CễNG TY TNHH BAYER VIT NAM CNG HO X HI CH NGHA VIT NAM c lp T Hnh phỳc BO CO TểM TT NH GI RI RO CA THC VT BIN I GEN I VI SC KHO CON NGUI, VT NUễI BễNG KHNG SU V CHNG CHU THUC TR C GHB119 CễNG TY TNHH BAYER VIT NAM Thỏng 4, 2015 Ton b d liu h s ng ký ny c bo v bn quyn v khụng c s dng, chộp hay a cho bờn th ba nu khụng cú c s ng ý ca cỏc tỏc gi Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt PHN I THễNG TIN CHUNG T chc cỏ nhõn ng ký: - Cụng ty TNHH Bayer Vit Nam Ngi i din ca t chc: ễng David John Champion Tng Giỏm c u mi liờn lc ca t chc: TS ng Ngc Chi Giỏm c ng ký ht ging a ch: Lụ 118/4 Khu cụng nghip Amata, Thnh ph Biờn Hũa, Tnh ng Nai in thoi: (+84-8) 3845 0828 Fax: (+84-8) 3997 9206 Email: chi.dang@bayer.com Tờn thc vt bin i gen - Tờn thụng thng: Bụng Tờn khoa hc : Gossypium hirsutum Tờn thng mi: Tờn s kin chuyn gen: GHB119 Tớnh trng liờn quan n gen c chuyn: khỏng sõu v chng chu thuc tr c ammonium glufosinat Mó nhn din nht (nu cú): BCS-GHỉỉ5-8 Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt PHN II THễNG TIN V CY CH NHN GEN (a) Tờn khoa hc: Gossypium hirsutum (b) Tờn thụng thng: Bụng (c) V trớ phõn loi: Tc: H: Chi: Loi: Gossypieae Malvaceae Gossypium hirsutum Thụng tin v lch s canh tỏc v phỏt trin chn ging, c bit thụng tin liờn quan n cỏc tớnh trng cú th gõy tỏc ng bt hi n sc khe ngi v vt nuụi Bụng cú tờn khoa hc l Gossypium hirsutum,c trng nhiu thiờn niờn k nhiu ni trờn th gii Khong 90% lng bụng th gii l G hirsutum Bụng ch yu c s dng rng rói nh si ca nú X si c to trờn v ht vũng quanh thnh si chc mnh Ch cú M v mt vi quc gia khỏc phỏt trin cỏc sn phm thng mi t ht bụng Ht bụng thụ cú th c s dng lm thc n cho i gia sỳc dng khụ ht bụng, v ht hoc ht cú th c ch bin thu du, mt thnh phn chớnh c s dng lm thc phm X ngn (si ngn, linter) cũn li trờn v sau loi b si di cú th s dng cho c mc ớch thc phm/thc n chn nuụi hoc cỏc mc ớch khỏc Bụng thuc vo chi Gossypium, nm h Malvaceae hoc Mallow Cỏc chi khỏc ca h ny bao gm mp tõy, thc qu, hoa hng sharon, hay thm l cỏc cõy trng nh c tr li l nhng loi c xp vo nhúm c di cú nh hng xu ti bụng Ch cú chi Gossypium v mt vi chng thuc cỏc chi khỏc, c xỏc nh c tớnh thụng qua si lụng ht (seed hair)/cu trỳc lụng phỏt trin ngoi biu bỡ ca v ht Cú 50 loi thuc chi Gossypium, nhng ch s chỳng c s dng sn xut x vi thng mi (Fryxell, 1992) B Gossypieae cú im c trng: s to thnh phụi (phc hn so vi cỏc nhúm khỏc h Malvaceae) v s cú mt ca cỏc mch h khỏc bit nhiu phn ca cõy nhng c bit khụng cú lỏ mm Cỏc mch h c bit l cỏc tuyn cha gossypol, khỏc bit v c im sinh thỏi v thnh phn hoỏ hc õy l nhng c im c trng cho b ny (Fryxell, 1979) Bụng cú ngun gc l cõy g lõu niờn, sau ú c thun hoỏ v chuyn thnh cõy trng thu hoch hng nm v thng khụng mang cỏc c tớnh c di Cõy bụng c trng t ht Nu vng mt cỏc cụn trựng h tr th phn, bụng cú th t th phn, nhng cỏc tỏc nhõn th phn cú mt, vic th phn chộo s din