EFFECT OF FOLIAR APPLICATION OF SELENIUM ON NUTRIENT CONCENTRATION AND YIELD OF COLORED- GRAIN WHEAT IN CHINA

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1 EFFET OF FOLIR PPLITION OF SELENIUM ON NUTRIENT ONENTRTION ND YIELD OF OLORED- GRIN WHET IN HIN XI, Q. 1 YNG, Z. P. 1 XUE, N. W. 2 DI, X. J. 1 ZHNG, X. 1 GO, Z. Q. 1* 1 ollege of griulture, Shnxi griulturl University, Tigu, hin 2 Shool of Life Sienes, Dtong University, Dtong, hin *orresponing uthor e-mil: gozhiqing1964@126.om (Reeive 8 th Nov 218; epte 25 th Jn 219) strt. fiel experiment ws onute to evlute the effiieny of folir pplition of ifferent onentrtions of Se fertiliztion (37.5, 56.25, 75., 93.75, n g h -1 of Se) t the erly grin filling stge on nutritionl qulity of olore-grin whet. The results showe tht Se plys role in influening the nutrients onentrtion of olore-grin whet. When it ws more thn 75. g h -1, the Se onentrtion in the ommon whet ws exessive. However, when it ws more thn 37.5 g h -1, the Se onentrtion in the olore-grin whet ws exessive, initing tht olore-grin whet ws likely to e more sensitive to Se umultion. Se onentrtion of olore-grin whet ws signifintly inrese uner the optiml folir pplition of Se onentrtion t 37.5 g h -1, erese strh onentrtion, without ffeting the yiel. Prtiulrly, the onentrtions of gliin n glutenin were signifintly inrese while oth lumin n gloulin were reue with the inrese Se onentrtion. itionlly, our reserh showe tht folir pplition Se inrese the onentrtion of iron (Fe), zin (Zn), ut erese the onentrtion of opper (u) n mngnese (Mn). More importntly, the intke of Se promote the proution of mino is n nthoynins. This inites the potentil roles Se plys on regulting nutrition vlue in olore-grin whet. Keywors: Se-enrih, totl protein, minerls, mylse, mylopetin Introution Hien hunger or mironutrient efiieny ours when the foo qulity oes not meet the requirements of essentil nutrient nee for the growth n evelopment of humns. The min use of hien hunger is the efiieny of minerls (for exmples, selenium, zin, iron, n ioine), lk of vitmins in iet, n lrge onsumption of stple foo (whet, rie, n mize) whih lk these nutrients (Zho et l., 216). Hien hunger is ffeting pproximtely 2 illion people roun the worl inluing 3 million people in hin (Toppe n homo, 215; Kenney et l., 23; Merly et l., 1994). Mny hroni iseses, inluing ietes, riovsulr isese, ner, n oesity were linke to the imlne or efiieny of nutrient intke. Selenium (Se) is n essentil tre element for humns n nimls. Numerous previous stuies hve proven it is enefiil to plnts, however, it is onsiere nonessentil for plnt growth n evelopment. Se plys n importnt role in the growth, evelopment, n mintenne of the immune system of humn n nimls (Shenkin, 26; Yv et l., 25; Spekmnn n Grune, 215). Se efiieny in the humn oy oul use out 4 kins of iseses, suh s Keshn n Kshin-ek iseses (Tn et l., 22; Zhng et l., 214; Fernnes n Gnin, 215; hen et l., 212; Kolesnikov et l., 215). In 1982, it ws reporte tht lmost 72% of regions in hin re fing the Se efiieny n out two-thirs of the popultion re experiening

