Resistance of Cowpea Genotypes to Zabrotes subfasciatus (Boheman, 1833) (Coleoptera: Chrysomelidae: Bruchinae)

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1 Journl of Agriulturl Siene; Vol. 5, No. 5; 2013 ISSN E-ISSN Pulished y Cndin Center of Siene nd Edution Resistne of Cowpe Genotypes to Zrotes sufsitus (Bohemn, 1833) (Coleopter: Chrysomelide: Bruhine) Aline Vin Kloss Toledo 1, Lúi d Silv Fontes 1, Dougls Rfel e Silv Bros 2, Pulo Roerto Rmlho Silv 3, Josynri Arújo Neves 4 & Ruty de Sous Melo 4 1 Deprtmento de Biologi, Lortório de Entomologi/CCN. CEP: Cmpus Universitário Ministro Petrônio Portell, Birro Ining, Teresin, PI, Universidde Federl do Piuí, Brsil 2 Deprtmento de Fitossnidde, Ru Dom Mnoel de Medeiros. s/n. Dois Irmãos. CEP: Reife-PE, Universidde Federl Rurl de Pernmuo, Brsil 3 Lortório de Fitossnidde, Deprtmento de Fitoteni. Centro de Ciênis Agráris, Cmpus d Soopo, CEP: Teresin-PI, Universidde Federl do Piuí, Brsil 4 Progrm de Pós-Grdução em Agronomi/Produção Vegetl; Cmpus d Soopo. CEP: ; Teresin, PI. Universidde Federl do Piuí, Brsil Correspondene: Dougls Rfel e Silv Bros, Deprtmento de Fitossnidde, Ru Dom Mnoel de Medeiros. s/n. Dois Irmãos. CEP: Reife-PE, Universidde Federl Rurl de Pernmuo, Brsil. E-mil: dougrs@hotmil.om Reeived: Jnury 25, 2013 Aepted: Mrh 12, 2013 Online Pulished: April 15, 2013 doi: /js.v5n5p83 URL: Astrt Cowpe (Vign unguiult (L.) Wlp.) when stored is minly ttked y ruhid Coleopterns. The ontrol of these pests is done primrily through hemils, however, with prolems relted to the seletion of resistnt insets, lterntive forms of ontrol re serhed. Seeking n lterntive ontrol, the ojetive this work ws to evlute the resistne of owpe genotypes to the ttk of Z. sufsitus. In the first seletion of genotypes were evluted 35 owpe genotypes, eing oserved the vriles: numer of eggs, numer of insets emerged nd weight loss (%). In this first seletion of owpe were seleted 10 genotypes. In the seond seletion of genotypes, the 10 most resistnt from the first seletion were used, ut with eight repetitions per genotype, esides evluting the egg viility (%) nd instntneous rte of popultion growth. The owpe genotypes showed signifint differenes for resistne to ttk y Z. sufsitus. The most resistnt to Z. sufsitus were BRS Truteu, 31 MNC03-720C-20, 26 MNC00-553D nd 37 MNC05-832B Keywords: weevil, Vign unguiult, stored grin insets, posthrvest 1. Introdution Cowpe (Vign unguiult (L.) Wlp.) is legume ultivted in semi-rid of Afri, Brzil nd United Sttes. In Brzil, the rop is very importnt in the North nd Northest, whih hve trdition in its ultivtion, trde nd onsumption. The rop presents inresing dvnement in the Brzilin Midwest, where owpe ultivtion hs een onduted in mehnized form, nd there is gret demnd for upright ultivrs (Roh et l., 2009). Among the lrgest produers in Brzil stnd out sttes: Amzons, Prá, Mrnhão, Piuí, Cerá nd Rio Grnde do Norte (Medeiros et l., 2007). In Brzilin Northest, the owpe rop hs gret importne to griulturl development in oth the eonomi nd nutritionl spets. It is the stple food in the diet of the poor, exerting soil funtion in supplying the nutritionl needs in this setion of the popultion (Teófilo et l., 2008), esides fixing hnd lor in the field. (Távor et l., 2003). The owpe grins hve good energy levels, with exellent protein ontent, 23-25% on verge (Amrl et l., 2005). It is rih in lysine nd other essentil mino ids, however, poor in sulfured mino ids, methionine nd ysteine. It onstitutes n exellent soure of niin nd lso ontins resonle quntities of hydrosolule vitmins suh s rioflvin, pyridoxine, folin, iron, zin nd phosphorus minerl elements (Silv et l., 2002). Thus, owpe is food tht meets si nutritionl needs of the low-inome popultion (Amrl et l., 2005). 83

