ANALISIS DE LA EXPRES! JON TRANSCRIPC }ONAL INDUCIDA BAJO CONDICIONES DE ESTRES BIOTICO Y ABIOTICO EN Capsicum chinense BG-3821 ANALYSIS OF TRANSCRIPTIONAL EXPRESSION INDUCED IN Capsien NDER CONDITIONS OF BIOTIC AND ABIQTIC STRI Altay Barret Pasha! Ma Crim 1, PévasPStea!, Lavina 4 anenio Garcia. Cubs, Gebya Sen Miguel de Alle sacultd de Invest Mores, (gerade sin, Cohoia Ri eSUMIEN EL ahjetivg dt pres chai fe anal Ia expres trans rip luis Ia dercaues une ears Mio » abd ica en plas te chile nadanero (Cmsteun earense 96-2921), Las estudioy dk eypeesiin ele genes de laa hao dlfevents condone de ese biitey y lsitiea son importartes pr ‘que sont uns estraegia past entender, a aivel molec, wena las plantas aetdan en fsmenos de reaiterss 2 paligenes y condones ubidticas Ue exuelnimto, lo que puede amplian ol conocininte sobre los mecaainmos molecules de defers 4 ests pes ae estrés «> bs plans, Mora mets transerlee faa se emplearon Sends moleculares de las EST RS® y R100, Aus san fesamentas diferencias ¥ posibiomeante inpliendos cou la respuesta de reste de C- Disenee BOR a tnkeeci fies par el sins Inasteco sna small det dks PHYVV), Se sampeobs la espesiicidad de ty exper tw de tas gems carson kitnets « KSI $ RIND ef past ce C. chigenve infection oom PITYV', ist como po fa cera Rowtderanas camper PR vesieneria ¥ el noun Fasarium slant. Planes infctadas con el gomicety Physyaliora caps? no indyjeron la expresion sila expres de etos ge nes os india medtine bx epleadtin de Side ssliico (ASI teelil Tasenoaln, eile « exis abiition Whidrien y téemicah oes Be estos genes. Asiana se eval 1 plata de Ce ohinoure Adicimalonent, so rein andlss de hbeidaeldu de APN compkmenticls «ADNel, para to cual se Selewonaron sania clanas de Danenssustestisas que contienen fragnnatas de gems de Cy chinense WSBT cya expresidn fue tgtiila en inixeiones de PLIY VY. Se demoste ‘aus kv expres de ls genes conresponaiantes 3 hos fragmeaen RS 9 ION no sa sf enn exneierneate por Is in i de PIV, ao atlecats por T eanpenris pr. yosearsria 1 F. solani, as cane por Ia aplicaion de dea scien met Jagnapate y eskeng. Kal ase de arregios ge aservaron Hifvencias en eb uived de expredn te varias EST para cada evalua, ouicn de es FResbiie: Nevietbee, 2008, Aprotuair Reviarbee 2007, Putco sum ARTICR! O em Agencies 42) 95.106, 2068 Abate ds J. Saugin Rea chinense BG-3821 friase Torres Pachow®, Mary M. Conedler-Ch evar Olver! y Rann G. Guenara- Gone? piri Bogan. teat Tenby de Cay, AVR Teenage OvIsssTE Cento, Campo experimen de &. Kin. 85, Fel saveewidad Audrina de Qvettan 3AGIO, Celaya, Guanajuin, Manion. “let agen Upidh da Bi Goria, Misi 6010 Apurvado Ps cere, Ga Aasteact af the preset work war tp analyze the raises intionst espuesonnn inde ander dfexe tyes of Wie aed ate stress Hava el peppsr Capoten ebiense BG 3401). The staes oF espresso af plant gees under erent coneltios OF bisie and abintle scree are faxportanl besavee ‘hoy ace a sirategs foe undersansing 3 mokeewar fev. Hew bas se in planet of tests otha au that srovth condiiens, which ‘way expand the knowledgs abuet inokchar defense anechanisar to this type of stress in plant. Far ¢raweriqional agus, molecskae proves of KS) ane RIVE EST'S were enployed. Bott ure deren fe sand pasty impticd in the resistunce vespense of C.ekiavane £H(71821 (8 jaferons by the pepper STvestteo yelss ven visas URYLY), Specifety of gene expressions correspondie vo RSI and RIL si contre fa C. ments we Hants infected by PUY, ssl the Naronadnas contpestrs pe. sesiatonia las hecle ite the Phoiopdahora capsisi carseat id nde indie ty expression of hese gums. Likewise, at was assessed if the expression ny and Farin setaut Tan of these genes i induced 19 C. chivense BG-3821 plants by ylcatias of slivlke aid (AS). avtienate methyl, ethane for abode stiese (lydrie and Addionally. uals ‘of cospplemontany DNA, hybridization (DNA) wae condvete for wc vari con frm subarccve banks were sect, containing €. eusense HKS-S820 tragmants, whase expeession sas inde i infetiyas of PHY WW. Tt wae dguinstrated that the gene expressions corresputing W RS0-and RIOU fragerents ‘are eo ons indeed spits hy PELYVY inkecten, bul aka hry AL ommpwris pe, vesieatiia, and smi, as well 98 pplication of sult sei. jesmanate mth a tise, In the analyns of arrangemcts, Aiffcruiey i die expression level of several EDT wae observe for eagh Key weds: agutinn puree osPalabras clave: Fusarium solani, Phytophuhore cape Xonthonomnes cangesris yw. wesicteia, chile, eaeés, PHYVY. IyTRODUCCION | caltivo del chile sufre pérdidas de 20 a 100% Feiss stress tia tn las que destacan las causadas por geminivirus (Cores-Pacheco et al, 1996; Godinez-Herninder. et aL., 2001). El virus huasteco vena amarilla del chile (PHYVV, antes PHY) es un geminivirus del género Begomovirus, tiene genoma bipariita, es transmitido por masquita blanca, infecta dicotiledéineas, inclu- yendo chile (Torres-Pacheco et af., 1996; AnayaLé- pez. et al., 2005). Dentro de los geminivirus mis dis tribuidos en los cultivos de chile en México estéa el virus nuasteco vena amarilia del chile (PHYVY) y el virus mosaico dorado del chile (PepGMY) (Anaya-Lé- pez ef al., 20038,b). En México se ha idemtificado y ccaracterizado fuentes de resistencia a estos geminivirus cen chile (Herninde2-Verdugo et al., 2001; Anaya-Lé- pez et ala, 2003a,b; Ansya-Lépez et al., 2005), Ura ccoleeta de chile habanero resistente a infecciones sim- ples y mixtas por los geminivirus PHY WV y PepGMV es la denominada BG-3821 del banco de germoplasma del Instituto Nacional de Investigaciones Foresteles, Agricolas y Pecuarias (INIFAP) perteneciente a la es- pecie C. chinense Jacq, (AnayacL6pez eal, 2003a,b; Anaya-Lépez er al., 2005). En la caracterizacién mo- lecular de la resisteacia se ka obienido seis bibliotecas de fragmentos de genes expresidos diferencialmente (EST) de estas plantas sometidas a infecciones. por geminiviras (Anaya-Lépez et al., 2005). Dentro de Ess estdn el R52 y el RIDO que presentan similitod con genes que codifican para proteinas tipo germina (GLP) reportados en otros sistemas como involucrs- ‘dos en mecanismos de resistencia en plantas & patoge- nos ¢ insectos hesbivoros (Schweizer er af., 1999; Low y Baldwin, 2006), Asimismo, se ha encontrado otros