Growing well. in ml. Nothing recorded. Upon data entry, recognized discrepancy with Camelina results for Soil types 1 and 3? Additional These do not appear to be accurate results? Perhaps transcribed from another soil type? Looks like Cam1 observations and SG 1 were switched. And Cam2 should be 2.1?

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 11- 86.1 105.9 77.3 101.0 1.0 21.5 2.9 30.5 30.9 24.3 22.5 37.4 Shoots MR/TB Jul recorded Lowest Shoots 7.5 15.2 12.0 9.2 N/A N/A N/A N/A N/A N/A N/A N/A with growth Watering Volumes 500 500 500 500 500 500 500 500 500 500 500 500 in ml. Additional observations Nothing recorded.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 13- 90.1 107.9 79.3 103.6 1.0 23.2 1.4 34.5 32.8 29.7 26.5 42.8 Shoots MR/TB Jul recorded Lowest Shoots 8.2 16.0 12.5 9.3 N/A N/A N/A N/A N/A N/A N/A N/A with growth Watering 500 500 500 500 500 500 500 500 500 500 500 500 Volumes in ml. Additional observations Nothing recorded.

Daily Record Keeping Watering 15- Volumes MR/TB Jul 250 250 250 250 250 250 250 250 250 250 250 250 in ml.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 16- Shoots MR/TB/EG Jul 93.0 116.0 85.2 111.3 1.4 25.4 4.3 32.5 35.5 32.6 32.3 45.7 recorded Lowest Shoots with 10.5 16.2 16.8 2.2 N/A N/A N/A N/A N/A N/A N/A N/A growth Watering Volumes 250 250 250 250 250 250 250 250 250 250 250 250 in ml. Additional observations. EG Took pictures.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 20- Shoots EG Jul recorded Lowest Shoots with No measurements taken, cuttings taken for sampling. growth Watering Volumes 250 250 250 250 250 250 250 250 250 250 250 250 in ml. Additional observations. EG Took pictures and took samples for plant tissue analysis Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 23- 47.6 57.8 49.3 83.8 0.0 21.8 0.0 54.4 16.8 22.8 20.6 51.3 Shoots TB Jul recorded Lowest Shoots 2.8 27.8 16.9 4.3 with growth Watering 500 500 500 500 500 500 500 500 500 500 500 500 Volumes in ml. Additional observations Nothing recorded.

with growth Watering 500 500 500 500 500 500 500 500 500 500 500 500 Volumes in ml. Additional observations Nothing recorded. Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 30- 64.9 82.3 62.7 115.0 0.0 25.1 0.0 45.8 26.5 26.8 30.3 65.9 Shoots EG Jul recorded Lowest Shoots 13.0 26.0 16.5 11.0 with growth Watering 250 250 250 250 250 250 250 250 250 250 250 250 Volumes in ml. Additional observations Miscanthus giganteus samples have black spots on leaves. Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 1- 71.5 88.2 72.5 115.5 0.0 26.3 0.0 62.3 33.5 29.5 31.3 67.8 Shoots EG Aug recorded Lowest Shoots 15.2 26.3 27.3 12.5 with growth Watering 250 250 250 250 250 250 250 250 250 250 250 250 Volumes in ml. Additional observations Nothing recorded. CamCon appears to have numbers reversed changed from 26.3 to 62.3. 4/13/13 eg

Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 5- 86.9 94.9 86.7 135.5 0.0 30.3 0.0 63.5 41.5 37.8 36.6 72.4 Shoots EG Aug recorded Lowest Shoots 15.4 27.4 27.3 12.4 with growth Watering 250 250 250 250 0 250 0 250 250 250 250 250 Volumes in ml. Additional observations Camelina Sativa 2 and control flowering. Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 7- Shoots MR Aug recorded Lowest Shoots

with growth Watering 500 500 500 500 500 500 500 500 500 500 500 500 Volumes in ml. Additional observations Nothing recorded. Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 8- 91.7 105.8 97.6 141.4 0.0 23.6 0.0 65.0 43.4 46.3 34.0 72.6 Shoots MR Aug recorded Lowest Shoots

with growth Watering 500 500 500 500 500 500 500 500 500 500 500 500 Volumes in ml. Additional observations Nothing recorded.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest Shoots EG 9-Aug recorded Lowest Shoots ` with growth Watering 500 500 500 500 500 500 500 500 500 500 500 500 Volumes in ml. Additional observations Nothing recorded. Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 97.5 122.1 102.9 151.3 0.0 35.3 0.0 68.9 48.3 48.5 50.3 72.5 Shoots EG 10-Aug recorded Lowest Shoots 16.5 2.7 35.2 10.3 with growth Watering Volumes in ml. Additional observations Nothing recorded. Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 92.0 117.5 122.3 151.3 0.0 24.4 0.0 72.3 50.9 51.7 51.9 72.0 Shoots EG 13-Aug recorded Lowest Shoots

with growth Watering 500 500 500 500 500 500 500 500 500 500 500 500 Volumes in ml. Additional observations Camelina Sativa control has lots of seeds.

with growth Watering Volumes in ml. Additional observations Water ran through for M2 and M3, Ccon, SG1, 2, 3. Ccon has new flowers. Ccon 2 fragile. Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 19- 107.5 129.1 120.7 154.1 0.0 0.0 0.0 76.4 53.3 54.8 52.3 72.9 Shoots EG Aug recorded Lowest Shoots

with growth Watering Volumes in ml. Additional observations Ccon seeds harvested, lady bugs recognized on plants. Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 22- 112.4 133.7 123.2 152.9 0.0 0.0 0.0 75.7 52.5 51.7 52.1 72.7 Shoots EG Aug recorded Lowest Shoots

with growth Watering 500 500 500 500 500 500 500 1000 500 500 500 500 Volumes in ml. Additional observations Ccon drying out. Given more water. Sgcon yellow. Pruned dried dead sections off.

Daily Record Keeping Scientist' Dat s initials e All Measurements are in cm Additiona Species Miscanthus giganteus Camelina sativa Pancium virgatum l remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 116. 134. 126. 153. 24- 0.0 0.0 0.0 76.7 54.0 54.2 52.5 73.7 Shoots 0 6 0 1 EG Aug recorded Lowest Shoots

with growth Watering 100 100 100 100 100 100 100 100 1000 1000 1000 1000 Volumes 0 0 0 0 0 0 0 0 in ml. Additional observations Water ran through on everything watered except the Ccon. Harvested seeds for Ccon. Daily Record Keeping

Scientist' Dat s initials e All Measurements are in cm Additiona Species Miscanthus giganteus Camelina sativa Pancium virgatum l remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 117. 136. 127. 153. 26- 0.0 0.0 0.0 41.5 52.8 53.5 54.3 65.2 Shoots 1 1 8 3 EG Aug recorded Lowest Shoots

with growth Watering Volumes in ml. Additional observations Nothing recorded. Daily Record Keeping

Scientist' Dat s initials e All Measurements are in cm Additiona Species Miscanthus giganteus Camelina sativa Pancium virgatum l remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 27- Shoots EG Aug recorded Lowest Shoots

with growth Watering Volumes in ml. Additional observations All good, moist soil. No measurements taken.

with growth Watering 500 500 500 1000 500 500 500 1000 500 500 500 1000 Volumes in ml. Additional observations Nothing recorded. Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 30- 120.3 136.5 131.3 154.0 0.0 0.0 0.0 43.3 53.4 55.0 55.1 71.8 Shoots EG Aug recorded Lowest Shoots

with growth Watering 1000 1000 1000 1000 0 0 0 1000 1000 1000 1000 1000 Volumes in ml. Additional observations Nothing recorded.

with growth Watering 1000 1000 1000 2000 0 0 0 1000 1000 1000 1000 1000 Volumes in ml. Additional observations Nothing recorded. Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 4- 122.7 139.2 134.9 152.3 0.0 0.0 0.0 43.2 51.7 54.3 55.7 73.4 Shoots EG Sep recorded Lowest Shoots

with growth Watering 1000 1000 1000 1000 0 0 0 500 500 500 500 500 Volumes in ml. Additional observations Daily Record Keeping

Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 6- 125.1 140.9 136.4 153.4 0.0 0.0 0.0 43.2 53.3 53.2 60.1 76.7 Shoots EG Sep recorded Lowest Shoots

with growth Watering 1000 1000 1000 1000 0 0 0 500 500 500 500 1000 Volumes in ml. Additional observations Ccon. Dying off, pruned back for new growth. (for seeds naturally replant)

with growth Watering 1000 1000 1000 2000 0 0 0 500 500 500 500 500 Volumes in ml. Additional observations Ccon. Seeds self-germinated. Prepped soil for trials (2,3,& 4) Daily Record Keeping Scientist's initials Date All Measurements are in cm Additional Species Miscanthus giganteus Camelina sativa Pancium virgatum remarks Sample # = 1 2 3 C 1 2 3 C 1 2 3 C Highest 12- 129.1 143.1 136.7 152.4 0.0 0.0 0.0 51.7 52.7 54.7 64.1 79.3 Shoots EG Sep recorded Lowest Shoots

with growth Watering 1000 1000 1000 2000 0 0 0 1000 1000 1000 1000 1000 Volumes in ml. Additional observations Took plant tissue samples from leaves, roots along with final soil samples. Held in deep freeze. Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 14- 76.4 84.1 72.5 124.3 0.0 0.0 0.0 32.4 30.2 37.5 30.1 60.3 EG Sep Trial 1 10.0 10.0 10.0 10.0 Trial 2 10.0 10.0 10.0 10.0 100 seeds placed in each sample pot. Trial 3 10.0 10.0 10.0 10.0 Trial 4 Trial 1 Watering 1000 1000 1000 2000 0 0 0 1000 1000 1000 1000 1000 Volumes in ml. Trial 2, 3, &4 100 100 100 100 100 100 100 100 100 100 100 100 watering in ml. Post cut measurement reading. Trial 1 complete for study. Kept alive for breeding and future study Additional through trials 2, 3, & 4. Pots for Trials 2, 3, & 4 prepped and ready for planting. 100 seeds planted in each observations of the Camelina Sativa and Switchgrass pots.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 16- EG Sep Trial 1 Trial 2 Trial 3 Trial 4 Watering 1000 1000 1000 2000 0 0 0 1000 1000 1000 1000 1000 Volumes in ml. Trial 2, 3, &4 100 100 100 100 100 100 100 100 100 100 100 100 watering in ml. Additional observations No visible change for newly planted samples. Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 16- 80.7 83.6 73.2 126.1 0.0 0.0 0.0 34.0 21.5 38.5 30.7 60.1 EG Sep Trial 1 Trial 2 Trial 3 Trial 4 Watering Volumes in ml. Additional observations Old growth dead in Ccon. Reseeded specimens are thriving. Sgcon. Mostly cut back. Daily Record Keeping

Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 18- 84.9 87.6 76.7 127.8 0.0 0.0 0.0 8.8 25.7 39.4 30.3 59.3 EG Sep Trial 1 10.3 10.6 10.3 11.5 0.0 37.0 4.0 53.0 0.0 0.0 0.0 0.0 Trial 2 10.3 10.3 10.0 11.3 0.0 52.0 2.0 38.0 0.0 0.0 0.0 0.0 Trial 3 11.1 10.5 10.2 11.9 0.0 55.0 10.0 82.0 0.0 0.0 0.0 0.0 Trial 4 Trial 1 watering 1000 1000 1000 1000 0 0 0 1000 1000 1000 1000 1000 volumes in ml. Trial 2, 3, &4 250 250 250 250 250 250 250 250 250 250 250 250 watering volumes in ml. Additional observations Placed catch pans for watering. Numbers for new samples indicated # of germinated seeds.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 21- 89.6 90.7 79.3 125.5 0.0 0.0 0.0 7.2 25.5 38.7 29.8 60.5 EG Sept Trial 1 10.8 10.9 10.5 11.3 6.0 59.0 12.0 83.0 0.0 0.0 0.0 11.0 Trial 2 10.3 10.5 9.0 11.7 4.0 68.0 21.0 45.0 0.0 0.0 0.0 13.0 Trial 3 10.8 10.1 10.2 11.9 4.0 73.0 36.0 3.0cm 0.0 0.0 0.0 3.0 Trial 4 Trial 1 watering 1000 1000 1000 1000 0 0 0 1000 1000 1000 1000 1000 volumes in ml. Trial 2, 3, &4 500 500 500 500 500 500 500 500 500 500 500 500 watering volumes in ml. Additional observations Steve Hans watered while I was attending to my wife and birth of Korynn on 9/23/12 :-)

Daily Record Keeping

Scientist' Dat s initials e All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots recorde Sample # = 1 2 3 C 1 2 3 C 1 2 3 C d 1- 108. 108. 122. 98.1 0.0 0.0 0.0 16.5 32.4 42.3 46.1 62.1 EG Oct 5 1 5 Trial 1 14 6.8/73 5.3/34 11.1 10.5 10.5 11.5 8# 45# 78# 31# 13.3 # # # Trial 2 15 5.8/62 15 6.8/90 10.3 10.7 9.5 11.7 15# 53# 52# 12.3 # # # # Trial 3 17 9.5/95 10.8 10.8 10.6 11.5 4# 6.4/58 10# 82# 43# 9.8 # # Trial 4 Trial 1 100 100 100 100 100 watering 1000 1000 1000 0 0 0 1000 0 0 0 0 0 volumes in ml. Trial 2, 3, &4 50 50 500 500 500 500 500 500 500 500 500 500 watering 0 0 volumes in ml. Additional observations # sign indicates the number of germinated seeds.

