Temesgen Belayneh Monika Komon-Zelazowska Christian P. Kubicek & Irina S. Druzhinina

1. Indigenous Hypocrea/Trichoderma species as efficient agents for control of coffee wilt disease in Ethiopia Institute of Chemical Engineering Research Area Applied Biochemistry and Gene Technology Group of Fungal Evolution and Biodiversity Temesgen BelaynehMonika Komon-ZelazowskaChristian P. Kubicek& Irina S. Druzhinina

2. Ethiopia as a hotspot for the global diversity a country situated in the Horn of Africa special country in terms of landraces and biological diversity Many of the plant species are still untouched and still unspoiled Of the several species of coffee, Ethiopia is the center of origin and diversity of Coffee arabica L. it accounts for 75% of coffee exports in the world.

3. Coffea arabica L. in Ethiopia Coffee is one of the most important agricultural commodities in the world, worth up to US$ 14 billion annually The economy of Ethiopia is based on agriculture and it highly depends on coffee production, therefore coffee is critical to the Ethiopian financial system Coffee belongs to the botanical family Rubiaceae, which has some 500 genera and over 6,000 species. Most are tropical trees and shrubs which grow in the lower storey of forests

4. Varieties of Coffea arabica L.: Typica and Bourbon Coffee plant is a large bush with dark-green oval leaves

5. Coffee wilt desease (tracheomycosis) Currently, the Ethiopian national average of coffee yield is about 472 Kg ha-1. That is much below the research results of 1500-2000 Kg ha-1. Such a substantial loss is to large extend determined by the wide spread of coffee wilt diseases which are usually caused by Fusarium species

6. Coffee wilt desease (tracheomycosis) Before its occurrence in Ethiopia these fungi wereearlier reported to be well known pathogen of other coffee species in the West and Central Africa attacking mostly Coffee excelsa plantations especially in Cameroon and later C. canephra (Robusta coffee) in Ivory Coast and Zaire. Its occurrence in Ethiopia was identified in the early 1970’s when Kranz and Mogk (1973) isolated pure culture from dying coffee trees.

7. Ethiopian Plant Protection Research Center Established in 1974 by the former USSRwith the science and technology development agreement

8. Ethiopian Plant Protection Research Center Sections: Mycology Bacteriology Virology Nematology Entomology Departments and Biocontrol is a section within biotechnology

9. MSc Temesgen Balayneh

10. Coffee project Ethiopian coffee ceremony

11. Coffee wilt disease and type of forestry introduction of individual plants of C. arabica in the natural forest ecosystem making of artificial coffee „plantations“ by desruption of original forest, cultivation of fast growing shadow trees and subsequent introduction of coffee plants

12. Resistance to coffee wilt disease (hypothesis) due to presence of native antagonists in rhizosphere of C. arabica due to symbiotic fungi (Glomus spp. and ?? Trichoderma ???) due to multidimentional interactions between coffee plant, symbionts and rhizosphere microbionts

13. Goals of the research to identify the causative agent of coffee wilt disease (Fusarium spp.) to study the native biodiversity of Trichoderma in Ethiopia to test the potential of different Trichoderma isolates from rhizosphere of Coffea arabica L. to control Fusarium

14. SNNP Jimma Harerga Wellega Road side survey • With 10 to 15 km stop of car • Collected in plastic bags • Stored at -4°C until processing • Pilot project for diversity study

15. Fusarium diversity strains of Fusarium causing coffee wilt disease were isolated directly from infected roots based of tef1 sequence analysis using FUSARIUM-SEQ (Geiser et al., 2004) we identified them as Gibberella fujikuroi/ Fusarium oxysporum species complex:

16. Trichoderma biodiversity • Generally, we collected 60 soil samples - Different coffee growing areas - Diff. Cultivation:- Forest, Semi- forest and “Plantation” - Alt. Range:- 1300 - 2400 m.a.s.l. • With the visual and microscopic identification - Found 45 samples of Trichoderma spp.

17. Trichoderma biodiversity All Trichoderma strains have been identified based on ITS1 and 2 and tef1 sequence analyses using TrichOKEY and TrichoBLAST tools, respectively (www.isth.info)

18. Trichoderma biodiversity

19. Trichoderma diversity in C. arabica rhizosphere

20. Trichoderma biodiversity in Ethiopia (coffee growing forest) New species 1, Sect. Longibrachiatum, related to T. longibrachiatum - H. orientalis species complex Harerga SNNP New species 2, Sect. Pachybasium, related to T. tomentosum - T. cerinum from T. harzianum species complex Jimma New species 3 Sect. Trichoderma, related to T. asperellum Welega Jimma New species 4 Sect. Pachybasium, related to T. spirale Jimma New species 5 Sect. Pachybasium, related to T. helicum Jimma

21. Trichoderma biodiversity in Ethiopia (coffee growing forest) Trichoderma biodiversity in Ivory Coast (coccoa plantations)

22. in vitro tests of antagonistic potential experimental strategies Fusarium – Trichoderma confrontations on plates Ability of Trichoderma to overgrow Fusarium cultures P1 strain of Trichodermaatroviride as the reference

23. in vitro tests of antagonistic potential strong weak no response moderate

24. T. oblongisporum (Jimma)

25. T. koningiopsis Samuels et al., ms submitted to Stud. Myc.

26. T. atroviride (Jimma)

27. T. sp. 1(Jimma and SNNP)

28. T. sp. 2(Welega and SNNP)

29. Conclusions In all four areas coffee wilt disease is caused by strains of Fusariumoxysporum species complex. The biodiversity of Fusarium is low. The first road side survey study has revealed 10 known and 5 potentially new species of Trichoderma inhibiting the rhizosphere of Coffea arabica. The biodiversity of Trichoderma is very high. 25 out of 45 (55%) Trichoderma strains have strong potential to prevent the growth of Fusarium oxysporum.

30. Future perspectives

31. Aknowledgements The research was supported by Ethiopia: PPRC scholarship to Temesgen Balayneh and partly by FWF project

32. Thank you for your attention!