2 Journl of Agriulturl Siene Vol. 5, No. 5; 2013 The owpe stored is minly ttked y ruhid eetles. In the sufmily Bruhine, the insets re ssoited with grins of mny plnts, onsisting in fmily of griulturl importne. The Mexin en weevil Zrotes sufsitus (Bohemn, 1833) is originting of the New World, eing distriuted in tropil nd sutropil regions of Centrl nd South Ameris (Dendy & Credlnd, 1991; Hines, 1991), Afri, Mediterrnen nd Indi (Oliveir & Vendrmim, 1999). The eggs of this inset re deposited on the surfe of the grins nd the lrve develop inside, using onsiderle weight loss, redution in germintion, deresed nutritionl qulity nd ommeril devlution (Oliveir & Vendrmim, 1999). Furthermore, the metolism of the lrve uses n inrese of the temperture in the storge ontiners, whih fvors the development of fungi ple of produing toxins (suh s fltoxin) dngerous to the helth of the onsumer (Amevoin et l., 2007). The use of hemil pestiides is ommon preventive mesure to protet stored grin from inset dmge. Mny pestiides re effetive t reltively low doses nd my offer long-term protetion, rnging from 6 to 12 months (Athnssiou et l., 2004). However, some of these pestiides, espeilly orgnophosphtes hve high toxiity to mmmls, nd the residues left n use helth prolems euse they re onventionl neurotoxins tht ffet the humn nervous system (Athnssiou et l., 2009). Thus, the use of vrietl resistne ginst the ttk of Z. sufsitus hs een the sujet of sientifi reserh y offering n lterntive mesure of ontrol. Some studies show tht is vile for ontrol Z. sufsitus in owpe (Vign unguiult) nd ommon en (Phseolus vulgris), the use of genetilly resistnt ultivrs (Mzzoneto & Boiç Junior, 1999; Boiç Júnior et l., 2002; Bros et l., 2011). These reserhes represent diret enefit to the produer y reduing posthrvest losses. Thus, the ojetive of this study ws to selet owpe genotypes resistnt to ttk of Z. sufsitus. 2. Mterils nd Methods 2.1 Rering of Insets The insets were rered for severl genertions in owpe (Sempre Verde v) grins, these were pked in glss ontiners (1.5 L) losed with perforted plsti lids, lined on the inside with thin loth, llowing gs exhnge. The rering of insets nd experiments were onduted t 27 ± 2ºC, 70 ± 10% reltive humidity nd 12 h photophse. 2.2 Cowpe Genotypes Were used 35 owpe genotypes from Ative Germplsm Bnk of Emrp Meio-Norte: BRS Truteu, BR17-Gurguei, TE67-304G-12, Monteiro, BRS-Nov Er, BRS-Uruuqur, BRS-Prguçu, BRS-Guri, BRS-Milenio, BRS-Mrtoá, BRS-Rouxinol, Cnpuzinho nº10, Inhum nº12, Pingo de ouro-1-2 nº13, Pulistinh nº15, 35 TVX C, 38 Vin Bln, 39 Clifornilkeye, 40 BRS-Guri, 21 MNC99-537F-1, 22 MNC99-537F-4, 23 MNC99-541F-5, 24 MNC99-541F-8, 25 MNC99-542F-5, 26 MNC00-553D , 27 MNC00-553D , 28 MNC99-557F-2, 29 MNC01-627F-14-2, 30 MNC01-627F-14-5, 31 MNC03-720C-20, 32 MNC03-720C-31, 33 MNC03-731C-21, 34 MNC03-732C-5, 36 MNC05-784B-38-2 e 37 MNC05-832B Experimentl Protool The grins were pked in plsti gs t -5 C (in freezer) to eliminte possile ltent infesttions. Before instlling the experiments, the grins were removed