Laboratory of Plant Physiology and Molecular Biology, University of Turku

Projects of plant-fungus interactions, biodiversity and biological control

Project leader:
Dr. Tapani Yli-Mattila, Adjunct Professor of Mycology, Finnish Coordinator Nordic network project "New emerging mycotoxins and other metabolites in toxigenic fungi of Northern Europe" http://fou02.planteforsk.no/NordforskNetworkMycotox/
phone: +358-2-3336587
e-mail: tymat@utu.fi

 

National coordinators of the Nordic network project in Turku in 2005.

Current topics:

1. Molecular identification and detection of plant pathogenic and toxigenic Fusarium species and strains.

2. Molecular evolution, biodiversity and taxonomy studies of Fusarium and Alternaria species complexes based on rDNA and beta-tubulin sequences, RFLP-PCR, isozymes and vegetative compatibility

3. Molecular identification and detection of fungal, bacterial and mite strains used in biological control and cultivated plants

4. The use of molecular markers in biodiversity and ecological studies

5. Cultivation of edible fungi

 

1. Molecular identification and detection of plant pathogenic and toxigenic Fusarium species and strains.

(Joint project with MTT, EVIRA, All-Russian Plant Protection Institute in St. Petersburg, USDA in Peoria and other Nordic participants of the Nordic network project)

 

 

 

 

 

 

 


Oats (left), spring wheat (middle) and barley (right) infected by Fusarium

 

Participants and collaborators

Sari Paavanen-Huhtala (post-doctoral scientist)
Asko Hannukkala (Agricultural Research Centre, Jokioinen, MTT)
Veli Hietaniemi (Agricultural Research Centre, Jokioinen, MTT)
Päivi Parikka (Agricultural Research Centre, Jokioinen, MTT)
Marika Jestoi (EVIRA, Helsinki)
Tatiana Gagkaeva (All-Russian Plant Protection Institute, St. Petersburg, Russia)
Olga Gavrilova (All-Russian Plant Protection Institute, St. Petersburg, Russia)
Mark Levitin (All-Russian Plant Protection Institute, St. Petersburg, Russia)
Kerry O’Donnell (USDA, Peoria, USA)

Sonja Klemsdal (Bioforsk, AAS, Norway)

Project description

Wheat, barley, oats and rye are important components in food and feed processing. All these cereals are known to be commonly colonised by Fusarium mould species, which are able to produce a range of mycotoxins. It is possible that some of the toxigenic Fusarium strains are specifically adapted to the Finnish climate during the long isolation period (several centuries) of Finnish agriculture.

The most common Finnish species F. avenaceum, which has been considered relatively ineffective in producing mycotoxins, has recently been found to produce moniliformin and enniatins. The first report on the natural occurrence of beauvericin in Fusarium infected rye and in F. avenaceum strains isolated from it was from Finland from the yield of the rainy year 1998. During the results of the year 2000, which was also rainy but warmer than the year 1998, the trichotehecene content and the amount of Fusarium infected seeds were higher in barley than during the year 1998. In 2001 even higher levels of deoxynivalenol (DON) were found in oats and spring wheat, especially in the western part of Central Finland, which was correlated with the presence of F. graminearum in this area. Also in 2002-2005 high DON levels were found in oats and they were connected to high F. gramienarum DNA levels. In 2003-2005 high HT-2 and T-2 levels were found in oats. The highest HT-2/T-2 levels were found in 2005 and they were connected to high F. langsethiae/F. sporotrichioides  DNA levels.  F. avenaceum is usually the dominant species especially in barley and wheat, but in spring wheat F. culmorum was at the same level in 2002. High F. avenaceum DNA levels are connected to high enniatin and moniliformin levels. 

We have developed species-specific primers for molecular detection, identification and quantification of Fusarium species and strains. During the years 1998-2003 and 2005-2007 grain samples were already collected from Finland and in 2000,  2002- 2005 and 2008 also from Northwestern Russia. The Finnish and Russian Fusarium isolates from the samples were studied by different morphological, molecular (e.g. RAPD-PCR, UP-PCR, diagnostic PCR, RFLP-PCR) and toxicological methods in order to find markers for toxigenic strains. Molecular methods were also used in studying the relationships between timothy and rye cultivars from different parts of the world. Pathogenicity tests of different Fusarium species were carried out on barley and spring wheat in 2002-2004 in Finland and in oats in Russia in 2007-2008. We have also developed real-time Taqman primers and probes, by which we can follow the survival of most harmful toxigenic and plant-pathogenic Fusarium strains and species directly in an environment and estimate their amount in grain samples and fields. By this way we can find out, how they are spreading in Finland and Northern Europe, study the correlation between Fusarium DNA and mycotoxin levels and select resistant varieties.

