phytoremediation microbial endophytes katie fishburn and jared bodecker

Phytoremediation Microbial Endophytes

Katie Fishburn and Jared Bodecker

Endophytes

Microbes that live in plant tissue that increase plant biomass and resistance to pathogens

Of the 300000 known species of plants there is at least one plant-associated endophyte (Strobel G et al)

Can remediate pollutants just as good as rhizospheric microbes in some cases better

There are clear advantages of endophytes compared to the closely associated rhizospheric cousins

Known endophytes in PR

Endophyte vs Rhizospheric

Population control is easier than soil microbes due to lack of competition (protected inside plant tissue)

Degradation of pollutants will be localized to inside the plant being contained from leaking in the soil and air environment

Plant species-specific selection of necessary bacteria for a specific pollutant

Microbial sampling is easier (plant tissue)

Applications endophyte-phytoremediation

Phytoremediation Organics

24-D (herbicide) degradationIsolates from poplar inoculated into pea plants planted in soils with 13 mg of 24-D

Control high levels of 24-D remain

Inoculated plants no 24-D remains in test soils

PAH tolerance in willow and poplarUsed also as carbon source by microbes

TNT RDX HMX degradation by methylobacterium found in hybrid poplar

Mineralized 60 of RDX and HMX in 2 months


Control high levels of 24-D remainInoculated plants no 24-D remains in test soils

2 4-D Degradation

Psuedomonas putida mini Tn5 (GFP)

Root tissue

Stem tissue









Backdrop picture courtesy of NASA

Poly Aromatic Hydrocarbons










Methylobacterium spp Strain BJ001

Phytoremediation Inorganics

Brassica chinensisMucor isolate

Cd and Pb toleranceCd and Pb more water soluble more bioavailable

Brassica chinensis

Thlaspi goesingense

Thlaspi goesingenseMethylobacterium and Shpingomonas

Ni accumulation and tolerancesiderophores and ethylene relieve

Plant Yield and Growth Promotion

Enhanced nutrientmineral availability and uptake

Phosphate nitrogen

Phosphate solubilization

Siderophore productionMinerals made more bioavailable solubilized by siderophore

Osmotic adjustment stomatal regulation

Plant Health and Protection

Lessenprevent effects of pathogens on plants

Fungal bacterial viral diseases less harmful after inoculation of endophytes

Endophytes trigger induced systemic resistance (ISR)

Immune-like response by plants

IndustrialMedical Applications

Pseudomonas Burkholderia Bacillus Antibiotics anti-cancer compounds antifungal antiviral insecticidal immunosuppressant agents

Making a (6) Million $ Endophyte

Engineering plant-associated microbes

Horizontal Gene Transfer

Improving already efficient phytoremediation

By-products of degradation can even more toxic when released into the air

When in planta you have natural reduction of volatile toxins

Improving Toluene Degradation

Normal plant degradation of toluene creates toxic by-products that are volatized into the environment

Burkholderia cepacia G4 (rhizoshperic) with toluene-degrading qualities from pTOM plasmid (toluene-ortho-monooxygenase)

B cepacia VM1330 (endophyte) with the pMON plasmid

Enhanced Toluene-resistance

Control- Yellow Lupine (no inoculate)

BU0072- YL endophyte strain

G4- YL rhizospheric strain (pTOM)

VM1330- YL transformed endophyte strain (pTOM)

Growth Index (Fresh Weight)

Total Weight (grams)

Reduced Toluene-volatization

Improved degradation in VM1330 and Yellow Lupine

Which lowers phytotoxicity and evapotranspiration

Sub-phytotoxic levels used in experiments (100mgL)

Lupine endophytes in Poplar

Control- Poplar (no innoculate)VM1466- P endophyte (pTOM)BU61- P rhizospheric (pTOM)

Decreased Toluene-volatization


Opposite of vertical transfer (parental or ancestral)

Genetic material is transferred from another organism to another organism without being an offspring

Common routes of transferTransformation Uptake of endogenous DNA (naturelab)

Transduction Delivery via vector (bacteriophage)

Conjugation a physical connection between two organisms delivers DNA from one to the other

ReferencesKieran J Germaine Xuemei Liu Guiomar Garcia Cabellos Jill P Hogan David Ryan amp David N Dowling (2006) Bacterialendophyte-enhancedphytoremediationofthe organochlorine herbicide 24 -

dichlorophenoxyacetic acid FEMS

Benoit Van Aken Jong Moon Yoon and Jerald L Schnoor (2003) Biodegradation of Nitro-Substituted Explosives 246-Trinitrotoluene Hexahydro-135-Trinitro-135-Triazine and

Octahydro-1357-Tetranitro-135-Tetrazocine by a Phytosymbiotic Methylobacterium sp Associated with Poplar Tissues (Populus deltoides 1113096 nigra DN34) APPLIED AND ENVIRONMENTAL MICROBIOLOGY Jan 2004 p 508ndash517 Vol 70 No 1