Daily Record Keeping Scientist Dat 's initials e All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots recorde Sample # = 1 2 3 C 1 2 3 C 1 2 3 C d 3- 111. 110. 103. 125. 33. 0.0 0.0 0.0 11.8 45.8 46.3 54.3 EG Oct 3 9 1 9 2 Trial 1 28 6.3/71 14 5.3/48 11.1 10.8 10.7 12.1 7.3/31# 20# 39# 5.3/38# # # # # Trial 2 39 6.5/82 15 5.3/32 5.1/27 10.3/48 10.6 10.2 10.5 12.1 7.1/95# 42# # # # # # # Trial 3 19 6.8/93 12 10.1/100 14.6/51 10.6 9.8 10.3 11.3 51# 5.8/51 42# # # # # # Trial 4 Trial 1 waterin Not needed still moist g volume s in ml. Trial 2, 3, &4 50 50 waterin 500 500 500 500 500 500 500 500 500 500 0 0 g volume s in ml. Additional observations C3 had withering past seed emergence.

Daily Record Keeping Scientist' Dat s initials e All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots recorde Sample # = 1 2 3 C 1 2 3 C 1 2 3 C d 8- 120. 117. 113. 135. 0.0 0.0 0.0 16.1 37.8 50.6 50.1 55.0 EG Oct 3 3 2 7 Trial 1 20 4.4/25 8.2/52 4.2/27 10.3 11.6 10.6 11.6 7.3 0.0 8.8 17.1 # # # # Trial 2 25 4.1/45 6.8/40 4.9/31 10.3 10.4 11.1 11.8 7.1 0.0 9.1 16.2 # # # # Trial 3 15 4.5/62 8.2/61 5.1/39 11.2 9.9 10.3 11.8 7.8 4# 10.2 14.3 # # # # Trial 4 Trial 1 100 100 watering 1000 1000 1000 2000 0 0 0 1000 1000 1000 0 0 volumes in ml. Trial 2, 3, &4 10 10 10 100 100 100 100 100 100 100 100 100 watering 0 0 0 volumes in ml. Additional observations

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 11- 121. 121. 117. 137. 0.0 0.0 0.0 17.1 40.1 44.9 50.5 43.4 EG Oct 2 1 3 3 Trial 1 10.9 11.7 10.4 11.8 10# 7.5 5# 10.3 4.5 11.6 5.1 21.4 Trial 2 11.1 10.9 11.1 12.2 18# 7.3 0.0 9.2 5.1 7.4 5.3 18.3 Trial 3 11.2 10.1 11.1 11.4 12# 8.4 0.0 10.2 6.4 11.3 6.2 15.4 Trial 4 Trial 1 watering 1000 1000 1000 1000 0 0 0 1000 1000 1000 1000 1000 volumes in ml. Trial 2, 3, &4 500 500 500 500 500 500 500 500 500 500 500 500 watering volumes in ml. Additional observations Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 15- 123. 124. 121. 142. 26. 39. 46. 50. 0.0 0.0 0.0 50.8 EG Oct 3 2 5 4 2 1 1 1 Trial 1 18. 17. 10.7 11.7 13.5 12.2 12# 7.3 0.0 6.1 6.8 31.9 3 3 Trial 2 3.8/16 21. 10. 10.9 13.2 10.8 11.8 9.8 0.0 8.1 7.4 21.6 # 7 2 Trial 3 12. 23. 16. 11.4 10.4 10.8 20.1 15# 3# 8.4 8.1 21.1 3 4 3 Trial 4 Watering 50 all trials 500 500 500 500 500 500 500 500 500 500 500 0 volumes in ml. Additional observations Did not water Camelina trial soils 1, 2 & 3, they are dead and will be removed from the experiment.

Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 22- 125.2 123.2 134.9 145.3 0.0 0.0 0.0 29.2 42.3 48.6 58.7 45.7 EG Oct Trial 1 16.1 12.4 23.8 16.2 7# 10.1 0.0 28.7 7.9 19.6 8.7 40.8 Trial 2 10.9 23.1 10.8 25.1 4.3/12# 12.1 0.0 25.4 9.8 13.3 14.6 31.8 Trial 3 11.2 10.4 11.2 34.9 11# 10.3 3# 28.3 10.8 18.8 10.5 18.8 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 26- 124.9 126.5 140.3 145.3 0.0 0.0 0.0 38.2 43.9 49.2 59.6 53.9 EG Oct Trial 1 25.2 18.9 31.3 30.5 2# 7.2 0.0 35.7 10.7 20.1 10.2 46.1 Trial 2 10.4 31.9 16.8 37.2 4.0/10# 13.1 0.0 31.6 11.2 16.4 13.4 39.2 Trial 3 11.3 10.9 10.1 31.7 13# 0.0 3# 32.1 15.6 18.2 13.1 19.6 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations MT4S2, M43S3, CT2S2, CT4S2, dead or dying; Ccont trial 1, 3 &4 flowering recognized.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 29- 127.1 128.4 141.5 145.4 0.0 0.0 0.0 41.7 42.2 48.5 59.5 45.3 EG Oct Trial 1 32.7 22.3 35.7 35.7 0.0 6.1 0.0 39.9 11.2 19.5 11.5 49.5 Trial 2 10.7 36.8 23.1 42.4 4.5/11# 13.4 0.0 39.1 18.1 16.5 16.1 40.8 Trial 3 11.2 10.7 11.1 50.2 4.1/10# 0.0 1# 33.5 16.2 19.3 12.5 20.5 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 3- 126.1 132.7 144.6 146.1 0.0 0.0 0.0 53.1 45.1 52.3 61.1 39.2 EG Nov Trial 1 40.2 30.9 40.6 45.7 0.0 3# 0.0 46.4 12.1 24.2 13.4 57.3 Trial 2 10.6 42.3 33.5 48.2 8.1/8# 13.8 0.0 46.8 11.2 19.6 16.5 42.4 Trial 3 11.3 10.5 10.2 60.1 5.2/15# 0.0 0.0 35.1 19.5 23.6 14.3 19.4 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Daily Record Keeping

Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 6- 128.8 133.1 144.3 146.5 0.0 0.0 0.0 59.3 43.6 54.3 62.8 34.2 EG Nov Trial 1 41.8 35.9 43.1 50.2 0.0 0.0 0.0 50.8 12.5 28.5 15.1 57.9 Trial 2 9.1 42.5 39.4 56.1 10.1 17.3 0.0 50.1 12.4 17.8 16.3 42.5 Trial 3 12.1 11.2 10.2 66.1 8.9 0.0 0.0 26.1 19.7 25.9 15.3 19.3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 9-

EG Nov Trial 1 Trial 2 Trial 3 Trial 4 Watering 50 all trials 500 500 500 500 500 500 500 500 500 500 500 0 volumes in ml. Additional observations Watered only. Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 12- 130. 132. 148. 151. 56. 47. 56. 61. 0.0 0.0 0.0 35.1 EG Nov 6 9 7 1 5 1 5 6 Trial 1 54. 17. 32. 13. 49.8 39.3 43.8 53.9 0.0 0.0 0.0 59.4 6 5 3 3 Trial 2 23. 52. 17. 18. 15. 17.6 52.2 51.8 62.5 15.1/4# 0.0 52.9 5 3 5 2 1 Trial 3 42. 21. 28. 19. 11.5 10.5 10.5 71.2 15.2/6# 0.0 0.0 19.2 1 8 4 6 Trial 4 Watering 50 all trials 500 500 500 500 500 500 500 500 500 500 500 0 volumes in ml. Additional observations Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded EG 15-Nov Trial 1 Trial 2 Trial 3 Trial 4 Watering 50 all trials 500 500 500 500 500 500 500 500 500 500 500 0 volumes in ml. Additional observations Watered only.

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 18- 130.8 135.2 149.3 151.1 0.0 0.0 0.0 58.3 48.3 56.8 61.5 37.5 EG Nov Trial 1 65.3 48.5 44.6 53.5 0.0 0.0 0.0 57.4 20.4 33.1 16.2 62.1 Trial 2 28.4 57.8 54.8 67.8 23.2 26.3 0.0 48.5 18.5 20.1 23.5 55.3 Trial 3 10.4 10.6 10.3 78.9 21.3 0.0 0.0 26.4 25.2 34.1 18.5 24.3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 20-

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 30-

EG Nov Trial 1 Trial 2 Trial 3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Watered only. Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded EG 3-Dec 127.9 139.1 150.3 154.4 0.0 0.0 0.0 59.4 48.5 57.1 60.9 35.8 Trial 1 78.8 60.7 44.7 53.7 0.0 0.0 0.0 65.4 26.6 48.1 17.8 74.2 Trial 2 50.2 57.8 65.2 75.4 37.8 29.4 0.0 59.1 27.1 26.5 27.3 70.2 Trial 3 0.0 0.0 0.0 79.4 32.5 0.0 0.0 27.3 29.7 43.5 22.1 22.1 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional Observe 1 thriving camelina w/ switchgrass in SGT3cont (volunteer?) growing very well. Perhaps the observations added support of switchgrass? Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded EG 5-Dec 130.7 137.5 153.5 153.1 0.0 0.0 0.0 40.5 48.2 57.2 60.9 36.0 Trial 1 78.9 61.0 44.3 52.5 0.0 0.0 0.0 66.2 30.5 48.9 18.1 74.7 Trial 2 54.9 58.5 66.1 84.0 38.1 30.9 0.0 60.1 31.1 31.6 29.2 71.5* Trial 3 0.0 0.0 0.0 79.9 35.2 0.0 0.0 28.1 27.9 42.7 21.9 25.3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations *SG also supporting a volunteer Camelina at 87.0 cm

100

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 10-

EG Dec Trial 1 Trial 2 Trial 3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Watered only. Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 11- 128.9 138.2 152.4 156.6 0.0 0.0 0.0 63.4 49.1 47.8 60.7 35.7 EG Dec Trial 1 79.2 64.9 49.7 54.5 0.0 0.0 0.0 62.9 36.1 51.7 19.8 74.8 Trial 2 68.8 59.9 68.9 87.4 48.1 35.2 0.0 46.8 27.9 36.5 28.4 72.7 Trial 3 0.0 0.0 0.0 80.8 42.5 0.0 0.0 23.8 33.9 50.7 23.4 28.7 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 16-

EG Dec Trial 1 Trial 2 Trial 3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Watered only.

101

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 20- 142.8 131.2 165.2 153.5 0.0 0.0 0.0 40.3 52.6 54.5 61.3 35.1 EG Dec Trial 1 80.1 69.5 52.1 54.5 0.0 0.0 0.0 63.5 45.0 63.1 19.5 75.5 Trial 2 73.5 60.5 74.3 99.8 37.5 34.3 0.0 55.3 40.1 59.5 31.5 75.5 Trial 3 0.0 0.0 0.0 81.3 28.3 0.0 0.0 21.5 39.1 47.0 25.8 29.5 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Soil and tissue samples to be taken in the next few days. Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 23-

EG Dec Trial 1 Trial 2 Trial 3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Watered only.

102

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 31- 168.5 136.0 185.9 152.3 0.0 0.0 0.0 8.5 48.2 56.2 61.8 29.6 EG Dec Trial 1 80.6 79.1 60.3 52.8 0.0 0.0 0.0 64.7 51.3 67.1 27.1 77.3 Trial 2 87.4 67.1 76.5 112.8 36.1 30.3 0.0 24.5 46.9 52.1 33.5 75.3 Trial 3 0.0 0.0 0.0 78.8 30.2 0.0 0.0 59.1 48.1 63.2 30.8 17.5 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional Soil and tissue samples were taken from all applicable trials for Miscanthus and Camelina samples and observations held in deep freeze for analysis. Daily Record Keeping

Scientist's initials Date All Measurements are in cm Species #NAME? Camelina Sativa Switchgrass Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded EG 1-Jan Trial 1 Trial 2 Trial 3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional Soil and tissue samples were taken from all applicable trials for Switchgrass samples and held in deep observations freeze for analysis. Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded EG 5-Jan Trial 1 Trial 2 Trial 3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Watered only.