2 efiieny of Se intke. Thus, the iofortifition tehnology for inresing Se onentrtion in foo rop hs een pi more n more ttentions y mny reserhers (M et l., 28; Khnm n Pltel, 216). Fertiliztion is short term n n effetive metho for improving mironutrient onentrtions in foo rop (Zho n MGrth, 29; Welh, 28). Se fertiliztion is pplie in soil n/or s folir pplition (Wng et l., 217). Folir fertiliztion is the most effetive n sfest wy to inrese the sorption of essentil mironutrients in grin. Whet is the most populr erel rops worlwie n is stple foo for nerly hlf of the popultion in hin where more thn 85% of whet is onsume s flour-erive prouts (Shewry, 29; Li, 26). Therefore, inresing Se nutrition in whet grin is of the utmost importne when it omes to improving the nutritionl sttus of people in hin n other ountries (Liu, 27). In Hungry, olore-grin whet ultivrs re less known in ontrst to Est-fri, New Zeln, n, n hin, for exmple, where suh forms our in ultivte vrieties (Vrg et l., 213). Whet vrieties with speil olore-grin suh s lue n purple grins h een proven to e rih in protein, mino is, n tre elements or minerls (Zeven, 1991). It is well known tht nthoynins re funtioning s ntioxints n, in ition, they lso hve nti-rinogeni effets. Similrly, Se hs nti-rinogeni pilities. euse of these helth enefits, the omintion of olore-grin whet n Se is more enefiil to the evelopment of foo inustry. Thus, olore-grin whet is onsiere to e enefiil to the humn oy ue to their high nutritionl vlue n hs eome speil rw mteril for foo inustry (He et l., 212). In ition, it ws reporte tht olore-grin whet ws riher in Se thn ommon whet, suggesting prtil wy to solve Se efiieny (Shi, 26). t present, there is little known out the enhnement of tre elements in oloregrin whet n Se-enrihe olore-grin whet hs not een fully utilize. The purpose of this stuy is to evlute the effet of iofortifition of Se in olore-grin whet in Se efiient region of Northern hin. In the present stuy, fiel experiment ws onute to (i) investigte the uptke of Se onentrtion in grins from three ifferent vrieties of olore-grin winter whet in the mile re of Shnxi provine in hin, (ii) evlute the effet of folir pplitions of Se on yiel, protein onentrtion, the onentrtion of Se n other minerls, strh n mino i onentrtion of the grin, n (iii) exmine the reltionship etween Se n nthoynin. Mterils n methos Fiel site esription The fiel experiment ws rrie out from Septemer 216 to June 217 in n experimentl site t Shnxi griulturl University, Tigu (37 42 N, E), Shnxi provine, hin (Fig. 1). The ltitue of fiel experiment ws m, elong to the wrm temperte zone ontinentl limte, verge nnul preipittion ws 45 mm, verge temperture ws 1 n verge frost-free perio is The experimentl fiel ws plin n wter ln, plnting one rop yer. There were four istintive sesons. Soil smples were ollete from the surfe lyer with epth of -2 m n the initil nutrient inies of the smpling soil were nlyze s follows: orgni mtter: 12.6 g kg -1 ; totl nitrogen: 1.8 g kg -1 ; lkli-hyrolyzle nitrogen: 53.6 mg kg -1 ; ville phosphorus: 9.6 mg kg -1 ; exhngele potssium:

3 mg kg -1 ; totl Se:.171 mg kg -1 n ville zin: mg kg -1. ove mentione inies were mesure using the methos of potssium ihromte volumetri, Kjelhl etermintion, lkli-hyrolyzle prolifertion, soium ironte extrtion, mmonium ette flme photometri, hyrie genertion tomi fluoresene spetrometry, n onentrtions of DTP-extrtle mironutrients respetively. Figure 1. Mp of experimentl region in hin Experimentl esign Two ftors split plot esign ws use to optimize Se folir fertiliztion ontent of olore-grin whet y tking ultivrs s min ftor n Se onentrtions s suplot ftor. Three winter whet ultivr Shnnong 1 (purple), Shnnong (lue), n Shnnong 129 (white) were use in the experiment, mong whih Shnnong 1 n Shnnong were olore-grin whet ultivr, while Shnnong 129 ws ommon whet. Shnnong 1 hs plnt height of 6 ~ 7 m, ompt plnt type, net tiller, high heing rte, long wn, purple see ot n rpi grouting. Shnnong hs plnt height of 8 ~ 9 m, ompt plnt type, net tiller, high heing rte, long wn, purple see ot n rpi grouting. Shnnong 129 hs plnt height of 7 ~ 8 m, ompt plnt type, net tiller, high heing rte, long wn, white see ot n rpi grouting. Se ws e in the form of N2SeO3 t rtes of (ontrol), 37.5 g h -1, g h -1, 75. g h -1, g h -1, n g h -1 respetively. The six onentrtions of N2SeO3 were ilute with istille wter. hnhel ompression spryer ws use to spry folir fertilizer uniformly n out 75 L h -1 solution ws sprye one. Whet sees were sown on Septemer 3, 216 t sowing ensity of plnt h -1 n row sping of 2 m. t the grin filling stge of winter whet (My 2, 217), eh ultivr ws sprye with Se. No rinfll ourre fter the spry. The plot sprye with the sme mount of istille wter ws tken s the ontrol. The hrvest time ws June 16, 217. Eh tretment ws repete three times. There were 54 plots in totl. re of eh plot ws 4 m 2 (2 m 2 m) n totl

