Journal of Integrative Agriculture 2017, 16(0): Available online at ScienceDirect. , ZHU Dan-shi

Transcription

1 Journl of Integrtive griulture 17, 1(): ville online t SieneDiret RESERCH RTICLE Effets of thimine on Trihotheium nd lternri rots of muskmelon fruit nd the possile mehnisms involved GE Yong-hong 1,, 3, LI Cn-ying 1,, 3, LV Jing-yi 1,, 3 1,, 3, ZHU Dn-shi 1 College of Food Siene nd Engineering, Bohi University, Jinzhou 1113, P.R.Chin Food Sfety Key L of Lioning Provine, Jinzhou 1113, P.R.Chin 3 Ntionl & Lol Joint Engineering Reserh Center of Storge, Proessing nd Sfety Control Tehnology for Fresh griulturl nd quti Produts, Jinzhou 1113, P.R.Chin strt The effets of thimine ginst pink nd lk spot rots used y Trihotheium roseum nd lternri lternt nd modultion on the metolism of retive oxygen speies (ROS) nd phenylpropnoid pthwy were investigted in this pper. In vitro test indited tht thimine signifintly inhiited myeli growth nd spore germintion of T. roseum nd. lternt. Thimine t 1 mmol L 1 effetively inhiited lesion development of muskmelon fruit inoulted with T. roseum or. lternt, enhned prodution rte of O nd H O ontent, tivities of tlse (CT), sorte peroxidse (PX) nd glutthione redutse (GR) in muskmelon fruit. Thimine lso ffet phenylpropnoid pthwy in muskmelon fruit y inresing the tivities of phenyllnine mmoni lyse (PL) nd peroxidse (POD), the ontent of totl phenoli ompounds, flvonoids nd lignin. These results suggest tht the effets of thimine on pink nd lk spot rots in muskmelon fruits re ssoited with its diret fungitoxi ginst the pthogens nd the modultion of O nd H O prodution, eliminting enzymes nd phenylpropnoid pthwy. Keywords: indued resistne, thimine, muskmelon fruit, ntimiroil tivity, phenylpropnoid pthwy 1. Introdution Muskmelon (Cuumis melo L.) fruit is one of the most importnt hortiulturl rops ultivted in the northwest of Chin (Ren et l. 1). However, fruit re esily infeted y Trihotheium roseum nd lternri lternt whih use pink nd lk spot rots in the fruit nd leds to onsiderle Reeived 1 Septemer, 1 epted 5 Feury, 17 Correspondene GE Yong-hong, E-mil: geyh1979@13.om 17, CS. ll rights reserved. Pulished y Elsevier Ltd. doi: 1.11/S (1)84-8 posthrvest losses in Chin (Ge et l. 8). pplitions of syntheti fungiides, suh s imzlil, iprodione, zoxystroin, effetively ontrol posthrvest diseses of muskmelon (hroni et l. 199; M et l. 4). However, fungiide usge hs eome limited due to the development of fungiide resistnt pthogens, potentil effets on humn helth nd environment, nd the need for new lterntive methods hve een proposed (Tripthi nd Duey 4; Bi et l. 9). One of the potentil methods of ontrolling disese is the indued resistne in plnts nd fruit (Ku 1). Severl litertures hve showed tht vitmins, suh s rioflvin (vitmin B ), mendione sodium isulphite (vitmin K 3 ) nd thimine (vitmin B 1 ) n indue disese resistne in plnts or fruit (Dong nd Beer ; Borges et l. 4; hn et l. 5; Yin et l. 1). Prehrvest pplition of thimine