ỏng k (Oosterhuis v Jernstedt, 1999) Vic trng bụng b hn ch v mt a lý hn bt k cõy trng no khỏc mt s chõu lc bi nú ch cú th phỏt trin c cỏc vựng cú di 180 ngy lnh giỏ nm (180 frost-free days per year) Bụng cn lng nc ma trung bỡnh ớt nht l 50 cm/nm, nhng ma nhiu v thi tit cú m ln giai on cui quỏ trỡnh phỏt trin d lm Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt gim nng sut cng nh tng nguy c tn cụng ca sõu bnh Ti thi im thu hoch, thi tit cn khụ rỏo, ma s lm bin mu si v nh hng ti cht lng x vi Bụng c trng v thu hoch M mt cỏch c gii hoỏ Cỏc k thut trng bao gm phng phỏp ti tiờu, luõn canh cõy trng, cỏc loi thuc tr c c s dng kim soỏt c di Vic kim soỏt tt c di cú th giỳp tng nng sut bụng, ú hu ht cỏc vựng trng M u s dng thuc tr c Ngi trng bụng cú th s dng thuc tr c cú ph tỏc dng rng ti nhiu loi c di, nhng khụng nh hng ti cõy bụng, cú tớnh kinh t v v thõn thin vi mụi trng Nhiu loi thuc tr c cú ph tỏc dng rng nhng cú th tiờu dit bụng c s dng lng khuyn cỏo i vi c Da trờn cỏc c im tớnh trng, cú th ỏnh giỏ c mc phự hp ca mt ging bụng mi s dng cho mc ớch thng mi húa Vic chn lc cỏc ging mi ban u da vo cỏc d liu ca cõy trng b m Cỏc nh chn to ging ỏnh giỏ cỏc ging bụng mi chn to qua nhiu ch tiờu ti cỏc thi k sinh trng khỏc ton b quỏ trỡnh phỏt trin ging giai on sm ca quỏ trỡnh sinh trng, cỏc nh to ging ỏnh giỏ s cõy tn ti v khe ca cõy ging Khi cõy bc vo thi k chớn, kh nng khỏng sõu v bnh c tip tc ỏnh giỏ Ht bụng thu hoch c phõn tớch cỏc ch tiờu nh nng sut, di v chc ca si bụng Trong mt s trng hp, cõy trng c ci bin nõng cao mt s thnh phn c bit, v cỏc nh chn to ging cõy trng thng chỳ ý phõn tớch cỏc thnh phn ú Thụng tin v s an ton ca cõy nhn gen bao gm c cỏc v c tớnh v tớnh d ng (bao gm c thụng tin i vi cỏc loi cõy trng cú h hng cựng loi m cú th s dng lm nn di truyn nhn gen tng t) Bụng khụng c coi l cõy trng gõy hi, gõy d ng v gõy bnh cho ngi, nhiờn nú to cỏc cht khỏng dinh dng nh gossypol v axit bộo cyclopropenoid (CPFA) Quỏ trỡnh ch bin ht bụng cú th bao gm cỏc cụng on: tỏch si, tỏch v, nghin, ộp tm, chit, ty mu, ty mựi thu du bộo thc phm Sn phm ph ca quy trỡnh ch bin cú th s dng lm thc phm cho ngi nh x ngn linter (si ngn), hay thc n chn nuụi nh v hay khụ du bụng (Freyssinet, 2007; OECD, 2009) Hỡnh th hin chi tit hn quy trỡnh sn xut cỏc sn phm t bụng Du ht bụng c sn xut u tiờn Hoa K nh mt loi du ht Du thụ cú cha khong 2% thnh phn phi-glyxerit nhng c loi b gn nh hon ton quỏ trỡnh ch bin Cỏc thnh phn ny gm cú phytoalexin, axit bộo cyclopropenoid (CPFA), phospholipid, sterols, resins, carbohydrate v cỏc sc t Thnh phn ỏng chỳ ý nht ca terpenoid phytoalexin l gossypol Quỏ trỡnh ch bin du loi tr hu ht gossypol, quỏ trỡnh kh mựi s loi b hu ht cỏc CPFA Khụ du ht bụng, v, x ngn l cỏc sn phm ph ca ngnh cụng nghip du ht bụng Trong nhng sn phm ny, khụ du ht bụng l nhiu nht v c sn xut bng cụng ngh ộp v s dng dung mụi tỏch