4 re ws 216 m 2. efore sowing, 75 kg h -1 of NPK ompoun fertilizer (N: P2O5: K2O = 18%: 22%: 5%) ws pplie s se fertilizer. Other fiel mngement mesures (suh s irrigtion n wee mintenne) were rrie out s ommon prties (Fig. 2). Shnnong129 Shnnong1 Shnnong31244 D E Figure 2. Smpling sites for inies nlysis t Shnxi provine, hin Se n minerls nlysis Whet ws hrveste t full mturity stge to etermine the yiel s well s the ontents of Se n minerls in grins. fter etermintion of the yiel, the otine grin smples were wshe refully n rpily with istille wter, n then rie t 65 for 1 h, grine n sieve. The otine flour smples were use to investigte the ontents of totl Se, minerls, protein, n the omponents. The flour smples were igeste y using HNO3-H2O2 mixture in n limentry furne retion system (LWY84, hin), n then the onentrtion of Se n minerls were mesure y using n inutively ouple plsm mss spetrometer (IP-MS, gilent 77x; gilent Tehnologies, US) (Fig. 2). Protein n strh nlysis The flour protein n its omponents were mesure using Kjelhl nitrogen metho (He, 1985). mylsein n mylopetin were etermine y ul wvelength metho. The sum of the mylsein n mylopetin is the totl strh. mino is n nthoynin nlysis The hyrolyze smples were nlyze oring to the ntionl foo sfety stnr of the hin (G ). The grin were rie, rushe n sieve (<.15 mm), whih ws weighe n put in the hyrolysis tue. First, 1 ml Hl n phenol (3-4 rops) were e. Seon, the hyrolysis tue ws put into the refrigernt (3-5 min) n tthe to the sution pipe of the vuum pump n fill nitrogen (fill

5 nitrogen three times). fter tht the hyrolysis tue ws sele n put in thermostt (11, 22 h). The hyrolyste ws filtere into 5 ml volumetri flsk n onstnte to the sle. The filtrte ws put into the test tue. In test tue enrihment pprtus eompression rying in 48 heting environment. The rie resiue ws issolve y wter. Then the solution ws vuume n rie gin n until the top up. Finlly, the uffer solution of ph 2.2 soium itrte ws e to the test tue. The solution is to e mesure pssing through.22 nm memrne. mino i smples were seprte y ion exhnge hromtogrphy n mesure using the utomti mino i nlyzer (moel iohrom 3+, Engln). The nthoynin nlysis using ph ifferentil metho (Siriwohrn et l., 24) (Fig. 2). Sttistil nlysis The experiment ws li out in two ftors split plot esign with three prllel mesurements. The t were sujete to seprte nlysis of vrine (NOV) for eh ultivr, n the Tukey s multiple rnge test t p <.5 ws use to etermine ifferenes etween tretment mens. The Person orreltion proeure n generl line regression moel ws use to evlute the reltionship etween grin Se n nthoynin. SS softwre (SS 8., US) ws use for ll nlyses. Results Grin yiel n Se onentrtion To evlute the effets of folir pplition of Se fertilizer to the grin yiel n Se onentrtion in grins, we onute the NOV nlyses on the grin yiel, onentrtion of the grin totl Se n grin Se umulnt. The grin yiel n Se nlysis rrie out on the experimentl fiel with the olore-grin whet showe folir pplition of Se fertilizer t ifferent onentrtions i not signifintly ffet the yiel of olore-grins (p >.5, Tle 1). The grin yiel of Shnnong 1 n Shnnong were higher thn Shnnong 129. Nevertheless, there were signifint effets of the totl Se n Se umulnt of three whet vrieties (p <.5). The totl Se n Se umulnt of three whet vrieties were inrese signifintly with the inrese the onentrtion of folir Se fertilizer. y the folir pplition of the sme onentrtions of Se t grin filling stge of ifferent olore-grin whet, results of the totl Se n Se umulnt showe olore-grin whet (Shnnong n Shnnong 1) > ommon-grin whet (Shnnong 129). ompre to the grin Se onentrtion in the untrete ontrol, the onentrtion of e Se inrese y up to 18-fol t the mximum Se pplition of Shnnong 129. However, it inrese up to 1-fol n 8-fol in the olore-grin whet (Shnnong n Shnnong 1) respetively. It inite tht the whet grin h strong ility to onentrte exogenous Se uner the onition of folir pplition. lthough there ws inrese etween the totl Se onentrtion n the mount of Se pplie in the grin, the utiliztion rte of Se erese with the inrese of the onentrtion of folir Se fertilizer. When the onentrtion of folir Se fertilizer ws 37.5 g h -1, utiliztion rte of Se rehe the mximum, with the pttern of (Shnnong n Shnnong 1) > ommon-grin whet (Shnnong 129) isplye in three whet vrieties. oring to the Ntionl Foo Sfety Stnr in hin, Se onentrtion in Seenrihe whet ws presrie in the rnge of mg kg -1 (G/T ,