2 *** et l. Journl of Integrtive griulture 17, 1(): indued resistne in perl millet, grpevine, rie nd rley ginst Slerospor grminiol, Plsmopr vitiol, Rhizotoni solni nd Rhoplosiphum pdi (Pushplth et l. 11; Bhugun et l. 1; Boukri et l. 1; Boukri et l. 13; Hmd nd Jonsson 13). In vitro test indited tht thimine n inhiit spore germintion nd myeli growth of. lternt (Yin et l. 1). Posthrvest thimine dipping tretment effetively inhiited lesion development in sin per fruit inulted with. lternt (Yin et l. 1). These results suggested tht deresed disese development y thimine ws losely relted to modultion of vrious physiologil, hemil nd moleulr host-defense responses inluding genertion of retive oxygen speies (ROS), pthogenesis-relted (PR) genes expression, phenolis nd lignin umultion, llose deposition, hypersensitive responses nd diret inhiition the pthogen growth ( hn et l. 7; Bhugun et l. 1; Boukri et l. 1; Yin et l. 1; Boukri et l. 13). However, the thimine effiieny in disese ontrol nd the physiologil nd hemil hnges indued in muskmelon fruit re still unknown. Therefore, the ojetives of this study were to investigte the effets of thimine on the ontrol of the posthrvest diseses used y T. roseum nd. lternt in muskmelon fruit, to evlute the ntifungl tivity of thimine ginst T. roseum nd. lternt in vitro, to determine the produetion rte of O nd H O, eliminting enzymes tivities, nd to investigte the enzymes tivities nd seondry metolites of phenylpropnoid pthwy in the muskmelon fruit.. Mterils nd methods.1. Fruit nd tretment Muskmelon fruit (Cuumis melo L. v. Jinhongo) were hrvested t 4 d fter full lossom in the field in Lioning, Chin. Fruit without mehnil injuries or diseses were lened, disinfeted, nd susequently immersed in, 5, 5, 1 mmol L 1 thimine solutions for 1 min. fter ir-drying, fruit were kept in rtons, nd stored t room temperture (± C, RH 7 8%) for the following experiments. Three replites per tretment were mde, eh replite ontining fruits, nd the entire experiment ws performed twie... Pthogen nd inoultion T. roseum nd. lternt were originlly isolted from dey fruit nd ultured on potto dextrose gr (PD) t 5 C. Spore suspensions were prepred y flooding the 7-dy-old ulture pltes with 4 5 ml of sterile distilled wter. The inoulums were diluted to spores ml 1 nd onfirmed using hemoytometer. Inoultions were rried out 4 h fter thimine tretment ording to Ren et l. (1). ll fruit were sterilized with 7% ethnol round the equtor of eh fruit, nd then four wounds were mde with disseting needle (3-mm deep 3-mm dimeter). volume of μl of spore suspensions ws injeted into eh wound. Fruit were ir-dried, then kept in rtons nd inuted t room temperture. The lesion dimeter ws reorded 7 d fter inoultion..3. Effets of thimine on spore germintion nd myeli growth of T. roseum nd. lternt in vitro To ssess the effets of thimine on spore germintion of T. roseum nd. lternt, µl liquots of spore suspensions (1 1 5 spores ml 1 ) were dded into 5 ml potto dextrose lended with, 5, 5, 1 mmol L 1 thimine, nd then inuted in Petri dishes t 5 C. fter 4,, 8 h of inution, the germintion rtes were ssessed y oserving 1 spores per tretment replite under light mirosope. Three replites per tretment were mde, nd the experiment ws performed twie. Effets of thimine on myeli growth of T. roseum nd. lternt were ssessed ording to the method of Li et l. (9). Myeli disks (5-mm in dimeter) from 7-dy-old ultures were pled in the entre of Petri dishes (9-mm in dimeter) with ml of PD ontining different onentrtions of thimine (, 5, 5, 1 mmol L -1 ), nd then inuted t 5 C. The myeli growth ws determined y mesuring the olony dimeter 7 d fter inoultion. Three replites per tretment were mde, nd the experiment ws performed twie..4. Smples olletion pproximtely 3. g of flesh tissue ws tken from 3 8 mm elow the skin round the equtor of fruit,, 4,, 8 d fter 1 mmol L -1 thimine nd distilled wter tretment. Eh smple ws pked