Chemotype specific primers and multilocus genetyping with Luminex flow cytometer have been used for identification of Fusarium isolates at species and chemotype level.

Publications:

Yli-Mattila, T., Paavanen, S., Hannukkala, A., Parikka, P., Tahvonen, R. & Karjalainen, R. 1996. Isozyme and RAPD-PCR analyses of Fusarium avenaceum strains in Finland. - Plant Pathol. 45:126-134.
Yli-Mattila, T., Mironenko, N.V., Alekhina, I.A., Hannukkala, A., Hyvönen, J. & Bulat, S.A. 1998. Identification of Fusarium avenaceum isolates by RAPD-PCR and UP-PCR. - Proceedings of COST 823 Workshop Advances in The Detection of Plant Pathogens by polymerase chain reaction, C. Budejovice, June 20-21 1996 (ed. by C. Manceau and J. Spak): 114-123, Luxenbourg, Office for Official Publications of the European Communities, ISBN 92-827-9197-1.
Paavanen-Huhtala, S., Hyvönen, J., Bulat, S.A. & Yli-Mattila, T. 1999. RAPD-PCR, isozyme,rDNA RFLP and rDNA sequence analyses in identification of Finnish Fusarium oxysporum isolates. - Mycol. Res. 103: 625-634.
Paavanen-Huhtala, S., 2000. Molecular-based assays for the determination of diversity and identification of Fusarium and Gliocladium fungi. PhD Thesis, Annales Universitatis Turkuensis AII 139, University of Turku, Finland, pp. 198.
Konstantinova, P., Karadzhova, J., Yli-Mattila, T. & van den Bulk, R. 2001 Molecular detection of Alternaria alternata Keisler and Fusarium Spp. in barley seeds and comparison with routine testing assays. - Proceedings book of Kromeriz meeting Sustainable Systems of Cereal Crop Protection against Fungal Diseases as the Way of Reduction of Toxin Occurrence in Food Webs. (Edited by L. Tvaruzek): 187-194, European Commission Activities, Agricultural Research Institute Kromeriz, LTD, ISBN 80-902545-3-5.
Yli-Mattila, T., Paavanen-Huhtala, S., Bulat, S.A., Alekhina, I.A. & Nirenberg, H.I. 2002. Molecular, morphological and phylogenetic analysis of Fusarium avenaceum/F.arthrosporioides/F. tricinctum species complex - a polyphasic approach. - Mycological Research 106: 655-669. 