103

Daily Record Keeping Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina Sativa Switchgrass Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 10-

EG Jan Trial 1 Trial 2 Trial 3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Watered only. Daily Record Keeping

Scientist's initials Date All Measurements are in cm Species Miscanthus giganteus Camelina sativa Pancium virgatum Highest Shoots Sample # = 1 2 3 C 1 2 3 C 1 2 3 C recorded 14-

EG Jan Trial 1 Trial 2 Trial 3 Trial 4 Watering all trials 500 500 500 500 500 500 500 500 500 500 500 500 volumes in ml. Additional observations Watered only.

104

Field Data Sheets:

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106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

Appendix C, LABORATORY RESULTS

Initial Soil Sample Results

Initial Native Plant Tissue Sample Results

Final Soil and Plant Tissue Sample Results for Trial 1

Final Soil and Plant Tissue Sample Results for Trials 2 – 4

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Initial Soil Sample Results [ALS Environmental]

125

126

127

128

129

130

131

Initial Native Plant Tissue Sample Results [ALS Environmental]

132

133

134

135

136

137

138

Final Soil and Plant Tissue Sample Results for Trial 1 [ALS Environmental]

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

Final Soil and Plant Tissue Sample Results for Trials 2 – 4 [Microbac Laboratory Services]

166

167

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174

175

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187

Appendix D, Calculations

Percent Germination Calculations

Mass-Volume Calculations

Approximate plant tissue mass calculations

Mass-Balance Calculations

188

Calculation Results:

Percent Germination Calculations:

[(Total Number of seeds sewn) - (Number of germinated seeds)] / (Total number seeds sewn) = (Number of non-germinated seeds)

[1 - (Number of non-germinated seeds)] * 100% = % Germination

(Nssewn – Nsgerminated) / (Nssewn) = Nsnon-germinated

(1 - Nsnon-germinated) * 100 % = % Germination

Mass-Volume Calculations:

Soil mass calculations:

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (clay) ρSoil1 = 2100 kg/m

3 3 Msoil1 = 0.0102 m * 1800 kg/m = 18.36 kg

3 3 Msoil2 = 0.0102 m * 2000 kg/m = 20.40 kg

3 3 Msoil3 = 0.0102 m * 2100 kg/m = 21.42 kg

3 3 Vsoil = 3 gallons * ( 3.785 liters/gallon) * (1 m / 1000 L) * (0.90) = 0.0102 m

189

Approximate plant tissue mass calculations:

Growth Areaplant = Areapot

Dpot = 10 in * (1 ft / 12 in) * (1 m / 3.281 ft) =

0.254 m

2 Areapot = (πD ) / 4

(π (0.254)2 ) / 4 =

0.051 m2

Trial 1, length of growth

Θgrowth-1 = 38 days * ( 1 year / 365 days) =

0.104 year

Trials 2 – 4, length of growth

Θgrowth-2-4 = 108 days * ( 1 year / 365 days) =

0.296 year

Mplants = Growth rateplants * (Growth Areaplants) * ( Growth Timeplants)

Camelina sativa:

MCamelina-1 = 2134.5 kg/ha/year (averaged from 1638, 3106, 1987, 3320, 1096, 1660 kg/ha) (Vakulabharanam, 2010)

2134.5 kg/ha/yr * (1 ha /10,000 m2 ) ( 0.051 m2 ) (0.104) =

0.0011 kg

2 2 MCamelina-2 = 2134.5 kg/ha/yr * (1 ha /10,000 m ) ( 0.051 m ) (0.295 year) =

0.0032 kg

2 2 MCamelina-3 = 2134.5 kg/ha/yr * (1 ha /10,000 m ) ( 0.051 m ) (0.295 year) =

0.0032 kg

190

Miscanthus giganteus:

Mplants = 8.2 metric ton/acre/year (averaged from 6.6 and 9.8 dry ton/acre/year) (Wang et al., 2013)

2 2 MMiscanthus-1 = 8.2 t/acre/year * (1000 kg / 1 metric ton) (1 acre / 4046.863 m ) (0.051 m ) (0.104 year) =

0.011 kg

2 2 MMiscanthus-2 = 8.2 t/acre/year * (1000 kg / 1 metric ton) (1 acre / 4046.863 m ) (0.051 m ) (0.295 year) =

0.030 kg

2 2 MMiscanthus-3 = 8.2 t/acre/year * (1000 kg / 1 metric ton) (1 acre / 4046.863 m ) (0.051 m ) (0.295 year) =

0.030 kg

Panicum virgatum:

Mplants = 6 metric tons/acre/year (Jensen et al., 2005)

2 2 MPanicum-1 = 6.0 t/acre/year * (1000 kg / 1 metric ton) (1 acre / 4046.863 m ) (0.051 m ) (0.104 year) =

0.008 kg

2 2 MPanicum-2 = 6.0 t/acre/year * (1000 kg / 1 metric ton) (1 acre / 4046.863 m ) (0.051 m ) (0.295 year) =

0.022 kg

2 2 MPanicum-3 = 6.0 t/acre/year * (1000 kg / 1 metric ton) (1 acre / 4046.863 m ) (0.051 m ) (0.295 year) =

0.022 kg

191

Mass-Balance Calculations:

Aluminum

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(Mout)]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Trial 1 Camelina sativa Soil 1: [(12,800 mg/kg dry * (0.0102 m3) * (1800 kg/m3)] = Σ [((5840 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = 0.128 kg or 127.79 g Soil 2: [(8,200 mg/kg dry * (0.0102 m3) * (2000 kg/m3)] = Σ[((7710 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (146 mg/kg dry * 0.0011 kg)] = = 0.010 kg or 10.00 g Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) *(2100 kg/m3)] = Σ[((7020 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011kg)] = = 0.124 kg or 123.81 g Miscanthus giganteous Soil 1: [(12,800 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((6390 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.011 kg)] = = 0.118 kg or 117.69 g Soil 2 [(8,200 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((6700 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) (<33.1 mg/kg dry * 0.011 kg)] = = 0.031 kg or 30.60 g

192

Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((4380 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (34.1 mg/kg dry * 0.011 kg)] = = 0.180 kg or 180.36 g Panicum virgatum Soil 1: [(12,800 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((8650 mg/kg dry) ) * (0.0102 m3) * (1800 kg/m3)) (1220 mg/kg dry * 0.008 kg)] = = 0.076 kg or 76.18 g Soil 2: [(8,200 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((5840 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (173 mg/kg dry * 0.008 kg)] = = 0.048 kg or 48.14 g Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((4120 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (807 mg/kg dry * 0.008 kg)] = = 0.186 kg or 185.92 g Trial 2 Camelina sativa Soil 1: [(12,800 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((8360 mg/kg dry) ) * (0.0102 m3) * (1800 kg/m3)) + (2800 mg/kg dry * 0.0032 kg)] = = 0.082 kg or 81.52 g Soil 2: [(8,200 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((5190 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (3790 mg/kg dry * 0.0032kg)] = = 0.061 kg or 61.39 g Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((3830 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 0.192 kg or 192.14 g Miscanthus giganteous Soil 1: [(12,800 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((8360 mg/kg dry) ) * (0.0102 m3) * (1800 kg/m3)) + (<15.7 mg/kg dry * 0.030 kg)] = = 0.082 kg or 81.52 g Soil 2: [(8,200 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((5190 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<21.7 mg/kg dry * 0.030 kg)] = = 0.061 kg or 61.40 g

193

Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((3830 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (<27.4 mg/kg dry * 0.030 kg)] = = 0.192 kg or 192.14 g Panicum virgatum Soil 1: [(12,800 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((8360 mg/kg dry) ) * (0.0102 m3) * (1800 kg/m3)) + (<16.0 mg/kg dry * 0.022 kg)] = = 0.082 kg or 81.52 g Soil 2: [(8,200 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((5190 mg/kg ) * (0.0102 m3) * (2000 kg/m3)) + (27.7 mg/kg dry * 0.022kg)] = = 0.061 kg or 61.40 g Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((3830 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (741 mg/kg dry * 0.022kg)] = = 0.192 kg or 192.12 g Trial 3 Camelina sativa Soil 1: [(12,800 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((8360 mg/kg dry) ) * (0.0102 m3) * (1800 kg/m3)) + (1860 mg/kg dry * 0.0032kg)] = = 0.082 kg or 81.51 g Soil 2: [(8,200 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((5190 mg/kg dry ) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 0.061 kg or 61.40 g Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((3830 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 0.192 kg or 192.14 g Miscanthus giganteous Soil 1: [(12,800 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((8360 mg/kg dry) ) * (0.0102 m3) * (1800 kg/m3)) + (<26.1 mg/kg dry * 0.030 kg)] = = 0.082 kg or 81.52 g

194

Soil 2: [(8,200 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((5190 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<20.5 mg/kg dry * 0.030 kg)] = = 0.061 kg or 61.40 g

Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((3830 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (<27.1 mg/kg dry * 0.030 kg)] = = 0.192 kg or 192.13 g Panicum virgatum Soil 1: [(12,800 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((8360 mg/kg dry) ) * (0.0102 m3) * (1800 kg/m3)) + (57.7 mg/kg dry * 0.022 kg)] = = 0.082 kg or 81.52 g Soil 2: [(8,200 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((5190 mg/kg dry) ) * (0.0102 m3) * (2000 kg/m3)) + (<16.2 mg/kg dry * 0.022 kg)] = = 0.061 kg or 61.40 g Soil 3: [(12,800 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((3830 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (293 mg/kg dry * 0.022 kg)] = = 0.192 kg or 192.13 g

195

Aluminum Total Percent MassIn Massout Mass of input Trial Plant Plant Balance, Total Soil Soil Total Cin Vin ρin ρout tissue tissue excess in Cout Vout Out Cout Mout positive, g T1S1 12800 0.0102 1800 235.0 5840 0.0102 1800 0 0.0011 107.222 127.786 Camelina 54.38 T1S2 8200 0.0102 2000 167.3 7710 0.0102 2000 146 0.0011 157.284 9.996 Camelina 5.98 T1S3 12800 0.0102 2100 274.2 7020 0.0102 2100 0 0.0011 150.368 123.808 Camelina 45.16 T1S1 12800 0.0102 1800 235.0 6390 0.0102 1800 33.7 0.011 117.321 117.687 Miscanthus 50.08 T1S2 8200 0.0102 2000 167.3 6700 0.0102 2000 33.1 0.011 136.680 30.600 Miscanthus 18.29 T1S3 12800 0.0102 2100 274.2 4380 0.0102 2100 34.1 0.011 93.820 180.356 Miscanthus 65.78 T1S1 12800 0.0102 1800 235.0 8650 0.0102 1800 1220 0.008 158.824 76.184 Panicum 32.42 T1S2 8200 0.0102 2000 167.3 5840 0.0102 2000 173 0.008 119.137 48.143 Panicum 28.78 T1S3 12800 0.0102 2100 274.2 4120 0.0102 2100 807 0.008 88.257 185.919 Panicum 67.81 T2S1 12800 0.0102 1800 235.0 8360 0.0102 1800 2800 0.0032 153.499 81.509 Camelina 34.68 T2S2 8200 0.0102 2000 167.3 5190 0.0102 2000 3790 0.0032 105.888 61.392 Camelina 36.70 T2S3 12800 0.0102 2100 274.2 3830 0.0102 2100 0 0.0032 82.039 192.137 Camelina 70.08 T2S1 12800 0.0102 1800 235.0 8360 0.0102 1800 15.7 0.03 153.490 81.518 Miscanthus 34.69 T2S2 8200 0.0102 2000 167.3 5190 0.0102 2000 21.7 0.03 105.877 61.403 Miscanthus 36.71 T2S3 12800 0.0102 2100 274.2 3830 0.0102 2100 27.4 0.03 82.039 192.137 Miscanthus 70.08 T2S1 12800 0.0102 1800 235.0 8360 0.0102 1800 16 0.022 153.490 81.518 Panicum 34.69 T2S2 8200 0.0102 2000 167.3 5190 0.0102 2000 27.7 0.022 105.877 61.403 Panicum 36.71 T2S3 12800 0.0102 2100 274.2 3830 0.0102 2100 741 0.022 82.055 192.121 Panicum 70.07 T3S1 12800 0.0102 1800 235.0 8360 0.0102 1800 1860 0.0032 153.496 81.512 Camelina 34.68 T3S2 8200 0.0102 2000 167.3 5190 0.0102 2000 0 0.0032 105.876 61.404 Camelina 36.71 T3S3 12800 0.0102 2100 274.2 3830 0.0102 2100 0 0.0032 82.039 192.137 Camelina 70.08 T3S1 12800 0.0102 1800 235.0 8360 0.0102 1800 26.1 0.03 153.490 81.518 Miscanthus 34.69 T3S2 8200 0.0102 2000 167.3 5190 0.0102 2000 20.5 0.03 105.877 61.403 Miscanthus 36.71 T3S3 12800 0.0102 2100 274.2 3830 0.0102 2100 27.1 0.03 82.039 192.137 Miscanthus 70.08 T3S1 12800 0.0102 1800 235.0 8360 0.0102 1800 57.7 0.022 153.491 81.517 Panicum 34.69 T3S2 8200 0.0102 2000 167.3 5190 0.0102 2000 16.2 0.022 105.876 61.404 Panicum 36.71 T3S3 12800 0.0102 2100 274.2 3830 0.0102 2100 293 0.022 82.045 192.131 Panicum 70.08

