Response of Soybean Cultivars to Bean pod mottle virus Infection

Transcription

1 Response of Soyen Cultivrs to Ben pod mottle virus Infetion Amy D. Ziems nd Loren J. Giesler, Deprtment of Plnt Pthology, nd George L. Gref, Agronomy nd Hortiulture Deprtment, University of Nersk, Linoln 68583; Mrgret G. Redinugh, United Sttes Deprtment of Agriulture Agriulturl Reserh Servie, Wooster, OH; nd Jen L. Vh, SueAnn Berry, Lurene V. Mdden, nd Anne E. Dorrne, Deprtment of Plnt Pthology, Ohio Stte University, Wooster ABSTRACT Ziems, A. D., Giesler, L. J., Gref, G. L., Redinugh, M. G., Vh, J. L., Berry, S. A., Mdden, L. V., nd Dorrne, A. E Response of soyen ultivrs to Ben pod mottle virus infetion. Plnt Dis. 91: Ben pod mottle virus (BPMV) hs eome inresingly ommon in soyen throughout the north-entrl region of the United Sttes. Yield loss ssessments on southern soyen germplsm hve reported redutions rnging from 3 to 52%. Currently, no soyen ultivrs hve een identified with resistne to BPMV. The ojetive of this study ws to determine the impt of BPMV infetion on soyen ultivrs representing rod rnge of northern soyen germplsm y ompring inoulted nd noninoulted soyen plnts in pired row studies. In ll, 30 nd 24 ultivrs were evluted in Nersk (NE) in whih soyen plnts were inoulted t the V3 to V4 growth stge. Eleven ultivrs from puli nd reeding lines were inoulted t the VC nd R5 to R6 growth stges in Ohio (OH). Disese severity, yield, nd perent seed ot mottling were ssessed t oth lotions, wheres protein nd oil ontent lso were ssessed t NE. Yield nd perent seed ot mottling ws signifintly redued following inoultion t the VC (OH) nd V3 to V4 (NE) growth stges. In ddition, seed oil nd protein omposition were impted in 1 of the 2 yers of the study. This study demonstrtes tht sustntil yield losses n our in soyen due to BPMV infetion. In ddition, protein nd oil my e ffeted depending on the environment during the prodution seson. Ben pod mottle virus (BPMV) (genus Comovirus, fmily Comoviride) first ws identified in soyen (Glyine mx L. Merr.) in 1951 in Arknss (28) nd hs used gronomi prolems sine this first finding in Mississippi (22) nd North Crolin (28). In the north-entrl region of the United Sttes, the first report of BPMV ws during 1968 in Iow (23); more reently, the virus hs een found in the mjority of the sttes in this region (4,5,8,17,18,21). From 1999 to 2004, inresed inidene of BPMV in the northentrl region ws ssoited with inresed popultions nd movement of en lef eetles (BLB; Cerotom trifurt), the primry virus vetor (8). Corresponding uthor: L. J. Giesler E-mil: lgiesler1@unl.edu Mention of trde nmes or ommeril produts in this pulition is solely for the purpose of providing speifi informtion nd does not imply reommendtion or endorsement y the University of Nersk, Ohio Stte University, nd the United Sttes Deprtment of Agriulture. Aepted for pulition 4 Jnury doi: / PDIS This rtile is in the puli domin nd not opyrightle. It my e freely reprinted with ustomry rediting of the soure. The Amerin Phytopthologil Soiety, BPMV infetion ws reported to redue soyen yields of southern ultivrs etween 3 nd52% (6), with the most severe redutions ourring when virus infetion ours erly in soyen development (14). BPMV produes further finnil losses for soyen produers euse it uses inresed seed ot mottling, whih is n irregulr pttern or streking of the hilum ssoited with BPMV infetion (18,38). The effets on yield nd seed ot mottling re inresed when there is dul infetion of BPMV nd Soyen mosi virus, whih uses yield losses >80% (26). Other symptoms ssoited with BPMV infetion re mild hloroti mottling of folige to severe mosi, lef rugosity, terminl nerosis, nd deth (6,27,29,36). Also, dely in soyen mturtion to use symptom known s green stem hs een reported (6,27,29,36); however, it hs een shown reently tht green stem is independent of BPMV infetion when rndom plnts were tested for BPMV t growth stge R6 (13). Current mngement reommendtions for reduing BPMV infetion inlude the pplition of insetiides to mnge BLB popultions to redue the potentil for virus movement (15). Host resistne hs een explored ut, urrently, no soyen lines hve een identified with resistne to BPMV (29,31,39). Trnsgeni soyen lines expressing the BPMV ot protein re resistnt to BPMV infetion (3,24); however, this resistne hs not een inorported into ommeril soyen ultivrs (8). The gol of this reserh ws to exmine the response of soyen ultivrs ville in the north-entrl region of the United Sttes to BPMV infetion y evluting symptom expression nd effets on gronomi trits in Nersk nd Ohio. Preliminry results from these studies hve een pulished (25,40). MATERIALS AND METHODS Ohio evlution of soyen ultivrs to erly nd lte infetion with BPMV. Seven ultivrs nd four reeding lines (designted olletively s ultivrs; Tle 1) were plnted in Wooster silt lom soil t the Ohio Agriulturl Reserh nd Development Center (OARDC), Snyder Frm, ner Wooster in Ten ultivrs were plnted in different field t the sme lotion in No fertilizer ws pplied to the field prior to or during the studies. Pre-emergene heriides in 2001 onsisted of lhlor (Mirotex, 4.6 liters/h; Monsnto Compny, St. Louis) plus linuron (Lorox DF, 2.2 kg/h; Griffin LLC, Vldost, GA) nd surftnt (2.3 liters/h) on the dy of plnting. The post-emergene heriide entzon nd ifluorfen (Storm; United Phosphorus In., Trenton, NJ) ws pplied (1.7 liters/h) on 2 July The insetiide permethrin (Poune, 559 g [.i.]