6 hin). When it ws more thn.3 mg kg -1, the high onentrtion oul e unhelthy for humns. The Se onentrtion in the grin of ommon whet ws over the ove rnge when folir pplition of Se onentrtion greter thn 75. g h -1. However, when folir pplition of Se onentrtion ws more thn 37.5 g h -1, the Se onentrtion in the olore-grin whet ws exessive. The results inite tht olore-grin whet ws more sensitive to Se umultion. Tle 1. Grin yiel n Se onentrtion of olore-grin whet ffete y the ifferent onentrtions of Se fertiliztion Tretments Selenium (g h -1 ) Inies ultivr Yiel (t h -1 ) Grin totl Se onentrtion (mg kg -1 ) Grin Se umulnt (mg h -1 ) Se utiliztion rte (%) Shnnong Shnnong Shnnong Shnnong e Shnnong e Shnnong 1.62f.285e Shnnong e Shnnong e Shnnong f e Shnnong Shnnong Shnnong Vlues were mens of three replites. Different line lowerse letters (,,,, e, f) inite signifint ifferenes mong Se tretments, t p <.5 in Tukey test. Different olumn upperse letters (,, ) inite signifint ifferenes mong us, t p <.5 in Tukey test Minerls onentrtion To further unerstn the effets of ifferent onentrtions of Se on olore-whet grins, the onentrtion of minerl elements in olore-grin whet re nlyze n shown in Figure 3. The onentrtion of minerl elements in olore-whet grins ws higher thn tht in ommon whet, initing tht the nutritionl vlue of olore-whet ws higher thn tht of ommon whet. fter leves were sprye with Se onentrtions of 37.5 g h -1, the onentrtion of Iron (Fe) in Shnnong 1 n Shnnong were signifintly inrese y 1.2-fol n.7-fol, respetively (p <. 5), the onentrtion of Zin (Zn) in Shnnong 1 n Shnnong were signifintly inrese y 1.2-fol n.7-fol, respetively (p <.5). The onentrtion of Fe n Zn erese ut it ws still hiher thn the ontrol vlue with the inrese of Se onentrtion in folir fertilizer. This inites tht the hnge of Fe n Zn in whet grin presents tren, whih ws low onentrtion promote n high onentrtion inhiite y Se pplition. However, the supply of Se woul inhiit the proution of opper (u) n Mngnese (Mn). The erese onentrtion rnge of u n Mn of ll whet were the sme, initing tht the onentrtion vritions of u n Mn of whet grins were only ffete y Se onentrtion other thn whet

7 Zn onentrtion (mg kg -1 ) Mn onentrtion (mg kg -1 ) Fe onentrtion (mg kg -1 ) u onentrtion (mg kg -1 ) vrieties. Overll, the results showe tht the sensitivity of minerl onentrtion to Se in ommon whet ws lower thn tht of olore-grin whet. The sensitivity of Se showe ifferent in two olore-grin whet, n purple whet more sensitive thn the lue one () () Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) 2 Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) () () Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Figure 3. Minerls in grins of olore-grin whet ultivr uner folir pplition of Se. X xes show the onentrtions of Se. Y xes show the onentrtions of ifferent minerls. Vlues were mens of three replites. The sufigures (-) show the onentrtions of Fe, u, Zn n Mn, respetively. Different line lowerse letters (,,, ) inite signifint ifferenes mong Se tretments, t p <.5 in Tukey test. Different olumn upperse letters (,, ) inite signifint ifferenes mong ultivrs, t p <.5 in Tukey test. onentrtion of totl protein, protein omponents The onentrtions of totl protein n protein omponents showe signifint hnges within olore-grin whet (Fig. 4, p <.5). Overll, totl protein onentrtion ws signifintly lower in Shnnong 129 thn in Shnnong without Se (p <.5). The onentrtion of totl protein inrese with the inrese the onentrtion of folir Se fertilizer. The onentrtion of lumin n gloulin were inrese, however, whih onentrtion Se inrese mplitue grully erese with the inrese the onentrtion of folir Se fertilizer with the folir pplition of 37.5 g h -1 whih erese t higher onentrtions of Se, while gliin n glutenin onentrtion s were inrese in ll whet ultivr with inrese onentrtions of Se (p <.5). The highest gliin n glutenin onentrtion ws foun t the highest

8 Totl protein onentrtion (g kg -1 ) Gliin onentrtion (g kg -1 ) Glutenin onentrtion (g kg -1 ) lumin onentrtion (g kg -1 ) Gloulin onentrtion (g kg -1 ) onentrtion of Se. Glutenin onentrtion of olore-grin whet (Shnnong 1, Shnnong 31244) ws signifintly higher thn ommon whet (Shnnong 129); however, the onentrtion of lumin ws highest in ommon whet (Shnnong 129). This inites tht the effet of folir Se ws signifintly ifferent for ultivr in terms of the protein omponents n totl protein onentrtion () () Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) () () Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) (e) e Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Figure 4. onentrtions of totl protein n protein omponents in grins of olore-grin whet ultivr uner folir pplition of Se. X xes show the onentrtions of Se. Y xes show the onentrtions of ifferent onentrtions of totl strh n ifferent strh omponents. Vlues were mens of three replites. The sufigures (-e) show the onentrtions of lumin, gloulin, gliin, glutenin n totl protein, respetively. Different line lowerse letters (,,, ) inite signifint ifferenes mong Se tretments, t p <.5 in Tukey test. Different olumn upperse letters (,, ) inite signifint ifferenes mong ultivrs, t p <.5 in Tukey test.

