Deoxynivlenol umultion nd fusrium hed light severity in winter whet fter spry-inoultion with mixture or single isoltes of Fusrium grminerum L. Tmuri-Ilini nd A. W. Shfsm. Ridgetown Cmpus, University of Guelph, Ridgetown, Ontrio, NP C, Cnd Corresponding uthor: PH: (59) 67-557, emil: ltmuri@ridgetown.uoguelph. OBJECTIVES ) to ompre ggressiveness of four F. grminerum isoltes nd their mixture sed on FHB severity nd DON umultion in grin, fter spry-inoultion of winter whet ultivrs with known FHB resistne nd ) to test the influene of isoltes, whet ultivr, yer nd their intertions on level of FHB symptoms nd DON umultion to ensure tht there ws no isolte-speifi resistne from different whet ultivrs with respet to DON umultion. INTRODUCTION Fusrium hed light (FHB), used y Fusrium grminerum (Shwe), is n importnt whet disese. In Cnd FHB is used primrily y Fusrium grminerum (Shwe) [teleomorph: Gierell zee Shw. (Peth)]. Aprt from yield nd qulity losses, the most serious onern ssoited with FHB infetion is the ontmintion of the hrvested rop with myotoxins. Deoxynivlenol (DON) is the myotoxin most ommonly reovered from whet grin in Cnd. In Ontrio, losses in winter whet prodution from yield nd qulity redutions, were more thn $ million CAD in 996 (Shfsm, ). Host speifi strins hve not een demonstrted nd resistne to FHB in whet is generlly onsidered horizontl (re-nonspeifi) (Vn Euwijk et l., 995). When reeding for FHB nd DON resistne, reeder often hs to rely on FHB symptoms rther thn to perform ostly DON nlysis. However, FHB severity nd DON ontent re not lwys well orrelted; we hve experiened orreltions, from s low s r=.8, to s high s r=.7, depending on the yer nd level of FHB resistne in ultivrs/lines tested in F. grminerum inoulted nurseries (unpulished dt). Resistne to FHB infetion nd DON ontent my e ontrolled y independent loi or genes (Somers et l., ; Tmuri-Ilini et l., ). MATERIALS AND METHODS Winter whet ultivrs were plnted in mid Otoer in, nd 5 t Ridgetown, Ontrio, Cnd in single rows, m long, sped 7.8 m prt, ontining 7 seeds eh. The experiments were rrnged in 5 x ftoril rndomized omplete lok design with four replitions. There were four ultivrs (FHB suseptile (S) AC Ron nd three FHB modertely resistnt (MR): Wisdom, Vienn nd AC Morley ) nd four isoltes of F. grminerum (DAOM788, DAOM, DAOM nd DAOM) nd mixture of the four. Spry-inoultions of eh row with suspension of 5-ml of mroonidi (5, spores/ml) of eh of four F. grminerum isoltes (-) nd their mixture (5) were done, when primry
whet heds were t 5% nthesis for eh ultivr (Zdoks Growth Stge, ZGS 65) (Zdoks et l., 97) using k-spryer. The suspension ws produed in liquid shke ulture using Bily s medium. F. grminerum isolte # (DAOM788) ws otined from Agriulture Cnd (ECORC, Ottw, Ontrio, Cnd) nd ws isolted from whet. Isoltes # (DAOM), (DAOM), nd (DAOM), were isolted from spring rley vrieties Chpis, AC Stephen, nd C-, respetively in t Elor Reserh Sttion, Guelph, Ontrio, Cnd y Tmuri-Ilini. After isoltion the isoltes were kept in liquid nitrogen nd they hve een sumitted to the Cndin olletion of fungl ultures in Ottw nd re ville s DAOM, DAOM nd DAOM. All the plots were misted dily eginning fter the first inoultions with out 7.5 mm of wter eh dy. The mist system ws engged until three dys fter the lst ultivr ws inoulted. In eh yer, ultivrs were ssessed for visul symptoms when the erly dough stge (ZGS 8) ws rehed. Disese levels were estimted s fusrium hed light