/h; FMC Corportion Ag Produts, Phildelphi) ws pplied on 16 July nd 15 August to limit the mount of inset lef feeding during Lmdyhlothrin (Wrrior, 209 g [.i.]/h; Syngent Crop Protetion, Greensoro, NC) ws pplied on 2, 16, nd 24 July nd 21 August Four 7.62-m rows (38.1 m prt) were plnted in no-till field in split plot design on 14 June 2001 nd 6 June 2003, with soyen ultivr s whole plot in ddition to inoultion time t the unifolite stge (VC) or R6 to R7. BPMV-inoulted nd noninoulted tretments were the suplots. The Ohio isolte of BPMV ws otined from soyen leves nd mintined in Hrosoy seedlings (4). Sequene of reverse-trnsription polymerse hin retion produts mplified from RNA1 (615 p) nd RNA2 (789 p) indite tht oth RNAs hve >98% identity with sugroup II viruses (10; G. L. Redinugh nd M. G. Redinugh, unpulished results). The Plnt Disese / June

2 unifolite lef ws inoulted with lefru inoultion t the unifolite stge (VC) on 28 June 2001 nd 24 June For lte-inoulted plots, the youngest fully expnded lef ws inoulted t the R6 to R7 stge on 6 Septemer 2001 nd 16 Septemer 2003 s desried y Louie et l. (19). To determine inoultion effiieny, 10 lef smples were olleted rndomly from eh plot on 21 Septemer 2001 nd 23 Septemer 2003 (VC inoultion) nd 26 Septemer 2001 nd 29 Septemer 2003 (R6 to R7 inoultion) nd tested using n F() 2 enzyme-linked immunosorent ssy (ELISA) similr to tht desried y Louie et l. (19) using BPMV ntiserum developed t OARDC (D. T. Gordon, unpulished results). Smples were onsidered positive for BPMV if the men sorne t 405 nm for two replites ws greter thn three times the men sorne of the helthy ontrols nd greter thn the men vlue of the helthy ontrols plus three times the stndrd devition of the ontrols (33). Visul ssessment of plots for BPMV symptoms, inluding hlorosis nd rugosity, ws done pproximtely 4 weeks post inoultion (30 July 2001 nd 24 July 2003) or t the R6 to R7 growth stge (5 Septemer 2001 nd 25 Septemer 2003) using visul sle of 1 to 5, where 1 = no symptoms, 2 = some hlorosis present, 3 = hlorosis evident on ll leves in upper Tle 2. Effet of V3 to V4 growth stge inoultion with Ben pod mottle virus (BPMV) on hlorosis nd rugosity expression in soyen ultivrs evluted in Nersk in 2003 Chlorosis Rugosity Cultivr I N I Med N Med I Rel N Rel Stine S Stine S Stine S NK/Syngent S28-W NK/Syngent S37-N NK/Syngent S32-G Myogen 5B283RR Myogen 5B288RR Myogen 5B311RR Lthm 917RR Lthm 967RR Lthm 1067RR LG Seeds C2625RR LG Seeds C2991RR LG Seeds C3110NRR Grst 2603RR Grst 2802RR Grst 3135RR Kup 299R Kup 279R Kup 267R Cropln RC Cropln RC Cropln RT AgriPro 2677RR AgriPro 2903RR AgriPro 3135RR Asgrow Asgrow Dekl I = BPMV-inoulted suplots nd N = noninoulted suplots. Rting sed on 0-to-10 sle, where 0 = 0% re lef hloroti nd 10 = 100% lef re hloroti. nopy, 4 = severe hlorosis with some Tretments were ompred using Fisher s proteted lest signifint differene (LSD; P 0.05); rugosity, nd 5 = severe hlorosis nd inoultion ultivr LSD (α = 0.05) = 0.9. rugosity on the uppermost set of leves. Rting sed on 1-to-5 sle, where 1 = flt leves nd 5 = severely rinkled leves. Med = medin nd Rel = reltive effet, the proility tht the rting of plnt from given ultivr is greter thn the rting of rndomly hosen plnt from popultion of ll ultivrs omined. Differenes of reltive tretment effets loosely represent the mgnitude of the differene of the distriution of two ultivrs (not just the entrl vlue). Tretments were ompred using Fisher s proteted LSD (P 0.05) lulted sed on the stndrd errors for reltive tretment effets for eh ultivr; inoul tion ultivr LSD (α = 0.05) = With the nonprmetri nlysis, there is different stndrd error for every reltive tretment effet; thus, this LSD is tully men LSD for ll possile pirs of ultivrs. During 2001, verge plnt height ws determined y mesuring 10 plnts in eh plot on 30 July. Yield ws determined for ll plots fter mehnil hrvest of the two rows on 5 Novemer 2001 nd 21 Otoer In 2001, 100-seed weights were determined following hrvest. In Tle 1. Effet of VC growth stge inoultion with Ben pod mottle virus on symptom expression of soyen ultivrs in Ohio Cultivr Type Med July Med Sept Rel July Rel Sept Med Sept Rel Sept Define Grin Flint Grin Nt Nt Generl Grin HC94-24 IR HC95-15 IR HF Breeding HS Breeding Resnik Grin Stlwrt Determinnt Tiffin Grin Willims82 Grin Nt Nt LSD (α = 0.05) Med = medin