in luminum foil individully nd frozen in liquid nitrogen immeditely, nd kept t 8 C until the iohemil determintion. There were three replites for enzyme ssys in eh tretment, nd the experiment ws repeted twie..5. Prodution rte of O nd H O ontent ssy Prodution rte of O ws determined ording to the method of Ren et l. (1). The retion mixture ontined ml of the superntnt, 1 ml 5 mmol L -1 phosphte uffer (ph 7.8), nd.5 ml hydroxylmine hydrohloride (1 mmol L -1 ) solution for 3 min t 5 C. Then 4-minoenzene sulfoni id (1 ml, 17 mmol L -1 ) nd 7 mmol L -1 α-nphthylmine

3 4 *** et l. Journl of Integrtive griulture 17, 1(): were dded for further 4 min. fter tht 4 μl of n-utnol ws dded into the retion mixture, nd n-utnol phse ws used for the determintion of O. The prodution rte of O ws expressed s ΔOD 53 min -1 g -1 FW. H O ontent ws ssyed y the method of Ren et l. (1). 1 ml of the superntnt ws re-entrifuged followed y the ddition of μl, % titnium tetrhloride, nd μl of onentrted mmoni solution to preipitte the titnium-hydro peroxide omplex. Preipitte ws wshed repetedly y old etone nd then dissolved in 3 ml of 1 mol L -1 H SO 4 nd then re-entrifuged. H O ontent ws monitored y tking the sorne t 41 nm nd expressed s µmol H O g -1 FW... Crude enzymes extrtion ll enzyme extrts were onduted t 4 C. Frozen smples (3. g) were ground in mortr on ie, using the following extrtion solutions: 5 ml of 5 mmol L -1 phosphte uffer (ph 7.5) ontining 1 g L -1 of polyvinylpyrrolidone (PVP),.1% Triton X-1 (v/v) for peroxidse (POD); 5 ml phosphte uffer (ph 7., 5 mmol L -1 ) ontining 5 mmol L -1 dithiothreitol (DTT) nd 8 g L -1 PVP for tlse (CT); 4 ml phosphte uffer (ph 7.5, 5 mmol L -1 ) ontining 1 mmol L -1 ethylenediminetetreti id (EDT) for sorte peroxidse (PX) nd glutthione redutse (GR); 5 ml,. mol L -1 sodium orte uffer (ph 8.7) ontining 5 mmol L -1 β-merptoethnol, mmol L -1 EDT nd 8 g L -1 of PVP for phenyllnine mmoni lyse (PL). The homogentes were entrifuged t g for min, nd the superntnts were ssyed for enzymti tivities..7. Enzyme tivity ssys CT tivity ws ssyed ording to Ren et l. (1) following the dispperne of H O t 4 nm. CT tivity ws expressed s ΔOD 4 g -1 FW. PX tivity ws determined s desried y Nkno nd sd (1987) with slight modifitions. The retion mixture inluded ml phosphte uffer (.1 mol L -1, ph 7.5), µl, 5 mmol L -1 sori id, 1 µl rude enzymes nd µl H O (1 mmol L -1 ). sorne of the solution ws mesured t 9 nm. PX tivity ws expressed s µmol s g -1 FW. GR tivity ws ssyed y the method of Foyer nd Hlliwell (197) with minor modifitions. The retion mixture inluded ml phosphte uffer (.1 mol L -1, ph 7.5), µl, 5 mmol L -1 redued glutthione (GSH), 1 µl rude enzymes nd 3 µl 4 mmol L -1 NDPH. sorne of the solution ws mesured t 34 nm. GR tivity ws expressed s µmol NDPH g -1 FW. PL tivity ws ssyed s desried y Kozukue et l. (1979) with some modifitions. The retion solution ontined 3 ml of L-phenyllnine nd 5 µl of rude enzyme extrt. The mixture ws inuted t 4 C for 1 h nd stopped y the ddition of. ml of mol L -1 HCl. sorne t 9 nm ws determined for PL. PL tivity ws expressed s U g -1 FW, where U=.1 ΔOD 9 min -1. POD tivity ws ssyed ording to the method of Liu et l. (5). The retion mixture ontined µl of rude enzyme extrt,.5 ml guiol (5 mmol L -1 ), nd µl of 5 mmol L -1 H O. The oxidtion rte of guiol ws monitored t 47 nm for min. POD tivity ws expressed s U g -1 FW, where U=.1 Δ OD 47 min Totl phenoli ompounds, flvonoids