Inoculum production for field experiments with artificial Fusarium inoculation M. Herrmann
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Transcript of Inoculum production for field experiments with artificial Fusarium inoculation M. Herrmann
Inoculum production for field
experiments with artificial
Fusarium inoculation
M. Herrmann
Material:
Oat cv. Jumbo or Pergamon, or other (wheat or triticale may also be suitable); about 15kg (3 kg per Fusarium isolate)
Fusarium isolates: (from the former BBA-
collection – now JKI)Species Substrate Substrate
Remarks Country Year of
Isolation Fusarium sporotrichioides var. minus Wollenweber
Avena sativa L. rotting halm base
Germany, Berlin
1965
Fusarium avenaceum (Corda : Fries) Saccardo
Hordeum vulgare
Seeds from variety: 'Frisia'
Germany 1981
Fusarium culmorum (W. G. Smith) Saccardo
Secale L. ear Germany 1994
Fusarium langsethiae Torp & Nirenberg
Avena grain Austria 1999
Fusarium graminearum Schwabe
Triticum aestivum L.
variety: 'Trimaran' (Triticale)
Germany 2003
Transfer the cultures to SNA-agar for multiplication of the different isolates.
Preparation of seeds for inoculum production
•Soaking a few minutes and boiling of oat seeds
for 15 min with water. •Transferring the boiled seeds with less water into
autoclaving bags and autoclaving it two times.•Cool off the bags to room temperature.•Under sterile conditions: Put some peaces of agar
with the Fusarium strain into the autoclaved seeds
in the bag. Do not close the bag completely, to
permit some gas exchange.
•Let the fungus grow under dark conditions and
room temperature until the whole seedlot is
completely occupied by the fungus.•Put the seeds with mycelium to cleaned basins
for a slowly drying (over a few weeks). Seeds
with mycelium should be covered by cloths, to
slow down speed of drying. •Mixing the samples by hand all two days.
•Dryed samples can be stored in refrigerator for
several years•Counting of spore density after washing the
inoculum substrate in de-ionised water and tween-20•Adjust the spore amount for each Fusarium isolate
to get a inoculum density of 1 Mio spores per ml
Spore counting and calculation in 2008:
Species Number of spores found for each isolate
Fusarium sporotrichioides 2.578.796.550
Fusarium avenaceum 120.111.728
Fusarium culmorum 8.686.408.502
Fusarium langsethiae 6.521.739.000
Fusarium graminearum 24.812.462
In Romania and Germany 60l inoculum (3 dates with each 20l) with about 300000 Sporen/ml
•Analysis of the mycotoxin content of the 2008 inoculum
mixture (eurofins, LC/GC; HPLC-MS/MS):
Vomitoxin (DON), NIV, 3-ADON, 15-ADON, DAS, FX,
<20 μg/ kg
Zeralenone (ZON) 6000 μg/ kg
HT-2 8500 μg/ kg
T2 80000 μg/ kg
Neosolaniol 600 μg/ kg
Are the Fusarium strains multiplied mycotoxin producer?
•Conclusions:
DON- and NIV-producing Fusarium strains need to be involved.
Inoculum production method have to be improved.
Many thanks to the sponsors:
The European Commission’s ( Council Regulation
(EC) no. 870/2004),
Peter Koelln KGaA,
Emco spol. s r. o.,
Gemeinschaft zur Förderung der privaten
deutschen Pflanzenzüchtung e.V. (GFP)
Meeting additionally supported by the Association of the
Friends and Promotors of the Julius Kühn-Institute
(GFF) and Peter Koelln KGaA