severity nd Rel = reltive effet in July or Septemer (Sept); Nt = not tested, due to very poor nd uneven stnds. Symptom severity ws determined using visul sle of 1 to 5, where 1 = no symptoms nd 5 = severe mottling nd lef rinkling on the uppermost set of leves. Reltive effet is the proility tht the rting of plnt from given ultivr ws greter thn the rting of rndomly hosen plnt from popultion of ll ultivrs omined. Differenes of reltive tretment effets loosely represent the mgnitude of the differene of the distriution of two ultivrs (not just the entrl vlue). Ohio soyen ultivrs nd reeding lines were indeterminnt grin type nd semideterminnt inset-resistnt (IR) reeding lines. Tretments were ompred using Fishers proteted lest signifint differene (LSD; P 0.05), lulted sed on the stndrd errors for reltive tretment effets for eh ultivr. With the nonprmetri nlysis, there is different stndrd error for every reltive tretment effet; thus, the LSD shown is tully men LSD for ll possile pirs of ultivrs. 720 Plnt Disese / Vol. 91 No. 6

3 ddition, 100-seed susmples from eh plot were ounted for the numer of seed exhiiting seed ot mottling. This vlue ws onverted to perent mottled seed. Nersk evlution of soyen ultivrs to erly infetion with BPMV. In ll, 30 (Tle 2) nd 23 (Tle 3) Roundup Redy soyen ultivrs representing the top mrket shres from 10 seed ompnies in Nersk were plnted in Kennee silt lom soil t the University of Nersk Linoln Est Cmpus Reserh Frm in Linoln during 2003 nd 2004, respetively. No fertilizer or heriide ws pplied to the field prior to or during the studies. Weed ontrol ws mnged through rouging. Wrrior (209 g [.i.]/h) ws pplied on 13 nd 29 July 2003 nd 19 July nd 17 August 2004 to suppress BLB popultions. Plots onsisted of three 0.8-m rows on 76-m sping. Plots were plnted in field tht ws disked prior to plnting in split-plot design on 25 My 2003 nd 19 My 2004, with soyen ultivr s the whole plots nd BPMV-inoulted or noninoulted plnts s the suplots. Whole plots were thinned to ommon plnt popultion of 6 to 23 plnts per 0.76 m mong whole plots t growth stge VC to V1 on 10 June 2003 nd 3 June A BPMV isolte (sugroup I; identified y S. Ghril, personl ommunition) originlly olleted from infeted soyen in Nersk ws used s the inoulum soure. The BPMV isolte ws mintined in infeted plnt mteril stored t 20ºC nd y seril trnsfer using mehnil inoultion to Phseolus vulgris L. v. Lndreth. The BPMV inoulum ws prepred from symptomti P. vulgris y lending infeted plnt tissue in M potssium phosphte uffer, ph 7.2 (tissue t 0.1 g/ml). The extrt ws filtered through otton flour sk towels to remove insolule plnt mteril, nd 28 g of Crorundum (Fisher Sientifi, 320 grit) ws dded per liter. Plnts were inoulted with the extrt using n irrush (Model 155 & 175; Bdger Air-Brush Co., Frnklin Prk, IL) t 75 psi. Four leflets per plnt were spryed until wter-soked spot ws pprent on the lef surfe. Inoultion ourred t growth stge V3 to V4 on 2 nd 3 July 2003 nd 7 nd 8 July To determine inoultion effiieny, four nd five new, fully expnded trifoliolte lef smples were olleted from eh plot on 18 August 2003 nd 4 August 2004, respetively, t growth stge R5 to R6. Lef smples were pled into dry 7.6-y 7.6-m mesh gs (Agdi, In.), nd 4 ml of phosphte-uffered sline with Tween (PBST, ph 7.2) ws dded to eh g nd ground using tissue homogenizer (Agdi, In.). Smples were tested using iotinvidin doule-ntiody sndwih ELISA ording to Krell et l. (15) using ntiodies provided y Dr. John Hill (Iow Stte University). Asorne vlues (405 nm) were determined using n OPTImx Tunle Miroplte Reder (Moleulr Devies Corportion, Sunnyvle, CA) nd were onsidered positive if the sorne exeeded twie the men plus 10% of the helthy ontrol (extrt from noninoulted soyen leves; 9). Symptom severity ws ssessed t growth stge R5 on 17 August 2003 nd 10 August 2004 using visul sle to estimte lef hlorosis (0 to 10) nd lef rugosity (1 to 5) s desried y Windhm nd Ross (37). The hlorosis sle used did inlude ny hlorosis ssoited with mosi pttern typil of plnt virus symptoms. Single-row plots were hrvested mehnilly on 17 Otoer 2003 nd 22 Otoer Following hrvest, seed smples were ssessed for moisture ontent, perent oil, nd perent protein ontent with ner-infrred spetrosopy (Infrte 1255 Food nd Feed Anlyzer; Foss, Eden Pririe, MN). All yields, perent oil, nd perent protein vlues were stndrdized to 13% moisture. To determine impt of seed ot mottling, 100-seed susmples from eh plot were ounted for the Tle 3. Effets of inoultion with Ben pod mottle virus (BPMV) t growth stge V3 to V4 on yield, perent mottled seed, perent oil ontent, nd perent protein ontent of soyen ultivrs evluted in Nersk in 2003 Yield (g/plot) Mottled seed (%) Oil ontent (%) Protein ontent (%) Cultivr I N Men I N Men I N Men I N Men Stine S Stine S Stine S NK/Syngent S28-W NK/Syngent S37-N NK/Syngent S32-G Myogen 5B283RR Myogen 5B288RR Myogen 5B311RR Lthm 917RR Lthm 967RR Lthm 1067RR LG Seeds C2625RR LG Seeds C2991RR LG Seeds C3110NRR Grst 2603RR Grst 2802RR Grst 3135RR Kup 299R Kup 279R Kup 267R Cropln RC Cropln RC Cropln RT AgriPro 2677RR AgriPro 2903RR AgriPro 3135RR Asgrow Asgrow Dekl Cultivr LSD I = BPMV-inoulted suplots nd N = noninoulted suplots. For perent mottling, 100-seed susmple from eh plot ws ounted for the numer of seed exhiiting seed ot mottling, nd this vlue ws onverted to perent mottled seed. LSD = lest signifint differene (α = 0.05). Plnt Disese / June

