Transcript Indigenous Hypocrea/Trichoderma species as efficient

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
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.
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
•Varieties of Coffea arabica L.:
Typica and Bourbon
Coffee plant is a large bush
with dark-green oval leaves
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
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.
Ethiopian Plant Protection
Research Center
Established in 1974 by the former USSR
with the science and technology development agreement
Ethiopian Plant Protection
Research Center
Sections:
Mycology
Bacteriology
Virology
Nematology
Entomology
Departments
and
Biocontrol is a section
within
biotechnology
MSc Temesgen Balayneh
Coffee project
Ethiopian coffee ceremony
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
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
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
Road side survey
SNNP
Jimma
•With 10 to 15 km stop of car
•Collected in plastic bags
Harerga
•Stored at -4°C until processing
•Pilot project for diversity study
Wellega
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:
Trichoderma biodiversity
• Generally, we collected 60 soil samples
- Different coffee growing areas
- Diff. Cultivation:- Forest, Semiforest and “Plantation”
- Alt. Range:- 1300 - 2400 m.a.s.l.
• With the visual and microscopic
identification
- Found 45 samples of Trichoderma
spp.
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)
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
1
1809
T. spirale
2
1822, 1839
T. hamatum
5
1810, 1811, 1814, 1826, 1827, 1835
T. koningiopsis
5
1813, 1816, 1821, 1823, 1831
T. asperellum
4
1819, 1820, 1829, 1838
T. atroviride
1
1832
T. longibrachatum
2
1817, 1889
T. orientalis
2
1815, 1837
9
1808, 1833, 1834, 1836, 1841, 1888, 1890,
1892, 1893
New species
Trichoderma diversity in
C. arabica rhizosphere
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
New species 3
Sect. Trichoderma,
related to T. asperellum
Welega Jimma
New species 4
Sect. Pachybasium,
related to T. spirale
New species 5
Sect. Pachybasium,
related to T. helicum
Jimma
Jimma
Jimma
Trichoderma biodiversity in Ethiopia
(coffee growing forest)
Trichoderma biodiversity in Ivory Coast
(coccoa plantations)
in vitro tests of antagonistic
potential
experimental strategies
• Fusarium – Trichoderma confrontations
on plates
• Ability of Trichoderma to overgrow Fusarium
cultures
• P1 strain of Trichoderma atroviride as the
reference
in vitro tests of antagonistic
potential
List of species:
Amount
Numbers from CPK collection
T. harzianum
5
1807, 1812, 1818, 1825, 1840
T. brevicompactum
1
1828
T. oblongisporum
1
1809
T. spirale
2
1822, 1839
T. hamatum
6
1810, 1811, 1814, 1826, 1827, 1835
T. koningiopsis
5
1813, 1816, 1821, 1823, 1831
T. asperellum
5
1819, 1820, 1829, 1838
T. atroviride
1
1832
T. longibrachatum
1
1817
T. orientalis
2
1815, 1837
New species 1
3
1808, 1833, 1892
New species 2
2
1836, 1893
New species 3
2
1841, 1890
New species 4
1
1888
New species 5
1
1834
strong
moderate
Jimma
weak
Harerga
SNNP
Wellega
no response
T. oblongisporum (Jimma)
T. koningiopsis
Samuels et al., ms submitted to Stud. Myc.
T. atroviride (Jimma)
T. sp. 1 (Jimma and SNNP)
T. sp. 2 (Welega and SNNP)
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.
Future perspectives
Aknowledgements
The research was supported by Ethiopia:
PPRC scholarship to Temesgen Balayneh
and partly by FWF project
Thank you for your attention!