EST como el R50, el RST y el R70, cuya funcida se desconoce. Se ha reporiadd que varios genes (entre ellos GLP) pueden ser inducides en plantas en res- puesta a varios tipos de estrés (Park ef af., 2004). La ‘expresién inducible a nivel transcripcional de algunos genes en plantas se ha relacionado fuertemente con el lipo de interacsion que tealiza con diversos microor- ganismos (compatible 0 incompatible) 0 condiciones abidticas (Park et al., 2004; Eichmann et al., 2005; Restrepo et al., 2006). Can base en lo anterior. se debe estudiar la especificidad de la expresién de los genes correspondientes a los EST identificados en la interaccién C. chinense BG-3821-geminivirus, con el objetivo de dilucidar su posible participacion en et me- canismo de respuesta de esa planta a diferentes tipos 6 VOLUSEN #2, NEMERO 1 Inrxopuctioy bili: pepper crop has losses of 20 10 100% ( due 10 diseases of viral etiology, among which those caused by geminivirus stand out Corres-Pacheco et al., 1996; Godiner-Heméinde7 er ai.,2001), The pepper Huasteco yellow vein virus (PHYVY, formerly PHY) is a geminivirus of genus Begonoviras, ithas bipartite genome, is transmitted by tie white fly and infects dicotyledones, including chili pepper (Torres-Pacheco er al., 1996; Anaya-Lépez @ al., 2005). Among the geminivituses most widely cistributed in chili pepper craps in México, there are Pepper Hvasteco yellow vein virus (PHYWV) and pepper golden mosaic virus (PepGMV) (Anays-Lépez f al., 20034,)). I Mexico, sources of resistance 10 these geminiviruses in chili pepper have been identifica and described (Heméndez-Verdugo et al., 2001; Anaye-Lépez et al., 2003a,b; Anava-Lépez et al., 2005), A collection of Havana chili pepper resistant to simple and mixed infections by geminivicuses PHYVV and PepGMV is the one called BG-3821 of the germplasm bank of the Instituto Nacional de Investigaciones Forestales, Agricolas y Pecuarias (INIFAP) (National Institue of Forest, Agriculture and Livestock Research), belonging to C. chinense Jaca spesies (Anaya-Lépex et al., 2003a,b; Anays-L6pe @t-al., 2005). In the molecular characterization of resistance, six library of differentially expressed gene fragments (EST'S) of these plants have been obtained subjected to infections by geminivirus (Anays-Lopez et cl., 2005), Among them, there are R52 and R100, ‘which present similarity with genes encoding germin- like proteins reported in other systems as involved in plant resistance mechanisms to pathogens and herbivorous insects (Schweizer ef al., 1999, Lou and Baldwin, 2006). Likewise, other EST"s have been found, like R5O, R57, and R70, whose function is rot known, IC has been reported that several genes (GLP among them) may be induced to plants in response {o varios types of stress (Park ef at. 2004), The inducible expression at transcriptional level af some genes in plants has been closely related to the type of interaction that i¢ performs with different (compatible and incompatible) microorganisins or abiotic conditions (Pack et al., 2004; Eichmann ef al., 2005; Restrepo et al, 2006). Based on the aforessid, specificity of gene expression corresponding to EST, identified in the interaction C. chinense BG-3821 with the geminivirus must be studied with the objective to clarify its possible participation in the response mechanism of this plant to different types of biodc (geminivicuses and other pathogens) and abiotic stress, as well & the possible route of signaling, which these genes followde esrés bictico (geminivitus y otros pawiyenos) y abiotico, asi como la posible ruta de senalizacion que sigan estos genes en la planta, Por tanta, el objetivo de este trabejo tue realizar andlisis de la expresion 2 nivel transcripcional de genes de C.chinense BG- 3821 inducida por diferentes tipos de estrés biético @PHYVY, Xanthomonas campestris yw Plytopluhora capsici y Fusarium solani) y abiético {térmico e hidrico), y mediante ta apticacion d sabetlico, metiljasmonato y etileno. Marertaces y Méronos Materia hiligico Seussron plants de chile de la especie Capscin chvense eg coleta BG-3821 provenieate del Bunep de germoplasma del Inst: ‘uto Nacional de Imesigacienes Ferestaes, Agricola y Pesuaras AINIFAP), Coneo de Investigncin de la Resién Ceateo. Carmo ex eriment) Bao. Uni de jeeecnolgia, Celaya, México. Com ponents Ay B del Gainivius PLYWY elonatos en el pli Blucseript (SK), La cepa de X, campestris py. vecatria fue eoporcioeade por el Dr. Angel Alice Solis del Inshute Potosino say Teenoligica IPICyT). Las epas de F soln’ y P. api se wmisran él separ del INIFAP, Unidad de Bioteemnlosia, 6 Investigacin Cie Incuceg de eats bidico La mtocologts pi las neewsoiones et ele de hy baeterka 2. campestris py. vesieatoria y el aamiveto P_ epic, foe ka epor ada per Shin ev af. (2002) La inoculci de F stent se reais eolecando res dscas (1 em dimero} del medio PDA cow siclio et bor. Todas ls pans tenian custo hos verdors alo: morn de i ilasiny 48 snatuvinon ius en inrernadera frente marzey abil de 2005, 29h fooperiado de 16 zy 8 hr oscurcal, y une temperatura ce 20 25 °C. De cada paiogena © verified su presencia en ls plantas imedladas, mediante PCR on oligonulesridos espesificos pace PHYWY (Anaya Lépez 7a 2005), reaeiém en panty recuperacidn en meso de cui X canvesris ps, vesiatoria, ¥ veeupeasion en medo de ealtva rapa dextnes| sar (DiGo) pars F salon, Asicionaonte, sa fy P capac dtectaron mediante PCR usandn sligomsclcidos sspxeitos (Rico Gueriero ef. 2008), Las exrauvienes de ABN ‘ hierar de acuerdo al provocalo de Dalayorta eal. (1983), ncn desir abistien Estes trie ‘Se eolnezon 20 plants de el ‘Lab Line nseurses, Melrose Park, I.) sos un ftoperiads de 16 blue y 8h oxcuridad a 40 °C. y otras 20 plantas se calosaron a 10 °C. Las pamas se mantuieron en abseraciin 48 h, A as 30, yun utara Boone, in the plant. Therefore, the aim of tis study was t0 conduct analyses of expression at transcriptional level of C. chinense BG-3821 genes, induced by different types of biotic (PHYWV, Xanthomonas campestris DX. vesieatoria, Phytophthora capsici, and Fusarium solani) and abiotic (thermal and hydric) stresss, and through application of salicylic acid, jasmonate methyl, and ethylene. MATERIALS AND METHODS Biologie material