chit cú th thng mi hoỏ cỏc sn phm cú hm lng gossypol thp, gii hn ca thnh phn gossypol phi nm di 0,04% hay (400 ppm) X ngn hu ht ch cha cellulose tinh X ngn cú cht lng cao nht c tinh sch bng cỏch x lý húa hc phõn gii, lm trng, v sy V cú hm lng ln cỏc cht x khụng tiờu húa c Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt Gossypol Bụng cú cha mt lng tng i cỏc cht terpenoid phytoalexin Cỏc phytoalexin l cỏc cht khỏng sinh, bụng cỏc cht ny c tớch tr cỏc tuyn sc t Cỏc cht ny úng vai trũ quan trng vic chng chu vi mt s loi bnh cú th gõy hi trờn cõy bụng Cỏc thnh phn terpenoid phytoalexin thụng dng cõy bụng gm cú gossypol, hemigossypol, desoxyhemigossypol, 2,7-dihydroxy cadalene, hemigossypolone v heliocides H1 v H2 Gossypol l cht ỏng chỳ ý nht cỏc cht terpenoid phytoalexin v c phõn lp u tiờn t cỏc tuyn sc t ca ht bụng Cht ny nhỡn chung l c t cho cỏc loi khụng phi ng vt nhai li v gõy bt dc c Gossypol cú th thy dng t hoc kt hp Nghiờn cu khuyn cao khong 18 mg gossypol t (tng ng vi 0,1%) l hm lng cao nht m bũ sa cú th c cho n hng ngy Cỏc nh khoa hc cng ó ch rng gossypol l cht cú th chng virus v chng li cỏc cht gõy ung th Cỏc axit bộo cyclopropenoid Bụng cha mt s axit bộo cyclopropenoid (CPFA) cú du Cỏc loi cú th nh lng c gm cú axit malvalic, sterculic v dihydrosterculic Cỏc CPFA tng im núng chy (melting point) cỏc cht bộo ng vt c cho n ht bụng v khụ du ht bụng C ch ca hin tng cú th l s c ch quỏ trỡnh kh bóo ho (desaturation) ca cỏc axit bộo no Trong g, thnh phn ny gõy nh hng xu l mt mu lũng trng v gim kh nng n ca trng, ú khụ du ht bụng b hn ch s dng ch dinh dng cho gia cm Cỏc CPFA cng cú liờn quan ti t l ung th gan cao cỏ hi c cho n ht bụng, mc dự ht bụng cú th nhim mycotoxin cú cha aflatoxin cng l tỏc nhõn gõy ung th cho cỏ hi cu vng (rainbow trout) Cỏc thnh phn khỏc Lỏ cõy bụng mang mt s cht flavonoid, cỏc tannin v anthocyanin Mt s lỏ bụng c thu hoch cựng vi qu bụng v b loi b quỏ trỡnh tỏch si ht (ginning) Trong mt s iu kin ngoi l, nh iu kin khụ hn, phn loi b sau tỏch si hay t bụng c s dng lm thc n chn nuụi cho gia sỳc ln Tuy nhiờn, vỡ lng s dng mc thp nờn flavonoid, tannin v anthocyanin khụng c coi l cỏc cht chng dinh dng hay cht c t nhiờn chớnh (OECD, 2009) Thụng tin v lch s s dng cõy ch lm thc phm, thc n chn nuụi (Mụ t chi tit v quỏn canh tỏc, chuyn, lu gi bo qun v cỏc iu kin c thự cn thit ch bin (nu cú) s dng thc phm, thc n chn nuụi, vai trũ, giỏ tr, thnh phn dinh dng ca cỏc b phn ca cõy s dng lm thc phm, thc n chn nuụi) Bụng, Gossypium spp c trng thu si t hng ngn nm Vic trng v sn xut vi si c phỏt trin c lp gia bỏn cu Nam v Bc Mt nhng ghi chộp lõu i nht v cụng nghip dt may thỡ cỏch õy 5000 nm cụng nghip vi si t bụng ó cú thung lng sụng Indus, thuc Pakistan Kho c Peru v Mexico ó phỏt hin nhiu mu vi bụng cú ti 4000 -7000 nm tui Vi bụng cng c tỡm thy nhiu nn minh Ai Cp c hay cỏc du tớch ca ngi da