4 numer of seed exhiiting seed ot mottling, nd this vlue ws onverted to perent mottled seed. The 100-seed susmple lso ws ssessed for overll seed qulity on 1-to-5 sle with 1 = 0% seed re mottled nd 5 = 76 to 100% seed re mottled (41). Sttistil nlysis. A nonprmetri mrginl-effets nlysis (1,30) ws used to determine the effets of ultivr on the ordinl folir symptom severity rtings in Ohio nd rugosity rtings in Nersk. Assessment time lso ws nlyzed in Ohio for 2001 dt. The so-lled reltive tretment effet ( sled version of the men rnk of severity; 1,30) ws lulted s the summry sttisti for ompring mong ultivrs. The reltive tretment effet is the proility tht the rting of plnt from given ultivr is greter thn the rting of rndomly hosen plnt from popultion of ll ultivrs omined. Differenes of reltive tretment effets loosely represent the mgnitude of the differene of the distriution of two ultivrs (not just the entrl vlue). Anlysis of vrine (ANOVA) ws used for the nlysis of disese inidene nd plnt height for plnts inoulted t growth stge VC in Ohio. Pro GLM of SAS (SAS Institute, Cry NC) ws used for the ANOVA. ANOVA lso ws used to test the effets of ultivr nd inoultion t eh inoultion time (erly nd lte) on hlorosis rting, perent mottled seed, yield, perent oil, nd perent protein ontent. In the nlysis of yield in Nersk, whole-plot plnt popultion served s ovrint vrile. The hlorosis rting, perent mottled seed, yield, perent oil, nd perent protein ontent in the noninoulted nd inoulted suplots were nlyzed using inoultion nd ultivr s min effets. All tretments were ompred y Fisher s proteted lest signifint differene (P < 0.05). Correltions etween the perent mottled seed, yield, perent oil, nd perent protein ontent were lulted using pointseril orreltion oeffiient s desried y Gions (7). RESULTS Effiieny of virus inoultions. In Ohio, 97% of the soyen plots inoulted t the VC growth stge were ll positive for BPMV, exept one plot of v. Stlwrt in 2001 nd one of v. Generl in 2003, wheres 52% of plots whih were inoulted t the R6 to R7 growth stge tested positive for BPMV y ELISA in eh yer. In Nersk, the inoultion effiieny for BPMV ws 96% in In 2004, inoultion effiieny deresed, with 44% of the lef mteril smpled testing positive for BPMV. Some plnts olleted from noninoulted plots in oth studies tested positive for BPMV. Even when folir symptoms were not present in ny of the noninoulted plots, BPMV ws deteted in 50 nd 14% of noninoulted plots t the VC nd R6 to R7 stges, respetively, during In 2003, BPMV ws deteted in 82 nd 2.5% of the noninoulted plots, in Ohio. For Nersk, 70 nd 29% of the noninoulted lef mteril tested positive for BPMV t growth stge R5 to R6 in 2003 nd 2004, respetively. Symptom severity. In the Ohio study, where BPMV sugroup II ws inoulted, symptom severity ws signifintly lower in plnts rted erly in the seson in 2001, nd the intertion etween rting dte nd ultivrs ws signifint (P < 0.05), sed on the nonprmetri mrginl-effets nlysis (30; Tle 1). In generl, the estimted reltive tretment effets were lower for July thn for Septemer, nd the orresponding medin severity rtings (not di- Tle 4. Anlysis of vrine for soyen ultivrs inoulted with Ben pod mottle virus t the unifolite nd R6 to R7 growth stges in Ohio nd for soyen inoulted t V3 to V4 growth stge in Nersk Mens squres Lotion, yer, soure df Yield Perent mottled seed Perent oil ontent Perent protein ontent Ohio 2001 Replition (Rep) * * Cultivr (C) * 2,335.81*** C Rep Inoultion (I) *** 9,077.82*** C I Growth stge (G) *** 15,319.11*** C G C (rep G) C I G *** 1,099.22*** 2003 Rep *** * C *** *** C Rep I *** 1,374.76*** C I G *** 3,468.91*** C G *** C (rep G) C I G *** *** Nersk 2003 Rep 3 27,165.12*** 1,188.30*** 5.71*** 1.60 C 29 8,199.05*** 1,371.70*** 1.94*** 8.14*** C Rep 87 5,338.88*** ** 0.83 I 1 133,084.72*** 3,059.61*** 15.22*** 71.53*** I C 29 1, Rep 5 20,190.22*** 20,79.62* 0.82** 12.21*** C 22 10,125.16*** 1,087.92** 1.71*** 4.47*** C Rep 114 2,064.11* * 0.85 I 1 7,357.65* * 1.97 I C Anlysis of vrine ws nlyzed using Pro GLM proedure in SAS (SAS Institute, Cry, NC). Vlues followed y *, **, nd *** indite signifine t P 0.05, P 0.01, nd P < , respetively. Dt on oil nd protein ontent were not olleted in Ohio. 722 Plnt Disese / Vol. 91 No. 6