Yli-Mattila, T., Paavanen-Huhtala, S., Parikka, P., Konstantinova, P., Gagkaeva, T., Eskola, M., Rizzo, A. 2002. Occurrence of Fusarium fungi and their toxins in Finnish cereals in 1998 and 2000. - Journal of Applied Genetics 43A: 207-214.
Jestoi, M., Rokka, M., Rizzo, A., Peltonen, K., Parikka, P. & Yli-Mattila, T., 2003. Moniliformin in Finnish grains: analysis with LC-MS/MS. – Aspects of Applied Biology 68: 211-216. Jestoi, M., Rokka, M., Yli-Mattila, T., Parikka, P., Rizzo, A. & Peltonen, K., 2004. The presence and concentrations of the Fusarium-related mycotoxins beauvericin, enniatins and moniliformin in Finnish grain samples. – Food Additives and contaminants: 21 (8): 794-802.
Yli-Mattila T, Paavanen-Huhtala S, Parikka P, Hietaniemi V, Jestoi M and Rizzo, A (2004) Toxigenic fungi and mycotoxins in Finnish cereals. In: Logrieco A and Visconti A (eds) An overview on toxigenic fungi and mycotoxins in Europe. (pp 83-100) Kluwer Academic Publishers, The Netherlands
Yli-Mattila, T. and Paavanen-Huhtala, S. 2007. Molecular identification and detection of plant pathogenic and toxigenic Fusarium fungi. In " Fungi Multifaceted Microbes (Ed. by B.N. Ganguli and S.K. Deshmukh) Anamaya Publishers, New Delhi- 110030, India. pp. 82-111. 
Yli-Mattila, T. Paavanen-Huhtala, S., Sarlin, T., Jestoi, M., Parikka, P., Rizzo, A., Haikara, T., Gagkaeva, T. ja Klemsdal, S. 2006. Kvantitatiivinen PCR-pikamääritysmenetelmä viljojen Fusarium-homeille - Maataloustieteen päivät 2006 abstraktit ( http://www.smts.fi/posterit2006.htm                                                     
Yli-Mattila, T., Parikka, P, Lahtinen, T., Rämö, S., Kokkonen. M., Rizzo, A., Jestoi, M. & Hietaniemi, V. 2008.  Kvantitatiivinen PCR-pikamääritysmenetelmä viljojen Fusarium-homeille. – Maataloustieteen päivät 2008 abstraktit. (http://www.smts.fi/mpol2008/index.htm).
 Sarlin, T,  Yli-Mattila T.,  Jestoi  M, Rizzo A., Paavanen-Huhtala S. and  Haikara A. 2006. Real-time PCR for quantification of toxigenic Fusarium species in barley and malt. European Journal of Plant Pathology 114: 371-380.
Yli-Mattila, T., Paavanen-Huhtala, S., Jestoi, M., Parikka, P., Hietaniemi, V., Gagkaeva, T., Sarlin, T., Haikara, A, Laaksonen, S.. & Rizzo, A. 2008. Real-time PCR detection and quantification of Fusarium poae, F. graminearum, F. sporotrichioides and F. langsethiae in cereal grains in Finland and Russia. – Archives of Phytopathology and Plant Protection 41: 243-260.
Jestoi, M., Paavanen-Huhtala, S., Parikka, P.. and Yli-Mattila, T. 2008. In vitro and in vivo mycotoxin production of Fusarium isolated from Finnish grains. – Arch. Phytopathol. Plant Prot. 41: 545-558.
Yli-Mattila, T., O’Donnell, K., Ward, T., and Gagkaeva, T. (2008). Trichothecene chemotype composition of Fusarium gramineaum and related species in Finland and Russia. – Journal of Plant Pathology 90:S60.
Yli-Mattila, T., Parikka, P, Lahtinen, T., Rämö, S., Kokkonen. M., Rizzo, A., Jestoi, M. & Hietaniemi, V. 2008.  Fusarium DNA levels in Finnish cereal grains. In “Current Advances in Molecular Mycology (Ed. by Y. Gherbawy) (in press).
 

 

2. Molecular evolution, biodiversity and taxonomy studies of Fusarium and Alternaria species complexes based on rDNA and beta-tubulin sequences, RFLP-PCR, isozymes and vegetative compatibility

 

Participants and collaborators

Sari Paavanen-Huhtala (post-doctoral scientist)
Mark Levitin (All-Russian Plant Protection Institute, St. Petersburg, Russia)
Tatiana Gagkaeva (All-Russian Plant Protection Institute, St. Petersburg, Russia)

Kerry O’Donnell (USDA, Peoria, USA)

Olga Gavrilova (All-Russian Plant Protection Institute, St. Petersburg, Russia)
Sonja Klemsdal (Bioforsk, AAS, Norway)
Ulf Thrane (Techn. Univ. of Denmark, Lyngby, Denmark)

Mohamed Hussein (South Valley University, Qena, Egypt)

Project description

The data from variable DNA sequences (ITS, IGS and beta-tubulin), RAPD-PCR, UP-PCR and isozymes have been used for studying taxonomy and molecular evolution of F. avenaceum/F. arthrosporioides/F. tricintum, F. oxysporum/F. redolens/F.commune and F. sporotrichioides/F. poae/F. langsethiae species complexes. The results will be compared to biochemical (including toxin data) and morphological characters of the Fusarium strains. RAPD- and UP-PCR methods were also used for studying taxonomy of Alternaria species. We have been involved in finding two new toxigenic Fusarium species, F. langsethiae and F. ussurinum.

Publications: 

Yli-Mattila, T., Hyvönen, J. 1996. RAPD-PCR analysis of Fusarium strains - cladistic evaluation of the results. - Sydowia 48: 184-195.

Yli-Mattila, T., Mironenko, N.V., Alekhina, I.A., Hannukkala, A. & Bulat, S.A. 1997. Universally primed polymerase chain reaction analysis of Fusarium avenaceum isolated from wheat and barley in Finland. - Agr. Food Sci. Finland 6: 25-36.

Yli-Mattila, T., Paavanen-Huhtala, S., Hannukkala, A., Alekhina, I.A., Hyvönen, J. & Bulat, S.A. 1997b. RAPD-PCR, UP-PCR, rDNA RFLP, rDNA sequence and isozyme analyses of Fusarium isolates in Finland.- Cer. Res. Comm. 25: 629-630.

