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Indigenous Hypocrea/Trichoderma species as efficient agents for control of coffee wilt disease in Ethiopia Institute of Chemical Engineering Research.

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Presentation on theme: "Indigenous Hypocrea/Trichoderma species as efficient agents for control of coffee wilt disease in Ethiopia Institute of Chemical Engineering Research."— Presentation transcript:

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 Belayneh Monika Komon-Zelazowska Christian 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. So now, I would like to say few words about the birthland of coffee. (The federal Democratic Republic of) Ethiopia is a nation situated in the Horn of Africa. Ethiopia is known as a special country in terms of landraces and biological diversity what is also known of this country that many of the plant sp……. of the….. it accounts…. The Federal Democratic Republic of Ethiopia is a country situated in the Horn of Africa. It has one of the most extensive known histories as an independent nation on the continent, or indeed in the world. …. “Ethiopia is a very special country in terms of landraces and biological diversity. Many of the plant species and genetic species in this country are still untouched and still unspoiled. “Or” the country is one of the global centers of biodiversity ... because of its heterogenous landmass, agro-ecological variations resulting in plant genetic diversity of landraces and other biodiversity resources.” ? Ethiopia is one of the East African nations with an estimated population of 77 million. ?? (probably not needed) The variety of life on Earth, its biological diversity is commonly referred to as biodiversity. The number of species of plants, animals, and microorganisms, the enormous diversity of genes in these species, the different ecosystems on the planet, such as deserts, rainforests and coral reefs are all part of a biologically diverse Earth. 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 is one of the….. - More than 80 countries, including Ethiopia, cultivate coffee. The economy of Ethiopiais based…… It is also worth to mention that : Coffea arabica L. was first described by Linnaeus in The best known varieties are 'Typica' and 'Bourbon' but from these many different strains and cultivars have been developed. The average arabica plant is a large bush with dark-green oval leaves Arabica coffee is often susceptible to atack by pests and diseases, therefore resistance is a major goal of plant breeding programms. Arabica coffee is grown throughout Latin America, in Central and East Africa, in India and to some extent in Indonesia). The export of coffee contributes more than 60 % of the country’s foreign currency 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 …………. Coffee is a tropical plant which requires very specific environmental conditions for commercial cultivation. Temperature, rainfall, sunlight, wind and soils are all important, but requirements vary according to the varieties grown. (very nice story ) The story of how coffee growing and drinking spread around the world is one of the greatest and most romantic in history. It starts in the Horn of Africa, in Ethiopia, where the coffee tree probably originated in the province of Kaffa. There are various fanciful but unlikely stories surrounding the discovery of the properties of roasted coffee beans. One story has it that an Ethiopian goatherd was amazed at the lively behaviour of his goats after chewing red coffee berries. What we know with more certainty is that the succulent outer cherry flesh was eaten by slaves taken from present day Sudan into Yemen and Arabia, through the great port of its day, Mocha, now synonymous with coffee. Coffee was certainly being cultivated in Yemen by the 15th century and probably much earlier than that. Because C. arabica is self-pollinating, these varieties tended to remain genetically stable. However, spontaneous mutations showing desirable characteristics have been cultivated in their own right, as well as being exploited for cross-breeding  purposes. Some of these mutants and cultivars are described below ….. The coffee bean is the seed of the coffee tree, but when stripped of its outer layers it becomes infertile. At first coffee was mainly sold by lemonade vendors and was believed to have medicinal qualities. Na koncu tego slajdu powiedziec o szacowaniu w produkcji kawy_z Temesgena report or wikipedia, etc. potem przejsc do coffee wilt disease 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 Coffee plant is a large bush with dark-green oval leaves
Varieties of Coffea arabica L.: Typica and Bourbon Coffee plant is a large bush with dark-green oval leaves Coffea arabica L. was first described by Linnaeus in The best known varieties are 'Typica' and 'Bourbon' but from these many different strains and cultivars have been developed. The average arabica plant is a large bush with dark-green oval leaves Arabica coffee is often susceptible to attack by pests and diseases, therefore resistance is a major goal of plant breeding programs.

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 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 At this time, coffee farmers are facing many difficulties. These include pests and diseases, and quality problems due to poor processing, poor infrastructure and poor commercialization. All of which lead to higher costs and/ or low prices. (dalej slajd) Fusarium is not oryginally from Ethiopia. it comes from neighbour countries (czytaj Tem report)

6 Coffee wilt desease (tracheomycosis)
Before its occurrence in Ethiopia these fungi were earlier 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. dopowiedziec: usually, affected plants show loss of turgor followed by the quick death and root discoloration normally noticed before foliage (listowie) symptomps. Fusarium is not oryginally from Ethiopia. it comes from neighbour countries (czytaj Tem report)