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Arsenic, As

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Trial 1 Camelina sativa Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((27.5 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -242.352 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((13.0 mg/kg dry) * (0.0102 m3) *(2000 kg/m3)) + (<10.0 mg/kg dry * 0.0011 kg)] = = 181.549 mg Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((33.9 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 * 0.0011 kg)] =

= -92.106 mg

Miscanthus giganteous Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((16.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<5.0 mg/kg dry * 0.011 kg)] = = -44.119 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)]] = Σ[((11.8 mg/kg dry) * (0.0102 m3) *(2000 kg/m3)) + (<4.9 mg/kg dry * 0.011 kg)] = = 205.986 mg

197

Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((43.2 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (<4.7 mg/kg dry * 0.011 kg)] = = -291.364 mg Panicum virgatum Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((17.0 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<6.9 mg/kg dry * 0.008 kg)] = = -49.627 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((11.8 mg/kg dry) * (0.0102 m3) *(2000 kg/m3)) + (<6.0 mg/kg dry * 0.008 kg)] = = 205.992 mg Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((36.2 mg/kg dry ) * (0.0102 m3) * (2100 kg/m3)) + (<7.5 mg/kg dry * 0.008 kg)] = = -141.432 mg Trial 2 Camelina sativa Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((21.4 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<22.6 mg/kg dry * 0.0032 kg)] = = -130.428 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((7.13 mg/kg dry) * (0.0102 m3) *(2000 kg/m3)) + (<11.4 mg/kg dry * 0.0032 kg)] = = 301.272 mg Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((14.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 334.152 mg Miscanthus giganteous Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((21.4 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<3.91 mg/kg dry * 0.030 kg)] = = -130.473 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((7.13 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<5.40 mg/kg dry * 0.030 kg)] = = 301.146 mg

198

Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((14.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<6.84 mg/kg dry * 0.030 kg)] = = 333.947 mg Panicum virgatum Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((21.4 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<3.98 mg/kg dry * 0.022 kg)] = = -130.444 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((7.13 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<3.91 mg/kg dry * 0.022 kg)] = = 301.222 mg Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((14.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<3.65 mg/kg dry * 0.022 kg)] = = 334.072 mg Trial 3 Camelina sativa Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((21.4 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<21.8 mg/kg dry * 0.0032 kg)] = = -130.426 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((7.13 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + ( NA * 0.0032 kg)] = = 301.308mg Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((14.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + ( NA * 0.0032 kg)] = = 334.152 mg Miscanthus giganteous Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((21.4 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + ( <6.50 mg/kg dry * 0.030 kg)]= = -130.551 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((7.13 mg/kg dry) * (0.0102 m3) *(2000 kg/m3)) + (<5.10 mg/kg dry * 0.030 kg)] = = 301.155 mg

199

Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((14.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<6.72 mg/kg dry * 0.030 kg)] = = 333.950 mg Panicum virgatum Soil 1: [(14.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((21.4 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<4.05 mg/kg dry * 0.022 kg)] = = -130.445 mg Soil 2: [(21.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((7.13 mg/kg dry) * (0.0102 m3) *(2000 kg/m3)) + (<6.01 mg/kg dry * 0.022 kg)] = = 301.176 mg Soil 3: [(29.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((14.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<3.89 mg/kg dry * 0.022 kg)] = = 334.066 mg

200

Arsenic MassIn Massout Total Mass Balance, Trial Plant Plant Total Soil Soil Total excess C V ρ ρ tissue tissue in in in in C V out Out positive, Percent out out C Mout out mg of input T1S1 14.3 0.0102 1800 262.55 27.5 0.0102 1800 0 0.0011 504.9 -242.352 Camelina -92.31 T1S2 21.9 0.0102 2000 446.76 13 0.0102 2000 10.3 0.0011 265.21 181.549 Camelina 40.64 T1S3 29.6 0.0102 2100 634.03 33.9 0.0102 2100 0 0.0011 726.14 -92.106 Camelina -14.53 T1S1 14.3 0.0102 1800 262.55 16.7 0.0102 1800 5 0.011 306.67 -44.119 Miscanthus -16.80 T1S2 21.9 0.0102 2000 446.76 11.8 0.0102 2000 4.9 0.011 240.77 205.986 Miscanthus 46.11 T1S3 29.6 0.0102 2100 634.03 43.2 0.0102 2100 4.7 0.011 925.4 -291.364 Miscanthus -45.95 T1S1 14.3 0.0102 1800 262.55 17 0.0102 1800 6.9 0.008 312.18 -49.627 Panicum -18.90 T1S2 21.9 0.0102 2000 446.76 11.8 0.0102 2000 6 0.008 240.77 205.992 Panicum 46.11 T1S3 29.6 0.0102 2100 634.03 36.2 0.0102 2100 7.5 0.008 775.46 -141.432 Panicum -22.31 T2S1 14.3 0.0102 1800 262.55 21.4 0.0102 1800 22.6 0.0032 392.98 -130.428 Camelina -49.68 T2S2 21.9 0.0102 2000 446.76 7.13 0.0102 2000 11.4 0.0032 145.49 301.272 Camelina 67.43 T2S3 29.6 0.0102 2100 634.03 14 0.0102 2100 0 0.0032 299.88 334.152 Camelina 52.70 T2S1 14.3 0.0102 1800 262.55 21.4 0.0102 1800 3.91 0.03 393.02 -130.473 Miscanthus -49.70 T2S2 21.9 0.0102 2000 446.76 7.13 0.0102 2000 5.4 0.03 145.61 301.146 Miscanthus 67.41 T2S3 29.6 0.0102 2100 634.03 14 0.0102 2100 6.84 0.03 300.09 333.947 Miscanthus 52.67 T2S1 14.3 0.0102 1800 262.55 21.4 0.0102 1800 3.98 0.022 392.99 -130.444 Panicum -49.68 T2S2 21.9 0.0102 2000 446.76 7.13 0.0102 2000 3.91 0.022 145.54 301.222 Panicum 67.42 T2S3 29.6 0.0102 2100 634.03 14 0.0102 2100 3.65 0.022 299.96 334.072 Panicum 52.69 T3S1 14.3 0.0102 1800 262.55 21.4 0.0102 1800 21.8 0.0032 392.97 -130.426 Camelina -49.68 T3S2 21.9 0.0102 2000 446.76 7.13 0.0102 2000 0 0.0032 145.45 301.308 Camelina 67.44 T3S3 29.6 0.0102 2100 634.03 14 0.0102 2100 0 0.0032 299.88 334.152 Camelina 52.70 T3S1 14.3 0.0102 1800 262.55 21.4 0.0102 1800 6.5 0.03 393.1 -130.551 Miscanthus -49.72 T3S2 21.9 0.0102 2000 446.76 7.13 0.0102 2000 5.1 0.03 145.61 301.155 Miscanthus 67.41 T3S3 29.6 0.0102 2100 634.03 14 0.0102 2100 6.72 0.03 300.08 333.950 Miscanthus 52.67 T3S1 14.3 0.0102 1800 262.55 21.4 0.0102 1800 4.05 0.022 392.99 -130.445 Panicum -49.68 T3S2 21.9 0.0102 2000 446.76 7.13 0.0102 2000 6.01 0.022 145.58 301.176 Panicum 67.41 T3S3 29.6 0.0102 2100 634.03 14 0.0102 2100 3.89 0.022 299.97 334.066 Panicum 52.69

201

Barium, Ba

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Trial 1 Camelina sativa Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((139 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = - 660.96 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((113 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<17.2 mg/kg dry * 0.0011 kg)] = = -634.46 mg Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((120 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -803.25 mg Miscanthus giganteous Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((98.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (50.8 mg/kg dry * 0.011 kg)] = = 78.39 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((153 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (14.6 mg/kg dry * 0.011 kg)] = = -1450.60 mg

202

Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((148 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<7.9 mg/kg dry * 0.011 kg)] = = -1403.10 mg Panicum virgatum Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((93.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (26.9 mg/kg dry * 0.008 kg)] = = 170.53 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((67.8 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (13.7 mg/kg dry * 0.008 kg)] = = 287.53 mg Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((98.9 mg/kg dry) * (0.0102 m3) * ( 2100 kg/m3)) + (16.3 mg/kg dry * 0.008 kg)] = = -351.42 mg Trial 2 Camelina sativa Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((101 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (87.8 mg/kg dry * 0.0032 kg)] = = 36.44 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((56.5 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (81.9 mg/kg dry * 0.0032 kg)] = = 517.90 mg Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((65.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 370.57 mg Miscanthus giganteous Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((101 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (36.9 mg/kg dry * 0.030 kg)] = = 35.61 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((56.5 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (12.1 mg/kg dry * 0.030 kg)] = = 517.80 mg

203

Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((65.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (18.7 mg/kg dry * 0.030 kg)] = = 370.01 mg Panicum virgatum Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((101 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (10.7 mg/kg dry * 0.022 kg)] = = 36.48 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((56.5 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<7.84 mg/kg dry * 0.022 kg)] = = 517.99 mg Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((65.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (17.4 mg/kg dry * 0.022 kg)] = = 370.18 mg Trial 3 Camelina sativa Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((101 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (43.8 mg/kg dry * 0.0032kg)] = = 35.32 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((56.5 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 518.16 mg Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((65.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 370.57 mg Miscanthus giganteous Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((101 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (25.5 mg/kg dry * 0.030 kg)] = = 35.95 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((56.5 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (20.0 mg/kg dry * 0.030 kg)] = = 517.56 mg

204

Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((65.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (14.1 mg/kg dry * 0.030 kg)] = = 370.14 mg Panicum virgatum Soil 1: [(103 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((101 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (12.9 mg/kg dry * 0.022 kg)] = = 36.44 mg Soil 2: [(81.9 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((56.5 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<8.10 mg/kg dry * 0.022 kg)] = = 517.98 mg Soil 3: [(82.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((65.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<12.0 mg/kg dry * 0.022 kg)] = = 370.30 mg

205

Barium MassIn Massout Total Mass Trial Plant Plant Balance, Total Soil Soil Total C V ρ ρ tissue tissue excess in in in in C V out Out Percent out out positive, mg Cout Mout of input T1S1 103 0.0102 1800 1891.1 139 0.0102 1800 0 0.0011 2552.0 -660.96 Camelina -34.95 T1S2 81.9 0.0102 2000 1670.8 113 0.0102 2000 17.2 0.0011 2305.2 -634.46 Camelina -37.97 T1S3 82.5 0.0102 2100 1767.2 120 0.0102 2100 0 0.0011 2570.4 -803.25 Camelina -45.45 T1S1 103 0.0102 1800 1891.1 98.7 0.0102 1800 50.8 0.011 1812.7 78.39 Miscanthus 4.15 T1S2 81.9 0.0102 2000 1670.8 153 0.0102 2000 14.6 0.011 3121.4 -1450.60 Miscanthus -86.82 T1S3 82.5 0.0102 2100 1767.2 148 0.0102 2100 7.9 0.011 3170.2 -1403.10 Miscanthus -79.40 T1S1 103 0.0102 1800 1891.1 93.7 0.0102 1800 26.9 0.008 1720.5 170.53 Panicum 9.02 T1S2 81.9 0.0102 2000 1670.8 67.8 0.0102 2000 13.7 0.008 1383.2 287.53 Panicum 17.21 T1S3 82.5 0.0102 2100 1767.2 98.9 0.0102 2100 16.3 0.008 2118.6 -351.42 Panicum -19.89 T2S1 103 0.0102 1800 1891.1 101 0.0102 1800 87.8 0.0032 1854.6 36.44 Camelina 1.93 T2S2 81.9 0.0102 2000 1670.8 56.5 0.0102 2000 81.9 0.0032 1152.9 517.90 Camelina 31.00 T2S3 82.5 0.0102 2100 1767.2 65.2 0.0102 2100 0 0.0032 1396.6 370.57 Camelina 20.97 T2S1 103 0.0102 1800 1891.1 101 0.0102 1800 36.9 0.03 1855.5 35.61 Miscanthus 1.88 T2S2 81.9 0.0102 2000 1670.8 56.5 0.0102 2000 12.1 0.03 1153.0 517.80 Miscanthus 30.99 T2S3 82.5 0.0102 2100 1767.2 65.2 0.0102 2100 18.7 0.03 1397.1 370.01 Miscanthus 20.94 T2S1 103 0.0102 1800 1891.1 101 0.0102 1800 10.7 0.022 1854.6 36.48 Panicum 1.93 T2S2 81.9 0.0102 2000 1670.8 56.5 0.0102 2000 7.84 0.022 1152.8 517.99 Panicum 31.00 T2S3 82.5 0.0102 2100 1767.2 65.2 0.0102 2100 17.4 0.022 1397.0 370.18 Panicum 20.95 T3S1 103 0.0102 1800 1891.1 101 0.0102 1800 43.8 0.032 1855.8 35.32 Camelina 1.87 T3S2 81.9 0.0102 2000 1670.8 56.5 0.0102 2000 0 0.032 1152.6 518.16 Camelina 31.01 T3S3 82.5 0.0102 2100 1767.2 65.2 0.0102 2100 0 0.032 1396.6 370.57 Camelina 20.97 T3S1 103 0.0102 1800 1891.1 101 0.0102 1800 25.5 0.03 1855.1 35.95 Miscanthus 1.90 T3S2 81.9 0.0102 2000 1670.8 56.5 0.0102 2000 20 0.03 1153.2 517.56 Miscanthus 30.98 T3S3 82.5 0.0102 2100 1767.2 65.2 0.0102 2100 14.1 0.03 1397.0 370.14 Miscanthus 20.95 T3S1 103 0.0102 1800 1891.1 101 0.0102 1800 12.9 0.022 1854.6 36.44 Panicum 1.93 T3S2 81.9 0.0102 2000 1670.8 56.5 0.0102 2000 8.1 0.022 1152.8 517.98 Panicum 31.00 T3S3 82.5 0.0102 2100 1767.2 65.2 0.0102 2100 12 0.022 1396.8 370.30 Panicum 20.95

206

Cadmium, Cd

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Not detected in any of the lab analyses.