nd lignin ontents ssy Totl phenoli ompounds nd flvonoids ontents were determined ording to the method of Wng et l. (11). The ontents of totl phenoli ompounds nd flvonoids were expressed s OD 35 g -1 FW nd OD 8 g -1 FW, respetively. Lignin ontent ws determined s desried y Wng et l. (11). The lignin ontent ws expressed s OD 8 g -1 FW..9. Sttistil nlysis ll sttistil nlyses were performed using SPSS 17. (SPSS In., Chigo, IL, US). Fisher s lest signifint differenes (LSD) t P<.5 were determined to ompre differenes etween mens. Dt re presented s the stndrd error of mens. 3. Results 3.. Effets of thimine on spore germintion nd myeli growth of T. roseum nd. lternt Thimine t 5, 5, 1 mmol L -1 signifintly inhiited myeli growth of T. roseum nd. lternt in vitro, with highest onentrtion hve greter inhiition (Fig. 1- nd B). The inhiitory rte of T. roseum nd. lternt treted with 1 mmol L -1 thimine ws 55.5 nd 5.7%, respetively. ll onentrtions of thimine signifintly inhiited spore germintion of T. roseum nd. lternt, with higher onentrtions hving greter inhiition (Fig. 1-C nd D). fter 8 h inution, thimine t 1 mmol L -1 signifintly inhiited spore germintion of T. roseum nd. lternt to rte of 41 nd 5% Effets of thimine on pink nd lk spot rots development in muskmelon fruit Posthrvest pplition of thimine signifintly deresed

4 *** et l. Journl of Integrtive griulture 17, 1(): Colony dimeter (m) 8 4 C Spore germintion (%) Control d d B Colony dimeter (m) D 1 Spore germintion (%) 8 4 d d Control Conentrtion of thimine (mmol L 1 ) 4 8 Inution time (h) Fig. 1 Effets of different onentrtions of thimine tretment on spore germintion nd myeli growth of Trihotheium roseum ( nd C) nd lternri lternt (B nd D) t 5 C for 7 dys inution. Brs with different letters re signifintly (P<.5) different ording to lest signifint differenes (LSD). lesion dimeter in muskmelon fruit inoulted with T. roseum or. lternt ompred to the ontrol fruit (Fig. - nd B). The redution ws enhned y inresing thimine onentrtions. fter inution t ± C for 7 d, fruits treted with thimine t 1 mmol L -1 showed the lest lesion dimeter whih ws 3.1 nd 38.7% lower thn the ontrol fruit inoulted with T. roseum or. lternt, respetively. Lesion dimeter (m) Effets of thimine tretment on O nd H O prodution nd ntioxidtive enzymes tivity Prodution rte of O mintined t reltively low level in the ontrol fruit, nd ws signifintly lower thn tht in thimine-treted fruit (Fig. 3-). The highest level of O prodution rte ws determined t d fter tretment whih ws 4.% higher thn tht in the ontrol fruit. H O ontent ws inresed t first d fter tretment, peked t 4 d fter tretment nd then deresed, ut thimine tretment signifintly inresed the ontent of H O in muskmelon fruit (Fig. 3-B). The tivity of CT in the ontrol fruit kept t low level throughout the ssy time. Thimine tretment enhned CT tivity, nd peked t d fter tretment whih ws 5.5% higher thn tht in the ontrol fruit (Fig. 4-). PX tivity ws indued y thimine tretment in muskmelon fruit (Fig. 4-B). The highest tivity of PX ws determined t d fter tretment, whih ws 4.9% higher B Lesion dimeter (m) Control Conentrtion of thimine (mmol L 1 ) Fig. Effets of different onentrtions of thimine tretment on lesion development in muskmelon fruit inoulted with Trihotheium roseum () nd lternri lternt (B) storge t room temperture 7 dys fter tretment. Brs with different letters re signifintly (P<.5) different ording to lest signifint differenes (LSD).