5 retly used in the nlysis) likewise were lower for July thn for Septemer. At the lter rting dte, there were severl differenes found from the July ssessment. There ws no ler greement in the reltive tretment effets of the ultivr for the two ssessments. There ws signifint effet of ultivr on the reltive tretment effets of severity rtings in 2003 (Tle 1), lthough most ultivrs were not different from eh other. Plots inoulted t growth stge R6 to R7 exhiited only mild hlorosis (dt not shown). Inoultion signifintly (P < 0.001) redued plnt height in plots inoulted t VC ompred with the noninoulted ontrol plots for oth yers (dt not shown). In Nersk, where BPMV sugroup I ws inoulted, there ws not signifint intertion of ultivr nd inoultion or ultivr lone (P < 0.05) on rugosity rtings sed on the nonprmetri mrginl effets nlysis (30) of the dt in 2003 (Tle 2). There ws signifint inrese in the reltive tretment effets for hlorosis rtings mong ultivrs inoulted with BPMV in 2003 ompred with not eing inoulted (Tle 2), with AgriPro 2903RR exhiiting the most hlorosis. However, in 2004, no signifint differenes in symptoms were found (dt not shown). Rugosity rtings rnged from 1.0 to 1.2 for oth inoulted nd noninoulted plots, while hlorosis rtings rnged from 1.0 to 1.3 for noninoulted plots nd from 1.0 to 1.7 for inoulted plots (dt not shown). Effet of BPMV infetion on yield, seed omposition, nd seed qulity. In Ohio, BPMV inoultion produed signifint redutions in yield (Tle 4). Suplots inoulted t VC hd 22 nd 29% redution in yield reltive to noninoulted suplots, wheres plnts inoulted t growth stge R6 to R7 showed 0.5 nd 5.5% redution in yield in 2001 nd 2003, respetively. (Tle 4). In ontrst, only Resnik nd Generl (2003) showed signifintly lower yield when inoulted t R6 to R7. Nonetheless, BPMV inoultion t the VC growth stge signifintly redued yield in oth yers ross ultivrs (Tle 5). In Nersk, there ws not signifint ultivr tretment intertion for yield in the nlysis of vrine for either yer (Tles 3, 6, nd 7). As in Ohio, BPMV inoultion signifintly redued yield in oth yers ross ultivrs y 11 nd 3% in 2003 nd 2004, respetively (Tle 5). BPMV inoultion ffeted seed omposition for soyen ultivrs evluted in Nersk in 2003, with differenes etween noninoulted nd inoulted plots in protein nd oil ontent. There ws no signifint ultivr tretment intertion (Tle 4). In 2004, there ws no signifint effet on perent protein ontent. However, there ws signifint derese in oil ontent in seed from inoulted plots. For 2003 nd 2004, there ws n overll redution of perent oil ontent y 0.5% in inoulted plots, with proportionl inrese in protein ontent of 0.9% in the sme plots (Tle 6). Seed ot mottling ws oserved in plots of inoulted nd noninoulted soyen in ll yers nd lotions. In Ohio, the proportion of mottled seed ws signifintly higher for ll entries exept Tiffin nd HC95-24 in suplots inoulted t VC in 2001 (Tle 4). In suplots inoulted t R6 to R7, there ws no signifint inrese in seed ot mottling mong entries etween noninoulted nd inoulted; how- Tle 6. Men tretment effets of noninoulted nd inoulted t the VC nd V3 to V4 growth stge with Ben pod mottle virus on soyen ultivrs in Ohio nd Nersk, respetively Lotion, Mottled seed Oil ontent Protein yer Tretment Yield (%) (%) ontent (%) Ohio 2001 Noninoulted BPMV-inoulted P vlue < < Noninoulted BPMV-inoulted P vlue < < Nersk 2003 Noninoulted BPMV-inoulted P vlue < < < < Noninoulted BPMV-inoulted P vlue For perent mottling, 100-seed susmple from eh plot ws ounted for the numer of seed exhiiting seed ot mottling, nd this vlue ws onverted to perent mottled seed. Yield presented in kg/plot. Yield presented in g/plot. Tle 5. Yield nd perent mottled seed for soyen ultivrs evluted in Ohio inoulted with Ben pod mottle virus (BPMV) t the unifolite nd R6 to R7 growth stges in the field in two-row plots (7.6 y 19 m) during 2001 nd Yield (kg/plot) Mottled seed (%) Yield (kg/plot) Mottled seed (%) Unifolite R6 R7 Unifolite R6 R7 Unifolite R6 R7 Unifolite R6 R7 Cultivr I N I N I N I N I N I N I N I N Define Flint Generl HF HS Resnik Tiffin Stlwrt HC HC Willims Nt Nt Nt Nt Men Two-wy LSD NS Three-wy LSD I = BPMV-inoulted suplots, N = noninoulted suplots, nd Nt = not tken, due to very poor nd uneven stnds. For perent mottling, 100-seed susmple from eh plot ws ounted for the numer of seed exhiiting seed ot mottling, nd this vlue ws onverted to perent mottled seed. Tretments were ompred using Fisher s proteted lest signifint differene (LSD; P 0.05; α = 0.05); two-wy intertion etween inoulted nd noninoulted nd three-wy intertion mong ultivr inoultion growth stge; NS = not signifint. Plnt Disese / June