from the freezer, pled in plsti reipients overed with thin tissue nd kept in the lortory for six dys to ome into equilirium moisture ontent (Lim et l., 2002). The tretments onsisted of grins of eh genotype (10 g) infested with five ouples Z. sufsitus (0-48h old), left for seven dys to oviposition. The grins were pled in plsti reipients (300 ml) with trnsprent lid, lined on the inside with thin loth. The experiments were evluted for 40 dys First Seletion of Genotypes The experimentl design ws ompletely rndomized with four replitions for eh genotype under no-hoie test. To evlute the resistne were oserved following prmeters: numer of eggs, numer of insets emerged nd weight loss (%) Seond Seletion of Genotypes The experimentl design ws ompletely rndomized with eight replitions for eh genotype under no-hoie test. However, were used the most resistnt genotypes from the first seletion. To evlute the resistne were oserved following prmeters: numers of eggs, numer of insets emerged, weight loss (%), egg viility (%) nd instntneous rte of popultion growth. For lulting this ws used the eqution: ri= [Ln (Nt/N0)/ΔT], 84

3 Journl of Agriulturl Siene Vol. 5, No. 5; 2013 where Nt is the finl numer of dults; N0 is the initil numer of dults trnsferred nd Δt is the hnge in time (Wlthll & Strk, 1997). The positive vlue of ri indites popultion growth inrese; ri = 0 mens tht the popultion is stle; nd negtive vlue of ri indites popultion deline to extintion (Strk & Bnks, 2003). 2.4 Sttistil Anlysis The dt were nlyzed y nlysis of vrine (ANOVA), while the signifint mens were ompred y Sott-Knott test (Sott & Knott, 1974) t the 5% level of signifine through the sttistil progrm Sisvr 5.0 (Ferreir, 2011). 3. Results nd Disussion 3.1 First Seletion of Genotypes The vlues otined for the numer of eggs present signifint differene etween them, showing tht genotypes BRS Truteu nd 31 MNC03-720C-20 otined the lowest oviposition in reltion to the others (Tle 1). The most resistnt genotypes otined on verge 61.93% less eggs thn the most suseptile. Tle 1. Numer of eggs, numer of insets emerged nd weight Loss on 35 owpe genotypes ttked y Z. sufsitus Genotypes Numer of Eggs Numer of Insets Emerged Weight Loss (%) BRS Truteu d 2.7 d 31 MNC03-720C d 2.8 d BRS-Milenio d 5.0 d 26 MNC00-553D d Monteiro d 5.3 d 36 MNC05-784B d 37 MNC05-832B d 4.3 d 32 MNC03-720C d 34 MNC03-732C MNC01-627F d 39 Clifornilkeye BRS-Mrtoá MNC03-731C BRS-Nov Er MNC01-627F MNC00-553D BRS-Uruuqur d 28 MNC99-557F d 23 MNC99-541F Vin Bln BR17-Gurguei MNC99-541F BRS-Rouxinol MNC99-537F MNC99-542F BRS-Guri TE97-304G BRS-Prguçu TVX C BRS-Guri Cnpuzinho nº MNC99-537F Pingo de ouro-1-2 nº Pulistinh nº Inhum nº *Mens followed y the sme letter in the olumn do not differ signifintly y Sott-Knott test t 5% proility. 85

4 Journl of Agriulturl Siene Vol. 5, No. 5; 2013 Mzzoneto nd Boiç Junior (1999) seeking lterntive methods for the ontrol of Z. sufsitus in ens, onduted tests with nd without hoie using genotypes: Goino Preoe, Onix, Dimnte Negro, Ipr MD 808, Preto 143, 2357, 2306, 2174, 2044, 2041, 2037, 133, 115, 2037, 2374E 2395 nd onluded tht genotype Preto 143 showed resistne to non-preferene for oviposition on free hoie test, nd the genotypes 2374, 2395, 2174, 133 e 155, showed resistne of the non-preferene for feeding nd/or ntiiosis. In this study, genotypes BRS Truteu nd 31 MNC03-720C-20 presented lower numer