Indian khu vc Tõy Nam nc M t hng trm nm trc cụng nguyờn Ht bụng, sn phm nụng nghip thụ trc õy phn ln c loi b, c chuyn thnh thc phm cho ngi, ng vt, phõn bún cho thc vt, si cho sn xut Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt tm lút ni tht v cellulose cho nhiu ngnh cụng nghip khỏc t sn xut thuc n ti bo mch chip mỏy vi tớnh Ngoi bụng l thc phm ca t nhiờn v cõy trng to si Mc dự si bụng l sn phm cú giỏ tr nht lnh vc vi nhng cng phi chỳ ý rng cõy trng ny cng l ngun cung cp du thc vt quan trng iu ny cú ngha, bụng cng l cõy trng s dng cho mc ớch thc phm Bụng c sn xut chớnh Trung Quc, M, n , Pakistan v Uzbekistan, vi tng sn lng chim gn 75% sn lng ton th gii Phng phỏp ch bin, giỏ tr v thnh phn ca cỏc sn phm ph t ch bin Lch s s dng an ton lm thc phm v thc n chn nuụi Ch cú qu nang ca bụng l c dựng thu si hoc lm thc phm/thc n chn nuụi Phn cũn li ca cõy c s dng lm phõn bún ti rung trng sau quỏ trỡnh phõn hu (OECD, 2009) Ht bụng l ngun cung cp du bộo v protein rt tt bi hm lng cao cỏc thnh phn ny ht cng nh cht lng axit bộo v axit amin ca nú Bn sn phm chớnh t quỏ trỡnh ch bin ht bụng l du bộo, khụ du, v ht v x ngn (linter) T l cỏc sn phm cụng nghip ch bin ht bụng M vũng nm gm cú: 16,14% du, 25,15% v , 46,00% khụ du bụng, 8,51% x ngn v 4,21% cht thi (Jones v King, 1993) Ngy bụng ti cỏc nc cụng nghip phỏt trin c thu hỏi mt cỏch c gii hoỏ Sau tỏch bụng, nang qu bụng thng c ộp c hc, chuyn vo dõy chuyn ch bin c gi l quy trỡnh ta bụng Trong quỏ trinh ta nang qu bụng, si cho cụng nghip dt may c tỏch riờng t ht Ht ó c tỏch bụng s c gi l ht tỏch si (fuzzy seed) s c s dng ch bin hoc dựng trc tip lm thc n gia sỳc Du bụng l ngun du thc vt quan trng, õy l mt cỏc loi du bộo cha nhiu axit cha bóo ho nht; cựng vi du ci, ngụ, bụng, hoa rum , hng dng, du bụng cng cú cht lng cao bi hm lng vitamin E v bn ca nú c s dng lm du rỏn X ngn ca ht (cottonseed linter fiber), bao gm trờn 99% l cht x cng c dựng lm thc phm sau c x lý hoỏ hc phõn gii, lm trng, v sy khụ Sn phm ny dng bt trng, khụng mựi, mu v bn hoỏ hc vy khụng phn ng vi cỏc thnh phn khỏc gõy bin mựi, mu cho thc phm Do vy, nú cú th c dựng nhiu thc phm nh bỏnh, nc trn salat, n nhanh hoc sn phm ch bin t tht Khụ du ht bụng Phng ỏn ch bin du bộo nh hng ti tớnh hu dng ca khụ du bụng (cottonseed meal) So vi cỏc phng phỏp ch bin khỏc, vic ộp vớt ti gim hm lng gossypol t do, gim Nit ho tan, tng hm lng cht bộo khụ bụng v tiờu tn nng lng S dng cho nuụi g, hm lng du cao cú th nh hng ti s mt mu ca trng quỏ trỡnh bo qun (do s cú mt ca axit bộo cyclopropenoid); i vi gia sỳc, hm lng du cao to nhiu nng lng cho vt nuụi Phng phỏp chit bng dung mụi trc tip s to khụ du bụng cú hm lng gossypol t v nit ho tan cao Nu da trờn quan im v cht lng protein thỡ khụ du bụng sn xut theo phng ỏn ny rt tt cho ng vt cú d dy n (ng vt nhai li) Phng phỏp ch bin s dng dung mụi tỏch chit cú ộp s b to sn phm