Combined Effects of Blue and Ultraviolet Lights on the Accumulation of Flavonoids in Tartary Buckwheat Sprouts

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1 Originl rtile Setion: Foo Chemistry Pol. J. Foo Nutr. Si., 216, Vol. 66, No. 2, pp DOI: /pjfns Comine Effets of n Ultrviolet Lights on the Aumultion of Flvonois in Trtry Bukwhet Sprouts Hongin Ji*, Wen Tng, Xioli Zhou, Ying Wu Deprtment of Foo Siene n Tehnology, Shool of Perfume n Arom Tehnology, Shnghi Institute of Tehnology, 1 Hiqun Ro, Fengxin Distrit, Shnghi 21418, Chin Key wors: lue light, /, trtry ukwhet sprouts, flvonois The effets of lue n (365 nm)/ (254 nm) or their omintions on the levels of totl flvonois, rutin, queretin, phenyllnine mmonilyse (PAL), hlone isomerse (CHI), rutin egring enzymes (RDEs) n 1,1-iphenyl-2-pirylhyrzyl (DPPH) ril svenging tivity in trtry ukwhet sprouts were investigte in this stuy. The totl flvonois ontent in the trtry ukwhet sprouts irrite with lue light followe y () rise y 1%, ompre with the opposite omintion sequene (). However, lue light i not show the sme results when omine with, n their omintions on the umultion of totl flvonois were still lower thn tht of /. Key enzymes (PAL,CHI n RDEs) revele signifint orreltion with totl flvonois in trtry ukwhet sprouts. INTRODUCTION Trtry ukwhet in the form of nooles, flour n steme un is ommonly onsume foo in Asin ountries. In reent yers, trtry ukwhet (Fgopyrum esulentum) hs reeive inresing ttention ue to its unnt nutritive ompouns. Trtry ukwhet sprouts re new vegetle prouts tht hve lso ttrte lot of ttention euse they hve not only high levels of flvonois [Kim et l., 24] ut lso some enefiil funtions on loo pressure [Nkmur et l., 213]. Flvonois s lrge lss of seon metolites re wiely istriute in mny plnts inluing trtry ukwhet n isply ntioxint tivity in plnts [Piett, 2]. Flvonois re synthesize y the phenylpropnoi metoli pthwy in whih the mino i phenyllnine is use to proue 4-oumroyl-CoA tlyze y the enzyme phenyllnine mmonilyse (PAL) [Koes et l., 1994]. Chlone isomerse (CHI), seon key enzyme, meites the phenylpropnoi pthwy for the synthesis of flvonois [Winkel-Shirley, 21]. Mny ftors suh s lights [Cortell & Kenney, 26] n rre erth elements [Ling et l., 26] hve effets on the umultion n istriution of flvonois. Wng et l. [213] reporte tht the totl flvonois ontent in trtry ukwhet inrese y ing the itives (Al 3+, Cu 2+, n Zn 2+ ) when their onentrtion ws in ertin rnge n erese when their onentrtion ws eyon ertin rnge. It ws lso reporte tht in the llus-ulture Mlv * Corresponing Author: Tel./Fx: ; E-mil: jihongin88@yhoo.om (Hongin Ji) neglet ells, UV irrition inrese flvonois n nthoynin ontent signifintly ompre with rkness [Khtmi & Ghnti, 211], n lue light inrese rutin n queretin ontent in trtry ukwhet sprouts [Tsurung et l., 213]. Ultrviolet s ommon nturl environment ftor ffete the morphologil growth, iohemil n physiologil responses of plnts [Clwell et l., 1998; Zhng et l., 23]. It ws tritionlly ivie into three wvelengths: (2 28 nm), UV-B (28 32 nm), (32 4 nm). light ws nother importnt environmentl ftor involve in wie rnge of plnt proesses suh s phototropism, photomorphogenesis, stomtl opening, n photosynthesis [Whitelm & Hlliy, 27], n it oul le to higher plnt iomss [Mtsu et l., 28]. Severl stuies inite tht the