Bulat, S.A., Alekhina, I.A., Kozlova, E.D. & Yli-Mattila, T. 1997. Molecular taxonomy of Fusarium fungi by means of ribotyping, rDNA sequencing and UP-PCR analysis. - Cer. Res. Comm. 25: 565-570.

Yli-Mattila, T., Paavanen-Huhtala, S., Bulat, S.A., Alekhina, I.A. & Nirenberg, H.I. 2002. Molecular, morphological and phylogenetic analysis of Fusarium avenaceum/F.arthrosporioides/F. tricinctum species complex - a polyphasic approach. - Mycological Research 106: 655-669.

Gagkaeva, T., Levitin, M. & Yli-Mattila, T. 2002. Vegetative compatibility and incompatibility of nitrate nonutilizing mutants in Fusarium avenaceum. - Journal of Applied Genetics 43A: 45-54.

Yli-Mattila, T., Mach, R., Alekhina, I.A., Bulat, S.A., Koskinen, S., Kullnig-Gradinger, C.M., Kubicek, C.P. & Klemsdal, S.S. 2004. Phylogenetic relationsip of Fusarium langsethiae to Fusarium poae and F. sporotrichioides as inferred by IGS, ITS, b-tubulin sequence and UP-PCR hybridization analysis. - International Journal of Food Microbiology 95: 267-285.

Konstantinova, P. & Yli-Mattila, T. 2004. IGS-RFLP analysis and development of molecular markers for identification of F. poae, F. pulverosum, F. sporotrichioides and F. kyushuense - International Journal of Food Microbiology 95: 321-331.

Schmidt, H., Adler, A., H., Holst-Jensen, A., Klemsdal, Logrieco, A., Mach, R., Nirenberg, H., Thrane, U., Torp, M., Vogel, R.F., Yli-Mattila, T. & Niessen, L. 2004. An integrated taxonomic study of Fusarium langsethiae, F. poae and F. sporotrichioides based on the use of composite datasets. International Journal of Food Microbiology 95: 341-349.

Yli-Mattila, T., Paavanen-Huhtala, S., Parikka, P., Konstantinova, P. and, Gagkaeva, T. 2004. Molecular and morphological diversity of Fusarium species in Finland and northwestern Russia – European Journal of Plant Pathology 110: 573-585.

Gagkaeva, T. & Yli-Mattila, T. 2004. Genetic diversity of populations of Fusarium graminearum in Europe and Asia – European Journal of Plant Pathology 110: 551-562.

Gannibal, P. & Yli-Mattila, T. 2005. Cultural and molecular differentiation of small-spored Alternaria species associated with poaceae. - Mycology and Phytopathology 39: 13-23.

Yli-Mattila, T., Paavanen-Huhtala, S.,  Parikka, P.,  A., Jestoi, M., Klemsdal, S. and Rizzo, A.,  2006. Genetic variation, real-time PCR, metabolites and mycotoxins of Fusarium avenaceum and related species. -  Mycotoxin Research 22: 79-86.
Gannibal, P. and Yli-Mattila, T. 2007. Morphological and UP-PCR analyses for differentiation of Alternaria infectoria species-group. – Mycology and Phytopathology 41: 313-322 .
 

3. Molecular identification and detection of fungal, bacterial and mite strains used in biological control and cultivated plants

 

(Joint project with MTT and All-Russian Plant Protection Institute)

Participants and collaborators

Sari Paavanen-Huhtala (post-doctoral scientist)
Galina Mitina (All-Russian Plant Protection Institute, St. Petersburg, Russia)
Galina Kalko (All-Russian Plant Protection Institute, St. Petersburg, Russia)
Brian Fenton (Scottich Crop Research Institute, Dundee, Scotland)

Project description

Gliocladium catenulatum and Nectria pityroides strains, which have been developed in Finland for biological control of plant pathogenic fungi, will be studied in naturally and artificially infected cereals. We have already developed specific DNA primers for identification of both strains. In addition we shall study the use of different metabolite bioprepatives and biopreparatives of alive cultures of Bacillus, Streptomyces and Trichoderma strains developed by our Russian collaborators. We are also studying molecular identification and detection of entomopathogenic Verticillium lecanii and Beauveria bassiana strains used in biological control of insects. We have also studied predatory mite strains of Euseius finlandicus and developed primers for strains used in biological control of harmful mite species. In plants we have studied genetic variation between timothy and rye cultivars.

Publications: 

Paavanen-Huhtala, S., Avikainen, H. & Yli-Mattila, T. 2000. Development of strain-specific primers for Gliocladium catenulatum strain used in biological control. - Eur. J. Plant Pathol. 106: 187-198.