of eggs, proly euse they hve ntiiosis resistne, sine the test ws performed without hoie. There ws signifint differene in the numer of insets emerged, eing tht BRS Truteu, 31 MNC03-720C-20, BRS-Milenio, Monteiro, 37 MNC05-832B nd 37MNC05-832B presented the lowest numer of insets emerged. These genotypes hd on verge insets emerged, hving n emergene 61.88% lower thn genotypes with higher numer of insets emerged. The genotypes TE97-304G-12, BRS-Prguçu, 35 TVX C, 40 BRS-Guri, Cnpuzinho nº10, 21 MNC99-537F-1, Pingo de ouro-1-2 nº13, Pulistinh nº15 nd Inhum nº12 showed the highest inset emergene (Tle 1). Bros et l. (2000) studied the stility of resistne Z. sufsitus in four genotypes of ommon en nd found tht the highest verge numer of dults emerged ws oserved in ultivrs Goino Preoe e Porrillo 70, differing from ARC4 e ARC1, genotypes with protein relin. Mirnd et l. (2002) oserved tht genotypes ARC1 e ARC4 presented numer of dults emerged signifintly lower thn those presented y genotypes ARC2 nd ARC3 nd genotypes without relin. In this reserh, ws oserved lower numer of insets emerged in genotype 31 MNC03-720C-20. This my hve ourred due to some dverse effet for inset development inside the grin, proly relted to ntiiosis used y this type of protein. In this study, 11 genotypes showed the lowest weight loss, eing BRS Truteu nd 31 MNC03-720C-20 less ttked, suggesting the possiility to hve ntiiosis resistne. The onsumption of grin is positively relted to the numer of insets emerged. Thus, the lrgest inset emergene reflets in inresed onsumption. The opposite n lso our due to the presene of sustnes ple of inhiiting the feeding ontent inside the grins y the lrve of Z. sufsitus. The presene of inhiitory sustnes feeding in weevil is reported in the literture, for exmple the relin whih onfers resistne to Z. sufsitus (Orini & Lr, 2000) nd trypsin inhiitors responsile for ntiiosis in some owpe genotypes (Gtehouse et l., 1979). The genotypes Cnpunzinho nº10, Pingo de ouro-1-2 nº 13 nd 35 TVX C were the most onsumed, demonstrting do not possess or hve low levels deterrents sustnes to insets. The resistne of BRS Truteu, 31 MNC03-720C-20, BRS-Milenio, 26 MNC00-553D , Monteiro, 36 MNC05-784B-38-2, 37 MNC05-832B-234-5, 32 MNC03-720C-31, 34 MNC03-732C-5 nd 29 MNC01-627F-14-2 n lso e explined y the ft they presented rough texture, whih my led to lower preferene for oviposition y the femles of Z. sufsitus. Lim et l. (2001) identifying owpe genotypes resistnt to Cllosoruhus multus (F.), oserved tht morphologil uses nd tegument texture ffet the preferene for oviposition. Nwnze et l. (1975) oserved higher oviposition of C. multus on smooth grin thn rough grin. 3.2 Seond Seletion of Genotypes Were oserved similrities in reltion to the first seletion, sine the genotypes showed signifint differene in the numer of eggs, numer of insets emerged nd weight loss (%). The genotypes BRS Truteu, 31 MNC03-720C-20, 26 MNC00-553D nd 37 MNC05-832B hd the lowest oviposition with 17.87, 31.12, nd eggs, respetively (Figure 1A). In these genotypes the verge of eggs ws whih represents 46.9% less eggs thn suseptile genotypes. Botelho et l. (2002) studying strins of Phseolus vulgris, oserved tht strin ARC 3 ws the lest oviposited, with 308 eggs. In this reserh, the owpe genotype with less eggs ws BRS Truteu, eing the most resistnt to Z. sufsitus in reltion to oviposition. 86

