PHYTOREMEDIATION
Introduction
 Phytoremediation is the use of living green plants for
  in situ risk reduction and/or removal of contaminants
  from contaminated soil, water, sediments, and air.
OVERVIEW
Phytoextraction
 Plant roots uptake metal contaminants
  from the soil and translocate them to
  their above soil tissues
 Once the plants have grown and
  absorbed the metal pollutants they are
  harvested and disposed off safely
 This process is repeated several times to
  reduce contamination to acceptable
  levels
 Hyper accumulator plant species are
  used on many sites due to their
  tolerance of relatively extreme levels of
  pollution
 Avena sp. , Brassica sp.
Contaminants removed:
 Metal compounds that have been
  successfully phytoextracted include zinc,
  copper, and nickel
Rhizofiltration
 It is concerned with the remediation of contaminated groundwater
 The contaminants are either adsorbed onto the root surface or are
  absorbed by the plant roots
 Plants used for rhizofiltration are acclimated to the pollutant
      • Plants are hydroponically grown in clean water
        rather than soil, until a large root system has
 1      developed


      • Water supply is substituted for a polluted
 2      water supply to acclimatize the plant


      • They are planted in the polluted area where
        the roots uptake the polluted water and the
 3      contaminants along with it


      • As the roots become saturated they are            Chernobyl - sunflowers
 4      harvested and disposed of safely
                                                          were grown in radioactively
                                                          contaminated pools
Phytostabilisation
 To immobilize soil and water contaminants from
  migration
 Mechanisms
     Phytochemical complexation in the root zone –
     precipitation
     Transport protein inhibition on the root membrane (B)
     Vacuolar storage in the root cells (C)
     Pb, Cu, Zn – Agrostis tenuis
Phytodegradation
 It is the degradation or breakdown
  of organic contaminants by internal
  and external metabolic processes
  driven by the plant

 Mechanisms:
  (A) Plant enzymatic activity
        oxygenases- hydrocarbons
        nitroreductases- explosives
   (B) Photosynthetic oxidation

 Used in breakdown of ammunition
  wastes, chlorinated solvents such
  as     TCE       (Trichloroethane),
  degradation of organic herbicides.
Contd..




External Process             Internal Process
Rhizodegradation
 It is the breakdown of organic contaminants in the soil by soil dwelling
  microbes which is enhanced by the rhizosphere‘s presence
 Rhizosphere = soil + root + microbes
 Symbiotic relation
 Also called:
       Enhanced rhizosphere biodegradation
       Phytostimulation
       Plant assisted bioremediation
 Sugars, alcohols, and organic acids act as carbohydrate sources for
  the soil microflora and enhance microbial growth and activity.
 Act as chemotactic signals for certain microbes.
 The roots also loosen the soil and transport water to the rhizosphere
  thus enhancing microbial activity
 Digest organic pollutants such as fuels and solvents, producing
  harmless products
Phytovolatilization
 Plants uptake contaminants which are
  water soluble and release them into the
  atmosphere as they transpire the water

 The contaminant may become modified
  along the way, as the water travels along
  the plant's vascular system from the roots
  to     the     leaves,     whereby     the
  contaminants                     evaporate
  or volatilize into the air surrounding the
  plant

 Poplar trees volatilize up to 90% of the
  TCE they absorb
 Selenium and Mercury - Arabidopsis
  thaliana L. and tobacco
Phytohydraulics

 The use of plants to control the migration of
  subsurface water through the rapid uptake of
  large volumes of water by the plants

 Plants - acting as natural hydraulic pumps

 A dense root network established near the
  water table can transpire up to 300 gallons of
  water per day
 This fact has been utilized to decrease the
  migration of contaminants from surface water
  into the groundwater (below the water table)
  and drinking water supplies
LEVEL OF POLLUTANT
                                                      Analytical methods
 Sampling                              Parameter    Media      Acceptable
 Field sampling quality assurance                              Test
         Avoiding contamination of                             Methods
          samples                       Petroleum Water &       Gas
         Equipment cleaning            Hydrocarbo soil         chromatogra
         Sample labels                 ns                      phy
Recent trend                            Volatiles    Water &    Infra-red
 Immunoassay testing and other         (including   soil       based
  technology may be used to provide     BTEX)                   methods
  on site screening.
 This work shall only be carried out   Semi-        water      EPA
  by properly trained personnel         volatiles               Standards
                                        (including
                                        PAH)
Measurement of aquatic biodegradation rates by determining
       heterotrophic uptake of radiolabeled pollutants.
             F K Pfaender and G W Bartholomew
• The heterotrophic uptake technique was modified to provide a rapid
  and simple technique for estimating the rates of biodegradation of
  organic pollutants under environmental conditions.
• The methodology is based on an evaluation of uptake into cells and
  subsequent respiration of radiolabeled organic substrates in short-
  term experiments.
• The resulting data can be used to calculate either turnover times or,
  if multiple concentrations of substrate are used, kinetic parameters.
• The procedure was applied to assess the biodegradation rates of m-
  cresol,     chlorobenzene,     nitrilotriacetic acid,     and   1,2,4-
  trichlorobenzene in fresh, brackish, and marine water samples from
  the coastal areas of North Carolina.
• Saturation kinetics for uptake were obtained with each of the
  compounds tested.
• Rates of metabolism were shown to be dependent on sample
  location and time of year.
Phytoremediation & Biotechnology

 Maximizing potential for phytoremediation

 GM approaches can be used to over express the enzymes involved in
  the existing plant metabolic pathways or to introduce new pathways
  into plants.