6 ever, BPMV inoultion used signifint inrese in the perentge of seed mottling ross entries (Tle 5). In Nersk, there ws signifint inrese in mottled seed in the inoulted plots ompred with the noninoulted plots in 2003 (Tles 3 nd 7). In 2004, there were more mottled seed in the inoulted plots ompred with noninoulted plots, ut these differenes were not signifint (Tle 7). In 2003, there were signifint differenes (P < 0.05) in seed qulity with noninoulted plots hving etter overll seed qulity rting (dt not shown). The iseril orreltions (Tle 8) indited tht the presene of BPMV hd n effet on yield nd other gronomi properties of soyen in three out of the four studies. In Ohio during 2001 nd 2003, yield nd perent mottled seed were signifintly (P < 0.001) lower in plnts with BPMV thn without. However, in 1 yer in Nersk (2003), yield ws tully higher (P < 0.001) when BPMV ws present thn when BPMV ws sent in the iseril orreltion; in the other yer in Nersk (2004), there ws no signifint effet. DISCUSSION Yields of soyen inoulted t the VC (Ohio) nd V3 (Nersk) growth stge with BPMV were signifintly lower thn noninoulted plots. Although there ws rnge in yield redution mong evluted soyen ultivrs nd yers, the overll redution ws 16.3% ross lotions nd yers. These yield redutions oserved in this study were not s high s previously reported (6,14). Although the noninoulted plots were symptom free, BPMV ws identified in the noninoulted ontrols in oth yers nd lotions; thus, this my e ontriuting ftor. However, it is prole tht vriility in yield mong the lrge numer of ultivrs used in this study msked redutions in yield tht n e deteted with single or few ultivrs. It lso is possile tht reeding progrms hve improved tolerne to BPMV sine those erlier studies. Breeding progrms do not typilly mke seletions for BPMV; however, y ulling out poorqulity seed nd lower-yielding lines, BPVM tolerne ould hve een seleted in yers with high BPMV inidene. Differenes in seed ot mottling were orrelted to BPMV infetion in this study. Krell et l. (16) found tht virus ntigen levels in seed were not orrelted to the mount of seed ot mottling; therefore, they suggested not using seed ot mottling s n inditor of soyen ultivr sensitivity to BPMV infetion. Other results indite tht the level of seed ot mottling is strongly influened y genotype nd other environmentl stresses, espeilly temperture (2,12,32,34,35). Thus, lthough seed ot mottling is strongly ssoited with BPMV infetion, it is importnt to ount for other vriles tht ffet seed ot mottling development when ssessing soyen ultivrs for tolerne or resistne to BPMV. The dt from Nersk demonstrte tht BPMV infetion ffets seed oil nd protein omposition. In urrent mrkets, high oil nd protein ontent re desired for interntionl trde of soyen produts. Tle 8. Biseril orreltions etween presene of Ben pod mottle virus (BPMV) on soyen yield, perent mottled seed, perent oil ontent, nd perent protein ontent in plots inoulted nd noninoulted with BPMV Lotion, yer Yield Perent mottled seed Perent oil ontent Perent protein ontent Ohio *** 0.188*** *** 0.349*** Nersk *** 0.048** 0.092*** * Correltions etween oth the perent seed ot mottling nd yield with the presene or sene of BPMV in the plots ws lulted using point-seril orreltion oeffiient s desried y Gions (7). Vlues followed y *, **, nd *** indite signifint differenes t P 0.05, 0.01, nd 0.001, respetively. For perent mottling, 100-seed susmple from eh plot ws ounted for the numer of seed exhiiting seed ot mottling, nd this vlue ws onverted to perent mottled seed. Tle 7. Effets of inoultion with Ben pod mottle virus (BPMV) t growth stge V3 to V4 on yield, perent mottled seed, perent oil ontent, nd perent protein ontent of soyen ultivrs evluted in Nersk in 2004 Yield (g/plot) Mottled seed (%) Oil ontent (%) Protein ontent (%) Cultivr I N Men I N Men I N Men I N Men Stine S Stine S NK/Syngent S32-G NK/Syngent S327R Myogen 5B288RR Myogen 5B311RR Myogen 5B242RR Lthm E2857R Lthm E2900R LG Seeds C2991RR LG Seeds C3031RR LG Seeds C3110NR Kup 299R Kup 279R Kup 2748R Cropln RT Cropln RT Cropln RT Asgrow Asgrow Dekl DKB Midwest Seed Genetis GR Midwest Seed Genetis GR Cultivr LSD (α = 0.05) I = BPMV-inoulted suplots nd N = noninoulted suplots. For perent mottling, 100-seed susmple from eh plot ws ounted for the numer of seed exhiiting seed ot mottling, nd this vlue ws onverted to perent mottled seed. LSD = lest signifint differene. 724 Plnt Disese / Vol. 91 No. 6

7 For exmple, premiums re pid in Asin mrkets for soyen seed tht ontin minimum of 19% oil nd 35% protein. Here, the BPMV infetion on soyen proessing is twofold nd ould either inrese or derese profits. For exmple, the 0.5% redution in oil ontent indued y BPMV ould negte prie premiums for produers. On the other hnd, the protein inrese ssoited with BPMV infetion (0.9%) ould inrese soyen protein ontent to the optiml level of 35% or higher nd inrese profits. Nonetheless, the overll derese in yield ssoited with BPMV infetion my negte ny inrese due to high protein ontent. The overll impt of BPMV on soyen ultivrs tht re eing mrketed s Proessor Preferred (high oil nd protein ontent) is unknown t this time. BPMV ws deteted in the noninoulted plots t oth lotions despite efforts to ontrol virus