Trial 1 Camelina sativa Soil 1: [(<0.52 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<0.55 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] =

= -23.13 mg

Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.55 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<3.4 mg/kg dry * 0.0011 kg)] = = -26.93 mg Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<0.57 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -30.42 mg Miscanthus giganteous Soil 1: [(<0.52 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<0.57 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<1.7 mg/kg dry * 0.011 kg)] = = -23.15 mg Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((< 0.46 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<1.6 mg/kg dry * 0.011 kg)] = = - 26.95 mg

207

Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<0.61 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<1.6 mg/kg dry * 0.011 kg)] = = -30.43 mg Panicum virgatum Soil 1: [(<0.52 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((< 0.53 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<2.3 mg/kg dry * 0.008 kg)] = = -23.15 mg Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.54 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<2.0 mg/kg dry * 0.008 kg)] = = - 26.94 mg Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<0.63 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<2.5 mg/kg dry * 0.008 kg)] = = -30.44 mg Trial 2 Camelina sativa Soil 1: [(<0.52 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[(( <1.78 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<9.08 mg/kg dry * 0.0032 kg)] = = -23.16 mg Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<1.80 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<4.56 mg/kg dry * 0.0032 kg)] = = -26.94 mg Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<2.00 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<8.21 mg/kg dry * 0.0032 kg)] = = - 30.44 mg Miscanthus giganteous Soil 1: [(<0.52 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[(( <1.78mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<1.57mg/kg dry * 0.030 kg)] = = - 23.18 mg Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<1.80 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<2.17 mg/kg dry * 0.030 kg)] = = -26.99 mg

208

Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<2.00 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<2.74 mg/kg dry * 0.030 kg)] = = -30.50 mg Panicum virgatum Soil 1: [(<0.52 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<1.78 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<1.60 mg/kg dry * 0.022 kg)] = = -23.17 mg Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<1.80 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<1.57 mg/kg dry * 0.022 kg)] = = -26.96 mg Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<2.00 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<1.47 mg/kg dry * 0.022 kg)] = = -30.45 mg Trial 3 Camelina sativa Soil 1: [(<0.52 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<1.78 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<8.77 mg/kg dry * 0.0032 kg)] = = -23.16 mg Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<1.80 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -26.93 mg Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<2.00 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -30.42 mg Miscanthus giganteous Soil 1: [(<0.52mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<1.78 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<2.61 mg/kg dry * 0.030 kg)] = = -23.21 mg

209

Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<1.80 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<2.05 mg/kg dry * 0.030 kg)] = = -26.99 mg Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<2.00 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<2.70 mg/kg dry * 0.030 kg)] = = -30.50 mg Panicum virgatum Soil 1: [(<0.52 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<1.78 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<1.65 mg/kg dry * 0.022 kg)] = = -23.17 mg Soil 2: [(<0.48 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<1.80 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<1.62 mg/kg dry * 0.022 kg)] = = -26.96 mg Soil 3: [(<0.58 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<2.00 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<2.41 mg/kg dry * 0.022 kg)] = = -30.47 mg

210

Cadmium Percent Mass Mass In out Total Mass of input Trial Balance, Plant Plant Total Soil Soil Total excess Cin Vin ρin ρout tissue tissue in Cout Vout Out positive, mg Cout Mout T1S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 0 0.001 32.68 -23.13 Camelina -242.31 T1S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 3.4 0.001 36.72 -26.93 Camelina -275.04 T1S3 0.58 0.0102 2100 12.42 2 0.0102 2100 0 0.001 42.84 -30.42 Camelina -244.83 T1S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 1.7 0.011 32.7 -23.15 Miscanthus -242.50 T1S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 1.6 0.011 36.74 -26.95 Miscanthus -275.18 T1S3 0.58 0.0102 2100 12.42 2 0.0102 2100 1.6 0.011 42.86 -30.43 Miscanthus -244.97 T1S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 2.3 0.008 32.7 -23.15 Panicum -242.50 T1S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 2 0.008 36.74 -26.94 Panicum -275.16 T1S3 0.58 0.0102 2100 12.42 2 0.0102 2100 2.5 0.008 42.86 -30.44 Panicum -244.99 T2S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 9.08 0.003 32.71 -23.16 Camelina -242.61 T2S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 4.56 0.003 36.73 -26.94 Camelina -275.15 T2S3 0.58 0.0102 2100 12.42 2 0.0102 2100 8.21 0.003 42.87 -30.44 Camelina -245.04 T2S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 1.57 0.03 32.73 -23.18 Miscanthus -242.80 T2S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 2.17 0.03 36.79 -26.99 Miscanthus -275.66 T2S3 0.58 0.0102 2100 12.42 2 0.0102 2100 2.74 0.03 42.92 -30.50 Miscanthus -245.49 T2S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 1.6 0.022 32.72 -23.17 Panicum -242.68 T2S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 1.57 0.022 36.75 -26.96 Panicum -275.35 T2S3 0.58 0.0102 2100 12.42 2 0.0102 2100 1.47 0.022 42.87 -30.45 Panicum -245.09 T3S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 8.77 0.003 32.71 -23.16 Camelina -242.60 T3S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 0 0.003 36.72 -26.93 Camelina -275.00 T3S3 0.58 0.0102 2100 12.42 2 0.0102 2100 0 0.003 42.84 -30.42 Camelina -244.83 T3S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 2.61 0.03 32.76 -23.21 Miscanthus -243.13 T3S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 2.05 0.03 36.78 -26.99 Miscanthus -275.63 T3S3 0.58 0.0102 2100 12.42 2 0.0102 2100 2.7 0.03 42.92 -30.50 Miscanthus -245.48 T3S1 0.52 0.0102 1800 9.547 1.78 0.0102 1800 1.65 0.022 32.72 -23.17 Panicum -242.69 T3S2 0.48 0.0102 2000 9.792 1.8 0.0102 2000 1.62 0.022 36.76 -26.96 Panicum -275.36 T3S3 0.58 0.0102 2100 12.42 2 0.0102 2100 2.41 0.022 42.89 -30.47 Panicum -245.25

211

Chromium, Cr

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Trial 1 Camelina sativa Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((83.6 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] =

= -1283.36 mg

Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((14.5 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<6.9 mg/kg dry * 0.0011 kg)] = = -71.41 mg Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((15.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -143.51 mg Miscanthus giganteous Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((11.8 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<3.4 mg/kg dry * 0.011 kg)] = = 34.85 mg Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((11.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<3.2 mg/kg dry * 0.011 kg)] = = -8.20 mg

212

Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((19.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<3.2 mg/kg dry * 0.011 kg)] = = -222.80 mg Panicum virgatum Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((12.2 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<4.6 mg/kg dry * 0.008 kg)] = = 27.50 mg Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((7.8 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<4.0 mg/kg dry * 0.008 kg)] = = 65.25 mg Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((12.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (19.7 mg/kg dry * 0.008 kg)] = = -64.42 mg Trial 2 Camelina sativa Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((12.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<45.3 mg/kg dry * 0.0032 kg)] = = 29.23 mg Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<22.7 mg/kg dry * 0.0032 kg)] = = -41.14 mg Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<8.21 mg/kg dry * 0.0032 kg)] = = -18.23 mg Miscanthus giganteous Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((12.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<7.84 mg/kg dry * 0.030 kg)] = = 29.14 mg Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<10.8 mg/kg dry * 0.030 kg)] = = 40.88 mg

213

Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<13.7 mg/kg dry * 0.030 kg)] = = -18.62 mg Panicum virgatum Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((12.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<7.98 mg/kg dry * 0.022 kg)] = = 29.20 mg Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<7.84 mg/kg dry * 0.022 kg)] = = 41.04 mg Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<7.32 mg/kg dry * 0.022 kg)] = = -18.37 mg Trial 3 Camelina sativa Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((12.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<43.7 mg/kg dry * 0.0032 kg)] = = 29.24 mg Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 41.21 mg Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -18.21 mg Miscanthus giganteous Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((12.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<13.0 mg/kg dry * 0.030 kg)] = = 28.99 mg

214

Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<10.2 mg/kg dry * 0.030 kg)] = = 40.90 mg Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<13.5 mg/kg dry * 0.030 kg)] = =-18.61 mg Panicum virgatum Soil 1: [(13.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((12.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<8.22 mg/kg dry * 0.022 kg)] = = 29.20 mg Soil 2: [(11.0 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<8.10 mg/kg dry * 0.022 kg)] = = 41.03 mg Soil 3: [(9.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<12.0 mg/kg dry * 0.022 kg)] = = -18.47 mg

215

Chromium Mass Mass In out Total Mass Trial Plant Plant Balance, Total Soil Soil Total C V ρ ρ tissue tissue excess in in in in C V out Out Percent out out C Mout positive, mg out of input T1S1 13.7 0.0102 1800 251.5 83.6 0.0102 1800 0 0.001 1535 -1283.364 Camelina -510.22 T1S2 11 0.0102 2000 224.4 14.5 0.0102 2000 6.9 0.001 295.8 -71.40759 Camelina -31.82 T1S3 9.1 0.0102 2100 194.9 15.8 0.0102 2100 0 0.001 338.4 -143.514 Camelina -73.63 T1S1 13.7 0.0102 1800 251.5 11.8 0.0102 1800 3.4 0.011 216.7 34.8466 Miscanthus 13.85 T1S2 11 0.0102 2000 224.4 11.4 0.0102 2000 3.2 0.011 232.6 -8.1952 Miscanthus -3.65 T1S3 9.1 0.0102 2100 194.9 19.5 0.0102 2100 3.2 0.011 417.7 -222.8032 Miscanthus -114.30 T1S1 13.7 0.0102 1800 251.5 12.2 0.0102 1800 4.6 0.008 224 27.5032 Panicum 10.93 T1S2 11 0.0102 2000 224.4 7.8 0.0102 2000 4 0.008 159.2 65.248 Panicum 29.08 T1S3 9.1 0.0102 2100 194.9 12.1 0.0102 2100 19.7 0.008 259.3 -64.4176 Panicum -33.05 T2S1 13.7 0.0102 1800 251.5 12.1 0.0102 1800 45.3 0.003 222.3 29.23104 Camelina 11.62 T2S2 11 0.0102 2000 224.4 8.98 0.0102 2000 22.7 0.003 183.3 41.13536 Camelina 18.33 T2S3 9.1 0.0102 2100 194.9 9.95 0.0102 2100 8.21 0.003 213.2 -18.233272 Camelina -9.35 T2S1 13.7 0.0102 1800 251.5 12.1 0.0102 1800 7.84 0.03 222.4 29.1408 Miscanthus 11.59 T2S2 11 0.0102 2000 224.4 8.98 0.0102 2000 10.8 0.03 183.5 40.884 Miscanthus 18.22 T2S3 9.1 0.0102 2100 194.9 9.95 0.0102 2100 13.7 0.03 213.5 -18.618 Miscanthus -9.55 T2S1 13.7 0.0102 1800 251.5 12.1 0.0102 1800 7.98 0.022 222.3 29.20044 Panicum 11.61 T2S2 11 0.0102 2000 224.4 8.98 0.0102 2000 7.84 0.022 183.4 41.03552 Panicum 18.29 T2S3 9.1 0.0102 2100 194.9 9.95 0.0102 2100 7.32 0.022 213.3 -18.36804 Panicum -9.42 T3S1 13.7 0.0102 1800 251.5 12.1 0.0102 1800 43.7 0.003 222.3 29.23616 Camelina 11.62 T3S2 11 0.0102 2000 224.4 8.98 0.0102 2000 0 0.003 183.2 41.208 Camelina 18.36 T3S3 9.1 0.0102 2100 194.9 9.95 0.0102 2100 0 0.003 213.1 -18.207 Camelina -9.34 T3S1 13.7 0.0102 1800 251.5 12.1 0.0102 1800 13 0.03 222.5 28.986 Miscanthus 11.52 T3S2 11 0.0102 2000 224.4 8.98 0.0102 2000 10.2 0.03 183.5 40.902 Miscanthus 18.23 T3S3 9.1 0.0102 2100 194.9 9.95 0.0102 2100 13.5 0.03 213.5 -18.612 Miscanthus -9.55 T3S1 13.7 0.0102 1800 251.5 12.1 0.0102 1800 8.22 0.022 222.3 29.19516 Panicum 11.61 T3S2 11 0.0102 2000 224.4 8.98 0.0102 2000 8.1 0.022 183.4 41.0298 Panicum 18.28 T3S3 9.1 0.01 2100 194.9 9.95 0.0102 2100 12 0.022 213.4 -18.471 Panicum -9.48