 Richard Meagher and colleagues introduced a new pathway
  into Arabidopsis to detoxify methylmercury, a common form of
  environmental pollutant to elemental mercury which can be volatilised
  by the plant
The genes originated in gram-negative bacteria


Mer B

Organomercurial Lyase

Methyl Mercury          Ionic Mercury

MerA

Mercuric Reductase

Ionic Mercury           Elemental Form
Arabidopsis plants were transformed with either MerA or MerB coupled with a
constitutive promoter

The MerA plants were more tolerant to ionic mercury, volatilised elemental
mercury, and were unaffected in their tolerance of methylmercury


The MerB Plants were significantly more tolerant to methylmercury and other
organomercurials and could also convert methylmercury to ionic mercury which
is approximately 100 times less toxic to plants


MerA MerB double transgenics were produced in an F2 generation. These plants
not only showed a greater resistance to organic mercury when compared to the
MerA, MerB, and wildtype plants but also capable of volatilizing mercury when
supplied with methylmercury.


The same MerA/MerB inserts have been used in other plant species including
tobacco(Nicotiana tabacum), yellow poplar(Liriodendron tulipifera)



Wetland species (bulrush and cat-tail) and water tolerant trees (willow and poplar)
have also been targetted for transformation.
Risk Assessment

Potential for the gene to recombine with other genes possibly
leading to the hyperaccumulation of non-contaminant
compounds

Reporter/marker genes may also escape into the environment

The GM plants may revert to a wild type genotype
Advantages of phytoremediation

It is more economically viable using the same tools and
supplies as agriculture
It is less disruptive to the environment
Disposal sites are not needed
Aesthetically pleasing than traditional methods
Avoids excavation and transport of polluted media thus
reducing the risk of spreading the contamination
It has the potential to treat sites polluted with more
than one type of pollutant
Disadvantages of phytoremediation
 Growing conditions required by the plant (i.e., Climate,
 geology, altitude, temperature)
 Tolerance of the plant to the pollutant
 Contaminants collected in senescing tissues may be released
 back into the environment in autumn
 Contaminants may be collected in woody tissues used as fuel
 Time taken to remediate sites far exceeds that of other
 technologies
 Contaminant solubility may be increased leading to greater
 environmental damage and the possibility of leaching
Phytoremediation