severity on sle of -9 where ws disese free nd 9 ws totl. The entire grin smple for eh ultivr ws hrvested in mid July in, 5 nd 6 nd finely ground through ROMER mill (Model A, Romer Ls, In. Union, MO). Deoxynivlenol (DON) ontent ws estimted from the three replitions with highest men FHB severity using quntittive fluorometri test-fluoroqun (Romer Ls, In, Union MO). PROC UNIVARIATE (SAS Institute In., ) ws used to test ANOVA ssumptions to determine if trnsformtions were needed. DON ontent ws trnsformed y log (x+.5). The dt ws nlyzed using PROC GLM (SAS Institute In., ) where yer, ultivr, isolte nd their intertions were soures of vrition. RESULTS AND DISCUSSION FHB severity nd DON umultion in this experiment depended on yer, isolte nd ultivr nd the intertion etween yer nd isolte nd yer nd ultivr, ut there ws no signifint intertion etween isoltes nd ultivrs (Tle ), suggesting tht vrition in resistne in these winter whet ultivrs ws independent of the vrition in ggressiveness of the F. grminerum isoltes tested. The results from the present study re lso in greement with Mesterhzy (997) who lso reported tht DON level in whet depends on oth ultivr nd isolte. FHB severity nd DON ontent in winter whet ultivrs fter inoultion with four isoltes nd their mixture during three yers re shown in Fig. nd Fig.. Aggressiveness of F. grminerum isoltes High isolte x yer intertion ws reported in the present study (Tle ). Averge FHB severity fter spry-inoultion with isoltes # (DAOM), # (DAOM) nd the mixture of ll (5) four ross the ultivrs ws higher in thn in 5 or 6 (Fig. ). Averge DON umultion ross the ultivrs ws lso higher in thn in 5 or 6 (Fig. ). In, fter spry-inoultion with isolte # (DAOM) the DON level ross the ultivrs ws signifintly higher thn fter inoultion with the three other isoltes or the mixture of ll four (Fig. ). The summer of 5 nd 6 ws hot nd dry nd ws not fvorle to DON umultion in winter whet in Ontrio. DON ontent mesured in survey of winter whet in Ontrio in 5 nd 6 rnged from. ppm to. ppm nd from
. ppm to. ppm, respetively, whih ws muh lower thn found in similr survey in, where the highest level of DON ws.9 ppm (Tmuri-Ilini et l., 6). FHB suseptiility nd DON umultion in winter whet ultivrs In nd 5, the verge FHB severity ws the highest for the FHB-suseptile v. AC Ron nd signifintly higher thn three FHB MR ultivrs (Fig. ). In 6, v. AC Ron hd signifintly higher FHB severity thn AC Morley nd Wisdom (Fig. ). AC Ron hd the highest verge DON level fter inoultion in ll three yers; while the reltive rnking of the three MR ultivrs ws slightly different in eh yer (Fig. ). In, Wisdom hd the lowest DON level (Fig. ); Wisdom nd Vienn umulted the lest DON in 5 (Fig. ). There ws no signifint differene in DON umultion mong MR ultivrs in 6 (Fig. ). In onlusion, it is utiously suggest tht perhps the seletion of isoltes my e less importnt when sreening for FHB symptoms thn when sreening for the propensity to umulte DON. Reltively similr FHB rtings etween yers resulted in very different DON levels etween yers. Although inonlusive the dt suggest tht mixture of isoltes might ontriute some stility in results over yers. This possiility would need further study with more ultivrs under more environments nd yers. It seems from the results tht high DON-produing isoltes would llow for differentition mong ultivrs whih differ in their propensity to produe DON, nd re reommended for sreening whet ultivrs for simultneous