5 *** et l. Journl of Integrtive griulture 17, 1(): Control Thimine 1 Control Thimine. prodution rte of O (ΔOD 53 min g 1 FW) 4 CT tivity (U g 1 FW) 8 4 B B 35 H O ontent (µmol H O g 1 FW) Dys fter tretment (d) PX tivity (U g 1 FW) C Fig. 3 Effets of thimine tretment on O prodution rte () nd H O ontent (B) in muskmelon fruit. Eh vlue is the men for three replites nd the vertil r indites the stndrd error. thn tht in the ontrol fruit. Thimine tretment lso signifintly inresed GR tivity in muskmelon fruit throughout the ssy time, nd peked t d fter tretment tht ws 8.8% higher thn ontrol fruit (Fig. 4-C). GR tivity (U g 1 FW) Dys fter tretment (d) 3.4. Thimine eliited the key enzymes tivities nd ntifungl ompounds ontents in phenylpropnoid pthwy PL tivity signifintly inresed in muskmelon fruit fter thimine tretment, nd peked t d fter tretment (Fig. 5-). POD tivity ws lso indued y thimine tretment, nd signifint differenes were determined t 4, nd 8 d fter tretment whih ws 45.5, 5.5 nd 1.4% higher thn tht in the ontrol fruit (Fig. 5-B). Content of totl phenoli ompounds inresed y thimine tretment, nd peked t 4 d fter tretment whih ws signifintly higher thn tht in the ontrol fruit (Fig. 5-C). Posthrvest thimine tretment signifintly inresed the ontent of flvonoids nd lignin (Fig. 5-D). The highest level of flvonoids ws oserved t 4 d fter tretment, whih ws 54.% higher thn tht in the ontrol fruit (Fig. 5-D). The lignin ontent in thimine-treted fruit rehed the mximum level t d fter tretment, whih ws 8% higher thn tht in the ontrol fruit (Fig. 5-E). Fig. 4 Chnges in tivity of tlse (CT; ), sorte peroxidse (PX; B) nd glutthione redutse (GR; C) in muskmelon fruit treted with thimine. Eh vlue is the men for three replites nd the vertil r indites the stndrd error. 4. Disussion Severl hemils hve een reported to indue disese resistne in plnts or fruit y either diret inhiition pthogen growth or/nd indution of disese to pthogen invsion (Wng et l. 1; Li W H et l. 1; Li Y C et l. 1; Yin et l. 1). Thimine hs een reently reported to indue resistne in sin per fruit through tivting defense-relted enzymes nd umultion of O nd H O (Yin et l. 1). The present study indited tht 5, 5, 1 mmol L -1 thimine signifintly inhiited spore germintion nd myeli growth of T. roseum nd. lternt. Our results were greed with Yin et l. (1) who found tht 5 mmol L -1 thimine inhiited spore germintion nd myeli growth of. lternt whih used lk spot of per fruit.

6 *** et l. Journl of Integrtive griulture 17, 1(): Control Thimine 5 B 5 PL tivity (U g 1 FW) C Totl phenoli ompounds ontent (OD 35 g 1 FW) Dys fter tretment (d) E Lignin ontent (OD 8 g 1 FW) POD tivity (U g 1 FW) 1 5 D Flvonoids ontent (OD 8 g 1 FW) Dys fter tretment (d) 4 8 Dys fter tretment (d) Fig. 5 Effets of thimine tretment on the tivity of phenyllnine mmoni lyse (PL; ) nd peroxidse (POD; B), the ontent of totl phenoli ompounds (C), flvonoids (D) nd lignin (E) in muskmelon fruit. Eh vlue is the men for three replites nd the vertil r indites the stndrd error. However, hn et l. (5) reported tht thimine t low onentrtions (<5 mmol L -1 ) did not inhiit the growth of Mgnporthe grise or Xnthomons oryze pv oryze. The differenes ould e explined tht the ntifungl tivity of thimine depends on using onentrtions or vry mong pthogen nd host speies. We lso found tht posthrvest pplition of thimine signifintly deresed lesion dimeter in muskmelon fruit inoulted with T. roseum or. lternt, nd inhiition effiy showed dose-dependent. Similr results were reported in sin per, rie, too, tomto, uumer, perl millet nd ridopsis ginst semi-iotrophi nd iotrophi pthogens (hn et l. 5; hn et l. 7; Liu et l. 8; Pushplth