7 Ethiopian Plant Protection
Research Center Established in 1974 by the former USSR with 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 Lately departments of biotechnology and biosystematics were also organized from which biocontrol is a section within biotechnology The Plant Protection Research Center (PPRC) was organized in Mycology, Bacteriology…… Biocontrol section has two working groups 1. Root Diseases control Trichoderma 2. Insect Pests control Bevaria and Metharizum

9 MSc Temesgen Balayneh Let‘s say few words about our co-worker form Ethiopia. Last year Temesgen was working with us for 6 months. During this time he applied for a doctoral research grant and now, in 3 weeks, he is coming to Vienna to make the study for three following years in Irina‘s group. Temesgen‘s activity in Ethiopia: - he is involved in 3 projects like biocontrol of : Fusarium solani on Faba bean, Phytophthora on citrus, and Fusarium on coffee, what is his main project and interest. - he is also involved in teaching with farmers, he is organizing a special training course for Development agents. Powiedziec cos o Temesg, o tym, ze uczy swoich ludzi, ma specjalne kursy o tym. On uczy farmerow, a ci z kolei ucza tubylcow  O tym, ze pracuja nad 3 projektami (slajdy) Ten projekt o kawie wymyslil on sam i ma support from Austrain Academic exchanges grants.

10 Coffee project Ethiopian coffee ceremony
Coffee project was proposed and written by Temesgen. 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 There are two ways of coffee breeding : - the one way can be made by introduction of individual plants of C. arabica in the natural forest ecosystem (expensive but environmentally friendly) - and the second way can be done by making of artifical coffee ‘‘plantations‘‘…….(cheap BUT environmentally hazardous).

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 ???) According to the preliminary observation, coffee trees cultivated according to the later mathod are more susceptible to wilt diseases. There are 3 hypothesis describing resistance to coffee wilt disease in natural forest. The resistance is probably due to presence of native antagonists in rhizosphere of Coffea arabica or due to.,…… or due to …… 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 Goals of the research: we wanted to identify the causative agent of coffee wilt disease we wanted to study….. we wanted to test the….. to test the potential of different Trichoderma isolates from rhizosphere of Coffea arabica L. to control Fusarium

14 Road side survey SNNP Jimma With 10 to 15 km stop of car
Collected in plastic bags Stored at -4°C until processing Pilot project for diversity study The first step was done by Temesgen. He has made the road sampling method. Harerga Wellega

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: 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) ITS- Internal Transcribed Spacer 1 and 2 tef1- translation elongation factor 1 alfa

18 Trichoderma biodiversity
List of species: Amount Strain numbers in CPK collection: T. harzianum 5 1807, 1812, 1818, 1825, 1840 T. brevicompactum 1 1828 T. oblongisporum 1809 T. spirale 2 1822, 1839 T. hamatum 1810, 1811, 1814, 1826, 1827, 1835 T. koningiopsis 1813, 1816, 1821, 1823, 1831 T. asperellum 4 1819, 1820, 1829, 1838 T. atroviride 1832 T. longibrachatum 1817, 1889 T. orientalis 1815, 1837 New species 9 1808, 1833, 1834, 1836, 1841, 1888, 1890, 1892, 1893

19 Trichoderma diversity in
C. arabica rhizosphere …significant part of this diagram is occupied by New Species.

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 we also did tests of antagonistic potential in vitro Our experimental strategies: we did confrontations Fus-Tricho on plates we tested the ability of Tricho to overgrow Fus cultures we took the P1 strain of T. atroviride as the reference P1 strain of Trichoderma atroviride as the reference

23 in vitro tests of antagonistic
potential List of species: Amount Numbers from CPK collection Jimma Harerga SNNP Wellega T. harzianum 5 1807, 1812, 1818, 1825, 1840 T. brevicompactum 1 1828 T. oblongisporum 1809 T. spirale 2 1822, 1839 T. hamatum 6 1810, 1811, 1814, 1826, 1827, 1835 T. koningiopsis 1813, 1816, 1821, 1823, 1831 T. asperellum 1819, 1820, 1829, 1838 T. atroviride 1832 T. longibrachatum 1817 T. orientalis 1815, 1837 New species 1 3 1808, 1833, 1892 New species 2 1836, 1893 New species 3 1841, 1890 New species 4 1888 New species 5 1834 we also did tests of antagonistic potential in vitro Our experimental strategies: we did confrontations Fus-Tricho on plates we tested the ability of Tricho to overgrow Fus cultures we took the P1 strain of T. atroviride as the reference strong moderate weak no response

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 Fusarium oxysporum 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 Coffea arabica L. is known to form complex associations with soil microflora, mainly with mycorrhizal, pathogenic and saprotrophic fungi. Therefore, the general goal of this study is to find out which mechanisms and factors control the relationship of coffee tree with fungi from these ecological groups. And to make a recommendation to coffee farmers on preferable methods of coffee cultivation and protection against phythopathogenic fungi.

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

32 Thank you for your attention!


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