WP 4 Field experiments

with artificial Fusarium

inoculation

Introduction

Fixed conditions

Methods and traits for discussion

Details of methods

AVEQ meeting CF 18.09.07

Salmon pink discoloration of husk und kernels after inoculation with

F. culmorum

AVEQ meeting CF 18.09.07

Field experiments with artificial

Fusarium inoculation

Aim

Evaluation/screening of genetic ressources for resistance to Fusarium ssp. and/or mycotoxin accumulation

These traits are quantitativ inherited and influenced by environment and other traits as plant height, lodging, panicle length, hull content etc.

AVEQ meeting CF 18.09.07

FIOR

SucajevaARIK

FCBR

WP4 Partners

AVEQ meeting CF 18.09.07

WP4 Field experiments with artificial Fusarium

inoculation

Fixed conditions

• 300 Accessions/a.

• 10 standard cultivars,

replicated 5 fold,

(augmented

design)

AVEQ meeting CF 18.09.07

Methods and traits

need to be discussed

- Design and number of replications and

Plot size (2 m2 or less)

- Inoculation method

- Fusarium ssp. for inoculation

- Method of harvesting and sample preparation

AVEQ meeting CF 18.09.07

Traits of evaluation, need to be discussed(Leaf index and number of kernel per spikelet were deleted)

1. heading date,

2. panicle density per m2,

3. crop height,

4. distance between panicle and flag leaf

5. panicle shape,

6. panicle length,

7. infection of panicles with Fusarium

8. lodging at harvest

9. thousand kernel weight (TGW),

10. test weight

11. Fusarium-infection of seeds

12. hull content (HUC)

AVEQ meeting CF 18.09.07

Design, plot size and number of replications

Augmented design or randomised block design ?

Details of methods

Augmented design at four climatically different locations

(Germany, Czech Republik, Romania, Italy)

• Comparisons with standard cv.s should give a good evaluation at least

susceptibility in accessions tested.

• Number of plots manageable and lower border effects in comparison with

smaller plots in a randomised design.

• Accessions grouped for crop height and heading date

AVEQ meeting CF 18.09.07

Inoculation method

• Replicated Spraying of spore suspension over the plots

with a portable sprayer (during or after flowering, 105-106 spores

per ml + 0.1ml/l Tween 20; 50ml/m2 , at 7 p.m., high air humidity, low

wind)

• alternatively/additionally: Scattering infected seeds

(6g/m2, before heading) within the plots (more natural)

• After inoculation and dry weather conditions: Irrigation, if available

Details of methods

AVEQ meeting CF 18.09.07

Inoculum:

Mycotoxin profile dependent from the Fusarium strain and Fusarium

species (no correlation to aggressiveness), high variability within and

between species

Tab. 1 according to Langseth et al. (1999)

Details of methods

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Fusarium species

Isolate NIV DON T-2; HT-2 Zea Monili-formin

culmorum 1163 14 - - 120

culmorum 1028 - 45 - 370

graminearum 1104 - 290 - 1960

poae F13 In other studies

- - 0,04

sporotrichioides 1079-15 - - 72 -

avenaceum 1035-18 - - - 0,03 1100

•Low host specialisation in F. graminearum, F.

culmorum and F. avenaceum

•Adaptation of Fusarium ssp. to local climatic conditions

and different aggressiveness.

Consequence for screening in WP4:

Usage of local defined mixtures of mycotoxin producing strains of F.

graminearum, F. culmorum, F. sporotricioides and F. avenaceum for

inoculation

disadvantage: competition between Fusarium strains results in reduced

aggressiveness and lower mycotoxin accumulation (Midaner et al. 2004)

Details of methods

AVEQ meeting CF 18.09.07

Traits of evaluation, need to be discussed and fixed

• heading date = day in June or July

• leaf index ??

• panicle density per m2, counting 1 m of two rows,

appropriate germination presumed

• crop height in cm (after flowering but before

lodging) Average height of the plants from ground to

top of panicle, excluding awns.

• panicle length in cm

Details of methods

AVEQ meeting CF 18.09.07

Infection of panicles with Fusarium:

Usually there is low or no visual scab, but it have to be

checked. 1=no infection ... 9=all spikelets and panicles

are infected

Lodging prior harvest: 1=no lodging…9= 100%lodging

Number of kernels per spikelet ???

Thousand kernel weight (TGW) (might be reduced due to

Fusarium infection) 3 x 100 unsorted kernel

Test weight in grams per hectoliter

Details of methods

AVEQ meeting CF 18.09.07

Percent Fusarium infected kernels by sorting 3 x 100 kernels (from TKW-measurement)

Three classes:1. sound

2. suspicious infected, grey tips

3. FDK= Fusarium damaged kernels, discoloured, smaller,

Details of methods

AVEQ meeting CF 18.09.07

influence on the assessment of Fusarium infection of seeds:

Color of the lemma as in BSA/IBPGR

Details of methods

AVEQ meeting CF 18.09.07

influence on the assessment of Fusarium infection of seeds:

Color of the lemma as in BSA/IBPGR

hull content (HUC):• Weigh out 100g seeds of equal moisture content,

• air-pressure dehuller, 8 at, 4 min,

• note the weight of kernels and hulls;

• score of undehulled kernels

• Additionally: score of proportion of discoloured groats

Details of methods

AVEQ meeting CF 18.09.07

Method of harvesting and sample preparation:Harvest with plot combine harvester with reduced air cleaning to prevent reduction of Fusarium damaged kernels (FDK)

AVEQ meeting CF 18.09.07

Details of methods