5 Journl of Agriulturl Siene Vol. 5, No. 5; Numer of eggs Egg viility (%) e d e Tr Cowpe genotypes A 0 Tr Cowpe genotypes B Figure 1. Effet of owpe genotypes in emryoni development nd oviposition of Z. sufsitus. A) Numer of eggs. B) Egg viility (%). Mens followed y the sme letter do not differ signifintly y Sott-Knott test t 5% proility The genotypes BRS Truteu nd 26 MNC00-553D llowed the lowest egg viility of Z. sufsitus (Figure 1B). Rieiro-Cost et l. (2007), seeking to verify the development of Z. sufsitus in genotypes with nd without relin, demonstrted tht genotypes s ARC1 nd ARC2 ontining this protein showed higher resistne, sine otined lower preferene for oviposition, nd low perentge of vile eggs. In this reserh, low egg viility oserved in the lest suseptile genotypes n lso e explined y presene of proteins tht n use negtive effets on emryoni development of insets. Sles et l. (2005) oserved 95% emergene of dults of Z. sufsitus in seeds of owpe suseptile, thus presenting egg viility ove 90%. In this reserh, the egg viility ws 76.41% in the genotypes 36 MNC05-784B-38-2, 32 MNC03-720C-31 nd 34 MNC03-732C-5, the most suseptile to oviposition. The men egg viility ws 43.03% in the genotypes BRS Truteu nd 26 MNC00-553D The genotypes BRS Truteu, 31 MNC03-720C-20, 26 MNC00-553D nd 37 MNC05-832B hd the lowest numer of insets emerged, with the inset emergene rnging from 7.50 to dults (Figure 2A). This low numer of insets emerged my e due to lrvl mortlity used y proteins with potentil insetiide suh s viilins present in some leguminous plnts. These proteins ind to hitinous strutures of the midgut (peritrophi memrne) interfering in the ssimiltion of nutrients, whih my use the inset deth (Amorim et l., 2008). Numer of insets emerged Tr Cowpe genotypes A Figure 2. Popultion growth of Z. sufsitus in owpe genotypes. A) Numer of insets emerged. B) Instntneous rte of popultion growth (ri). Mens followed y the sme letter do not differ signifintly y Sott-Knott test t 5% proility Instntneous rte of popultion growth (ri) 0,05 0,04 0,03 0,02 0,01 0,00-0,01 h Tr Tr f d g Cowpe genotypes e B 87

6 Journl of Agriulturl Siene Vol. 5, No. 5; 2013 Almost ll owpe genotypes llowed the popultion of Z. sufsitus ontinue growing, exept BRS Truteu whih showed negtive vlue of the instntneous rte of popultion growth (Figure 2B). Aording to Strk nd Bnks (2003) the popultion is delining nd will reh to extintion. This result showed tht genotype BRS Truteu ws effetive to ontrol Z. sufsitus. The genotypes 34 MNC03-732C-5 nd 29 MNC01-627F-14-2 hd the highest weight loss (%), nd therefore, were the most onsumed (Figure 3). These genotypes were 67.13% more onsumed thn BRS Truteu, 31 MNC03-720C-20, 26 MNC00-553D nd 37 MNC05-832B-234-5, whih hd n verge 18% weight loss. Shfique nd Chudry (2007) suggested tht the low inset popultion nd low weight loss of grin n e used s one ttriute of the resistne to insets Weight Loss (%) Tr Cowpe genotypes Figure 3. Weight Loss of owpe grin when sujeted to ttk Z. sufsitus. Mens followed y sme letter do not differ signifintly y Sott-Knott test t 5% proility Our results showed tht the relese of owpe genotypes resistnt to Z. sufsitus n e n importnt tool to help ontrol this pest. Furthermore, seletion of owpe genotypes resistnt to insets n lso e used s prmeter in plnt reeding progrms. 