et l. 11; Yin et l. 1). hn et l. (5) hve firstly reported tht thimine indued resistne in rie nd ridopsis through tivtion of host defense responses. Our findings indited tht thimine funtion s eliitor gent in muskmelon fruit euse of induing host defense responses without ny pthogen infetion. Thimine indued the umultion of retive oxygen speies t the erly stge fter tretment, nd retive oxygen speies served s signl to tivte the prodution of pthogenesis-relted proteins nd phenylpropnoid pthwy to enhne the resistne of muskmelon fruit to pthogen invsion. The present results indited tht pplition of thimine fter hrvest indued O prodution nd H O umultion in muskmelon fruit. Boukri et l. (1) found tht umultion of H O in grpevine ws losely relted with resistne ginst Plsmopr vitiol. Study in per fruit lso showed tht thimine indued H O prodution ws enefiil to disese resistne ginst. lternt (Yin et l. 1). Genertion of ROS is one of

7 8 *** et l. Journl of Integrtive griulture 17, 1(): the erliest ellulr events in plnts or fruit fter pthogen invsion or following eliitor tretment (Boukri et l. 1). H O hs severl funtions in host ginst pthogen infetion inluding diret inhiition pthogen growth, tivtion PR genes expression, ross-linking ell wll, triggering phytolexins umultion nd hypersensitive responses (Torres et l. ; Shetty et l. 8). The present study lso indited tht thimine inresed the tivity of CT, PX nd GR in muskmelon fruit. Study hs shown tht pplition of sori id improved ntioxidnt enzymes inluding CT, SOD of Pihi rii to enhne its ioontrol effiy on pple fruit (Li et l. 14). CT nd PX n svenge exess H O to eliminte the dmge to host tissues. GR keeps the lne etween redued glutthione (GSH) nd glutthione disulfide (GSSH) through sorte-glutthione (s-gsh) yle. Different result ws found in per fruit tht CT tivity ws inhiited y thimine tretment (Yin et l. 1). This differene suggested tht the H O svenging system in muskmelon fruit mye depend on oth CT nd s-gsh yle. Similr induing effets in fruit hve een reported fter tretment with other eliitors, inluding SM (Ren et l. 1), sodium silite (Li et l. 1). In ddition, POD n lso eliminte H O to void hrmful dmge to ells. The present study showed tht thimine tretment signifintly inresed PL tivity in muskmelon fruit. s rte-limiting enzyme in the phenylpropnoid pthwy, the inresing tivity of PL is ssoited with the iosynthesis of ntifungl ompounds inluding phenolis, flvonoids nd lignin (Vogt 1). pplition of thimine fter hrvest lso indued the inrese of totl phenoli ompounds, flvonoids nd lignin in the muskmelon fruit. In ddition, the umultion of phenolis my provide dequte sustrte to produe fungitoxi quinines t the presene of polyphenol oxidse (PPO) nd/or POD to diretly inhiit pthogen growth (Lttnzio et l. ). Phenolis hve lso een reported to e involved in the restrition of. lternt growth in per fruit (Yin et l. 1). Similr results hve een found in muskmelon fruit pre-hrvest or posthrvest treted with SM (Zhng et l. 11; Liu et l. 14). Higher levels of POD were found in muskmelon fruit fter treting with thimine. POD is lso involved in lignin umultion together with innmte-4-hydroxylse nd lse (Liu et l. 14). The umultion of lignin ould strengthen the ell wll nd t s n effetive rrier ginst pthogen penetrtion nd spred (Hemty et l. 9; Kärkönen nd Köutniemi 1). Indution of lignin umultion ws lso reported in muskmelon fruit fter dipping with SM (Liu et l. 14). Flvonoids re phenylpropnoid metolites, whih re synthesized from one of the rnhes of the phenylpropnoid pthwy nd signifintly ontriuted to nthoynin formtion (lln et l. 8). Flvonoids ontriute to resistne ginst pthogens ommonly found during the storge of fruit nd vegetles, nd lso hve ntimiroil iotivity (Pn nd Liu 11). In our experiment, thimine indued the deposition of flvonoids in muskmelon fruit. Studies in grpevine, sin per fruit demonstrted tht thimine tretment inresed flvonoids ontent y inresing the tivities of key enzymes in phenylpropnoid pthwy (Yin et l. 1; Boukri et l. 13). These results suggest tht inreses in tivities of PL, POD re losely relted to phenylpropnoid metolites umultion in fruit tissues. 