Qui peppee plants of Capsicum shinewe Jase, eTlesion BO-S82E species ware used FROM the germ bank of the Nasional Insute of Forest, Agrisulttel, and Livestock Research ((NIFAP), Research Cemer of the Central Region, Experiments Field Bai, Unit of Biaweehnalogy, Celaya, México. Ca A and B of tke PHYVW seminvinis, clone in the Bluest (SK) plasmid, X campestris pw. vesicatora stain was provided hy De. Angel Alpache Soli ofthe Insts Potosine of Sieetific Research aid Technology (PICT). The F.solue! an! P, capsid sontins were ken fram INIFAP strains collection, Unit ot Biotechnology, Indvction of bite stress ‘The suethedlogy for imeultions in chill pepper of compestis pv. vesienoia ta thet spores hy Shir ef at (2002), Incewston of F, soli ves pesfonmed placing thee disks (em dameter) of PDA mediuen wih myeetim of the Fungus, Every plant ha four res leaves at the moment of inaeuation and was Kept incubated in sxeenhouse ‘during Marc ed April 2006, with a photonering of 16 High ae sium ané P.capsiel omyeste was 8 dark and a temperate of 26.40.28 °C. The prewnee of uch paltogen nthe iveulae las was wri by PCR wih pest ‘ovjgomuehoudes for PHYWY (Anasa-Lope et, 2008), reason inthe plang, and eecwery in culture mediun for X campers ps vesicaoria, and recuvery in potaw dextrose agar (Dito culture medium for F sok. Addiiowally, F. colon’ and P.capsici w leveted by PCR using specific oligonuclearides (Rien. Guerrero +t fa, 005), The esirctions of DNA were made according to ho pritacol of Deliports et af, (1983 Ieclvction of abiotic stress “Thermal stress Theoaky popper phine ere placed in an isubatos (Biot orate, Lah Line siaments, Melrose Pan, I) with a photeperiod of 16h Tig and 8 h dskness, at 40 °C, and ance 29 plans were pecs (0 °C; the plans were kop in cbservtion for 8 h, ALB, the cise was gathered, cuning apical ard basil eaves feeeieg thes, ‘mediately in lige ninogen, and eenserved at ~80 °C uni het BARRERA PACHIECO a 7AGROCIENCIA, 1 et 15 de Fa, 2068 1 se reco el (gid eotand ls tos spicales y haales, con gelindsasinmetiaan =) °C tas ss ullizaciin, Los experiments se realzaron en es oeasiones inéependioes, Estes hideico Se reliro sein el procedimienso de Castile ef. 200) Para La eoicidn de exces de humedad 1s plas se incubseon con mavetes 9 se colociron et tm chara om e a sets 28 °C en una icudadrea Boonete, Lab Line Tnsruerss, Melerse Patk, ML.) ccm un foperies €e 16 9 Ine de agua du indepondisnts Aplcacin de eicitores abidtcos (ici sabilico, met jasmo- ato y etefon) Con fini de spevtar elementos para canover fa posible rts de slain on In a participa los gotescomrespondinter 1 los EST RSD y R100, se hizo ws aisis de su enpeosisn wenserip ional en sespuesta & atadentos eon seyulaores de exeeinieno ‘ei slitbeo, meni jasmrato y steno fetton) que son moe las ue pastspan et ata de transucién. ce Le sealizasion et respuesta de deensa en planas (Park ea, 2008). Laaplcacién de icido sien (SIGMA-ALDRICH, S. Louis, Mo, USA neti jnmonao (SIGMA-ALDRICH, St Los, Mo, USA y ETEFON {SIGMA ALDRICH, St. Lewis, Ma, USA) se hoo « at 2002), Aislamiento del ARN Ei tos las essos el ARN se eb a lis 72 hpost-nocalaia ime el preincato de RNEASY (QIAGEN, Hide, ems yy tune del ARN se anilias por eleeioforesis €2 els de agaross cm onmaidchi, La eumntificsciin y pure 9 Inicio espestotekometrcennte ciel relsion de worn (260280 mn, Anilisis ipo Northera Se realind segin Amys per ei al 2018). Aum cal las variates de eat etulio son cualtativa, se Beier tes siplcas Sndepenietes de os atamienies Preyarseidn y marcaje de sendas de ADNe « hibridacién de memibeanss Cow la final de explora si alguna ovror EST diferentes 6 R50 y RID, tambssn som inicio em pastas dC esinense 1HGE-3421 por algutos de tos faaores bidicos yabidticns evsluadon te est abo, se hieo un estidio de tnbridacion de rneylos det ADNG de 66 EST & ncompatine entre C. chimeave BG-3821 y PHYVV, Se iidaron bane obtenid por SSH en inerasion 98 VOLUMEN 22, SUMERO 1 ablation, The experiments were earl cut om thre independent Water stress 1 was conducted secerding 9 the procedure of Cestillo ef ‘at (2000). Por the contin of aisture excess, the plants were Inouteted in Hower poss ad placed on a tray with water excess a1 28 °C in an incubator (Diokonete, Lab Line lnsteuneats, Meltose Pe. 11) wih x phowoperiad of 16) ligt and Sh dark. The experiments were conduct on nse inp nat Application of aint dlctors (alicylis sok jo and etefon) With the purpose wo combate elements for reovgnizing te possible coure of signing, in which te genes corresponding to the EST's RSD and RIOD paricipate, an analysis of their teansriptional expression wae made fn esponee to wecxments with roeth regula (eletom, which are mbieules, parcpating in te wansduetion rues of signaling io plan defense response (Pak er af, 2008) The application of silieylis aid (SIGMA-ALDRICH, St. Lous, Mo, USA), jasmonate methyl (SIGMA-ALDRICH, St.Louis. Mo, USA) and BTEFON (SIGMA-ALDRICH, Si Louis, Ma, USR) seus mae socoring to Pare of al (20D!) sallytic acid, jasmonate methyl, and ethylene RNA otatton lh every ease, RNA was obtained al 72 h post incculaton by the RNEASY protocol (QUIAGEN, Hilden, Germany), Ineegrty and size of RNA were analyzed by electrophoresis in auanse fel with formalhyde, Quautfeation and. purgy were spect Photomtrially measored hy rlaion of ubsorbanee (261/280 nm) NNorthera type anatyis Ie was comuctedaevoring to Anaya- Lipo a. (2005). Even when the variables of this study are qaitine, thie independ replications ofthe treatments wens earrid t Preparation aed marking of DNA probes ‘aud nybriaization of membranes With the purpose exploring fan cther EST einen fume R50 anc RLOO a aso nue to C. chinense BG-3B2 plans by seme ofthe biotic an abiotic factors asiese in ths pager, a sty of hybrlization of aremgements of 86 ESTs from 2 bak bined by SSE was performed in inccmparile ineraation between C. chinense 86-3821 and PHYWY. Repicates te gene eotection| (0 ©. chinese BG-3821 by anlecion of PHY obtained by SSH were hybridized by sulhemn-ype analysis (Sambrook ea. 1989) with probes obtained fom eDNA. problem or contol, oF with theolen de In Biblioteca de Los gener de C:chinense BG-3H2E por ides por SSH, meciame ass ipo Soutters. (Sambrcok erat. 