216

Lead, Pb

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Trial 1 Camelina sativa Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((189 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -1896.59 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((22.5 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<6.9 mg/kg dry * 0.0011 kg)] = = -118.33 mg Soil 3: [(46.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((48.4 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -51.41 mg Miscanthus giganteous Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((96.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<3.4 mg/kg dry * 0.011 kg)] = = -202.00 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((22.2 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<3.2 mg/kg dry * 0.011 kg)] = = -112.24 mg

217

Soil 3: [(46.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((60.3 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<3.2 mg/kg dry * 0.011 kg)] = = -306.34 mg Panicum virgatum Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((86.9 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<4.6 mg/kg dry * 0.008 kg)] = = -22.07 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((18.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<4.0 mg/kg dry * 0.008 kg)] = = -34.71 mg Soil 3: [(46.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((71.3 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<5.0 mg/kg dry * 0.008 kg)] = = -541.97 mg Trial 2 Camelina sativa Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((79.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<45.3 mg/kg dry * 0.0032 kg)] = = 117.27 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((21.2 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<22.7 mg/kg dry * 0.0032 kg)] = = -91.87 mg Soil 3: [(46.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((35.7 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<8.21 mg/kg dry * 0.0032 kg) = = 220.60 mg Miscanthus giganteous Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((79.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<7.84 mg/kg dry * 0.030 kg)] = = 117.27 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((21.2 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<10.8 mg/kg dry * 0.030 kg)] = = -92.12 mg

218

Soil 3: [(46.0 mg/kg dry) * (0.0114 m3) * (2100 kg/m3)] = Σ[((35.7 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<13.7 mg/kg dry * 0.030 kg)] = = 220.22 mg Panicum virgatum Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((79.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<7.98 mg/kg dry * 0.022 kg)] = = 117.33 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((21.2 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<7.84 mg/kg dry * 0.022 kg)] = = -89.93 mg Soil 3: [(46.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((35.7 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (8.28 mg/kg dry * 0.022 kg)] = = 220.44 mg Trial 3 Camelina sativa Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((79.3mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<43.7 mg/kg dry * 0.0032 kg)] = = 117.36 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((21.2 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -89.76 mg Soil 3: [(46.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((35.7 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 220.63 mg Miscanthus giganteous Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((79.3 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<13.0 mg/kg dry * 0.030 kg)] = = 117.11 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((21.2 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<10.2 mg/kg dry * 0.030 kg)] = = -90.07 mg

219

Soil 3: [(46.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((35.7 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<13.5 mg/kg dry * 0.030 kg)] = = 220.22 mg Panicum virgatum Soil 1: [(85.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((79.3mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<8.22 mg/kg dry * 0.022 kg)] = = 117.32 mg Soil 2: [(16.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((21.2 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<8.10 mg/kg dry * 0.022 kg)] = = -89.94 mg Soil 3: [(46.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((35.7 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<12.0 mg/kg dry * 0.022 kg)] = = 220.36 mg

220

Lead Mass Mass In out Total Mass Trial Plant Plant Balance, Total Soil Soil Total C V ρ ρ tissue tissue excess in in in in C V out Out Percent out out C Mout positive, mg out of input T1S1 85.7 0.0102 1800 1573 189 0.0102 1800 0 0.001 3470 -1896.588 Camelina -120.54 T1S2 16.7 0.0102 2000 340.7 22.5 0.0102 2000 6.9 0.001 459 -118.328 Camelina -34.73 T1S3 46 0.0102 2100 985.3 48.4 0.0102 2100 0 0.001 1037 -51.408 Camelina -5.22 T1S1 85.7 0.0102 1800 1573 96.7 0.0102 1800 3.4 0.011 1775 -201.997 Miscanthus -12.84 T1S2 16.7 0.0102 2000 340.7 22.2 0.0102 2000 3.2 0.011 452.9 -112.235 Miscanthus -32.94 T1S3 46 0.0102 2100 985.3 60.3 0.0102 2100 3.2 0.011 1292 -306.341 Miscanthus -31.09 T1S1 85.7 0.0102 1800 1573 86.9 0.0102 1800 4.6 0.008 1596 -22.069 Panicum -1.40 T1S2 16.7 0.0102 2000 340.7 18.4 0.0102 2000 4 0.008 375.4 -34.712 Panicum -10.19 T1S3 46 0.0102 2100 985.3 71.3 0.0102 2100 5 0.008 1527 -541.966 Panicum -55.00 T2S1 85.7 0.0102 1800 1573 79.3 0.0102 1800 45.3 0.003 1456 117.359 Camelina 7.46 T2S2 16.7 0.0102 2000 340.7 21.2 0.0102 2000 22.7 0.003 432.6 -91.873 Camelina -26.97 T2S3 46 0.0102 2100 985.3 35.7 0.0102 2100 8.21 0.003 764.7 220.600 Camelina 22.39 T2S1 85.7 0.0102 1800 1573 79.3 0.0102 1800 7.84 0.03 1456 117.269 Miscanthus 7.45 T2S2 16.7 0.0102 2000 340.7 21.2 0.0102 2000 10.8 0.03 432.8 -92.124 Miscanthus -27.04 T2S3 46 0.0102 2100 985.3 35.7 0.0102 2100 13.7 0.03 765.1 220.215 Miscanthus 22.35 T2S1 85.7 0.0102 1800 1573 79.3 0.0102 1800 7.98 0.022 1456 117.328 Panicum 7.46 T2S2 16.7 0.0102 2000 340.7 21.1 0.0102 2000 7.84 0.022 430.6 -89.932 Panicum -26.40 T2S3 46 0.0102 2100 985.3 35.7 0.0102 2100 8.28 0.022 764.9 220.444 Panicum 22.37 T3S1 85.7 0.0102 1800 1573 79.3 0.0102 1800 43.7 0.003 1456 117.364 Camelina 7.46 T3S2 16.7 0.0102 2000 340.7 21.1 0.0102 2000 0 0.003 430.4 -89.760 Camelina -26.35 T3S3 46 0.0102 2100 985.3 35.7 0.0102 2100 0 0.003 764.7 220.626 Camelina 22.39 T3S1 85.7 0.0102 1800 1573 79.3 0.0102 1800 13 0.03 1456 117.114 Miscanthus 7.44 T3S2 16.7 0.0102 2000 340.7 21.1 0.0102 2000 10.2 0.03 430.7 -90.066 Miscanthus -26.44 T3S3 46 0.0102 2100 985.3 35.7 0.0102 2100 13.5 0.03 765.1 220.221 Miscanthus 22.35 T3S1 85.7 0.0102 1800 1573 79.3 0.0102 1800 8.22 0.022 1456 117.323 Panicum 7.46 T3S2 16.7 0.0102 2000 340.7 21.1 0.0102 2000 8.1 0.022 430.6 -89.938 Panicum -26.40 T3S3 46 0.0102 2100 985.3 35.7 0.0102 2100 12 0.022 765 220.362 Panicum 22.36

221

Mercury, Hg

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Trial 1 Camelina sativa Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.36 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = 0.37 mg Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((0.072 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<0.35 mg/kg dry * 0.0011 kg)] = = 1.80 mg Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.22mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -.021 mg Miscanthus giganteous Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.46mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<0.17 mg/kg dry * 0.011 kg)] = = -1.47 mg Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((0.12 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<0.17 mg/kg dry * 0.011 kg)] = = 0.81 mg

222

Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.30 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<0.16 mg/kg dry * 0.011 kg)] = = -1.93 mg Panicum virgatum Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.39 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<0.19 mg/kg dry * 0.008 kg)] = = -0.19 mg Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((0.098 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<0.20 mg/kg dry * 0.008 kg)] = = 1.26 mg Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.14mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<0.25 mg/kg dry * 0.008 kg)] = = 1.50 mg Trial 2 Camelina sativa Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.302 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<45.3 mg/kg dry * 0.0032 kg)] = = 1.29 mg Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.0478 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 2.29 mg Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.0616 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<0.0524 mg/kg dry * 0.0032 kg)] = = 3.18 mg

Miscanthus giganteous Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.302 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<0.0501 mg/kg dry * 0.030 kg)] = = 1.43 mg

223

Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.0478 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<0.0537 mg/kg dry * 0.030 kg)] = = 2.29 mg Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.0616 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<0.299 mg/kg dry * 0.030 kg)] = = 3.17 mg Panicum virgatum Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.302 mg/kg dry ) * (0.0102 m3) * (1800 kg/m3)) + (<0.0661 mg/kg dry * 0.022 kg)] = = 1.43 mg Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.0478 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<0.0344 mg/kg dry * 0.022 kg)] = = 2.29 mg Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.0616 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.0489 mg/kg dry * 0.022 kg)] = = 3.18 mg Trial 3 Camelina sativa Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.302 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 1.43 mg Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.0478 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 2.29 mg Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.0616 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 3.18 mg Miscanthus giganteous Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.302 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<0.109 mg/kg dry * 0.030kg)] = = 1.43 mg

224

Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.0478 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<0.0785 mg/kg dry * 0.030 kg)] = = 2.29 mg Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.0616 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<0.0472 mg/kg dry * 0.030 kg)] = = 3.18 mg

Panicum virgatum Soil 1: [(0.38 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((0.302 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<0.0368 mg/kg dry * 0.022 kg)] = = 1.43 mg Soil 2: [(0.16 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.0478 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<0.0366 mg/kg dry * 0.022 kg)] = = 2.29 mg Soil 3: [(0.21 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((0.0616 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<0.247 mg/kg dry * 0.022 kg)] = = 3.18 mg