hlone synthse (CHS), key enzyme for flvonois iosynthesis, ws involve in synergisti effet of UV n lue light [Jenkins, 1997]. Ohl et l. [1989] reporte tht lue light inrese the response to susequent UV-B pulse, n it le to the inrese of CHS impression. However, there re only few stuies on the effet of lue light n / on levels of flvonois n flvonois iosynthesis relte enzymes in trtry ukwhet sprouts. Hene, we exmine the effets of the omine effets of lue n (356 nm)/ (254 nm) lights or their omintions on flvonois of trtry ukwhet sprouts. MATERIAL AND METHODS Mteril n hemil regents Drie trtry ukwhet sees (Fgopyrum esulentum) lle HeiFeng NO.1 in the experiments were otine from Copyright y Institute of Animl Reproution n Foo Reserh of the Polish Aemy of Sienes 216 Author(s). This is n open ess rtile liense uner the Cretive Commons Attriution-NonCommeril-NoDerivs Unuthentite Liense ( Downlo Dte 1/3/18 3:43 AM

2 94 Effet of Light on Flvonois of Trtry Bukwhet Sprouts A/C A 8 The irrition intensity of the light soures is shown in Tle 1. The sees were ultivte in rkness for 6 h to llow them to germinte, n then were irrite y / n lue light or their omintions for 12 h. Their omintions were shown s follows: + light (+BL), light+ (BL+), + light (), light+ (). For exmple, +BL: ttry ukwhet sprouts were irrite with for 6 h n followe y lue light for nother 6 h. Drkness ws set s ontrol Determintion of sprouts length Three ys lter, sprouts from 1 replite plnts per tretment were mesure from the surfe of the ottom to the top of the sprouts. FIGURE 1. Irrition equipment for trtry ukwhet sprouts (units in mm). A/C shows swith of / n lue shows swith of lue light. TABLE 1. Intensity of irrition from ifferent light. Soure Intensity (W/m 2 ) (rkness) BL BL Determintion of totl flvonois The etermintion of totl flvonois ws se on the metho of Stokov et l. [29] with some moifitions. One grm of freeze rie trtry ukwhet sprouts ws immerse in 5 ml 6% (v/ v) ethnol, n put into 7 o C wter th for 6 h, n then entrifuge t 3 g for 1 min. The superntnt ws the extrt smple. Then, 1 ml of smple extrt ws trnsferre to 1 ml volumetri flsk n 2 ml of.1 mol/l luminum hlorie solution n 3 ml of 1 mol/l potssium ette were e. The mixture ws ilute to sle (1 ml) with 6% (v/v) ethnol, mixe well n kept in rkness for 3 min. The sorne t 42 nm of the mixture fter retion ws mesure. Rutin ws use to plot stnr urve (shown s follows), n totl flvonois were expresse s rutin equivlents. A 42 = (mg/ml) rutin.28, R 2 =.993 Shnxi provine, Chin. Chemil regents s rutin ( purity >99%) n queretin (purity >99%) were purhse from Sigm-Alrih (USA) n the other regents were purhse from Sinophrm C hemil Regent Co., Lt. Germintion proeures Trtry ukwhet sees were sterilize with 1.25% (w/v) soium hypohlorite t room temperture for 3 min to eliminte miro-orgnisms, n then wshe three times with istille wter. Before germintion, sees were soke in tp wter t 25±1 o C in rtifiil limte inutor for 12 h. Then, 2 g sees were spre into eh try (3 m 4 m) with three piees of filter pper. The sees were inute t 98% RH n 3±1 o C for 3 ys. After germintion, the sprouts were refully wshe with istille wter n remove thes soon s possile n store t -8 o C until their use. Some sprouts were rie t -8 o C for 24 h, groun into power n psse through 2-mesh sreen