5. Conlusion Our dt demonstrte tht thimine ould diretly inhiit the growth of T. roseum nd. lternt nd tivte the phenylpropnoid pthwy, prodution of ROS nd enhned ntioxidtive enzymes tivities in muskmelon fruit. This suggests tht posthrvest pplition of thimine ould e promising in reduing dey nd utiliztion of syntheti fungiides to ontrol posthrvest diseses in fruit nd vegetles. However, further studies t the moleulr level re needed to investigte the tion of thimine. knowledgements This study ws supported y the Ntionl Nturl Siene Foundtions of Chin (31454), the Dotorl Initil Funding of Bohi University, Chin (sqd145). Referenes hroni Y, Copel, Dvidson H, Brki-Goln R Fungiide pplition inwter nd in wx for dey ontrol in Gli melons. New Zelnd Journl of Crop nd Hortiulturl Siene,, hn I P, Kim S, Lee Y H. 5. Vitmin B1 funtions s n tivtor of plnt disese resistne. Plnt Physiology, 138, 5. hn I P, Kim S, Lee Y H. Suh S C. 7. Vitmin B1-indued priming is dependent on hydrogen peroxide nd the NPR1 gene in ridopsis. Plnt Physiology, 143, lln C, Hellens R P, Ling W. 8. MYB trnsription ftors tht olor our fruit. Trends in Plnt Siene, 13, Bhugun R N, Joshi R, Shukl, Pndey M, Kumr J. 1. Thimine primed defense provides relile lterntive to systemi fungiide rendzim ginst sheth light disese in rie (Oryz stiv L.). Plnt Physiology nd Biohemistry, 57, Bi Y, Li Y C, Ge Y H, Wng Y. 9. Indued resistne in melons y eliitors for the ontrol of posthrvest diseses. Posthrvest Pthology,, Borges, Perez B, Flon M F. 4. Indued resistne to fusril wilt of nn y mendione sodium isulphte

8 *** et l. Journl of Integrtive griulture 17, 1(): tretments. Crop Protetion, 3, Boukri H, Poutrud, Wh M, Clyeux C, Bltenwek- Guyot R, Steyer D, Mri C, Mliki, Soustre-Gougnolle I. 13. Thimine modultes metolism of the phenylpropnoid pthwy leding to enhned resistne to Plsmopr vitiol in grpevine. BMC Plnt Biology, 13, Boukri H, Wh M, Chong J, Bertsh C, Mliki, Soustre- Gougnolle I. 1. Thimine indued resistne to Plsmopr vitiol in grpevine nd eliited host defense responses, inluding HR like-ell deth. Plnt Physiology nd Biohemistry, 57, Dong H, Beer S V.. Rioflvin indues disese resistne in plnts y tivting novel signl trnsdution pthwy. Phytopthology, 9, Foyer C H, Hlliwell B The presene of glutthione nd glutthione redutse in hloroplsts: proposed role in sori id metolism. Plnt, 133, 1 5. Ge Y H, Bi Y, Li X, Li M. 8. Indueds resistne ginst Fusrium nd pink rotsy ienzolr-s-methyl in hrvested muskmelon (v. Yindi). griulturl Siene in Chin, 7, Hmd M, Jonsson L M V. 13. Thimine tretments llevite phid infesttions in rley nd pe. Phytohemistry, 94, Hemty K, Cherk C, Somerville S. 9. Host pthogen wrfre t the plnt ell wll. Current Opinion in Plnt Biology, 1, Kärkönen, Köutniemi S. 1. Lignin iosynthesis studies in plnt tissue ultures. Journl of Integrtive Plnt Biology, 5, Kozukue N, Kozukue E, Kishiguhi M Chnges in the ontents of phenoli sustnes, phenyllnine mmonilyse (PL) nd tyrosine mmonilyse (TL) ompnying hilling injury of eggplnt fruit. Sienti Hortiulture, 11, Ku J. 1. Conepts nd diretions of indued systemi resistne in plnts nd its pplition. Europen Journl of Plnt Pthology, 17, 7 1. Lttnzio V, Lttnzio V M