1989) eon sondas otters de ADNe problem 6 testgo, @ bien eon los fragments de los KSTS RSO (17 pb, nimero de weeesidn DQSBETSE) 4 RIOD (6S ph ames de accesin DQG™7385), sein el eso, Estas sands se generaron insorporande, AUTP.LLshaiescena mane am peace rie rerio (Gene (mages CDP-Str Rendon prnse labuliag meu: Amersbitn PhasnsicaBioweh Ins, Piseataeay, NE, USA) La d= tescion de fi some Se hiro mediante el conjugate anttorescetna- fosfaasealealina y el vexcive de deteecién CDF-Sie (Amersham Pharmacia Biotec Ia Piscitaway. NI, USA). Para consti os sretlos de clones se us um dispositive mulicopiador eo tins Homa de vaca (Hoefer PR G48, Amersham iow! Resurrapos ¥ Discusion Sintomas de plantas de C. chinense BG-3821 los diferentes tipos de estrés Se evalu Ia expresién de sintomas en plantas de ©. chinese BG-3821 inoculadss de forma indepen diente con el PHYVY, X. campestris pw.vesivatoria, F. solani y P. capsici. En la Figura 1 se muestra (a presencia del PHYVY detcetade por PCR en las plaa- tas inoculadas. Las plantas se mostraron asintometicas este Virus como se esperaha, segiin lo reportado por Godiney-Hernindes ef af. (2001). Fn. la Figura 2 se de C. chiseme BGT, Pandy Carril 1-4 confir~ rmacién de la presencia de) PAYWY cn C. chisense BG-3821, Panel B. Carri, t marcador de tanaio joleeular (QETM)s 2 y 8 testigys negatives. La lech ‘naa $50 pb. C* (estigo postien), carr mestrando ‘on aampiicacon para st PAYVW a partir del ADN dona del womponcate & de ote views Figure |. Confiemation cf PHVVW presence in C._chisenve RG-3S21_ pleats, Pavel A, Track [4 confirmetion af PHYWY prowence in C, chinease BG-3S21 pias Panel B. Track I marker of molecular size (MTS: 2 sand 3 negative conteals. The arraw indicates 35) pb. C= {positive caatol, track showing an aeaphification for PHYVY from cloned DNA of the A component of this viens EST fragments HS (035 pm, acess These protes were gens secession mumbur BQKGITSA or RIOD numer 0977355), areucdiag to the ee 4 incorporating dU19-11-toreseein by es CDP-Suz Rando prime lbeling ode: Amerstamn Pharsitcia Biowch Ine, Fiseatanay, NB, USAY The detection of the prabe was made by anhuerecedn-powphatase and the CDP-Sar ektetion etacive 1 routine prowl (Gene conjugate altaine- ‘Asner Phas to eomsrust clove etraagement, duplioaor device, comnested un ia Bice lo, Poaivay, NI USA. Inorder 2 vunn pump was wed (Hoefer PR 548, Ametstam bioseences Buckinghamshire, UK) Resuvts anv Discussion nptoms of C. ehinense BG-2821 plants at different types of stress The expression of symptoms was assessed in C. chinense BG-3821 plants independently inoculated with PHYVV, X. campestris pv. vesicatoria, F. solani, and P. capsici. Figure 1 shows The presence of PHYVV detected by PCR jn the inocula Figure |. Plants appeared asymptomatic to this virus as expected according to what was reported by Godline7: Hemandez et al, 2001). In Figure 2, neeresis caused by inoculation of X. campestris py. vesicatoria is observed; the plants presented hypersensitive response jn the incculation zone (stem and lea). F solani presence in the plants was proved by re-isolation of (ed plants, is shown in Lesien por XK. canpestis igure 2-Canfemaclin de la presencia de anomenas ‘ampesirs pv. vesieaionia. Panel A, lesions weer dues tipicns cn tall mostradas por X. exmperi po. vest ‘corneas plantay eC. chinenve BG-3801, Pans BR asreamsionte de lesonee neerteas musta jas Contfemation of presence of Xanthomonas campestris py. vesieatoria, Panel A, (pial necrotic lesions ou Stem, shown by X, caapesiis py. vesiatoria in C ‘hnease BG-3821 plans. Pana B, dose-up of nerotie Tesons shown on BARRERA PACHECD e at omobserva la necrosis causitdat por la inocutacién de X, campestris pr. vesicatoria; las plantas wostraron una resoussthipersesileen la zona de inoculcion (allo y hoja}. La presencia de #. solant en las plantas comprobd mediante reaistamiiento del hongo en medio PDA y por PCR, un mes postinsculacién ya que las plantas de C. chinense BG-3821 no mostraron.sinto mas para F. solani al menos hasia el tiempo mencio- ado, Finalmente, P. capsiet tue muy agresivo y en 3 6 postinioculacién las plantas murieron, por lo que al segundo dia post-inoculacién se tomé Ja muestra respectiva Fstos resaltados sugieren que la recolecta BG 3821, dems de ser resistente a PHYVY, también lo os a cepas de X. campestris py. vesicatoria y F. solani usedas en este trabajo, Ast, es posible suponer que al- gummos de los genes cuye expresidn es aumentada por el PHYYVY en [as plantas de C. chinense BG-3821, pue- den tambien serlo por X, campestris py. vesicatoria y F. solani. pero no por la cepa de P. cupsie’, eomo se na sugerida para las respuestas de defensa de plantas a virus, hactcrias, honges y oomivetos (Lee et ol, 2001; Park ef af., 2004). Las plantas de C. chinense BG-3821 no soportaron el estrés térmica a 10 *C ya que a las § h Jas plantas estaban muy estresadas; por tanto, a las 24 h se hizo la recuperacién del tejido. A 40 °C las plantas.no presentaran altersciones a simple vista, siempre y eusndo tuvieran suficiente agua, el tejido se recolects a las 24 b para su andlisis, Para el estiés hidrico las plantas se mantuvieron 2 28 °C y las sometidas a exceso de agua no presentaron esteés aparente; sin embargo, las plantas sometides a deficit de agua, presentaron sefiales de marchiter (flacidez de Ja planta) a los 10 d de incubactén, Aniilisis de Ia expresién de RSO y R100 en plantas de C. chinenve BG-3821 sometidas a estrés En la Figura 3 se muestran los resultados del and- isis tipo northern realizado para la expresidn de los genes correspondientes a los EST R50 y RIGO. Para RSD se observ la induccién especifica por PHYY y ademés una leve induecién por infeeciones eon F. solani Figura 3, panel A, niimero 5), Para R100 se confirm la induccion especifica por el PHYVY. y ademas induccidn por intecciones con X, campestris pv. vesicatoria (Figura 3, panel B, miimero 3). Tanto RSD (Figura 3, panel A) como R100 (no mostrado} no fueron indueidas por iniecoiones por P. capsicis ademas, ninguno ée los genes para RSO y R100 fueron inducidls en las condiciones de estrés abictico evalae das (Figure 3). Los resultados anteriores indican que al menos los genes correspondientes a las ESTs R50 y R100 no soo | vou the fungus in PDA medium and by PCR one month post-inoculation, since the C. chinense BG-3821 plants did not show symptoms for F. solani, at least until the time mentioned. Finally, P.capsicd was very aggressive and within 3d postineculation the plants died; therefore, the respective sample was taken on the second day after inoculation ‘These results suggest that the BG-3821 collection, besides being resistant 10 PHYVY, is also resistant to the strains of X. campestris pw. vesivatoria and F. solani, utilized in this study. So, it can be assumed that some of the genes, whose expression is increesed by PHYWV in C. chinense BG-3871 plants, may also do so by X. campestris py. vesicatoria and F. solani but not by the strain of P. cupsici, ay suggested for the defense response of plants towards viruses, bacteria, fungi, and oomycetes (Lee er al., 2001: Park ef al., 2004). C. chinease BG-3821 plants did not withstand thermal stress at 10 °C, since at 8 h ehe planis were highly stressed: therefore, at 24 h tissue recovery was performed. Just looking at the plants, at 40 °C they did not show alterations, provided thet they had enough water; the tissue was gathered at 24 h to be analyzed. For water stress, the plants were kept at 28 °C, and those subjected to water excess did not show apparent stress: nevertheless, the plants subjected 10 lack of water presented signs of wilt {plant withering) at 10 ¢ after incubstion Analysis of RS0 and R10 expression in C. chinense BG-3821 plants subjected to stress ‘The results ofthe northern type analysis, conducted for the expression of genes corresponding to RSO andl R100 EST, are shown in Figure 3, For R50 the specific induction by PHYVV was observed, besides a light induction by infections with F- soland (Figure 3. panel A, number 5), For R100 the specific induction by PHYVY was confirmed, and furthermore, ‘induction by infections with X. campestris py. vesicatoria (Figure 3, panel B. number 3). RSO (Figure 3, panel ‘Ay as well 28 R100 (not shown) were not induced by infections with P. capsici; besides, none of the genes for R50 and RICO was induced under conditions of evaluated abiotic stress (Figure 3) ‘The previous results indicate that at least the genes comespanding to the R50 and R100 EST's are induced to the BG-3821 plants not only by PHYVY but also by the strains of F, solani and X. campestris pv. vesicatoria, Only etiological agen's, against which the plants showed incompatible interaction, could ‘induce one or both studied genes, since the strain of P. capsici used, agamst which C. chinense BG- 3821 was susceptible, did not induce the expressionANALISIS DE LA EXPRESION TRANSCRIPCIONAL INDUCIDA HAJO CONDICIONES DE ESTRES BIOTICO Y ABIGTICO EN Canam cineme s6ko son inducidos por PHYWY cn las plantas de BG- 3821, sino tambien por las cepas de F, solant y X. campestris pv. vesicatoria, Silo agentes eticlogicns, contra los cuales las plantas presentaron interaccién incompatible, pudieron inducir alguno o ambos genes estudiados ya que la cepa de P. capsici usada contra la cual C. chinense BG-3821 fue susceptible, no indujo Ja expresién de ninguno de los genes evaluedes. Estos resultados sugieren que algunas de las rutas de sefal- zacion de la interacckon plania-patogeno respectiva son compartidas, como se ha demostrado en otras plantas de chile infectadas por virus, bacteries, oomicetos u hongos (Lee et al., 2001; Shin ef al., 2002; Kim et al., 2006a,b). Adem, las condiciones evaluacas de estrds abidtico (térmico e hidrico) en estes plantas, no indujeron ia expresidin transcripcional de estos genes, Park et ai. (2004) aislaron y caracterizaron un gen que coditica para una proteina tipo germina (GLP) ¢ partir de plantas de C, annua ev, Bugang resistente a patotipo PO del virus mostico del tebaco (TMV) y # una cepa de X. campesiris pv. vesicatoria. Ellos ‘encontraron que este gen era especificamente indw sido en imeracciones incompatibles pero no en com- patibles, as{ como en respuesta al watamiento con moléculas involucradas en rutas de sefalizacién re- lacionadas con defensa en plantas (écido salicilico, metil jasmonato y etileno). Este fue el primer repor- te que involuera a una GLP en la respuesta de una plonta «virus fitopatigenos. Dicha proteina fue Ie base para crear una nueva familia de protefnas rela- vionadas con patogenicidad (PR-16), La clona R100 estudiada en el presente trabgjo presents homologia con una proteina tipo germina; ademas fue incucide por un geminivirus y también por X. campestris py. Vesicatoria, Por ello se puede sugerir que este gen debe ser considerado como un posible miembro de Ia Familia PR-16, En cuanto al R50 (Figura 3), aunque no se iden tifico en la base de datos homologia con algtin gen involucrado en mecanismos de resistencia en plantas, Dresent6 un pairda de exoresién donde se indujo por PHYWV y por F. solani. Por tanto, se debe conside. rar como un gen que posiblemente esté involucrade cn algiin mecenismo de resistencia de C. chinense BG-3821 contra estos patdgenos. Respecto a le nula Induccion de los genes RSV y RIOD ante Ios tipos de esirés abistico evaluados, posiblemente estos genes ‘no son parte de una ruta de respuesta importante de la planta a estos tipos de estrés, a diferencia de lo sugerido para genes como piruvato cinasa 1 de C. annuum, que si responde a estrés bistico por TMV y aplicaciones de reguladores de crecimiento como dcido salicilico, etileno y metil jasmonato (Kim et al., 20062) of any of the evaluated genes. These results suggest that some of the routes of signaling of plant-pathogen interaction, respectively, are shared, as has been demonstrated in other chili pepper plants infected bby viruses, bacteria, oomycetes, or fungi (Lee et al.2001: Shin et ai.,2002: Kim et af.. 2006a,b), Furtrermore, the assessed conditions of abictic stress Figura 3. Expresin inducible de genes RS0 (Panel A) y RIGO (Pavel 8) en plantas de C. chinense B-8821 érestdas can estrés bistico » ablotlos, Panel Az muestras de ARN obtenidas de plantas de C. ckivense DG-3821 Infestadas por ol PHYVY (I) y tetigo @)yinfociadas por X. campers pv. vsietoria (3) y testigo (0 Feetadas por F. solani (5) y testi (6; infectalas por epee) 3a) Les 412 cre ‘a muestras de ARN de