sieve. Lights irrition tretments light fluoresent lmps (15 W) were purhse from Toshi n UV lmps (16 W, GL 946) were purhse from Shnghi Chojing siene n Tehnology Co. Lt. The irrition equipment ws esigne oring to Tsurung et l. [213] with some moifitions n it is shown in Figure 1. Assy for PAL tivity etermintion The proeure of extrtion n mesurement ws performe oring to Chen et l. [23] n Assis et l. [21] with some moifitions. Fresh ukwhet sprouts (1. g) without test (see ot) were grou n with mortr n pestle t o C in 5 ml.1 mol/l orte uffer (ph 8.8) ontining 1 mmol/l ethylene imine tetreti i (EDTA), 5 mmol/l merptoethnol, 1% (w/v) polyvinylpyrrolione (PVP) n 1% (v/v) glyerol, n then entrifuge t 12, g, 4 o C fo r 2 min. The superntnt ws use s resoure of rue enzyme. Totl protein mount ws etermine oring to Brfor metho with ovi ne serum lumin s stnr (A 595 =.219 ovine serum lumin (μg/ml)-.28, R 2 =.9991). A.5 ml liquot of rue enzyme extrt ws e to 3 ml.1 mol/l orte uffer (ph 8.8), n 1.5 ml of sustrte (.2 mol/l L-phenyllnine). The mixture ws inute in wter th t 3 o C for 1 h. PAL tivity ws estimte y mesuring the vrition of the sorne t 29 nm (A 29 ) per hour. Vrition of.1 A 29 ws efine s one unit (U) n speifi enzyme tivity ws efine s one unit per mi lligrm of protein. Assy for CHI tivity etermintion The etermintion of CHI tivity ws rrie out oring to the metho of Lister et l. [1996] n Wng et l. Unuthentite Downlo Dte 1/3/18 3:43 AM

3 H. Ji et l. 95 [213] with some moifitions. Sprouts were groun with mortr n pestle t o C in 5 ml, ph 8. soium phosphte uffer, n then entrifuge t 1, g, 4 o C for 2 min. The superntnt ws use s resoure of rue enzyme. A.75 ml liquot of rue enzyme liqui ws e to the mixture ontining 2 ml of.5 mol/l Tris-HCl uffer (ph 7.4), 7.5 mg/ml of ovine serum lumin (BAS), 5 mmol/l of KCN n 5 μl of 1 mg/ml hyroxylte hlone. The retion ws onute t 3 o C for 3 min. The vrition of sorne t 381 nm (A 381 ) ws use to estimte CHI tivity. Vrition of.1 A 381 ws efine s one unit (U) n speifi enzyme tivity ws efine s one unit per milligrm of protein. Assy for RDEs tivity etermintion The RDEs tivity ws etermine oring to the proeure of Chen et l. [211] with some moifition. One grm of trtry ukwhet sprout power ws extrte in 5 ml of.2 m ol/l ette uffer (ph 4.) t 4 C for 3 h, n then ws entrifuge t 1, g, 4 C for 2 min. The superntnt ws the rue enzyme extrt n ws store t 4 C until its use..4 ml of enzyme extrtion rete with.15 ml of enzyme sustrte (.1 g of rutin issolve in methnol, wh ih ws ilute 1 times y eti i uffer solution) t 5 o C wter th for 3 min n.8 ml methnol ws immeitely use to stop the retion fter 3 min. Then, it ws ilute y 5 ml of iluents liqui (16.8 ml,.2 mol/l ette uffer ws e to 1 ml volumetri flsk, n ilute to the sle y methnol). The sorne of smple ws mesure t selete wvelengths (343.3 nm n 372 nm). The vrition of sorne of.1 t nm n 372 nm ws efine s one unit (U) n the RDEs t ivity ws U/mg ry mtter. DPPH ril sv enging tivity DPPH ri l svenging tivity (RAS) ws mesure oring the metho esrie y Tkht et l. [211] n Nkjim et l. [29] with some moifitions. Briefly, 5 ml of the previous ly prepre 6% (v/v) e thnol extrt ws e to 5 ml, 2 μmol/l DPPH in ethnol. The mixture ws inute t room temperture in rkness for 3 min, n the sorne ws mesure t 517 nm (A T ). Pure DPPH