9 Totl strh onentrtion (g kg -1 ) mylse onentrtion (g kg -1 ) mylopetin onentrtion (g kg -1 ) onentrtion of totl strh n its omponents The onentrtion of totl strh n its omponents showe signifint hnges within olore-grin whet (Fig. 5, p <.5). Overll, the onentrtion of mylse ws signifintly lower in Shnnong 129 n Shnnong thn in Shnnong 1 without Se (p <.5). The highest onentrtion of totl strh n its omponents ws foun t the Se onentrtion of 37.5 g h -1 uner folir pplition. The onentrtion of totl strh, mylse, n mylopetin first inrese then erese with the inrese the onentrtion of folir Se fertilizer. This inites tht the effet of folir Se ws signifintly ifferent for ultivr in terms of the strh omponents n totl strh onentrtion () () Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) () Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Figure 5. onentrtions of totl strh n ifferent strh omponents in grins of oloregrin whet ultivr uner folir pplition of Se. X xes show the onentrtions of Se. Y xes show the onentrtions of ifferent onentrtions of totl strh n ifferent strh omponents. Vlues were mens of three replites. The sufigures (-) show the onentrtions of mylse, mylopetin n totl strh, respetively. Different line lowerse letters (,, ) inite signifint ifferenes mong Se tretments, t p <.5 in Tukey test. Different olumn upperse letters (,, ) inite signifint ifferenes mong ultivrs, t p <.5 in Tukey test. onentrtion of mino is n nthoynin The onentrtion of mino is ws higher in olore-grin whet thn in ommon grin whet without Se tretment (Fig. 6). In the present stuy, smll ose of Se h positive effet on the onentrtion of the totl mino is.

10 Vline onentrtion (g kg -1 ) lnine onentrtion (g kg -1 ) Phenyllnine onentrtion (g kg -1 ) Threonine onentrtion (g kg -1 ) Lysine onentrtion (g kg -1 ) Methionine onentrtion (g kg -1 ) Isoleuine onentrtion (g kg -1 ) Leuine onentrtion (g kg -1 ) () () Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) () () Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) (e) (f) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) (g) (h) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Shnnong 129 Shnnong Shnnong 1 Se onentrtion (g h -1 ) Figure 6. onentrtions of mino is of olore-grin whet ffete y ifferent onentrtions of Se fertiliztion. X xes show the onentrtions of Se. Y xes show the onentrtions of ifferent mino is. Vlues were mens of three replites. The sufigures (-p) show the onentrtions of Ile, Leu, Lys, Met, Phe, Thr, Vl, l, rg, sp, Glu, Pro, Gly, His, Ser n Tyr, respetively. Different line lowerse letters (,, ) inite signifint ifferenes mong Se tretments, t p <.5 in Tukey test. Different olumn upperse letters (,, ) inite signifint ifferenes mong ultivrs, t p <.5 in Tukey test.

