WP 4 Field experiments with artificial Fusarium inoculation

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

WP 4 Partners FCBR ARIK FIOR AVEQ meeting CF 18. 09. 07 Sucajeva

WP 4 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 m 2 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 m 2, 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

Details of methods Design, plot size and number of replications Augmented design or randomised block design ? 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

Details of methods Inoculation method • Replicated Spraying of spore suspension over the plots with a portable sprayer (during or after flowering, 105106 spores per ml + 0. 1 ml/l Tween 20; 50 ml/m 2 , at 7 p. m. , high air humidity, low wind) • alternatively/additionally: Scattering infected seeds (6 g/m 2, before heading) within the plots (more natural) • After inoculation and dry weather conditions: Irrigation, if available AVEQ meeting CF 18. 09. 07

Inoculum: Details of methods 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) Fusarium species Isolate NIV DON T-2; HT-2 Zea culmorum 1163 14 - - 120 culmorum 1028 - 45 - 370 graminearum 1104 - 290 - 1960 poae F 13 In other studies - - sporotrichioides 1079 -15 - - 72 avenaceum - - - AVEQ meeting CF 18. 09. 07 1035 -18 Moniliformin 0, 04 0, 03 1100

Details of methods • 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 WP 4: 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) AVEQ meeting CF 18. 09. 07

Details of methods Traits of evaluation, need to be discussed and fixed • heading date = day in June or July • leaf index ? ? • panicle density per m 2, 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 AVEQ meeting CF 18. 09. 07

Details of methods 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 AVEQ meeting CF 18. 09. 07

Details of methods 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, AVEQ meeting CF 18. 09. 07

Details of methods influence on the assessment of Fusarium infection of seeds: Color of the lemma as in BSA/IBPGR AVEQ meeting CF 18. 09. 07

Details of methods influence on the assessment of Fusarium infection of seeds: Color of the lemma as in BSA/IBPGR hull content (HUC): • Weigh out 100 g 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 AVEQ meeting CF 18. 09. 07

Details of methods 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