solution ws set s ontrol n its sorne t 517 nm ws A The DPPH ril svenging tivity (RAS) ws lulte s follows: RAS(%)=(A -A T ) 1/A HPLC nlysis for rutin n queretin The ontent of rutin n queretin in trtry ukwhet sprouts ws nlyze y using quntittive SHIMADZU HPLC system, onsisting of SHIMADZU 2AD pumps, SHIMADZU SIL-2A uto smpler n SHIMADZU SPD-M2A etetor. Smples were seprte on reverse phse Phenomenex C 18 olumn ( mm, 5 μm prtile size) n ll the smples were psse through.22 μm millipore filter. Aetonitrile n.5% (v/v) phosphori i were use s solvents, pumpe t flow rte of 1 ml/min for rutin etermintion n.8 ml/min for queretin etermintion. The UV etetion ws performe t 256 nm. Quntifitions of rutin n queretin t ifferent onentrtions were use to plot stnr urve. Sttistil nlysis All the t were expresse s mens ± SD t three replites of eh smple. Anlysis of vrine n signifint ifferene nlysis vi one-wy ANOVA followe y Dunn metho (P<.5) were onute y using SPSS 18. softwre. RESULTS AND DISCUSSION Effet of light omintions on growth of trtry ukwhet sprouts The length of ontrol n irrite sprouts ws mesure fter 72 h n the results re shown in Figure 2. light signifintly inrese length of sprouts, n wrfe the sprouts. light in omintion with or le to higher length of sprouts thn n lone. Thus, it ws ovious tht lue light enhne the growth of trtry ukwhet sprouts. Effet of light omintions on the levels of totl flvonois n PAL, CHI, RDEs tivity Totl flvonois ontent in trtry ukwhet sprouts ws gretly sensitive to lue light n / or their omintions (Figure 3). The totl flvonois ontent in sprouts irrite with, n lue light inrese respetively 56.1%, 51.2%, 24.2% ompre with rkness. The highest ontent of totl flvonois ws oserve in the trtry ukwhet sprouts irrite with (47.52 mg/g ry mtter) or (46.2 mg/g ry mtter). / inrese in higher extent the totl flvonois ontent thn lue light lone or omine with /. As severl previous stuies reporte, oth / [Khtmi & Ghnti, 211] n lue light [Ser et l., 1998] in- Lenght of sprouts (m) f BL+ e +BL e g FIGURE 2. Effet of irrition with ifferent light omintions on the length of trtry ukwhet sprouts. Vertil rs inite the stnr evition. BL+: irrition with lue light followe y ; +BL: irrition with followe y lue light; : irrition with lue light followe y ; : irrition with followe y lue light. Different letters show signifint ifferene (P<.5). Unuthentite Downlo Dte 1/3/18 3:43 AM

4 96 Effet of Light on Flvonois of Trtry Bukwhet Sprouts TABLE 2. Effet of light omintions on PAL, CHI n REDs tivity in trtry ukwhet sprouts. PAL (U/mg protein) CHI (U/ mg protein) RDEs (U/mg DW) ± ± ±.11 BL ±7.26 e 55.47± ±.26 +BL ± ± ±.33 Content of totl flvonis (mg/g ry mtter) ±7.11 e 6.26± ± ± ± ± ± ± ±.75 (365nm) ± ±2,75 e 27.19±1.31 (254nm) ±9.41 e 97.92±1.98 e 28.22±.41 Dt presente re mens±stnr evition of the men for three inepenent experiments. Different letters show signifint ifferenes (P<.5). BL+ +BL FIGURE 3. Effet of irrition with ifferent light omintions on the totl flvonois ontent in trtry ukwhet sprouts. Vertil rs inite the stnr evition. BL+: irrition with lue light followe y ; +BL: irrition with followe y lue light; : irrition with lue light followe y ; : irrition with followe y lue light. Different letters show signifint ifferene (P<.5). rese the umultion of flvonois. However, the