T, Crdinli.. Role of phenolis in the resistne mehnisms of plnts ginst fungl pthogens nd insets. Phytohemestry, 37, 3 7. Li C L, Zhng H Y, Yng Q Y, Mhunu G K, Zhng X Y, Zhu S Y. 14. sori id enhnes oxidtive stress tolerne nd iologil ontrol effiy of Pihi rii ginst posthrvest lue mold dey of pples. Journl of griulturl nd Food Chemistry,, Li W H, Bi Y, Ge Y H, Li Y C, Wng J J, Wng Y. 1. Effets of posthrvest sodium silite tretment on pink rot disese nd oxidtive stress-ntioxidtive system in muskmelon fruit. Europen Food Reserh nd Tehnology, 34, Li Y C, Sun X J, Bi Y, Ge Y H, Wng Y. 9. ntifungl tivity of hitosn on Fusrium sulphureum in reltion to dry rot of potto tuer. griulturl Sienes in Chin, 8, Li Y C, Yin Y, Bi Y, Wng D. 1. Effet of rioflvin on posthrvest disese of si per nd the possile mehnisms involved. Phytoprsiti, 4, 1 8. Liu H X, Jing W B, Bi Y, Luo Y B. 5. Posthrvest BTH tretment indues resistne of peh (Prunus persi L. v. Jiuo) fruit to infetion y Peniillium expnsum nd enhnes tivity of fruit defense mehnisms. Posthrvest Biology nd Tehnology, 35, 3 9. Liu L Z, He H L, Pn J S, Du H, Ci R. 8. The primry reserh of indued resistne to uumer powdery mildews y vitmin B1 tretment. t phytophysiologi Sini, 35, Liu Y Y, Ge Y H, Bi Y, Li C Y, Deng H W, Dong B Y. 14. Effet of posthrvest ienzolr-s-methyl dipping on phenylpropnoid pthwy metolism in muskmelon (Cuumis melo L.) fruits. Sienti Hortiulture, 18, M L Y, Bi Y, Zhng Z K, Zho L, n L, M K Q. 4. Control of pre-nd posthrvest min diseses on melon vriety Yindi with pre-hrvest zoxystroin sprying. Journl of Gnsu griulturl University, 1, (in Chinese) Nkno Y, sd K Purifition of sorte peroxidse in spinh hloroplsts: Its intivtion in sorte-depleted medium nd retivtion y monodehydrosorte rdil. Plnt Cell Physiology, 8, Pn Y G, Liu X H. 11. Effet of enzo-thidizole-7- rothioi id S-methylester (BTH) tretment on the resistnt sustne in posthrvest mngo fruits of different vrieties. fri Journl of Biotehology, 1, Pushplth H G, Sudish J, Geeth N P, mruthesh K N, Shetty S H. 11. Thimine seed tretment enhnes LOX expression, promotes growth nd indues downy mildew disese resistne in perl millet. Biologi Plntrum, 55, Ren Y L, Wng Y F, Bi Y, Ge Y H, Wng Y, Fn C F, Li D Q, Deng H W. 1. Posthrvest BTH tretment indued disese resistne nd enhned retiveoxygen speies metolism in muskmelon (Cuumis melo L.) fruit. Europen Food Reserh nd Tehnology, 34, Shetty N P, Jørgensen H J L, Jensen J D, Collinge D B, Shetty H S. 8. Roles of retive oxygen speies in intertions etween plnts nd pthogens. Europen Journl of Plnt Pthology, 11, 7 8. Torres T, Jones J D G, Dngl J L.. Retive oxygen speies signling in response to pthogens. Plnt Physiology, 141, Tripthi P, Duey N K. 4. Exploittion of nturl produts s n lterntive strtegy to ontrol posthrvest fungl rotting of fruit nd vegetles. Posthrvest Biology nd Tehnology, 3, Vogt T. 1. Phenylpropnoid iosynthesis. Moleulr Plnt, 3, -. Wng J J, Bi Y, Zhng Z K, Zhng H Y, Ge Y H. 11. Redution of ltent infetion nd enhnement of disese resistne in muskmelon y prehrvest pplition of hrpin. Journl of griulture nd Food Chemistry, 59, Wng Y, Bi Y, Li Y C, Yin Y, Ge Y H. 1. ntifungl tivity of sodium silite on Trihotheium roseum in vitro. t Hortiulture, 877, Yin Y, Bi, Y, Li Y C, Wng Y, Wng D. 1. Use of thimine for ontrolling lternri lternt posthrvest rot in sin per (Pyrus retshneideri Rehd. v. Zosu). Interntionl Journl of Food Siene nd Tehnology, 47, Zhng Z K, Bi Y, Ge Y H, Wng J J, Deng J J, Xie D F, Wng Y. 11. Multiplepre-hrvest tretments with ienzolr- S-methyl redue ltent infetion nd indue resistne in muskmelon fruit. Sienti Hortiulture, 13, (Mnging editor SHI Hong-ling)