4. Conlusions The owpe genotypes most resistnt to Zrotes sufsitus were BRS Truteu, 31 MNC03-720C-20, 26 MNC00-553D nd 37 MNC05-832B The genotype BRS Truteu hs gret potentil to ontrol Zrotes sufsitus euse it ws the most resistnt in ll prmeters evluted. Aknowledgement We thnk Emrp Meio-Norte for giving the owpe genotypes used in the experiments. Referenes Amrl, J. A. B., Beltrão, N. E. M., & Silv, M. T. (2005). Zonemento Agríol do Feijão-Cupi no Nordeste Brsileiro Sfr 2005/ Estdo d Prí. Comunido Ténio, 253, Cmpin Grnde, PB. Amevoin, K., Snon, A., Aposs, M., & Glitho, I. A. (2007). Biologil ontrol of ruhids infesting owpe y the introdution of Dinrnus slis (Rondni) (Hymenopter: Pteromlide) dults into frmer's stores in West Afri. Journl of Stored Produts Reserh, 43, Amorim, T. M. L., Medo, L. L. P., Uho, A. F., Oliveir, A. S., Pitng, J. C. M., Medo, F. P., Sles, M. P. (2008). Proteolyti Digestive Enzymes nd Peritrophi Memrnes during the Development of Plodi interpuntell (Lepidopter: Pirlide): Trgets for the tion of Soyen Trypsin Inhiitor (SBTI) nd Chitin-Binding Viilin (EvV). Journl of Agriulturl nd Food Chemistry, 56,

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8 Journl of Agriulturl Siene Vol. 5, No. 5; 2013 Orini, M. A. G., & Lr, F. M. (2000). Antiiosis effets of wild en lines ontining relin on Bemisi ti (Genn.) iotype B (Homopter: Aleyrodide). Anis d Soiedde Entomológi do Brsil, 29(3), Rieiro-Cost, C. S., Pereir, P. R. V. S., & Zukovski, L. (2007). Desenvolvimento de Zrotes sufsitus (Boh.) (Coleopter: Chrysomelide, Bruhide) em genótipos de Phseolus vulgris L. (Fee) ultivdos no Estdo do Prná e ontendo relin. Neotropil Entomology, 36(4), Roh, M. M., Crvlho, K. J. M., Freire Filho, F. R., Lopes, A. C. de A., Gomes, R. L. F., & Sous, I. d S. (2009). Controle genétio do omprimento do pedúnulo em feijão-upi. Pesquis Agropeuári Brsileir, 44(3), Sles, M. P., Andrde, L. B. S., Ary, M. B., Mirnd, M. R. A., Teixeir, F. M., Oliveir, A. S.,... Xvier-Filho, J. (2005). Performne of en ruhids Cllosoruhus multus nd Zrotes sufsitus (Coleopter: Bruhide) rered on resistnt (IT81D-1045) nd suseptile (Epe 10) Vign unguiult seeds: Reltionship with trypsin inhiitor nd viilin exretion. Comprtive iohemistry nd physiology Prt A: moleulr & integrtive physiology, 142(4), Sott, A. J., & Knott, M. A. (1974). A luster nlysis method for grouping mens in the nlyses of vrine. Biometris, 30, Shfique, M., & Chudry, M. A. (2007). Sreening of rie genotypes for resistne to storge insets. Pkistn Journl of Zoology, 29(1), Silv, J. E., Resk, D. V. S., & Shrm, R. D. (2002). Alterntiv gronômi pr o iossólido produzido no Distrito Federl. I - Efeito n produção de milho e dição de metis pesdos em ltossolo no errdo. Revist Brsileir de Ciênis do Solo, 26, Strk, J. D., & Bnks, J. E. (2003). Popultion-level effets of pestiides nd other toxints on rthropods. Annul Review of Entomology, 48, Távor, F. J. A. F., Nogueir, S. L., & Pinho, J. L. N. (2001). Arrnjo e populção de plnts em ultivres de feijão-de-ord om diferentes rterístis de op. Revist Ciêni Agronômi, 32(1/2), Teófilo, E. M., Dutr, A. S., Pitimeir, J. B., Dis, F. T. C., & Bros, F. S. (2008). Potenil fisiológio de sementes de feijão upi produzids em dus regiões do Estdo do Cerá. Revist Ciêni Agronômi, 39(3), Wlthll, W. K., & Strk, J. D. (1997). Comprison of two popultion-level eotoxiologil endpoints: the intrinsi (rm) nd instntneous (ri) rtes of inrese. Environmentl Toxyology nd Chemistry, 16(5),