225

Mercury Percent Mass Mass Total Mass In out of input Trial Balance, Plant Plant excess Total Soil Soil Total Cin Vin ρin ρout tissue tissue positive, in Cout Vout Out Cout Mout mg T1S1 0.38 0.0102 1800 6.98 0.36 0.0102 1800 0 0.0011 6.61 0.367 Camelina 5.26 T1S2 0.16 0.0102 2000 3.26 0.072 0.0102 2000 0.35 0.0011 1.47 1.795 Camelina 54.99 T1S3 0.21 0.0102 2100 4.50 0.22 0.0102 2100 0 0.0011 4.71 -0.214 Camelina -4.76 T1S1 0.38 0.0102 1800 6.98 0.46 0.0102 1800 0.17 0.011 8.45 -1.471 Miscanthus -21.08 T1S2 0.16 0.0102 2000 3.26 0.12 0.0102 2000 0.17 0.011 2.45 0.814 Miscanthus 24.94 T1S3 0.21 0.0102 2100 4.50 0.3 0.0102 2100 0.16 0.011 6.43 -1.930 Miscanthus -42.90 T1S1 0.38 0.0102 1800 6.98 0.39 0.0102 1800 0.19 0.008 7.16 -0.185 Panicum -2.65 T1S2 0.16 0.0102 2000 3.26 0.098 0.0102 2000 0.2 0.008 2.00 1.263 Panicum 38.70 T1S3 0.21 0.0102 2100 4.50 0.14 0.0102 2100 0.25 0.008 3.00 1.497 Panicum 33.29 T2S1 0.38 0.0102 1800 6.98 0.302 0.0102 1800 45.3 0.0032 5.69 1.287 Camelina 18.45 T2S2 0.16 0.0102 2000 3.26 0.0478 0.0102 2000 0 0.0032 0.98 2.289 Camelina 70.13 T2S3 0.21 0.0102 2100 4.50 0.0616 0.0102 2100 0.0524 0.0032 1.32 3.179 Camelina 70.66 T2S1 0.38 0.0102 1800 6.98 0.302 0.0102 1800 0.0501 0.03 5.55 1.431 Miscanthus 20.50 T2S2 0.16 0.0102 2000 3.26 0.0478 0.0102 2000 0.0537 0.03 0.98 2.287 Miscanthus 70.08 T2S3 0.21 0.0102 2100 4.50 0.0616 0.0102 2100 0.299 0.03 1.33 3.170 Miscanthus 70.47 T2S1 0.38 0.0102 1800 6.98 0.302 0.0102 1800 0.0661 0.022 5.55 1.431 Panicum 20.51 T2S2 0.16 0.0102 2000 3.26 0.0478 0.0102 2000 0.0344 0.022 0.98 2.288 Panicum 70.10 T2S3 0.21 0.0102 2100 4.50 0.0616 0.0102 2100 0.0489 0.022 1.32 3.178 Panicum 70.64 T3S1 0.38 0.0102 1800 6.98 0.302 0.0102 1800 0 0.0032 5.54 1.432 Camelina 20.53 T3S2 0.16 0.0102 2000 3.26 0.0478 0.0102 2000 0 0.0032 0.98 2.289 Camelina 70.13 T3S3 0.21 0.0102 2100 4.50 0.0616 0.0102 2100 0 0.0032 1.32 3.179 Camelina 70.67 T3S1 0.38 0.0102 1800 6.98 0.302 0.0102 1800 0.109 0.03 5.55 1.429 Miscanthus 20.48 T3S2 0.16 0.0102 2000 3.26 0.0478 0.0102 2000 0.0785 0.03 0.98 2.287 Miscanthus 70.05 T3S3 0.21 0.0102 2100 4.50 0.0616 0.0102 2100 0.0472 0.03 1.32 3.177 Miscanthus 70.64 T3S1 0.38 0.0102 1800 6.98 0.302 0.0102 1800 0.0368 0.022 5.55 1.431 Panicum 20.51 T3S2 0.16 0.0102 2000 3.26 0.0478 0.0102 2000 0.0366 0.022 0.98 2.288 Panicum 70.10 T3S3 0.21 0.0102 2100 4.50 0.0616 0.0102 2100 0.247 0.022 1.32 3.173 Panicum 70.55

226

Selenium, Se

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Trial 1

Camelina sativa Soil 1: [(<2.6 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((4.0 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -25.70 mg oil 2: [(<2.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((< <2.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<17.2 mg/kg dry * 0.0011 kg)] = = -6.14 mg Soil 3: [(6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((6.6 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = 4.28 mg Miscanthus giganteous Soil 1: [(<2.6 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((3.4 mg/kg dry) * (0.0102 m3) * (1800 kg/m3) + (<8.4 mg/kg dry * 0.011 kg)] = = -14.78 mg Soil 2: [(<2.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<2.3 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<8.1 mg/kg dry * 0.011kg)] = = 1.95 mg

227

Soil 3: [(6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((9.5 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) +(<7.9 mg/kg dry * 0.011 kg)] = = -57.92 mg Panicum virgatum Soil 1: [(<2.6 mg/kg dry)* (0.0102 m3) * (1800 kg/m3)] = Σ[( <2.7 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<11.5 mg/kg dry * 0.008 kg)] = = -1.93 mg Soil 2: [(<2.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<2.7 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<10.0 mg/kg dry * 0.008 kg)] = = -6.20 mg Soil 3: [(6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((11.0 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<12.6 mg/kg dry * 0.008 kg)] = = -90.07 mg Trial 2 Camelina sativa Soil 1: [(<2.6 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<8.87 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<45.3 mg/kg dry * 0.0032 kg)] = = -115.26 mg Soil 2: [(<2.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<22.7 mg/kg dry * 0.0032 kg)] = = -134.31 mg Soil 3: [( 6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -67.47 mg Miscanthus giganteous Soil 1: [(<2.6 mg/kg dry )* (0.0102 m3) * (1800 kg/m3)] = Σ[((<8.87 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<7.84 mg/kg dry * 0.030 kg)] = = -115.35 mg Soil 2: [(<2.4 mg/kg dry )* (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + ( <10.8 mg/kg dry * 0.030 kg)] = = -134.56 mg

228

Soil 3: [(6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<13.7 mg/kg dry * 0.030 kg)] = = -67.89 mg Panicum virgatum Soil 1: [(<2.6 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<8.87 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<7.98 mg/kg dry * 0.022 kg)] = = -115.29 mg Soil 2: [(<2.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<7.84 mg/kg dry * 0.022 kg)] = = -134.40 mg Soil 3: [(6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<7.32 mg/kg dry * 0.022 kg)] = = -67.47 mg Trial 3 Camelina sativa Soil 1: [(<2.6 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<8.87 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (43.7 mg/kg dry * 0.0032 kg)] = = -115.26 mg Soil 2: [(<2.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -134.23 mg Soil 3: [(6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -67.88 mg Miscanthus giganteous Soil 1: [(<2.6 mg/kg dry )* (0.0102 m3) * (1800 kg/m3)] = Σ[((<8.87 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<13.0 mg/kg dry * 0.030 kg)] = = -115.51 mg Soil 2: [(<2.4 mg/kg dry )* (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<10.2 mg/kg dry * 0.030 kg)] = = -134.54 mg

229

Soil 3: [(6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<13.5 mg/kg dry * 0.030 kg)] = = -67.88 mg Panicum virgatum Soil 1: [(<2.6 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<8.87 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<8.22 mg/kg dry * 0.022 kg)] = = -115.30 mg

Soil 2: [(<2.4 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<8.98 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<8.10 mg/kg dry * 0.022 kg)] = = -134.41 mg Soil 3: [(6.8 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[(<9.95 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<12.0 mg/kg dry * 0.022 kg)] = = -67.74 mg

230

Selenium Mass Mass In out Total Mass Trial Plant Plant Balance, Total Soil Soil Total C V ρ ρ tissue tissue excess in in in in C V out Out Percent out out C Mout positive, mg out of input T1S1 2.6 0.0102 1800 47.74 4 0.0102 1800 0 0.001 73.44 -25.704 Camelina -53.85 T1S2 2.4 0.0102 2000 48.96 2.7 0.0102 2000 17.2 0.001 55.1 -6.139 Camelina -12.54 T1S3 6.8 0.0102 2100 145.7 6.6 0.0102 2100 0 0.001 141.4 4.284 Camelina 2.94 T1S1 2.6 0.0102 1800 47.74 3.4 0.0102 1800 8.4 0.011 62.52 -14.780 Miscanthus -30.96 T1S2 2.4 0.0102 2000 48.96 2.3 0.0102 2000 8.1 0.011 47.01 1.951 Miscanthus 3.98 T1S3 6.8 0.0102 2100 145.7 9.5 0.0102 2100 7.9 0.011 203.6 -57.921 Miscanthus -39.77 T1S1 2.6 0.0102 1800 47.74 2.7 0.0102 1800 11.5 0.008 49.66 -1.928 Panicum -4.04 T1S2 2.4 0.0102 2000 48.96 2.7 0.0102 2000 10 0.008 55.16 -6.200 Panicum -12.66 T1S3 6.8 0.0102 2100 145.7 11 0.0102 2100 12.6 0.008 235.7 -90.065 Panicum -61.83 T2S1 2.6 0.0102 1800 47.74 8.87 0.0102 1800 45.3 0.003 163 -115.262 Camelina -241.46 T2S2 2.4 0.0102 2000 48.96 8.98 0.0102 2000 22.7 0.003 183.3 -134.305 Camelina -274.32 T2S3 6.8 0.0102 2100 145.7 9.95 0.0102 2100 0 0.003 213.1 -67.473 Camelina -46.32 T2S1 2.6 0.0102 1800 47.74 8.87 0.0102 1800 7.84 0.03 163.1 -115.352 Miscanthus -241.65 T2S2 2.4 0.0102 2000 48.96 8.98 0.0102 2000 10.8 0.03 183.5 -134.556 Miscanthus -274.83 T2S3 6.8 0.0102 2100 145.7 9.95 0.0102 2100 13.7 0.03 213.5 -67.884 Miscanthus -46.61 T2S1 2.6 0.0102 1800 47.74 8.87 0.0102 1800 7.98 0.022 163 -115.293 Panicum -241.52 T2S2 2.4 0.0102 2000 48.96 8.98 0.0102 2000 7.84 0.022 183.4 -134.404 Panicum -274.52 T2S3 6.8 0.0102 2100 145.7 9.95 0.0102 2100 7.32 0.022 213.3 -67.634 Panicum -46.43 T3S1 2.6 0.0102 1800 47.74 8.87 0.0102 1800 43.7 0.003 163 -115.257 Camelina -241.45 T3S2 2.4 0.0102 2000 48.96 8.98 0.0102 2000 0 0.003 183.2 -134.232 Camelina -274.17 T3S3 6.8 0.0102 2100 145.7 9.95 0.0102 2100 0 0.003 213.1 -67.473 Camelina -46.32 T3S1 2.6 0.0102 1800 47.74 8.87 0.0102 1800 13 0.03 163.2 -115.507 Miscanthus -241.97 T3S2 2.4 0.0102 2000 48.96 8.98 0.0102 2000 10.2 0.03 183.5 -134.538 Miscanthus -274.79 T3S3 6.8 0.0102 2100 145.7 9.95 0.0102 2100 13.5 0.03 213.5 -67.878 Miscanthus -46.60 T3S1 2.6 0.0102 1800 47.74 8.87 0.0102 1800 8.22 0.022 163 -115.298 Panicum -241.53 T3S2 2.4 0.0102 2000 48.96 8.98 0.0102 2000 8.1 0.022 183.4 -134.410 Panicum -274.53 T3S3 6.8 0.0102 2100 145.7 9.95 0.0102 2100 12 0.022 213.4 -67.737 Panicum -46.50

231

Silver, Ag

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Not detected in any of the lab analyses.

Trial 1

Camelina sativa Soil 1: [(<1.0 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<1.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = -1.84 mg Soil 2: [(<0.96 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<1.1 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<6.9 mg/kg dry * 0.0011 kg)] = = -2.86 mg Soil 3: [(<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<1.1 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = 2.14 mg Miscanthus giganteous Soil 1: [(<1.0 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<1.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3) + (<3.4 mg/kg dry * 0.011 kg)] = = -1.87 mg Soil 2: [(<0.96 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<0.92 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<3.2 mg/kg dry * 0.011kg)] =

232

= 0.78 mg

Soil 3: [(<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) +(<3.2 mg/kg dry * 0.011 kg)] = = - 0.04 mg Panicum virgatum Soil 1: [(<1.0 mg/kg dry)* (0.0102 m3) * (1800 kg/m3)] = Σ[( <1.1 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<4.6 mg/kg dry * 0.008 kg)] = = -1.87 mg Soil 2: [(<0.96 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<1.1 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<4.0 mg/kg dry * 0.008 kg)] = = -2.89 mg Soil 3: [(<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<1.3 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<5.0 mg/kg dry * 0.008 kg)] = = -2.18 mg Trial 2 Camelina sativa Soil 1: [(<1.0mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<4.44 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<22.7 mg/kg dry * 0.0032 kg)] = = 63.23 mg Soil 2: [(<0.96 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<4.49 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<11.4 mg/kg dry * 0.0032 kg)] = = 71.44 mg Soil 3: [(<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<4.98mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -80.97 mg Miscanthus giganteous Soil 1: [(<1.0 mg/kg dry )* (0.0102 m3) * (1800 kg/m3)] = Σ[((<4.44 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<3.92 mg/kg dry * 0.030 kg)] = = -63.27 mg

233

Soil 2: [(<0.96 mg/kg dry )* (0.0102 m3) * (2000 kg/m3)] = Σ[((<4.49 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + ( <5.41 mg/kg dry * 0.030 kg)] = = -71.56 mg Soil 3: [(<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<4.98 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<6.85 mg/kg dry * 0.030 kg)] = = -81.17 mg Panicum virgatum Soil 1: [(<1.0 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<4.44 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<3.99 mg/kg dry * 0.022 kg)] = = 63.24 mg Soil 2: [(<0.96 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<4.49 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<3.92 mg/kg dry * 0.022 kg)] = = 71.49 mg Soil 3: [(<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<4.98 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<3.66 mg/kg dry * 0.022 kg)] = = 81.05 mg Trial 3 Camelina sativa Soil 1: [(<1.0 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<4.44mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<21.9 mg/kg dry * 0.0032 kg)] = = 63.23 mg Soil 2: [(<0.96 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<4.49 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -71.40 mg Soil 3: [(<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<4.98 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = -80.97 mg Miscanthus giganteous Soil 1: [(<1.0 mg/kg dry )* (0.0102 m3) * (1800 kg/m3)] = Σ[((<4.44 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<6.52 mg/kg dry * 0.030 kg)] = = -63.35 mg