11 However, higher levels of Se supplementtion i not result in further inrese of the onentrtion regring the totl mino is. fter pplying lrger ose of Se, there ws smll erese in the totl ontent of mino is, ut the overll onentrtion of mino is ws still lrger thn the ontrol. There ws erese tren fter pplition Se onentrtion of 37.5 g h -1 in olore-grin whet, however, erese tren ws foun fter Se onentrtion of 75. g h -1 in ommon-grin whet. The sixteen mino is were etete (inluing seven essentil mino is n nine non-essentil mino is). ll essentil mino is were signifintly ffete exept leuine (Leu) (p <.5). The onentrtion of essentil mino is ws signifintly lower in Shnnong 129 thn in Shnnong 1 n Shnnong with Se. However, with the inrese of Se onentrtion in folir fertilizer, there were no signifint effets of the essentil mino is of three whet vrieties (p >.5). The results for non-essentil mino is re lso liste. The pplition of Se mrkely influene the onentrtion of non-essentil mino is exept rginine (rg) n tyrosine (Tyr) (p <.5). Menwhile, with the inrese of Se onentrtion in folir fertilizer, there were slo no signifint effets of the non-essentil mino is of three whet vrieties (p >.5). To exmine the reltionship etween nthoynins n Se, the onentrtion of nthoynins ws mesure in olore-grin whet (Fig. 7). We foun the synergisti effet etween nthoynins n Se in olore-grin whet, whih inite there ws likely to e gret Se-enrihment potentil in olore-grin whet. Figure 7. The orreltion etween nthoynin n folir pplition of Se in grins of olore-grin whet. X xes show the onentrtions of Se. Y xes show the onentrtions of ifferent minerls Disussion The ustomer s eptne of innovtive, new prouts is epenent on their feek to the sensory n funtionl properties of flour, inluing olor, flvor, nutritionl vlue, n overll eptility. In prtiulr, for flour, it is neessry to mintin the typil sensory n nutritionl hrteristis for ommeril suess in the evelopment of funtionl foos. The eviene tht Se enrihment of olore-grin whet improves the nutritionl spet of the flour without ltering its sensory properties is ritil for onsumers to ept Se-enrihe flour (Fio et l., 214). However, nutritionl nlyses of io-fortifie olore-grin whet flour hve not een stuie extensively. Our stuy fouse on the evlution of the effet of Se on the nutritionl

12 ontents of olore-grin whet n provie vlule prtie on the evelopment of new funtionl selenium-fortifie olore-grin whet. The soil Se-supplement, ress sees, soking sees, folir pplition of Se n exogenous Se supplementtion n inrese the mount of Se in griulturl prouts (Dusy et l., 29). mong ove folir pplition of Se hs the vntges of fst, effiient, low ost n low environmentl risk. Our finings suggeste the effetiveness of folir Se fertiliztion s suitle strtegy to improve nutritionl qulity of oloregrin whet, whih is onsistent with the previous stuies whih hve shown the greter effetiveness of folir Se tretment thn other Se supply methos in whet (Zhng et l., 21; Glinh et l., 215). Moreover, we foun tht ll of the Se tretments were highly effetive t inresing the Se onentrtion in the olore-grin whet, for three whet ultivrs, n tht the Se onentrtions in the grin inrese with inresing onentrtions of Se pplition oring to positive orreltion. This eviene is in greement with severl previous stuies in whih high orreltion vlues were oserve (roley et l., 21; Rorigo, Sntmrí, López-ellio n Poliones, 213). The Se-enrihe whet require Se onentrtion to e mg kg -1 (G/T , hin). Therefore, pproprite externl Se will e neessry to mke up the efiieny to evelop the Se-enrihe whet flour. In our result, when the folir pplition of Se t the optiml ose (37.5 g h -1 ), the onentrtion of Se in oloregrin whet h rehe the ove stnr. When the whet flour ove this stnr will not e goo for people to et. The sene of Se poisoning oserve in our stuy is n importnt onsiertion for frmers, euse this folir Se pplition oul not use ny reutions in yiel through wie rnge of Se onentrtions. There ws no signifint ifferene in the yiel of olore-grin whet y folir pplition of selenium fertiliztion, whih ws onsistent with the previous stuies, in whih pre-hrvest folir pplition of minerl elements i not ffet grin yiel of rops (hen et l., 22; Rorigo et l., 213). There ws muh reserh out the effet of folir pplition Se on ommon whet, ut little reserh hs een one on olore-grin whet. In the ontrol group of our stuy, the Se onentrtion in the ommon whet grin ws little lower thn in the olore-grin whet, whih onfirme high nutritionl vlue in the olore-grin whet. The onentrtion of Se n the umultion of Se inrese with the inrese of Se onentrtion; however, the inrese vries epening on whet vrieties. The Se onentrtion in olore-grin whet ws signifintly higher thn ommon grin whet whih might e use y the sorption, trnsformtion, n trnsport pity of the exogenous soium Se solution of olore-grin whet. The utiliztion effiieny of Se in whet is similr to other elements, tht is, uner reltively stle tehnil onitions, the Se onentrtion of whet inrese with the inrese of Se injetion, ut the utiliztion of Se goes own. This explins the high onentrtion of Se inhiite plnt growth ue to Se toxiity interfering with the plnt metolism. lerly, our results showe the enefiil role of Se t low onentrtions oul promote olore-grin whet growth, whih is onsistent with previous stuies (olrin et l., 216; Guerrero et l., 214). For the first time, we ompre the Se umultion etween ommon whet n olore-grin whet, n reporte the signifint inrese of minerl onentrtions in the olore-grin vrieties ompring to the ommon vriety, whih inite gret potentil to inrese the nutritionl ontent of olore-grin whet flour for the ommeril uses y the strtegy of Se folir spry.