vetors. Smpling to evlute infetion within suplots ws done lter in the seson nd ppers to e ssoited with the unsuessful ontrol of lef-feeding insets in the field plots. For the studies performed in Ohio, there ws evidene of some feeding y Mexin en eetles (Epilhn vrivestis Mulsnt). In Nersk, BLB, nded uumer eetles (Diroti ltet LeConte), spotted uumer eetles (D. undeimpuntt howrdi Brer), nd western orn rootworm eetles (D. virgifer virgifer LeConte), ll vetors of BPMV, were identified (6,20). Despite the inility to mintin noninoulted plots t 0% BPMV inidene, there were signifint differenes oserved etween inoulted nd noninoulted plots. In Ohio, BPMV inoultion t the VC growth stge resulted in 25.5% yield redution. A smller redution (2.5%) ws found in plnts inoulted t R6 to R7. These results re omprle with those of Hopkins nd Mueller (10), who demonstrted tht inoultion of vs. Brgg nd Lee 74 t growth stge V1 used 44.3 nd 13.7% yield redution, respetively. Interestingly, n intermedite redution in yield (7%) ws found for plnts inoulted t growth stge V3 to V4 in Nersk despite the differene in lotion nd BPMV strin type. The work y Windhm nd Ross (37) demonstrted tht inoultion of vs. Centennil nd Rnsom must our prior to growth stge V6 for signifint redutions in yield. The greter effet of erly virus inoultion on yield redution is ritil for developing mngement strtegies for produers. The gronomi effets of inoultion prior to the V6 growth stge lso my ply role in interprettion of survey results. In Nersk, survey onduted in 2001 nd 2002 showed tht BPMV infetion ws present in 54 nd 91% of the fields tested nd tht the within-field inidene rnged from 0 to 90% nd 0 to 100% in 2001 nd 2002, respetively (9). However, the plnt tissue ws olleted in lte July nd August t growth stge R2 to R5, signifintly eyond the V6 growth stge. Future surveys to determine the inidene of BPMV t soyen vegettive stges oupled with BLB popultion ssessments would provide informtion for mngement progrms ssoited with erly ontrol of BLB popultions nd provide etter ssessment of potentil impt on prodution. Current mngement strtegies to redue the impt of BPMV for soyen produers inlude the use of insetiide seed tretments (42) nd folir insetiide pplitions (15). The use of soyen ultivrs resistnt or tolernt to BPMV infetion would e the most prtil pproh for produers to mnge this prolem; however, geneti mteril tolernt to BPMV is not urrently ommerilly ville. However, the rnge of response oserved mong the soyen ultivrs used in this study suggests tht tolerne to BPMV my exist in soyen germplsm. Another possiility for BPMV mngement might e the use of soyen ultivrs tht re resistnt to inset feeding (11). By preventing feeding y vetors like BLBs, the spred nd inidene of BPMV ould e redued ACKNOWLEDGMENTS Funding for these studies ws reeived in prt from the Nersk Soyen Bord, North Centrl Soyen Reserh Progrm, Ohio Seed Improvement Assoition, Ohio Soyen Counil, nd the Ohio Agriulturl Reserh nd Development Center Reserh Enhnement Competitive Grnts Progrm. Slries nd reserh support were provided y Stte nd Federl funds pproprited to the University of Nersk nd The Ohio Agriulturl Reserh nd Development Center, The Ohio Stte University. The Nersk uthors wish to thnk the following seed ompnies: AgriPro, Asgrow, Cropln Genetis, Grst, Kup, Lthm, LG Seeds, Midwest Genetis, Myogen, Stine, nd Syngent for providing the soyen seed for this study; E. Colfx, K. Gillispie, M. Stmm, S. Wtson, T. Weissling, nd S. Werner for ssistne with virus inoultions; nd L. Corte nd D. White for field preprtion, plnting, nd hrvesting. The Ohio uthors wish to thnk R. Anderson for ssistne with virus purifition; J. At, J. Brnes, M. Jones, H. Ji, K. Bluemel, nd M. Veg-Snhez for ssistne with virus inoultions; D. Crpio nd E. Helliwell for ssistne with ELISAs; D. Crpio for ssistne with sutype hrteriztion; nd B. Brdll nd R. Berry for field preprtion, plnting, nd hrvesting. LITERATURE CITED 1. Brunner, E., nd Prui, M Nonprmetri methods in ftoril designs. Stt. Pp. 42: Cooper, R. L A mjor gene for resistne to seed ot mottling in soyens. Crop Si. 6: Di, R., Purell, V., Collins, G. B., nd Ghril, S. A Prodution of trnsgeni soyen lines expressing the en pod mottle virus ot protein preursor gene. Plnt Cell Rep. 15: Dorrne, A. E., Gordon, D. T., Shmitthenner, A. F., nd Gru, C. R First report of Ben pod mottle virus in soyen in Ohio. Plnt Dis. 85: Doughty, D. M., Lee, M. E., Kurtzweil, N. C., Boeroom, C. M., nd Gru, C. R Ourrene of soyen viruses nd ssoition with green stem of soyen in Wisonsin. (Astr.) Phytopthology 91:S Gergerih, R. C Comoviruses: Ben pod mottle omovirus. Pges in: Compendium of Soyen Diseses, 4th ed. G. L. Hrtmn, J. B. Sinlir, nd J. C. Rupe, eds. Amerin Phytopthologil Soiety, St. Pul, MN. 7. Gions, J. D Nonprmetri Methods for Quntittive Anlysis. Holt, Rinehrt nd Winston, New York. 8. Giesler, L. J., Ghril, S. A., Hunt, T. E., nd Hill, J. H Ben pod mottle virus: A thret to U.S. soyen prodution. Plnt Dis. 86: Giesler, L. J., nd Ziems, A. D Inidene of Alflf mosi virus, Ben pod mottle virus nd Soyen mosi virus in Nersk soyen fields. Plnt Helth Progress doi: /php hm. 10. Gu, H., Clrk, A. J., de S, P. B., Pfeiffer, T. W., nd Ghril, S. A Geneti diversity mong isoltes of Ben pod mottle virus. Phytopthology 92: Hmmond, R. B., Biermn, P., Levine, E. nd Cooper, R. L Field resistne of two soyen germplsm lines, HC95-15MB nd HC95-24MB, ginst en lef eetle (Coleopter: Chrysomelide), western orn rootworm (Coleopter: Chrysomelide), nd Jpnese eetles (Coleopter: Sride). J. Eon. Entomol. 