Phytoremediation

  • 1.
  • 2.
    Introduction  Phytoremediation isthe use of living green plants for in situ risk reduction and/or removal of contaminants from contaminated soil, water, sediments, and air.
  • 3.
  • 4.
    Phytoextraction  Plant rootsuptake metal contaminants from the soil and translocate them to their above soil tissues  Once the plants have grown and absorbed the metal pollutants they are harvested and disposed off safely  This process is repeated several times to reduce contamination to acceptable levels  Hyper accumulator plant species are used on many sites due to their tolerance of relatively extreme levels of pollution  Avena sp. , Brassica sp. Contaminants removed:  Metal compounds that have been successfully phytoextracted include zinc, copper, and nickel
  • 5.
    Rhizofiltration  It isconcerned with the remediation of contaminated groundwater  The contaminants are either adsorbed onto the root surface or are absorbed by the plant roots  Plants used for rhizofiltration are acclimated to the pollutant • Plants are hydroponically grown in clean water rather than soil, until a large root system has 1 developed • Water supply is substituted for a polluted 2 water supply to acclimatize the plant • They are planted in the polluted area where the roots uptake the polluted water and the 3 contaminants along with it • As the roots become saturated they are Chernobyl - sunflowers 4 harvested and disposed of safely were grown in radioactively contaminated pools
  • 6.
    Phytostabilisation  To immobilizesoil and water contaminants from migration  Mechanisms Phytochemical complexation in the root zone – precipitation Transport protein inhibition on the root membrane (B) Vacuolar storage in the root cells (C) Pb, Cu, Zn – Agrostis tenuis
  • 7.
    Phytodegradation  It isthe degradation or breakdown of organic contaminants by internal and external metabolic processes driven by the plant  Mechanisms: (A) Plant enzymatic activity oxygenases- hydrocarbons nitroreductases- explosives (B) Photosynthetic oxidation  Used in breakdown of ammunition wastes, chlorinated solvents such as TCE (Trichloroethane), degradation of organic herbicides.
  • 8.
    Contd.. External Process Internal Process
  • 9.
    Rhizodegradation  It isthe breakdown of organic contaminants in the soil by soil dwelling microbes which is enhanced by the rhizosphere‘s presence  Rhizosphere = soil + root + microbes  Symbiotic relation  Also called: Enhanced rhizosphere biodegradation Phytostimulation Plant assisted bioremediation  Sugars, alcohols, and organic acids act as carbohydrate sources for the soil microflora and enhance microbial growth and activity.  Act as chemotactic signals for certain microbes.  The roots also loosen the soil and transport water to the rhizosphere thus enhancing microbial activity  Digest organic pollutants such as fuels and solvents, producing harmless products
  • 11.
    Phytovolatilization  Plants uptakecontaminants which are water soluble and release them into the atmosphere as they transpire the water  The contaminant may become modified along the way, as the water travels along the plant's vascular system from the roots to the leaves, whereby the contaminants evaporate or volatilize into the air surrounding the plant  Poplar trees volatilize up to 90% of the TCE they absorb  Selenium and Mercury - Arabidopsis thaliana L. and tobacco
  • 12.
    Phytohydraulics  The useof plants to control the migration of subsurface water through the rapid uptake of large volumes of water by the plants  Plants - acting as natural hydraulic pumps  A dense root network established near the water table can transpire up to 300 gallons of water per day  This fact has been utilized to decrease the migration of contaminants from surface water into the groundwater (below the water table) and drinking water supplies
  • 13.
    LEVEL OF POLLUTANT Analytical methods  Sampling Parameter Media Acceptable  Field sampling quality assurance Test  Avoiding contamination of Methods samples Petroleum Water & Gas  Equipment cleaning Hydrocarbo soil chromatogra  Sample labels ns phy Recent trend Volatiles Water & Infra-red  Immunoassay testing and other (including soil based technology may be used to provide BTEX) methods on site screening.  This work shall only be carried out Semi- water EPA by properly trained personnel volatiles Standards (including PAH)
  • 14.
    Measurement of aquaticbiodegradation rates by determining heterotrophic uptake of radiolabeled pollutants. F K Pfaender and G W Bartholomew • The heterotrophic uptake technique was modified to provide a rapid and simple technique for estimating the rates of biodegradation of organic pollutants under environmental conditions. • The methodology is based on an evaluation of uptake into cells and subsequent respiration of radiolabeled organic substrates in short- term experiments. • The resulting data can be used to calculate either turnover times or, if multiple concentrations of substrate are used, kinetic parameters. • The procedure was applied to assess the biodegradation rates of m- cresol, chlorobenzene, nitrilotriacetic acid, and 1,2,4- trichlorobenzene in fresh, brackish, and marine water samples from the coastal areas of North Carolina. • Saturation kinetics for uptake were obtained with each of the compounds tested. • Rates of metabolism were shown to be dependent on sample location and time of year.
  • 15.
    Phytoremediation & Biotechnology Maximizing potential for phytoremediation  GM approaches can be used to over express the enzymes involved in the existing plant metabolic pathways or to introduce new pathways into plants.  Richard Meagher and colleagues introduced a new pathway into Arabidopsis to detoxify methylmercury, a common form of environmental pollutant to elemental mercury which can be volatilised by the plant
  • 16.
    The genes originatedin gram-negative bacteria Mer B Organomercurial Lyase Methyl Mercury Ionic Mercury MerA Mercuric Reductase Ionic Mercury Elemental Form
  • 17.
    Arabidopsis plants weretransformed with either MerA or MerB coupled with a constitutive promoter The MerA plants were more tolerant to ionic mercury, volatilised elemental mercury, and were unaffected in their tolerance of methylmercury The MerB Plants were significantly more tolerant to methylmercury and other organomercurials and could also convert methylmercury to ionic mercury which is approximately 100 times less toxic to plants MerA MerB double transgenics were produced in an F2 generation. These plants not only showed a greater resistance to organic mercury when compared to the MerA, MerB, and wildtype plants but also capable of volatilizing mercury when supplied with methylmercury. The same MerA/MerB inserts have been used in other plant species including tobacco(Nicotiana tabacum), yellow poplar(Liriodendron tulipifera) Wetland species (bulrush and cat-tail) and water tolerant trees (willow and poplar) have also been targetted for transformation.
  • 18.
    Risk Assessment Potential forthe gene to recombine with other genes possibly leading to the hyperaccumulation of non-contaminant compounds Reporter/marker genes may also escape into the environment The GM plants may revert to a wild type genotype
  • 19.
    Advantages of phytoremediation Itis more economically viable using the same tools and supplies as agriculture It is less disruptive to the environment Disposal sites are not needed Aesthetically pleasing than traditional methods Avoids excavation and transport of polluted media thus reducing the risk of spreading the contamination It has the potential to treat sites polluted with more than one type of pollutant
  • 20.
    Disadvantages of phytoremediation Growing conditions required by the plant (i.e., Climate, geology, altitude, temperature) Tolerance of the plant to the pollutant Contaminants collected in senescing tissues may be released back into the environment in autumn Contaminants may be collected in woody tissues used as fuel Time taken to remediate sites far exceeds that of other technologies Contaminant solubility may be increased leading to greater environmental damage and the possibility of leaching

Editor's Notes