effet of lue light on flvonois ws muh lower thn tht of ultrviolet (Figure 3). light showe ifferent influene on totl flvonois umultion when omine with or. light omine with inrese totl flvonois ontent signifintly (P<.5) when trtry ukwhet sprouts were irrite with lue light first (). Totl flvonois ontent of ws 4.15 mg/g ry mtter, higher thn tht of. This effet ws not oserve when lue light ws omine with (BL+). Ohl et l. [1989] reporte tht lue light irrite first followe y UV-B on prsley ells inrese the expression of CHS, key enzyme in flvonois iosynthesis. He expline tht the effet of lue light on the expression of CHS ws stle. Besies, Jenkins [1997] reporte tht lue light irrite efore UV-B resulte in n inrese of CHS expression. Tht effet ws not oserve when UV-B ws irrite efore lue light. These stuies together with the present investigtion suggest tht lue light oul generte signl tht enhne susequent response to UV-B n. This signl i not pper when the omintion BL+ ws use. Therefore, lue light in omintion of lue light n UV-B/ generte stle signl to enhne umultion of flvonois. light n / or their omintions lso hve n importnt influene on the relevnt enzymes involve in flvonois iosynthesis (Tle 2). light n / or their omintions inrese the tivity of these enzymes (PAL, CHI n RDEs) signifintly (P<.5) ompre with rkness. The effet of lue light in omintion with / on PAL, CHI n RDEs tivity in trtry ukwhet sprouts ws similr to tht oserve on totl flvonois ontent. Tht is, lue light in omintion of lue light n inrese PAL, CHI n REDs tivity when lue light ws irrite first () ut showe little influene when omine with (BL+). Cresy [1968] foun tht the tivity of PAL ws orrelte with the rte of flvonois synthesis. Wng et l. [213] reporte Content of rutin (mg/g ry mtter) BL+ +BL tht the effets of Al 3+, Cu 2+ n Zn 2+ on the totl flvonois uring germintion of trtry ukwhet were in line with the tivity of PAL n CHI. In the present work, the tivity of PAL n CHI ws in line with the vriety of totl flvonois ontent s well. Therefore, lue light in omintions oul influene flvonois ontent y hnging the tivity of PAL n CHI. Effet of light omintions on the levels of rutin n queretin As previous stuies reporte, the flvonoi rutin hs high ntioxint tivity [Yng et l., 28] n queretin is the egrtion prout of rutin, retion tlyze y RDEs. Stpleton & Wlot [1994] n Lois [1994] inite tht the umultion of flvonois inluing rutin n queretin prevent plnts from environmentl mge (suh s ultrviolet). In this stuy, lue light n /C or FIGURE 4. Effet of irrition with ifferent light omintions on the ontent of rutin in trtry ukwhet sprouts. Vertil rs inite the stnr evition. BL+: irrition with lue light followe y ; +BL: irrition with followe y lue light; : irrition with lue light followe y ; : irrition with followe y lue light. Different letters show signifint ifferene (P<.5). Unuthentite Downlo Dte 1/3/18 3:43 AM