Paavanen-Huhtala, S., 2000. Molecular-based assays for the determination of diversity and identification of Fusarium and Gliocladium fungi. PhD Thesis, Annales Universitatis Turkuensis AII 139, University of Turku, Finland, pp. 198.

Kekki, K, 2000. Identification of strain J76 and RAPD-PCR analysis of Nectria pityrodes isolates. - M.Sc. thesis, Univerisity of Helsinki, pp. 55 (in Finnish).

Yli-Mattila, T. Paavanen-Huhtala, S., Fenton, B. & Tuovinen, T. 2000. Molecular identification of predatory mites by RAPD-PCR and ITS sequences - Exp. Appl. Acar. 24:863-88.

Yli-Mattila, T. 2000. Identification of J1446 strain of Gliocladium catenulatum by RAPD-PCR, UP-PCR and by Specific primers. - A research report to Kemira Agro Company.

Mitina, G. & Yli-Mattila, T. 2002. RAPD-PCR, UP-PCR and rDNA sequence analyses of entomopathogenic Verticillium lecanii and Beauveria bassiana strains - Journal of Russian Phytopathological Society, 2002, Vol.3, p.7-17.

Yli-Mattila, T. 2002. Identification of strain K61 by RAPD-PCR and UP-PCR. - A research report to Kemira Agro Company.

Guo, Y-D, Yli-Mattila, T. and Pulli, S. 2003. Assessment of genetic variation in timothy (Phleum pretense L.) using RAPD and UP-PCR. – Hereditas 138: 101-113.

Rui, M., Yli-Mattila, T. and Pulli, S. 2004. Phylogenetic relationships among cultivars of spring and winter rye (Secale cereale L.) determined by RAPD-PCR markers – Hereditas 140: 1-12.

Gannibal, P. & Yli-Mattila, T. 2005. Cultural and molecular differentiation of small-spored Alternaria species associated with poaceae. - Mycology and Phytopathology 39: 13-23.

Mitina, M., Mikhailova L.A. and Yli-Mattila, T.  2008. RAPD-PCR, UP-PCR and rDNA sequence analyses of entomopathogenic fungus Verticillium lecanii and its pathogenicity towards insects and phytopathogenic fungi. – Archives of Phytopathology and Plant Protection 41: 113-128.

 

4. The use of molecular markers in biodiversity and ecological studies

(Joint projects with MTT)

Participants and collaborators

Sari Paavanen-Huhtala (post-doctoral scientist)
Päivi Parikka (Agricultural Research Centre, Jokioinen, MTT)
Asko Hannukkala (Agricultural Research Centre, Jokioinen, MTT)
Veli Hietaniemi (Agricultural Research Centre, Jokioinen, MTT)
Kaija Hakala (Agricultural Research Centre, Jokioinen, MTT)
Galina Kalko (All-Russian Plant Protection Institute, St. Petersburg, Russia)

Project description:

In these projects molecular markers have been used for studying biodiversity of Fusarium fungi in cereals and red clover diseases in Europe.

Publications 

Yli-Mattila, T., Kalko, G., Hannukkala, A., Paavanen-Huhtala, S. ja Hakala, K. 2006. Apilamätä, Fusarium-punahomeet ja juurilaho puna-apilan hävittäjinä. - Maataloustieteen päivät 2006 abstraktit (http://www.smts.fi/posterit2006.htm)  
Kalko, G. and Yli-Mattila, T. 2007. Detection of phytopathogenic Fusarium oxysporum by specific PCR – Mycology and Phytopathology  41: 79-88.                                                                                                                     
Yli-Mattila, T., Parikka, P, Lahtinen, T., Rämö, S., Kokkonen. M., Rizzo, A., Jestoi, M. & Hietaniemi, V. 2008.  Fusarium DNA levels in Finnish cereal grains. In “Current Advances in Molecular Mycology (Ed. by Y. Gherbawy) (in press).

 

5. Cultivation of edible fungi

Participants and collaborators:

Tapani Yli-Mattila (supervisor of the M.Sc. thesis of Ville Aukee)
Gary Mills (MycoSci, supervisor of the M.Sc. thesis of Ville Aukee)
Ville Aukee (M.Sc. student)

Project description

In this project the effect of environmental factors on fruit body formation in Morchella strains have been studied. In addition morphological and molecular methods are used for idenfification of fungal species competing with Morchelal on medium.

Publications

Aukee, V. 2004. Morel (Morchella esculenta) cultivation – a search for a correct soil treatment. M.Sc. Thesis, University of Turku.