94 (6): Hos, H. A., Hrtmn, G. L., Wng, Y., Hill, C. B., Bernrd, R. L., nd Pedersen, W. L Ourrene of seed ot mottling in soyen plnts inoulted with Ben pod mottle virus nd Soyen mosi virus. Plnt Dis. 87: Hos, H. A., Hill, C. B., Gru, C. R., Kovl, N. C., Wng, Y., Pedersen, W. L., Domier, L. L, nd Hrtmn, G. L Green stem disorder of soyen. Plnt Dis. 90: Hopkins, J. D., nd Mueller, A. J Effet of en pod mottle virus on soyen yield. J. Eon. Entomol. 77: Krell, R. K., Pedigo, L. P., Hill, J. H., nd Rie, M. E Ben lef eetle (Coleopter: Chrysomelide) mngement for redution of Ben pod mottle virus. J. Eon. Entomol. 97: Krell, R. K., Pedigo, L. P., Rie, M. E., Westgte, M. E., nd Hill, J. H Using plnting dte to mnge Ben pod mottle virus in soyen. Crop Prot. 24: Lnghm, M. A., Doxtder, D. C., Smolik, J. D., nd Sott, R. A Outrek of virl disese ffeting soyens [Glyine mx (L.) Merrill] in South Dkot. (Astr.) Phytopthology 89:S Lin, M. T., nd Hill, J. H Ben pod mottle virus: ourrene in Nersk nd seed trnsmission in soyens. Plnt Dis. 67: Louie, R., Redinugh, M. G., Gordon, D. T., At, J. J., nd Anderson, R. J Mize neroti strek virus, new mize virus with similrity to speies of the fmily Tomusviride. Plnt Dis. 84: Mry, T. R., Hos, H. A., Steinlge, T. A., Johnson, B. B., Pedersen, W. L., Spener, J. L., Levine, E., Isrd, S. A., Domier, L. L., nd Hrtmn, G. L Distriution of leffeeding eetles nd Ben pod mottle virus (BPMV) in Illinois nd trnsmission of BPMV in soyen. Plnt Dis. 87: Milrth, G. M., MLughlin, M. R., nd Goodmn, R. M Identifition of en pod mottle virus from nturlly infeted soyens in Illinois. Plnt Dis. Rep. 59: Pitre, H. N., Ptel, V. C., nd Keeling, B. L Distriution of en pod mottle disese Plnt Disese / June

8 of soyen in Mississippi. Plnt Dis. Rep. 63: Quiniones, S. S., Dunlevy, J. M., nd Fisher, J. W Performne of three soyen vrieties inoulted with soyen mosi virus nd en pod mottle virus. Crop Si. 11: Reddy, M. S. S., Ghril, S. A., Redmond, C. T., Dinkins, R. D., nd Collins, G. B Resistne to Ben pod mottle virus in trnsgeni soyen lines expressing the psid polyprotein. Phytopthology 91: Redinugh, M. G., Vh, J., Berry, S. A., nd Dorrne, A. E Comprison of erly nd lte inoultions of soyen with Ben pod mottle virus. (Astr.) Phytopthology 93:S Ross, J. P Intertion of the soyen mosi nd en pod mottle viruses infeting soyens. (Astr.) Phytopthology 53: Ross, J. P Effet of single nd doule infetion of soyen mosi nd en pod mottle viruses on soyen yield nd seed hrters. Plnt Dis. Rep. 52: Ross, J. P., nd Butler, A. K Distriution of en pod mottle virus in soyens in North Crolin. Plnt Dis. 69: Shwenk, F. W., nd Nikell, C. D Soyen green stem used y en pod mottle virus. Plnt Dis. 62: Shh, D. A,. nd Mdden, L. V Nonprmetri nlysis of ordinl dt in designed ftoril experiments. Phytopthology 94: Skotlnd, C. B Ben pod mottle virus of soyens. Plnt Dis. Rep. 42: Srinivsn, A., nd Arihr, J Soyen seed disolortion nd rking in response to low tempertures during erly reprodutive growth. Crop Si. 34: Sutul, C. L., Gillet, J. M., Morrissey, S. M., nd Rmsdell, D. C Interpreting ELISA dt nd estlishing the positive-negtive threshold. Plnt Dis. 70: Tkhshi, R., nd Ae, J Geneti nd linkge nlysis of low temperture-indued rowning in soyen seed ots. J. Hered. 85: Tkhshi, R., nd Asnum, S Assoition of T genes with hilling tolerne in soyens. Crop Si. 36: Wlters, H. J A virus disese omplex in soyens in Arknss (Astr.). Phytopthology 48: Windhm, M. T., nd Ross, J. P Phenotypi responses of six soyen ultivrs to en pod mottle virus infetion. Phytopthology 75: Zumeyer, W. J., nd Thoms H. R Pod mottle, virus disese of ens. J. Agri. Res. 77: Zheng, C., Chen, P., Hymowitz, T., Wikizer, S., nd Gergerih, R Evlution of Glyine speies for resistne to Ben pod mottle virus. Crop Prot. 24: Ziems, A. D., Giesler, L. J., nd Gref, G. L Commeril soyen vriety response to Ben pod mottle virus infetion. (Astr.) Phytopthology 95:S Ziems, A. D., Giesler, L. J. Gref, G. L., nd Lne, L. C Effet of Ben pod mottle virus on soyen seed qulity. (Astr.) Phytopthology 91:S Ziems, A. D., Giesler, L. J., nd Hunt, T. E Entomologil Soiety of Ameri Annul Meeting. Fort Luderdle, FL. Entomologil Soiety of Ameri, Lnhm, MD. 726 Plnt Disese / Vol. 91 No. 6