5 H. Ji et l. 97 Content of queretin (mg/g ry mtter) BL n. light n / or their omintions on DPPH ril svenging tivity showe the sme tren thn tht of rutin ontent. Some previous stuies inite tht mong phenoli ompouns, rutin h the highest ril svenging tivity [Wtne, 27] n it exhiite the strong DPPH ril svenging tivity [Yng et l., 28]. Therefore, DPPH ril svenging tivity ws gretly influene y rutin ontent. BL+ +BL FIGURE 5. Effet of irrition with ifferent light omintions on the ontent of queretin in trtry ukwhet sprouts. Vertil rs inite the stnr evition. BL+: irrition with lue light followe y ; +BL: irrition with followe y lue light; : irrition with lue light followe y ; : irrition with followe y lue light. Different letters show signifint ifferene (P<.5). DPPH ril svenging tivity (%) BL+ +BL FIGURE 6. Effet of irrition with ifferent light omintions on DPPH ril svenging tivity in trtry ukwhet sprouts. Vertil rs inite the stnr evition. BL+: irrition with lue light followe y ; +BL: irrition with followe y lue light; : irrition with lue light followe y ; : irrition with followe y lue light. Different letters show signifint ifferene (P<.5). their omintions signifintly inrese the levels of rutin (Figure 4) n queretin ompouns (Figure 5). The rutin ontents in sprouts irrite with n were n mg/g ry mtter, respetively, whih ws higher thn these of the others. The rutin ontent in irrition with lue light irrite first in omintion of lue light n () ws 13.5% higher thn tht of ( +BL), ut it showe no ifferene in the sprouts irrite with omintion of n lue light either. DPPH ril sveng ing tivity The ntioxint tivity of rutin ws etermine in this stuy s DPPH ril svenging tivity (Figure 6). DPPH ril svenging tivity in the n ws pproximtely 1.48 fol higher thn rkness. There were no ifferenes from the sttistil point of view etween irrition with,, n with the omintions BL+, CONCLUSION On the sis of the results otine, we n onlue tht lue light in omintion with () enhne the umultion of iotive ompouns suh s totl flvonois, rutin n queretin in trtry ukwhet sprouts while this effet ws not oserve when it ws omine with (BL+). ACKNOWLEDGEMENT This reserh ws finnilly supporte y Ntionl Nturl Siene Fountion of Chin ( ). REFERENCES 1. Assis J.S., Mlono R., Munoz T., Esrino M.I., Meroio C., Effet of high ron ioxie onentrtion on PAL tivity n phenoli ontents in ripening herimoy fruit. Posthrv. Biol. Te hnol., 21, 23, Clwell M.M., Bjorn L.O., Bommn J.F., Flint S.D., Kulnivelu G., Termur M., Tevini M., Effet of inrese solr ultrviolet rition on terrestril eosystem. J. Photoh. Photoi ol. B., 1998, 46, Chen K., Feng H., Zh ng M., Wng X., Nitri oxie llevi tes oxitive mge in the green lge Chlorell pyrenoios use y UV-B rition. Foli Miroiol., 2 3, 48, Chen P., Gu J.J., A rpi mesurement of rutin-egring enzyme tivity in extrt of trtry ukwhet sees. Foo Biopro. Proe ss., 211, 89, Cortell J.M., Kenney J.A., Effet of shing on umultion of flvonoi ompouns in (Vitis vinifer L.) pinot noir fruit n extrtion in moel system. J. Agr. Foo Chem., 26, 54, Cresy L.L., The in rese in phenyllnine mmoni-lyse tivity in strwerry lef isks n its orreltion with flvonoi synthesis. Phytohemistry, 1968, 3, Jenkins G.I., UV n lue light signl trnsution in Ariopsis plnt. Plnt Cell Environ., 1997, 2, Khtmi F., Ghnti F., Effets of UV irrition on ell viility, nthoynin, n flvonoi ontents of llus-ulture Mlv neglet ells. ICLST, 211, 3, Kim S.L., Kim S.K., Prk C.H., Introution n nutritionl evlution of ukwhet sprouts s new vegetle. Foo Res. Int., 24, 37, Koes R.E., Quttrohio F., Mol J.N.M., The flvonoi iosyntheti pthwy in plnts: funtion n evolution. BioEssys,1994, 16, Ling B., Hung X.H., Zhng G.S., Zhng F., Zhou Q., Effet of Lnthnum on plnts uner supplementry ultrviolet-b r- Unuthentite Downlo Dte 1/3/18 3:43 AM

