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Microbiological and ecological testing of Ambrosia artemisiifolia's endophyta

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Microbiological and ecological testing of Ambrosia artemisiifolia's endophyta

Mária Fehér

Supervisors:

László Körmöczi and

László Manczinger

Faculty of Science and Informatics, University of Szeged

Department of Ecology Hungary

(2)

The enemy…

(3)

Timeliness of the theme

• Ambrosia artemisiifolia (Common ragweed) causes global problem health and ecosystem

(Mihály és Botta-Dukát — 2004)

• Invasive, highly allergenic

• Symbiosis with

mycorrhizae – helps with invasion (Fumanal et. al.

— 2006) – What about

endophytic fungi?

(4)

The tested plant

• Common ragweed (Ambrosia artemisiifolia) - From United States of America to Europe in the 1800’s

• Generative reproduction

• Seeds: primer and secunder dormancy - breaks at low

temperatures (in laboratory:

12 weeks, 4 °C)

• Live with endophytes –

Preliminary research

(5)

History of Research – part 1

About mycorrhizal fungi

• Plants live with

arbuscular mycorrhizal fungi in symbioses

• Phylum

Glomeromycota

• Obligate biotroph

• Increased uptake of nutrients such as

phosporus and nitrogen.

(Juan et. al. — 2006)

(6)

History of Research – part 2

About Endophytic fungi

• Never inside the cells

• Hydrolytic enzymes:

polymers ->

monomers (Huaijun et. al. — 2004)

• Protect plants from insects and

herbivorous ->

secrete toxic alkaloids (Breen

1994, Bush, Wilkinson

& Schardl 1997).

• Mycorrhizae are not free-living, but

endophytes do

• Plants protect and feed them

• Endophytic fungi don’t causes negative

symptoms in plants (Suryanarayan et. al.

— 2009)

• Between the cells - in

the leaf

(7)

First period of the research

• Sample

collection:

• Bank of River Maros near Szeged

• natural habitat

• Szolnok

• urban habitat

http://maps.google.com

(8)

Fungi isolated during the first experiment

• Fusarium chlamydosporum

• Fusarium oxysporum ( Fusarium oxysporum var redolens )

• Fusarium solani

• Fusarium redolens

• Leptosphaerulina chartarum

• Absidia repens

• Mucor circinelloides

(9)

Fusarium

chlamydosporum

http://www.mold.ph

Fusarium oxysporum

http://www.reviberoammicol.com

http://www.pf.chiba-u.ac.jp

Fusarium solani

Mucor

circinelloides

http://genome.jgi-psf.org

http://www.livne.co.il http://www.doctorfungus.org

http://www.mold.ph/fusarium.htm

Leptosphaerulina chartarum

http://www.doctorfungus.org

http://www2.mst.dk

Fusarium

oxysporum var redolens

Absidia sp.

http://www.doctorfungus.org

http://www.caltexmoldservices.com

(10)

Relative enzyme activity

Fungi from Szolnok

0,00 0,50 1,00 1,50 2,00 2,50

Abs idia repens

Fusarium chlam ydosporum

Fus

arium oxysporum

Fusarium redolens Fus

arium solani

Leptosphaerulina chartarum

Mucor circinelloides

Species

Extinctions

Cellulase Xilanase β-1,3-glucanase Lipase

Cellobiohidrolase β-glukosidase Trypsin Chymotrypsin Exocitinase

Fungi from bank of River Maros

0,00 0,10 0,20 0,30 0,400,50 0,60 0,70 0,80 0,90

Absidia repen s

Fusarium chlamydosporum Fusarium oxy

sporum Fusarium

redo lens

Fusarium solani

Leptosphaerulina chartarum

Mucor circinelloides

Species

Extinctions

Cellulase Xilanase β-1,3-glucanase Lipase

Cellobiohidrolase β-glukosidase Trypsin Chymotrypsin Exocitinase

(11)

Outlook

• The role of endophytic fungi in the adaptability of ragweed

– Enzyme productions – Competition tests

– Species level identification

• Test 4 different soils on the arable field ,

abandoned fields

and grasslands

(12)

Sampling methods

• Areas:

• Arable fields

• Abandoned fields

• Grasslands

• Whole root samples (5-5 from each area) – microbiological tests

• Soil samples –

organic carbon, P, Ca,

K, Na, pH,

(13)

Questions

• What kind of endophytic fungi live in the root of common ragweed?

• Which enzymes are produced by endophytic fungi?

• What kind of fungi can be isolated from plants living in different habitats such as arable field, abandoned fields and grasslands?

• What is the quantitative distribution of fungi

in different habitats?

• How many fungi live

in the roots of the plants living in different habitats?

• Which nutrients are better for endophytic fungal?

• Which fungal are necessary for

Ambrosia and which fungal are better for Ambrosia?

(14)

References

• Fumanal et al. (2006) – Which role can arbuscular mycorrhizal fungi play in the facilitation of Ambrosia artemisiifolia L. invasion in France? — Mycorrhiza 17:25-35

• Mihály Botond és Botta-Dukát Zoltán (2004): Biológiai inváziók Magyarországon – Özönnövények — TermészetBÚVÁR Kiadó, Budapest

• Brantlee Spakes Richter et. al. (2002) - Assessment of arbuscular mycorrhizal fungal propagules and colonization from abandoned agricultural fields and semi-arid

grasslands in riparian floodplains — Applied Soil Ecology 20, 227–238

• Huaijun Michael Li, Jo Anne Crough and Faith C. Belanger (2004): Fungal endophyte N-acetylglucosaminidase expression in the infected host grass — Mycol. Res. 109 (3): 363–373

• T. S. Suryanarayan, N. Thirunavukkarasu, M. B. Govindarajulu, F. Sasse, R.

Jamsem, T. S. Murali (2009): Fungal endophytes and bioprospecting — Fungal Biology Review 23: 9-19

• Breen, J. P. (1994): Acremonium endophyte interactions with enhanced plant resistance to insects — Annual Review of Entomology 39: 401–423.

• Juan C. Santos, Roger D. Finlay and Anders Tehler (2006): Molecular analysis of arbuscular mycorrhizal fungi colonising a semi-natural grassland along a fertilisation gradient - New Phytologist 172: 159–168.

• Bush, Wilkinson & Schardl (1997): Bioprotective Alkaloids of Grass-Fungal Endophyte Symbioses - Plant Physiol. 114: 1-7

(15)

Thank you for your attention

Thanks to:

László Körmöczi László Manczinger Enikő Sajben-Nagy

Márta Zalatnai Zoltán Bátori László Erdős

TÁMOP-4.2.2/B-10/1-2010-0012 projekt

Judit Márton Ágnes Szepesi

Ágnes Méri

Nikoletta Geltsch

Tamás Marik

Gábor Reszler

(16)

The publication / presentation is supported by the European Union and cofunded by the European

Social Fund.

Project title: „Broadening the knowledge base and supporting the long term professional sustainability of

the Research University Centre of Excellence at the University of Szeged by ensuring the rising

generation of excellent scientists.”

Project number: TÁMOP-4.2.2/B-10/1-2010-0012

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