13 It is generlly onsiere tht high osges of Se my e toxi to plnts y ompeting with some essentil elements, suh s Mn, Zn, u n Fe (Kt-Penis n Penis, 211; Frgsov et l., 26), however, in some ses, the stimulting effets of Se on uptkes of u, Fe n Zn in plnts were lso oserve (Feng et l., 29). Intriguingly, folir pplition of Se h signifint effet on minerl onentrtions in whet grins. The olore-grin whet vrieties h signifintly higher minerl onentrtion thn the ommon whet. Our results showe folir pplition of Se promote the umultion of Fe n Zn n inhiite u n Mn umultion, similr to the previous suggestion y Feng et l. (29). s Se is inorporte into proteins s selenominois, the ifferent ptitues of these olore-grin vrieties to umulte Se might e linke to issimilrities in their protein ontent (rly n MPherson, 1992). If the protein ontent in whet grin is high, Se is minly ssoite with plnt proteins. Nossier et l. (211) foun selenium ws relte to protein funtion. Se ws further reporte to e involve in protein metolism s omponent in n RN hin in plnts, with the min physiologil funtion of trnsporting mino is to synthesize proteins (Grousi, 217; Shivon et l., 217). In our reserh, Se folir fertilizer showe signifint impt on protein omponents in grins of olore-grin whet vrieties too. The ontent of lumin n gloulin were signifintly reue ut the ontent of gliin n glutenin were signifintly inrese with the inrese the onentrtion of folir Se fertilizer. The signifint effet of the high Se onentrtion on the protein omponents of oloregrin whet ws oserve, with eing more prominent in the eeper olore vrieties. Furthermore, there ws no effet of Se fertiliztion on gluten qulity, similr to previous suggestions y roley et l. (21). In ition to the protein omponents, the Se fertiliztion h no effet on either gluten or ft prmeters, whih inite the folir spry of Se will not ffet other min orgni nutrient thn proteins in olore-grin whet. Furthermore, the strh is the min soure of whet extrt. In theory, mylose ws onverte into fermentle sugr n mylopetin ws onverte into fermentle sugr n low moleulr extrin uner the tion of enzyme. The possile reson for the eline of strh is tht the high ose of Se supply me the grin filling spee erese, thus preventing the trnsformtion of strh omponent. The signifint influene of mino i onentrtion in plnt tissues n e use y nutrition n fertiliztion, suh s Se (Munshi, 199). For exemple, methionine (Met) ws the limiting essentil mino i, n pplition of Se in our stuy ffete the Met ontent of olore-grin whet. Moreover, the pplition of Se mrkely influene the ontent of sprti i (sp) n sprgine (sn) whih is resulte from mition of sp. oth sp n sn re importnt in the trnsport n storge of nitrogen in plnt tissues (Le et l., 27). In ition, we foun the synergisti effet etween nthoynins n Se in olore-grin whet, whih inite there ws likely to e gret Se-enrihment potentil in olore-grin whet. In the future stuies, the following spets re expete to e fouse: (i) The physiologil regultion mehnism of Se in the proess of migrtion, istriution, n umultion in olore-grin whet plnts n the ntgonisti mehnism of Se n hevy metls in olore-grin whet plnts. (ii) Further uses of vne iotehnology on seleting n reeing new vrieties of Se-enrihe olore-grin whet.