6 98 Effet of Light on Flvonois of Trtry Bukwhet Sprouts ition: Effet of Lnthnum on flvonoi ontents in soyen seelings expose to supplementry ultrviolet-b rition. J. Rre Erth., 26, 24, Lister C.E., Lnster J.E., Wlker J.R., Developmentl hnges in enzymes of flvonoi iosynthesis in the skins of re n green pple ultivrs. J. Si. Foo Agr., 1996, 71, Lois R., Aumultion of UV-soring flvonois inue y UV-B rition in Amiopsis thlin L. Plnt, 1994, 194, Mtsu R.K., Ohshi-Kneko K., Fujiwr K., Kurt., Effets of lue light efiieny on limtion of light energy prtitioning in PSII n CO 2 ssimiltion pity to high irrine in spinh leves. Plnt Cell Physiol., 28, 49, Nkmur K., Nrmoto K., Koym M., Bloo-pressure-lowering effet of fermente ukwhet sprouts in spontneously hypertensive rts. J. Funt. Foos, 213, 5, Nkjim K., Yoshie-Strk Y., Ogushi M., Comprison of ACE inhiitory n DPPH ril svenging tivities of fish musle hyrolystes. Foo Chem., 29, 114, Ohl S., Hhlrok K., Shǎfer E., A stle lue light erive signl moultes ultrviolet light inue tivtion of the hlone synthse gene in ulture p rsley ells. Plnt, 1989, 177, Piett P.G., Flvonois s ntioxints. J. Nt. Pro., 2, 63, S tokov L., Mtejov E., Jnovsk D., Sykorov S., Comprison of nlytil methos for rutin etermintion in trtry ukwhet. Chem. Listy, 29, 13, Ser N., Aou-Zei H., Brkt S., Effet of rition qulity on phenyllnine mmoni-lyse n pigment ontent in the shoots of ro en (Vii f) seelings. Phyton (Horn, Austri), 1998, 38, Stpleton A.E., W lot V., Flvonois n protet mize DNA from the inution of ultrviolet rition mge. Plnt Physiol., 1994, 15, T surung Y., Tkhshi T., Ktsue T., Kuo A., Kurmitsu O., Ishiwt M., Mtsumoto S., Effets of UV-B irrition on the levels of nthoynin, rutin n ril svenging tivity of ukwhet sprouts. Foo Chem., 213, 141, Tkht Y., Ki Y., Tnk M., Nkym H., Yoshing M., Enlrgement of the vrines in mount n omposition of nthoynin pigments in sweetpotto storge roots n their effet on the ifferenes in DPPH ril-svenging tivity. Si. Hort-Amsterm, 211, 127, W tne M., An nthoynin ompoun in ukwhet sprouts n its ontriution to ntioxint pity. Biosi. Bioteh. Biohem., 27, 71, Wng L., Li X.D., Niu M., Wng R., Chen Z.X., E ffet of itives on flvonois, D-hiro-Inositol n trypsin inhiitor uring the germintion of trtry ukwhet sees. J. Cerel Si., 213, 58, Whitelm G., Hlliy K., Light n plnt evelopment. 27. Blkwell Pulishing, Oxfor, UK. 27. Winkel-Shirley B., F lvonoi iosynthesis. A olorful moel for genetis, iohemistry, ell iology, n iotehnology. P lnt Physiol., 21, 126, Yng J.X., Guo J., Yun J.F., In vitro ntioxint properties of rutin. LWT-Foo Si. Tehnol., 28, 41, Zhng M., An L., Feng H., Chen T., Chen K., Liu Y., Tng H., Wng C.H., The se mehnisms of nitri oxie s seon messenger of ultrviolet B in inhiiting mesootyl elongtion. Photohem. Photoiol., 23, 77, Sumitte: 3 August 214. Revise: 2 My n 13 June 215. Aepte 3 June 215. Pulishe on-line: 31 Mrh 216. Unuthentite Downlo Dte 1/3/18 3:43 AM

Title of Experiment: Author, Institute and address:

Title of Experiment: Author, Institute and address: Title of Experiment: Trsfetion of murine mrophge RAW264.7 ells with METAFECTENE PRO. Author, Institute n ress: Ptrizi Pellegtti n Frneso Di Virgilio. Deprtment of Experimentl n Dignosti Meiine, Setion