plantas de C. dhinense HG3821 sometts a exes ain ( °C, 10°C, cexceso y austnda de agua). C+, testigo pastvo de Inbeidaciéa (ADN de la clona RSD). Panel B, mismo ‘orden de tratamientos como en e pane A desde hs hess 14, Liness 7 a 10 corresponden a muestras de ARN de plantas de C.efinense BG-3821 sometiday ‘etris abiitico 40 °C, 10°°C, exceso y ausenci de a. igure 3. Taducitle expressions of genes R50 (Panel A) and 8100 (Panel B) in C. chinewse BG-2821 plants grown with biotic and abiotic strss. Panel A: samples of RNA. ‘obteined from C. chinense BG-3821 plants infected by PHYVY (1) and control (2); infected by X. campestais . vescatoria (3) and coatrol d); infected by Solan (5) and control (0; infected by P. capsict om ‘tem (7) and leat (8). Lines 8 co 12 correspond to RNA. samples of €. chinense BG-3821 plants subjected to abletie stress G40 °C, 10 °C, excess and absence of water). C+, postive hybridization conteel (DNA of ‘cone R50). Panel &, sare order of treatments 3s in panel A from lines 146. Lines 7 10 10 correspond 0 RNA samples of C. chinesse BG-8821 plants subjected to abiotie stress (40 °C, 10°C, excess and absence of water) DARRERA-FACHECO e woExpresin de RSO y R100 como respuesta a tratamientos con reguladores de crecimiento En la Figura 4 se muestran los resultados de lt eexpresién tmanscripcional de los genes corresponclien- tes & RSW y RIGO con respecte al tempo de aplica- cién de éido salie‘ticn en las plantas de C. ehinense 1BG-3821., Los mismos experiinentas se realizaront con aplicaciones de met jasmonato y etefon, pero se pre- sent6 silo una Teve induccion de Tes dos EST a las 6h yy después descpareci6 Ja seal de induceida. Respecio 4 Ia induecion por Acido salicilion, se chserv6 que saungue fend, comenzd desde las 6h post-splicaciin clas plantas para ambos EST (Figura 4). Para RSO el nivel de expresin fue ereviente desde 6 a 18 hy y se mantuvo en un nivel similar hasta tas a8 1 (iguca 4), Para R100 el nivel de expresion present6 ua ten- dencia ereciente de 62 30 hy a las 48 hmostrO ura Tigera disminucidn (Figura 4). Testas resultados sugieren bos genes es sipida y sostenida al tratamiento de las pans can dcido salicilico, y por tanto, se espera que sean genes que participan en respuestas de defensa de C. ciimease BG-3821 contra tos patogenos estudia- dos por la ruia de seflalizacidn del Acido salctic. Esto se sugiere com hase en resultados de interacciones phinir-paiigeno de plantas de C. enna con vis y bpacterias (Lee er al., 2001: Park et af., 2001: Kim et ail., 2606a,b). Estos resultados indiean que al menos i la inddaceién de am- ESTERS ESERINO gura 4, Andliss de expen de las ESTs RSGpo (Panel I en plants eC, cinene BG-382 tralacis ‘om icito 6 sia de hbedacién obtenid en snuest wa 6, 12, 18,24, My 48 h postaglicacton te asin apliecién de eido 0 IADN de cada elona, ‘de ARN rspectiva Seguin el panel Analyss o expression of ETS RSO (Panel A) ad R100 (Panel B) iC, ehivease By-3821 plants erate wit Fig sale uci. For both panels Lines 16, hybridization Sgn obtained ie semmping of RNA obtained st 6.12 18. 24020. and 48 post-appleation af sabesiic aid, Line 3, plant without salievie act application, Cr, pistive control (DNA af ei scoring ta the panel respective clone wr VOLUMEN 12, NUMERO 1 hermal aud hydric) in these pants did not induce the ‘transcriptional expression of these genes, Park ef al.(2004) isolated and described a gene encoding for @ germin-ike protein (GLP) from C. annum ov. Bugang plants. resistant to the PO pathotype of tobacco mosaic virus (TMY), and a strain of X. campesiris pv. vesicatoria. They found cally induced 10 incompatible interactions, but not © compatible ones, as well as in response to the treatment with molecules involved in signaling routes related t plant defense (salicylic acid, jasmonate methyl, and ethylene), This was the Firs report involving a GLP in a plant response to phytopathogenous vinases. Seid protein was the base for creating & new protein family relate w pathoxenicity (PR-10) The RIOD clone studied in the present paper presented homoloyy with a germin-like protein: hesides. it was induced by a geminivirus and also vesieatoria, Therefore, it may that this gene was spe x ene must be considered as a possible member of the PR-16 fami ‘As for R5O (Figure 3), though in the daca base, homology with any gene involved in plant resistance mechanisms was not identified, it presented an expression pattern, where it was induced by PHYVV and by F. solani. Pherefore, it musi be considered as 4 gene possibly involved in 4 resistance mechanism of ©. chinense BG-3821 against these pathogens. With respect 10 Zero induction of genes R50 and R100, given the assessed types of abiotic stress, these genes possibly do aot form part of an important route of plant response to these types of stress, unlike the ‘one suggested for genes like pyruvate kinase | of C. nnn, which does respond to biotic stress by TMV and applications of growth regulaiors such as salieylic acii, ethylene, and jasmonate meiiyl (Kim er a, 200063) R50 and R100 expressions as response to treatments with growth regulators Fi expression of genes corresponding w R50 and R100 with respect fo time of salicylic acid application to € chiense BG-3821 plants. The same experiments were carried out with applications of jasmonate methyl and ctefon, but only a slight induction of the two EST was presented st 6h , and afierwards the signal of induetion disappears. Regarding the indvetion by stlicylie acid it was observed Mat though teauons~ ib after application w the plants for bow EST (Figure 4). For R50 the expression level was growing from 6 to 18h, and it maintained a similar level up 10 48 we 4 shows the results of transcriptional in since 6Expresién de R50 y R100 como respuesta a tratamientos con reguladores de crecimiento En Ta Figura 4 se muestran los resultados de la fexpresién transeripeional tes a RSO y RIGO con respecto al tiempo de aplica- cciin de deide salicilioo en las plantas de C, ehinense BG-3821, Los miismos experinentos se realization con aplicaciones de rietil jasmonato y etefon, pero se pre- sentd sblo una leve induecidn de les dos EST a las 6 h vy después desaparecié la sonal de induceidn. Respecio fa Ta induccién