Heavy Metal Accumulation in Nectar of

Streptanthus polygaloides within

Green Lents Bioswale
________________________________________________________________________

Jordynn Alexander, Alivia DeGagne, Bassel Hamieh, SR Kanna, Marissa Reisen

Portland State University
Introduction
Plants are classified as evaders or accumulators of heavy
metals (Tangahu et. al 2011). Figure 1 (Tangahu et al. 2011)
illustrates the process by which plants uptake heavy metals.
The adaptive role of metal accumulation in nature by plants is
not yet understood (Meindl, Bain, Ashman 2014 - A; Martens
& Boyd 2001; Shaw 1990). Accumulation of metal in reproductive rewards is Figure 1: Diagram of metal accumulation
in plants detrimental for gamete formation, but deters predators, reinforcing the survivability and reproduction of the species (Meindl, Bain, Ashman 2014 - A; Martens & Boyd
2001).
Several studies have tried to understand the effect of
metal accumulation in plants (Meindl, Bain Ashman 2013 - B,
Meindl & Ashman 2013 -C). A 2013 study showed that
edaphic factors such as serpentine soil, known for the low Figure 1 - Diagram of metal
magnesium to calcium ratio, lack of essential nutrients and accumulation in plants.
high concentration of heavy metals such as nickel and chromium, led to changes in the chemical-composition and plant morphology in the plants
(Meindl, Bain, Ashman 2013 - B). Other studies show heavy metal content appear in the reproductive organs and rewards (Meindl, Bain, Ashman 2014 - A; Meindl, Bain, Ashman
2014 -D; Meindl & Ashman 2013 - C). In one study, Streptanthus polygaloides were exposed
to nickel for a short period of time. Visitors couldnt detect nickel in flowers prior to visitation, but visitation to high nickel concentration flowers decreased afterward, supporting the
theory that metal accumulation is an evolutionary mechanisms because it deters predation.
The short-term exposure to nickel saw an accumulation of the heavy metal in floral rewards:
nectar, but no change in size or volume (Meindl, Bain, Ashman 2014 - A; Meindl, & Ashman
2013 - C).
Metal in plants accumulate naturally
from polluted soil and water (Tangahu et al.
2011; Meindl & Ashman 2013 - C). This
occurs as a byproduct of urban living and
poor industrialization regulations. Therefore,
many cities have incorporated a phytoremediation technique to harness the potential of

plants as an accumulator of metals to clean

up pollution. Green Lents is an example of a
local organization that planted three bioswales throughout the neighborhoods of Portland and maintains them through the help of
the community. These bioswales are concentrated in the quadrant of SE on 10245 SE

Yukon, 10303 SE Yukon, and 10317 SE

Yukon. The grassy swales offer beauty to the
neighborhood and an effective methodology
to remove pollutants from runoff in Portland
(Green Lents 2007). Bioswales (depicted by
figure 2) are one such example, where native
plants are planted on sidewalks to remove
pollution and silt by absorbing the contaminated runoff absorbed by the soil of the
plants. Bioswales are typically built in naturally Figure 2: Process of Bioswale absorbing city runoff and filteration system (solution.ecovillage.org) existing swales such as
riparian zones and additional thick grasses,
organic debris or compost for filtration. The
water travels down a wide and shallow path,
to allow maximum infiltration of pollutants
from storm water runoff (United State Department of Agriculture 2007).

Figure 2 - Process of Bioswale absorbing city

runoff and filtration system (solution.ecovillage.org)

Runoff in cities which bioswales filter

typically contains heavy metals such as arsenic, cadmium, chromium, copper, nickel,
lead and mercury (Hughes et. al 2000). The
pollution is often regional specific due to
different industries and lifestyles. Runoff in
the Pacific Northwest contains mercury,
lead, copper and petroleum (Washington
State Department of Ecology 2011). As part
of the phytoremediation technique, bioswales will remove some of the pollutants in
the runoff. A study reported the average bioswale contain water pollution such as silt,
inorganic and organic compounds, and pathogens including heavy metals such as lead,

chromium, and cadmium (Xiao & McPherson 2009).

What kind of heavy metal is accumulated in the nectar of Streptanthus polygaloides of the Lents bioswale? In this experiment, we seek to measure the concentrations
of heavy metals of flower rewards, nectar, of
Streptanthus polygaloides in Lents bioswale.
Urban ecology studies living organisms and
their relationship with each other in the context of an urban setting. Metal concentrations
in nectar are a consequence of urban living
and should be investigated by urban ecologists.
Streptanthus polygaloides has been
used by the scientific community to determine heavy metals accumulate in the nectar
because of its hyper-accumulative properties
(Meindl & Ashman 2013 - C). Therefore,
Streptanthus polygaloides would be the ideal
model organism in our study because we
would like to extend the existing parameters
studied to include Lents bioswale. The concentration of heavy metals in ppm found in
bioswale nectar is expected to have a direct
correlation to outside pollutants. We predict
that in areas of increased urbanization, water
samples will indicate higher concentration of
heavy metals than those bodies of water further away from the urban environment. Cities have reported heavy levels of lead, chromium and cadmium in their runoff and bioswales have also correlated a large accumulation of these heavy metals (Hughes et. al
2000; Washington State Department of
Ecology 2011). The Pacific Northwest has
high levels of mercury, lead, copper and petroleum, therefore we predict to see a similar
correlation in bioswale and uptake of metal
concentrations in the flower rewards (Washington State Department of Ecology 2011).
Methods
Justification for approach: We will
be testing the accumulation of metals in the
nectar of an indicator species by extracting
and measuring the nectar in the flower. We
will be using this method because it is a way