14 onlusion It is the first time to report the Se-enrih olore-grin whet with the nutritionl nlyses n evlution upon ifferent Se onentrtions pplie in folir spry. The optiml fertiliztion of Se oul enhne nutritionl qulity of olore-grin ultivr inluing the onentrtions of protein, minerls n mino i without ffeting the grin yiel, gluten, n ft ontent, however, the strh onentrtion ws erese. itionlly, whet grin ws reporte to e enrihe with Se through exogenous Se uner the folir pplition of Se. Thus, the folir pplition of Se t the optiml ose (37.5 g h -1 ) is suggeste to e use for Se io-fortifition of olore-grin whet ultivr. Moreover, the purple grin whet is more Se-enrihe thn lue one. Further stuies re expete to unerstn the mehnisms of miro-nutrient uptke n trnsport to quire more effiient umultion of nutrients in olore-grin whet. Our stuy provies vlule prtie in nutrition nlyses of the olore-grin whet n the evelopment of nutrient-enrihe whet flour. knowlegements. The uthors wish to thnk the meiums n sitters who volunteere their time for this stuy. This stuy ws finnilly supporte y the Shnxi Sholrship ounil of hin from the Ministry of Finne, PR of hin (215-Key4), the the Key projet of Shnxi Key R&D Progrm of hin (2173D2111-2) n the Speil Pln of Sientifi Reserh for Shnxi griulture Vlley of hin (SXNGJSKYZX 2172) n the hin griulture Reserh System (RS ). REFERENES [1] rly, M. N. I., MPherson,. (1992): Selenium ontent of whet for re mking in Sotln n the reltionship etween glutthione peroxise (E ) levels in whole loo n re onsumption. ritish Journl of Nutrition 68(1): [2] olrin, P. F., e Figueireo, M.., Yng, Y., Luo, H., Giri, S., Hrt, J. J.,... Li, L. (216): Selenium promotes sulfur umultion n plnt growth in whet (Tritium estivum). Physiologi Plntrum 158(1): [3] roley, M. R., lok, J., lfor, J., rtwright, P., Foot, I., Firwether-Tit, S. J.,... Mehm, M.. (21): Selenium iofortifition of high-yieling winter whet (Tritium estivum L.) y liqui or grnulr Se fertilistion. Plnt n Soil 332(1-2): [4] hen, J. S. (212): n originl isovery: selenium efiieny n Keshn isese (n enemi hert isese). si Pifi Journl of linil Nutrition 21(3): [5] hen, L., Yng, F., Xu, J., Hu, Y., Hu, Q., Zhng, Y., Pn, G. (22): Determintion of selenium onentrtion of rie in hin n effet of fertiliztion of selenite n selente on selenium ontent of rie. Journl of griulturl n Foo hemistry 5(18): [6] Dusy, L., Ložek, O., Vrg, L. (29): The influene of selenium soil pplition on its ontent in spring whet. Plnt Soil Environ 55(2): [7] Frgsov,.; Pstierov, J.; Svetkov, K. J. P. S. (26): Effet of Se-metl pir omintions (, Zn, u, P) on photosyntheti pigments proution n metl umultion in Sinpis l L. seelings. Plnt Soil & Environment 52(1): [8] Feng, R.; Wei,.; Tu, S.; Wu, F. J. P. (29): Effets of Se on the uptke of essentil elements in Pteris vittt L. Plnt & Soil 325(1-2): [9] Fernnes,. P., Gnin, V. (215): Selenium ompouns s therpeuti gents in ner. iohimi et iophysi t ()-Generl Sujets 185(8):

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16 [31] Shi R L, Zhou Q, Zhng F S, et l. (26): Reserh progress on grin iron n zin nutrition n humn helth. Gungong Miroelement Siene 13(7): 1-8 (in hinese with English strt). [32] Siriwohrn, T., Wrolst, R. E., Finn,. E., Pereir,.. (24): Influene of ultivr, mturity, n smpling on lkerry (Ruus L. Hyris) nthoynins, polyphenolis, n ntioxint properties. Journl of griulturl n Foo hemistry 52(26): [33] Spekmnn,., Grune, T. (215): Epigeneti effets of selenium n their implitions for helth. Epigenetis 1(3): [34] Tn, J.., Zhu, W., Wng, W., Li, R., Hou, S., Wng, D., Yng, L. (22): Selenium in soil n enemi iseses in hin. Siene of the Totl Environment 284(1-3): [35] Toppe J, homo, V. (215): Seon Interntionl onferene on Nutrition (IN2): ontriution of Fish to Humn Nutrition. FO quulture Newsletter (53): 4. [36] Vrg, M., ánhiy, J., seuz, L., Mtuz, J. (213): The nthoynin ontent of lue n purple oloure whet ultivrs n their hyri genertions. erel Reserh ommunitions 41(2): [37] Wng, S., Li, M., Liu, K., Tin, X., Li, S., hen, Y., Ji, Z. (217): Effets of Zn, mronutrients, n their intertions through folir pplitions on winter whet grin nutritionl qulity. PloS One 12(7): e [38] Welh, R. M. (28): Linkges etween Tre Elements in Foo rops n Humn Helth. In: llowy,. J. (e.) Mironutrient Defiienies in Glol rop Proution. Springer, Dorreht, pp [39] Yv, S. K., Singh, I., Singh, D., Hn, S. D. (25): Selenium sttus in soils of northern istrits of Ini. Journl of Environmentl Mngement 75(2): [4] Zeven,.. (1991): Whets with purple n lue grins: review. Euphyti 56(3): [41] Zhng, Y., Shi, R., Rezul, K. M., Zhng, F., Zou,. (21): Iron n zin onentrtions in grin n flour of winter whet s ffete y folir pplition. Journl of griulturl n Foo hemistry 58(23): [42] Zhng, Z., Zhng, J., Xio, J. (214): Selenoproteins n selenium sttus in one physiology n pthology. iohimi et iophysi t ()-Generl Sujets 184(11): [43] Zho, F. J., MGrth, S. P. (29): iofortifition n phytoremeition. urrent Opinion in Plnt iology 12(3): [44] Zho, Q G, Yin, X.. (216): Funtionl griulture. The Siene Pulishing ompny (in hinese with English strt).