FHB resistne nd DON umultion. A highly pthogeni isoltes of F. grminerum, tht simultneously produe high level of FHB symptoms nd DON, or mixture of isoltes is reommended for reeding progrms trgeting FHB resistne nd redued DON prodution in winter whet. ACKNOWLEDGEMENTS This projet ws funded y prtnership etween the Cndin Adpttion Counil (CnAdpt) the Ontrio Whet Produers Mrketing Bord nd the Ontrio Ministry of Agriulture nd Food nd Rurl Affirs through the University of Guelph Contrt. Tehnil ssistne y Dine Pul nd Todd Phis is grtefully knowledged. REFERENCES Mesterhzy, A., 997. Breeding for resistne to Fusrium hed light of whet. In H.J. Duin, L. Gilhrist, J. Reeves, nd A. MN (eds), Fusrium Hed Blight: Glol Sttus nd Future Prospets. CIMMYT, Mexio, 79-85. SAS Institute In.. 8 th ed. SAS Institute In. Cry N.C. Shfsm, A. W,. Eonomi hnges imposed y myotoxins in food grins: se study of deoxynivlenol in winter whet. In DeVries, J. W. et l. (ed.), Myotoxins nd Food Sfety. 7-76. Kluwer Ademi/Plenum Pulishers. Somers, D. J., Fedk, G., nd Svrd, M.. Moleulr mpping of novel genes
ontrolling Fusrium hed light resistne nd deoxynivlenol umultion in spring whet. Genome. 6:555-56. Tmuri-Ilini, L., Fedk, G., nd Shfsm, A. W.. Study on deoxynivlenol (DON) nd Fusrium Hed Blight (FHB) resistne in F winter whet popultion. J. Appl. Genet. A: -. Tmuri-Ilini, L., Shfsm, A. W., nd Flk. D. 6. 5 survey for Fusrium hed light of winter whet in S.W. Ontrio. Cn. Plnt Dis. Surv. 86: 87 (www.pssp./pds.htm) Vn Euwijk, F. A., Mesterhzy, A., Kling, C. I., Rukenuer, P., Sur, L., Burstmyr, H., Lemmens, M., Keizer, L.C.P., Murin N., nd Snijders, C. H. A. 995. Assessing non-speifiity of resistne in whet to hed light used y inoultion with Europen strins of Fusrium ulmorum, F. grminerum nd F. nivle using multiplitive model for intertion. Theor. Appl. Genet. 9: -8. Zdoks, J. C., Chng, T. T., nd Konzk, C. F. 97: A deiml ode for the growth stges of erels. Weed Reserh. :5-.
Tle. Anlysis of vrine for the effet of yer, Fusrium grminerum isolte, winter whet ultivr, nd intertions on: ) FHB severity (-9) nd ) trnsformed (log (x+.5)) deoxynivlenol (DON) ontent (ppm) in winter whet. Ridgetown, ON, -, -5 nd 5-6. ) FHB severity (-9) Soure df Men squre F P>F Yer 6.5 9.6 <. Isolte.5.98 <. Cultivr.9.8.98 Yer*Isolte 8.9.8 <. Yer*Cultivr 6.9 9.95 <. Isolte*Cultivr.7.5.59 Yer*Isolte*Cultivr.57.68. ) DON umultion Soure df Men squre F P>F Yer 9. 7. <. Isolte.58 7.5 <. Cultivr 5.7.86 <. Yer*Isolte 8.7 8. <. Yer*Cultivr 6.5.9 <. Isolte*Cultivr..96.9 Yer*Isolte*Cultivr.95.9.567 5
.5 FHB severity (-9).5.5.5 d 5 6.5 5 Isoltes DON (ppm) 8 6 5 6 5 Isoltes Figure.. The effet of Fusrium grminerum isoltes (-) nd their mixture (5) ( + SE) on: ) FHB severity (-9) nd ) deoxynivlenol (DON) ontent (ppm) ross winter whet ultivrs. Ridgetown, ON. Mens within yers followed y the sme letter re not different ording to Fisher s proteted lest signifint differene test (P=.5). 6
FHB severity (-9).5.5.5.5 5 6 AC RON AC MORLEY VIENNA WISDOM Winter whet ultivrs 9 8 DON (ppm) 7 6 5 5 6 AC RON AC MORLEY VIENNA WISDOM Winter whet ultivrs Figure.. The effet of winter whet ultivrs ( AC RON, AC Morley, Vienn nd Wisdom ) on: ) FHB severity (-9) nd ) deoxynivlenol (DON) ontent (ppm) ( + SE) fter spry-inoultion ross Fusrium grminerum isoltes. Ridgetown, ON. Mens within yers followed y the sme letter re not different ording to Fisher s proteted lest signifint differene test (P=.5). 7