por Acido salicilicn, se observ que faungue tenue, comenzé desde las 6 h post-plicncién ton las plantas para ambos EST (Figura 4). Para R50) cl nivel de expresion fue creckeme desde 6a 18 fy y ‘Se mantuvo en un nivel similar hasta las 48 1 OF 4), Para R100 el nivel de expresicn presente una tea- dencia creciente de 6 a 30h y a las 48 h mosird una ligeta disminucidin (Figura 4) Exios resultados sugieren que la induceidn de am hos genes es sipida y sostenida al eracamiento de las los genes corresponclien plantas con acide salieilico, y por tanto, se espera que Sean genes que pariicipan etl respuesias de defensa de . chinense BG-3821 cantra las patogenos estudis dos por [a ruta de sefalizacién del acido saliflic. Esto se sugiere con base en resultados de interacciones planta-pardgeno de plantas de C hhuvterius (Lee et ai., 2001; Park ef ef., 2001; Kim or a., 2006a,b). Estes resaltados indican que al menos ESTRSD ESE RIOG Figura 4. Andis ds exprosin de los ESTs RSO(pan AV y RIDO (Panel Bom plantas de Chinen RCGeARDL trates ‘om se salieen. Fara ambos fneas a 6, soil de hiridaci6n cbtenida dtenidas.a 6, 18, 24 30 y Scio saleiico. Lived 7, planta sin apliaetin de sido ‘alien. C=, tesige positive (ADN de cada clona respect seguin el panel igure 4, analysis of expression of BSTS RO Fanet a) ane 100 (Wamel B) tC. efanense Bg-3821 plants created wih salicylic osid. For both panel: Lines 1-6, gbridizatinn ‘Sgn obtained in sampling of RNA ebeained at 6 12 1K. 2410, ana 48" pextapplieation of saesiic seid. Line 7. plant without salistic acid application. CH. pisive control (DNA of cach respective cane according tote panel. VOLUMIEN 42, NUMERO 1 (ermal and hydric) in these plants did not induce the transcriptional expression of these genes. Park ef al.Q0D4) isolated and described a gene encoding for a germin-like proiein (GLP) from ©. arm ev. Bugang plants, resistant to the PO pathowype of tozeco mosaic virus (TMY), and a strain of X. campestris pw, vesieaorin, They found thal this zene was specifically induced « incompatible interactions, but not to compatible ones, as well 3 in response (0 the treatment with molecules involved in signaling routes related to plant defense (salievlic acid, jasmonate methyl, and ethylene). This was the first report involving 4 GLP in a plant response 16 phytopathogenous viruses. Sid protein was the base for creating a new protein family related to pathogenicity (PR-10), The RLOO clone studied in the present paper presented homology with a germin-like protein: besides. it was induced by a geminivirus and also by X. campestris py. vesieatoria. Therefore, it may be suggested that this possible member of the PR-16 family As for RSO (Figure 3}, though in the daa base, homology with any gene involved sn plant resistance mechanisms was not identified, it presented an expression pattern, where it was induced by PHYVV and by F. solani, Therefore. it must be considered as 4 gene possibly involved in a resistance mechanism of ©. chinense BG-2821 against these pathogens. With resect 10 vero induction of genes R50 and R100, given the assessed types of abiotic stress, these genes possibly do not form part of an important route of plant response to these types of stress, unlike the ‘one suggested for genes like pyruvate kinase 1 of C. rene mast be considered as & annum, whieh does respond to hiote stress hy TMV and applications of growth regulacors such as salicylic acid, ethylene, and jasmouae meihyl (Kim er af, 20063) R50 and R100 expressions as response to treatments with growth regulators ire 4 shows the results of canseriptional xpression of genes corresponding to R50 and R100 ‘wilh respect (0 time of salieylie avid application 0 © chinense BG-3821 plants. ‘The same experiments were carried out with applications of jasmonate methyl and sefon, but only a slight induction of the two EST was presented at 6h, nd aferwards the signal of induction disappears, Regarding the induction by saieylie acid. it-way observed thiat tho Ih after application w the plants for bows EST Figure 4). For R9Q the expression level was growing from 6 [8h, and it mainiained a similar level up 10 48) tenuous it began since 6Jos wenes correspondicntes & los EST R50 y R100 ehen estar participando en la respuesta de detensa de C, chinense BG-3821 a los patogenos estudiados. mediante lz ruta de seializaci6n por el acide sulicitico principalmente, pero también por las rulas mediadas por metil jasmonato o ctileno. Las rutas de sefaliza- on plantas hacia algiin estiés bidtice 0 abidtico pueden 6 mo ser compartidas por To que los resultados mostrades en este trabajo son consistentes con otros reportes (Hong et al., 2005; Kim et af., 2005 a,b. ign para respuestas de defensa de hibridacién de arreglos de cDNA Sc evalus el perfil de expresién de 66 EST de C. ehinense BG-3821 en respuesta aX. campestris py vesicatoria, F. solani y aplicacion de deido salicilic. Se usaron como testigo negativo plantas sin tratar (Fi 5 y 6), Las clonas de R50 y R1OO se ineluyeron fo | 13 Be ta Fj wi | ‘Acid satietied Figura 8, Pate de hibidcién de snrglos ele DN Co chinense-PIVW. C Figure §, Hybriivzats probe used in each paitern of un bajo nivel de induce, CONCLUSIONES Los EST R50 y R100 fueron inducides en int racciones incompatibles entre C. chinense BG-3821 y PHYVY, X. campestris pv. vesieworia 0 F, solani, tmientras que no presentaren induccién en interaecio hes compatibles como la observada con P. eapsicl Asimismo, estos EST respondieron con una induccién répida al tratamiento con dcida saliciico, y en menor inteasidad 4 meti jasmonato o etileno, sugiriendo que wea | vonuMen «2, NeMERO 1 cated, The numbers indicate the number af eh ele of the hah, abiotic stress may or may be not shared, for whieh che results shown in this study are consistent with other reports (Hong. at af., 2008; Kim er al., 2006a,b). Hybridizs ‘ion analysis of CDNA arrangements ‘The expression protile of 66 EST of C. chinense BG-3821 was assessed in response t0 X. campestris pv vesicatoria, F. solani, and salicylic acid application Untreated plants were used as negative control (Figures 5 and 6). The clones of R50 and R100 vere included as postive control, and in all the vases, the expression pawern described in