Zujun Dengab Lixiang Caoc Haiwei Huanga Xinyu Jianga Wenfeng Wangc Yang Shia Renduo Zhangalowast (2010) Characterization of Cd- and Pb-resistant fungal endophyte Mucor sp CBRF59 isolated from rapes (Brassica chinensis) in a metal-contaminated soil Journal of Hazardous Materials

Rughia Idris1 Radoslava Trifonova1 Markus Puschenreiter2 Walter W Wenzel2 and Angela Sessitsch1 (2004) Bacterial Communities Associated with Flowering Plants of the Ni Hyperaccumulator Thlaspi goesingense APPLIED AND ENVIRONMENTAL MICROBIOLOGY May 2004 p 2667ndash2677

Robert P Ryan1 Kieran Germaine2 Ashley Franks1 David J Ryan2 amp David N Dowling2 (2007) Bacterial endophytes recent developments and applications FEMS

Germaine K Keogh E Borremans B et al (2004) Colonisation of poplar trees by gfp expressing bacterial endophytes FEMS Microbiol Ecol 48 109ndash118

Germaine K Liu X Cabellos G Hogan J Ryan D amp Dowling DN (2006) Bacterial endophyte-enhanced phyto-remediation of the organochlorine herbicide 24-dichlorophenoxyacetic acid FEMS Microbiol Ecol 57 302ndash310

Miller CM Miller RV Garton-Kenny D Redgrave B Sears J Condron MM Teplow DB amp Strobel GA (1998) Ecomycins unique antimycotics from Pseudomonas viridiflava J Appl Microbiol 84 937ndash944

Lodewyckx C Vangronsveld J Porteous F Moore ERB Taghavi S Mezgeay M amp van der Lelie D (2002) Endophytic bacteria and their potential applications Crit Rev Plant Sci 21 583ndash606

Lodewyckx C Taghavi S Mergeay M Vangronsveld J Clijsters H amp van der Lelie D (2001) The effect of recombinant heavy metal resistant endophytic bacteria in heavy metal uptake by their host plant Int J Phytoremediation 3 173ndash187

Taghavi S Barac T Greenberg B Borremans B Vangronsveld J amp van der Lelie D (2005) Horizontal gene transfer to endogenous endophytic bacteria from poplar improved phyto-remediation of toluene Appl Environ Microbiol 71 8500ndash8505

Strobel G Daisy B Castillo U amp Harper J (2004) Natural products from endophytic microorganisms J Nat Prod 67 257ndash268

Van Aken B Peres C Doty S Yoon J amp Schnoor J (2004) Methylobacterium populi sp nov a novel aerobic pink- pigmented facultatively methylotrophic methane-ultilising bacterium isolated from poplar trees (Populus deltoides x nigra DN34) Evol Microbiol 54 1191ndash1196

Xi C Lambrecht M Vanderleyden J amp Michiels J (1999) Bi-functional gfp-and gusA-containing mini-Tn5 transposon derivatives for combined gene expression and bacterial localization studies J Microbiol Methods 35 85ndash92

Phytoremediation

Endophytes





2 4-D Degradation

Phytoremediation Organics (2)



Slide 11


Brassica chinensis

Thlaspi goesingense










Slide 24

References

Endophytes

Microbes that live in plant tissue that increase plant biomass and resistance to pathogens

Of the 300000 known species of plants there is at least one plant-associated endophyte (Strobel G et al)

Can remediate pollutants just as good as rhizospheric microbes in some cases better

There are clear advantages of endophytes compared to the closely associated rhizospheric cousins

















2 4-D Degradation


Root tissue

Stem tissue
























Brassica chinensis

Thlaspi goesingense





Phosphate nitrogen

























































Phytoremediation

Endophytes





2 4-D Degradation




Slide 11


Brassica chinensis

Thlaspi goesingense










Slide 24

References

















2 4-D Degradation


Root tissue

Stem tissue
























Brassica chinensis

Thlaspi goesingense





Phosphate nitrogen

























































Phytoremediation

Endophytes





2 4-D Degradation




Slide 11


Brassica chinensis

Thlaspi goesingense










Slide 24

References
























Brassica chinensis

Thlaspi goesingense





Phosphate nitrogen

























































Phytoremediation

Endophytes





2 4-D Degradation




Slide 11


Brassica chinensis

Thlaspi goesingense










Slide 24

References

Thlaspi goesingense





Phosphate nitrogen

























































Phytoremediation

Endophytes





2 4-D Degradation




Slide 11


Brassica chinensis

Thlaspi goesingense










Slide 24

References



Phosphate nitrogen

























































Phytoremediation

Endophytes





2 4-D Degradation




Slide 11


Brassica chinensis

Thlaspi goesingense










Slide 24

References






















































Phytoremediation

Endophytes





2 4-D Degradation




Slide 11


Brassica chinensis

Thlaspi goesingense










Slide 24

References

phytoremediation microbial endophytes katie fishburn and jared bodecker

Documents

d control

plant tissue slide

hmx degradation

inoculated plants

hybrid poplar

pr slide

pea plants

jared bodecker slide