234

Soil 2: [(<0.96 mg/kg dry )* (0.0102 m3) * (2000 kg/m3)] = Σ[((<4.49 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<5.11 mg/kg dry * 0.030 kg)] = = -71.55 mg Soil 3: [(<1.2 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((<4.98 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<6.74 mg/kg dry * 0.030 kg)] = = -81.17 mg Panicum virgatum Soil 1: [(<1.0 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((<4.44 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (<4.12 mg/kg dry * 0.022 kg)] = = 63.25 mg

Soil 2: [(<0.96 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((<4.49 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (<4.06 mg/kg dry * 0.022 kg)] = = 71.49 mg Soil 3: [(<1.2mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[(<4.98 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (<6.03 mg/kg dry * 0.022 kg)] = = 81.10 mg

235

Silver

MassIn Massout Trial Plant Plant Total Mass Total Soil Soil Total C V ρ ρ tissue tissue Balance in in in in C V out Out Percent out out C Mout out of input T1S1 1 0.0102 1800 18.36 1.1 0.0102 1800 0 0.001 20.2 -1.836 Camelina -10.00 T1S2 0.96 0.0102 2000 19.58 1.1 0.0102 2000 6.9 0.001 22.45 -2.86359 Camelina -14.62 T1S3 1.2 0.0102 2100 25.7 1.1 0.0102 2100 0 0.001 23.56 2.142 Camelina 8.33 T1S1 1 0.0102 1800 18.36 1.1 0.0102 1800 3.4 0.011 20.23 -1.8734 Miscanthus -10.20 T1S2 0.96 0.0102 2000 19.58 0.92 0.0102 2000 3.2 0.011 18.8 0.7808 Miscanthus 3.99 T1S3 1.2 0.0102 2100 25.7 1.2 0.0102 2100 3.2 0.011 25.74 -0.0352 Miscanthus -0.14 T1S1 1 0.0102 1800 18.36 1.1 0.0102 1800 4.6 0.008 20.23 -1.8728 Panicum -10.20 T1S2 0.96 0.0102 2000 19.58 1.1 0.0102 2000 4 0.008 22.47 -2.888 Panicum -14.75 T1S3 1.2 0.0102 2100 25.7 1.3 0.0102 2100 5 0.008 27.89 -2.182 Panicum -8.49 T2S1 1 0.0102 1800 18.36 4.44 0.0102 1800 22.7 0.003 81.59 -63.23104 Camelina -344.40 T2S2 0.96 0.0102 2000 19.58 4.46 0.0102 2000 11.4 0.003 91.02 -71.43648 Camelina -364.77 T2S3 1.2 0.0102 2100 25.7 4.98 0.0102 2100 0 0.003 106.7 -80.9676 Camelina -315.00 T2S1 1 0.0102 1800 18.36 4.44 0.0102 1800 3.92 0.03 81.64 -63.276 Miscanthus -344.64 T2S2 0.96 0.0102 2000 19.58 4.46 0.0102 2000 5.41 0.03 91.15 -71.5623 Miscanthus -365.41 T2S3 1.2 0.0102 2100 25.7 4.98 0.0102 2100 6.85 0.03 106.9 -81.1731 Miscanthus -315.80 T2S1 1 0.0102 1800 18.36 4.44 0.0102 1800 3.99 0.022 81.61 -63.24618 Panicum -344.48 T2S2 0.96 0.0102 2000 19.58 4.46 0.0102 2000 3.92 0.022 91.07 -71.48624 Panicum -365.02 T2S3 1.2 0.0102 2100 25.7 4.98 0.0102 2100 3.66 0.022 106.8 -81.04812 Panicum -315.31 T3S1 1 0.0102 1800 18.36 4.44 0.0102 1800 21.9 0.003 81.59 -63.22848 Camelina -344.38 T3S2 0.96 0.0102 2000 19.58 4.46 0.0102 2000 0 0.003 90.98 -71.4 Camelina -364.58 T3S3 1.2 0.0102 2100 25.7 4.98 0.0102 2100 0 0.003 106.7 -80.9676 Camelina -315.00 T3S1 1 0.0102 1800 18.36 4.44 0.0102 1800 6.52 0.03 81.71 -63.354 Miscanthus -345.07 T3S2 0.96 0.0102 2000 19.58 4.46 0.0102 2000 5.11 0.03 91.14 -71.5533 Miscanthus -365.37 T3S3 1.2 0.0102 2100 25.7 4.98 0.0102 2100 6.74 0.03 106.9 -81.1698 Miscanthus -315.79 T3S1 1 0.0102 1800 18.36 4.44 0.0102 1800 4.12 0.022 81.61 -63.24904 Panicum -344.49 T3S2 0.96 0.0102 2000 19.58 4.46 0.0102 2000 4.06 0.022 91.07 -71.48932 Panicum -365.04 T3S3 1.2 0.0102 2100 25.7 4.98 0.0102 2100 6.03 0.022 106.8 -81.10026 Panicum -315.52

236

Sulfur, S

Total Massin = Total Massout

Total [(Concentrationin)*(Volumein )] = Total [(Concentrationout)*(Volumeout)]

Σi [Soil(CinVin ρsoil) ] = Σo [Soil(CoutVout ρsoil )] + Plant Tissue(CoutVout ρplant-tissue )]

Approximate density of the soils:

3 Soil 1 (sandy) ρSoil1 = 1800 kg/m

3 Soil 2 (gravel) ρSoil1 = 2000 kg/m

3 Soil 3 (silty) ρSoil1 = 2100 kg/m

Trial 1 Camelina sativa Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((793 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = 0.0040 kg or 3.98 g Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((545 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (6150 mg/kg dry * 0.0011 kg)] = = - 0.003 kg or - 2.74 g Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((4580 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0011 kg)] = = - 0.046 kg or - 45.84 g Miscanthus giganteous Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((782 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (2290 mg/kg dry * 0.011 kg)] = = 0.004 kg or 4.15 g Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((441 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (1770 mg/kg dry * 0.011 kg)] = = - 0.001 kg or - 0.63 g

237

Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((2540 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (2240 mg/kg dry * 0.011 kg)] = = - 0.002 kg or - 2.17 g

Panicum virgatum Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((754 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (1880 mg/kg dry * 0.008 kg)] = = 0.005 kg or 4.69 g Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((523 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (2540 mg/kg dry * 0.008 kg)] = = - 0.002 kg or - 2.31 g Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((2720 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (3550 mg/kg dry * 0.008 kg)] = = - 0.006 kg or - 6.03 g Trial 2 Camelina sativa Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((1040 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (6900 mg/kg dry * 0.0032 kg)] = = - 0.0010 kg or - 0.57 g Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((735 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (10100 mg/kg dry * 0.0032 kg)] = = - 0.007 kg or - 6.64 g Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((2600 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = - 0.003 kg or - 3.43 g Miscanthus giganteous Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((1040 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (1640 mg/kg dry * 0.030 kg)] = = - 0.001 kg or - 0.60 g

238

Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((735 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (1800 mg/kg dry * 0.030 kg)] = = - 0.007 kg or - 6.66 g Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((2600 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (1060 mg/kg dry * 0.030 kg)] = = 0.047 kg or 46.66 g Panicum virgatum Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((1040 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (1780 mg/kg dry * 0.022 kg)] = = - 0.001 kg or - 0.59 g Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((735 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (1880 mg/kg dry * 0.022 kg)] = = - 0.007 kg or - 6.65 g Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((2600 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (2050 mg/kg dry * 0.022 kg)] = = 0.047 kg or 46.65 g Trial 3 Camelina sativa Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((1040 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (10500 mg/kg dry * 0.0032 kg)] = = - 0.001 kg or - 0.58 g Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((735 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = - 0.007 kg or - 6.61 g Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((2600 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (0.00 mg/kg dry * 0.0032 kg)] = = 0.047 kg or 46.70 g Miscanthus giganteous Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((1040 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (1810 mg/kg dry * 0.030 kg)] = = - 0.001 kg or - 0.61 g

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Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((735 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (1500 mg/kg dry * 0.030 kg)] = = - 0.007 kg or - 6.66 g Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((2600 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (1160 mg/kg dry * 0.030 kg)] = = 0.047 kg or 46.66 g Panicum virgatum Soil 1: [(1010 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)] = Σ[((1040 mg/kg dry) * (0.0102 m3) * (1800 kg/m3)) + (2460 mg/kg dry * 0.022 kg)] = = - 0.001 kg or - 0.61 Soil 2: [(411 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)] = Σ[((735 mg/kg dry) * (0.0102 m3) * (2000 kg/m3)) + (1830 mg/kg dry * 0.022 kg)] = = - 0.007 kg or - 6.65 g Soil 3: [(2440 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)] = Σ[((2600 mg/kg dry) * (0.0102 m3) * (2100 kg/m3)) + (2190 mg/kg dry * 0.022 kg)] = = - 0.004 kg or - 3.48 g

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Sulfur

MassIn Massout Trial Plant Plant Total Mass Soil Soil Total C V ρ Total in ρ tissue tissue Balance, g in in in C Vout out Out Percent out C Mout out of input T1S1 Camelina 1010 0.01 1800 18.5436 793 0.01 1800 0 0.001 14.559 3.984 21.49 T1S2 Camelina 411 0.01 2000 8.3844 545 0.01 2000 6150 0.001 11.125 -2.740 -32.68 T1S3 Camelina 2440 0.01 2100 52.2648 4580 0.01 2100 0 0.001 98.104 -45.839 -87.70 T1S1 1010 0.01 1800 18.5436 782 0.01 1800 2290 0.011 14.383 4.161 Miscanthus 22.44 T1S2 411 0.01 2000 8.3844 441 0.01 2000 1770 0.011 9.016 -0.631 Miscanthus -7.53 T1S3 2440 0.01 2100 52.2648 2540 0.01 2100 2240 0.011 54.431 -2.167 Miscanthus -4.15 T1S1 Panicum 1010 0.01 1800 18.5436 754 0.01 1800 1880 0.008 13.858 4.685 25.27 T1S2 Panicum 411 0.01 2000 8.3844 523 0.01 2000 2540 0.008 10.690 -2.305 -27.49 T1S3 Panicum 2440 0.01 2100 52.2648 2720 0.01 2100 3550 0.008 58.291 -6.026 -11.53 T2S1 Camelina 1010 0.01 1800 18.5436 1040 0.01 1800 6900 0.003 19.116 -0.573 -3.09 T2S2 Camelina 411 0.01 2000 8.3844 735 0.01 2000 10100 0.003 15.026 -6.642 -79.22 T2S3 Camelina 2440 0.01 2100 52.2648 2600 0.01 2100 0 0.003 55.692 -3.427 -6.56 T2S1 1010 0.01 1800 18.5436 1040 0.01 1800 1640 0.03 19.144 -0.600 Miscanthus -3.24 T2S2 411 0.01 2000 8.3844 735 0.01 2000 1800 0.03 15.048 -6.664 Miscanthus -79.48 T2S3 2440 0.01 2100 52.2648 260 0.01 2100 1060 0.03 5.601 46.664 Miscanthus 89.28 T2S1 Panicum 1010 0.01 1800 18.5436 1040 0.01 1800 1780 0.022 19.134 -0.590 -3.18 T2S2 Panicum 411 0.01 2000 8.3844 735 0.01 2000 1880 0.022 15.035 -6.651 -79.33 T2S3 Panicum 2440 0.01 2100 52.2648 260 0.01 2100 2050 0.022 5.614 46.651 89.26 T3S1 Camelina 1010 0.01 1800 18.5436 1040 0.01 1800 10500 0.003 19.128 -0.584 -3.15 T3S2 Camelina 411 0.01 2000 8.3844 735 0.01 2000 0 0.003 14.994 -6.610 -78.83 T3S3 Camelina 2440 0.01 2100 52.2648 260 0.01 2100 0 0.003 5.569 46.696 89.34 T3S1 1010 0.01 1800 18.5436 1040 0.01 1800 1810 0.03 19.149 -0.605 Miscanthus -3.26 T3S2 411 0.01 2000 8.3844 735 0.01 2000 1500 0.03 15.039 -6.655 Miscanthus -79.37 T3S3 2440 0.01 2100 52.2648 260 0.01 2100 1160 0.03 5.604 46.661 Miscanthus 89.28 T3S1 Panicum 1010 0.01 1800 18.5436 1040 0.01 1800 2460 0.022 19.149 -0.605 -3.26 T3S2 Panicum 411 0.01 2000 8.3844 735 0.01 2000 1830 0.022 15.034 -6.650 -79.31 T3S3 Panicum 2440 0.01 2100 52.2648 2600 0.01 2100 2190 0.022 55.740 -3.475 -6.65

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