previous sections for these genes under every evaluated concition, was reflected again ‘Figure 6). Regarding the expression levels ofthe EST placed in the arrangements. it was evident that several of them, not being R50 ar R100, responded 1 the treatments by X. campesivis pv. vesicatoria and 10 the application of salicylic acid in C. chivense BG-3821 plants wid an induction a¢ different levels (Figure 5 and 6}. For the case of F.solané, a low indoction level was observed (Figure 5) ‘These resus may be explained based on the behavior of plants towards different types of stress asessed inthis section, where they presented a strong reaction of hypersensitivity on the ste of inoculation for inoculetion with X. campestris pv. vesicatora. Tn the inoculations with F. solani, the plants did not show evident symptomatology; therefore, their response may be considered light manifesting low level of inductionlay rutas Oe sefalizavién mediades por estos compues tos son importantes en el sistema estudiado, Varios EST diferentes a R50 y R100, que también provienen de una coleccidn de fragmentos de genes inducidos en C. chinence BG-3821 por infecciones con el PHYVV, -mostraron patrones de induccién en respuesta aa ino- eulacida con X. canpesiris pr. vesicatoria y le apli- cacién de acido salicilice, Bl hecho de que varios de los EST evaluados en este trabajo (diferentes 2 R50 y R100) fueron inducidos en plantas de C. chinense én interacciones incompatibles por infecciones por et PHYVY. X. campestris pv. vesicatoria y F. solani. sugiere que estos genes pueden estar invaiucrados en la respuesta de defensa a estos patégencs, por lo que se deben caracierizar mayor detalle posteriormente ‘Los resultados sugieren la presencia de una res- puesta sistémica adquirida en las plantas evaluadas, debido a que ef icido salicilioo result6 un fuerte in- ductor de la expresion de estos genes en interacciones incompatinles, de: la misma manera camo se ha repor lado en otros sistemas de interaccién planta-patégeno. Sin embargo, no s¢ puede descartar que algunos de los genes estudiados diferentes 4 R50 y RLOO tengant participacion por alguna otra ruta de sefalizacién adi- ional Con este trabajo se presentan varios genes candida- tos a estar invalucrados en mecanismas de resistencia en plantas de C. chinense a varios patégenos. Asi es fietible que al estudiar la regulacién de la expre- sion estos genes se encuentren rutas de sefalizacién comunes para la proteccion de estas plantas contra patogenos que pudieran ser tiles en el disefo futuro de estrateaias de proteccién de estas piatas (¥ quiz a infecciones por virus, hongos y bucterias Acrapecisnenros EL presente tabap se eeliz6 con apoyo dal Fonte misto CO- NACYT-Gobierno del Estado de Guang (FOMIX-GTO-2008 (CK0-32) y of apoyo 4 PRECT 9999 del INIPAP. Aerio Barer ¥ Auizol de Jess Jorain Ramos, aradecen el aposo de stra a CONACYT y CONCYTEG Lirrratura Crrapa J bs Y. Godiner-Berninder, C. 1. Maioe Si ner Olvera, R, G. Guevars-Gonziler, RL E. Rivers MM. Gonzilee-Chasira,y {. Tortes-Pacbeco 200%, Kentheacion de resistencia entra mfescines simples ‘nixtis port virus dl mosice doraah del che Pep) y virus nuastece de tile en plantas decile sane 1Capsicun rense acy. Red. Chapingo See Host. 9 225-234 Araya Lips, I. Ly M. Gonzalo Chivira, JL. Poss Homes 1. A. Garz6n Tiamido. 1. Torres Pacheco, RF. Rivers usimants, §, Herender-Verdugo, RG. Guevara Gone, CL Miier-Sinches, and C. Guevara Ober. 20036 Concustons EST R50 and R100 were induced in incompatible interactions between C. chinense BG-3821 and PHYWV, X. campestris pv. vesicatoria or F. solani, ‘whereas they did not present induction in compatible Interactions, as the one observed with P. capsic. Likewise, these EST responded with a quick induction to the treatment with salicylic acid, and at less imensity, to jasmonate methyl or ethylene, suggesting that the routes of signaling mediated by these compounds are important in the stadied system. Several EST. different to RSO and ROO, which are also from a eollection of gene fragments induced to C. chinense BG-3821 by Infections with PEYVY, showed induction pavers in resporse (0 the inoculation with X. campesiris py. vesicatoria and application of salicylic acid. The fact that various of the EST assessed in this study (different from R50 and R100) were induced to C. chinense plants in incompatible interactions through infections by PHYVY, X. campestris py. vesicatorie, and F solani, suggests that these genes may be involved in the defense response to these pathogens, for which they must subsequently he characterized at greater detail The results suggest the presence of an acquired systemic response in the assessed plants, due to the fact thet salicylic acid turned out to be a strong inductor of expression of these genes in incompatible interactions, im the same way as reported in other ystems of plant-pathogen interactions. However, it cannot be discarded that some of the studied genes, ciferent to R50 and R100, may participate through another additional route of signalin With this study, several genes are presented, candidates of being involved in resistance mechanisins in €. chinense plants (0 various pathogens. Thus, itis feasible that a¢ studying the regulabon of these gene expressions, common signaling routes may be found for the protection of these plants against pathogens, ‘which might be usefil in the future design of protection sirategies of these plants (and may be others) against infections by viruses, fungi, and bacteria En of the Exes version ofea Resistance fo gominvieus mixed infetios in Mexican wild neppers. Horscence 38: 251-25 Anayachopes, J. 1, B Péter-Mera, L TonresFactees, ©. MutorSinckez, L, Guevara OWvers, M. M, Gonzi Chavim, N, Ochoa Abjo, RF. Rivers Ductumante, and R. Gwevars-Gorziler 2008 Iniucl gone express by Pepper Insts vires in Capsicum ehiverse plants with resistance 10 seminsinas aftr, Can, Bact Pub. 27: 276-282. THARRERA PACHIECO erat wsCamo, J, M1 Rodrigo, 3. A Morgice, A. Zuniga, ond L. Franso. 2000 4 pea nuclear protein dat is induced by hydration belo a the vieiinSupareumly. Fae Poche 2156-2165, Dellapar, 8, L. 1, Word, and J.B. Hicks, 1983. 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