to display how the runoff that collects in bioswales is affecting the species, which grow
in it. It leads to the discussion of what kind
of impact a bioswale has, and by testing the
nectar we will see how using bioswales to
filter storm water which collects in our sewers can change the metal accumulation in
soil and the plants that inhabit it. We are
specifically testing the metal concentration
from the flower nectar with InductivelyCoupled Plasma Mass Spectrometry (ICPMS) technology because we believe it will
give a more accurate and precise measurement of the metals within the nectar. These
methods answer the posed questions and test
the hypothesis in the way that we use these
plants as active reflections of how the soil
can change the metal makeup in a plant. By
using a control (20 plants left growing in the
greenhouse; soil is not treated by any unnatural chemicals or metals), and by using a
plant, which can be tested for hyperaccumulated metals, we will be able to see
exactly how much a bioswale changes the
species and be able to make other conclusions about its impacts.
Preparation of Plants
We will be collecting seeds of the
species (Streptanthus polygaloides) in an
effort to extend the research of Meindl &
Ashman (2013) on the hyper-accumulation
of metals in these plants. However, instead
of testing the adaptation of these plants in an
open serpentine field, we will be testing the
reaction of the experiment in the Oregon
Green Lents bioswale (Lents Neighborhood,
Portland, Oregon 97266) (Green Lents
2007). The seeds will be collected in California, where the plant is native, and shipped
to Oregon for planting and growing. The
seeds will first be planted in Fafard organic
potting mix, set in flats inside the Research
Greenhouse at Portland State University beginning in April 2015, which is a good time
for the plants to being germination. Before
transplanting, we will use the tool Niton
XRF Analyzer to test the soils of both loca-

tions for metals (Thermo Scientific, 2015).

The negative control of our experiment will
be the soil of the greenhouse plants, which
will contain no contaminated metals. The
positive control will be the soil of the bioswale, naturally treated with contaminated
metals, which a soil test will explain the specific measure of. After germination and
growing time of twelve weeks, 20 flowering
plants of the species will be moved from the
greenhouse to the bioswale. As a control, an
additional 20 flowering plants will remain
growing in the greenhouse. The soil of the
bioswale is affected by water pollution of
urban groundwater and runoff. This will be
considered the soil treatment or tested variable in comparison to Meindl, Bain and
Ashmans experiment with a nickel treatment. The control will be watered with untreated water. After four weeks in the
ground, the tested variable plants will be
ready for observation of the nectar affected
by the bioswale treatment.
Collecting Data
Using micro-capillary tubes, we will
remove the standing Streptanthus polygaloides nectar from the style of each flower.
We will harvest the entire amount of nectar
present in each of the flower members from
the 20 controls (no metal contamination) and
20 flowers from the bioswale (naturally contaminated plants with metal) by inserting
micro-capillaries between the two staminodia and the floral scale (nectar data will be
taken during the first half of the staminate
phase to ensure that the data are comparable). For the metal measuring process, we
will pipet four drops of the nectar into 1 mL
of distilled water and dilute for all the samples collected from the individual plants, and
place into micro test tubes for a total of 40
labeled samples. The quantitative examination of studied elements (Cu, Se, Pb and Cd)
will be done using ICP-MS technology,
which combines a high-temperature Inductively Coupled Plasma source with a mass
spectrometer, which then converts the atoms

of the elements in the sample to ions and

separates them by the mass spectrometer
(U.S. Geological Survey 2013). In order to
use the ICP-MS technology, we will convert
the diluted nectar solution into an aerosol.
We will do so by using an analytical nebulizer, which uses oxygen, compressed air,
and ultrasonic power to break up the diluted
nectar solution into small aerosol droplets
(ICP-MS with Universal Cell Technology,
2012). Once we obtain the sample aerosol,
we will insert it into the ICP torch, which
analyzes and transfers the sample results of
different metal concentrations into its computer software program. After careful navigation through the software, we will recorded our data, specifically examining the levels
and type of metal that accumulated within
the plant. The above process will be repeated
with each of the 40 Streptanthus polygaloides samples.
Expected results
In areas of increased urbanization,
water samples will indicate higher concentration of metals than those bodies of water
further away from the urban environment.
The concentration of heavy metals in ppm
found in Lents bioswale nectar is expected to
have a direct correlation to outside pollutants. As shown in Figure 3, by gathering
multiple samples of the positive and negative
control variables, we can observe the increase or decrease of metal in ppm.
In order to determine quantifiable data we will measure the differences in metal
concentrations in each plant and conduct a ttest to analyze the average metal concentration from the samples. We expect the test
will show a significant difference between
the greenhouse constants and exemplify the

data collected for the bioswale plant (Meindl

& Ashman 2013 - C).
The runoff in Pacific Northwest contains mercury, lead, copper and petroleum
and bioswales typically contain silt, inorganic and organic compounds and pathogens
including heavy metals such as lead, chromium and cadmium (Hughes et al. 2000;
Washington State Department of Ecology
2013) . Therefore, we predict lead, the common heavy metal, will be in the nectar of the
bioswales plants. Overall we expect to see an
increase of lead concentration in ppm in
high-urbanized areas due to human interaction with the environment that create heavy
metal pollutants.
We expect accurate results through
repetition and efficient instruments that have
been tested to previously detect metal accumulation. Outcome can lead to questions
about how to control water pollution and
how treatments are emplaced in different
environments. By gathering evidence from
urban and suburban environments, we will
expand on this relationship.

Figure 3: This graph illustrates a predicted

outcome of testing for lead in Lents bioswales
and controls after calculating averages and
standard deviation.

Timeline: Url to Gannt Table - https://app.smartsheet.com/b/home

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