khụ du bụng cú c im trung gian gia phng phỏp tỏch du l ộp v s dng dung mụi Nú Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt cú hm lng gossyol t thp, ho tan nito cao hn mc trung bỡnh, hm lng cht bộo thp Loi khụ du bụng ny c s dng cho g trng (Ensminger v cng s, 1994) Ht bụng c ch bin thnh cỏc khụ du bụng cú cha hm lng protein khỏc 36%, 41%, 44%, v 48-50% Khụ du bụng cú cha 36% protein thu c t phng phỏp ộp bng trc vớt, ú phng phỏp chit dung mụi trc tip hoc chit dung mụi trờn ht bụng ó c ộp s b to khụ du bụng cú hm lng protein cao hn 44% (Ensminger v cng s, 1994) Ngoi mc ớch s dng cung cp protein cho cỏc sn phm thc phm/thc n chn nuụi, khụ du bụng cũn l ngun cung cp protein k thut lm cht kt dớnh Protein ny c ộp ựn to vt liu si s dng cho cụng nghip dt may Cỏc sn phm protein t ht bụng Trc õy, sn phm protein t ht bụng c s dng ch yu cho mc ớch phõn bún hoc lm thc n chn nuụi T u nhng nm 60, ngi ta bt u mi phỏt trin cỏc sn phm protein t ht bụng lm thc phm cho ngi Sn phm u tiờn c thng mi hoỏ l bt ht bụng s dng b sung vo bỏnh mỡ hay mụi trng nuụi cy vi sinh vt (trong nghiờn cu) Nam M, thc phm n dm cha hm lng protein cao cú tờn l Incaparinha c sn xut s dng cho tr thiu dinh dng õy l hn hp gia bt ngụ v ht bụng vi t l 60:40 (Bressani, 1997) X ngn ht bụng (Cottonseed linters) Ba loi x ngn chớnh c thu hi quỏ trỡnh tỏch si: cụng on ct th nht: 0.15-0.38 kg / 10 kg ht; cụng on ct th 2, 0.90-1.05 kg / 10 kg ht; v thu t cụng on nghin, 0.35-1.00 kg / 10 kg ht Sn phm t cụng on ct th nht c s dng to cỏc sn phm s dng cho phu thut, giy, bao gúi; sn phm t cụng on ct th s dng sn xut si tỏi sinh, phim nh, thuc n, cht do, giy, v sn phm thu t cụng on nghin s dng cụng nghip to cellulose hoỏ hc v cỏc sn phm nm lút (Cherry v Leffler, 1984) X ngn ht bụng - Linter, cha 90-95% cellulose, khụng cha lignin, cú hm lng sỏp thp, pectin, axit hu c, cỏc cht tro vụ c (Jones v King, 1993) Sau quỏ trỡnh ch bin pH kim v nhit cao, t t ht bụng cú th c s dng nh l mt sn phm cú hm lng cht x cao Cht x ny dựng cho thc phm cú cha trờn 99% hm lng x tng (Freyssinet, 2007) V ht V l phn bờn ngoi ht cú tỏc dng bo v ht, phn ny phi c tỏch l phn bờn ht cho quỏ trỡnh tỏch chit du S cú mt v ht khụ ht lm gim hiu sut thu hi du cú th hp ph mt lng du t Khong 1% ca v ht tớnh theo lng l du v 3% l protein V cú cha hm lng cht x cao, c bit l cellulose v lignin (NCPA, 2000; Jones v King, 1993) V ht bụng c s dng lm thc n thụ cho chn nuụi hay cú th dựng lm nhiờn liu, vt liu cỏch nhit, lm m t, s dng du khoan, b sung vo cht cha phenol tng bn, cung cp cellulose cho cỏc loi x tỏi sinh v cng l ngun to xylose v furfural (Cherry v Leffler, 1984) Lecithin Photpholipit Trong cỏc loi du t ht núi chung, tr u tng, thỡ ht bụng cú hm lng photpho lipit l cao nht Phõn on cha lng photpho lipit thu t quỏ trỡnh tinh ch du Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt bộo c x lý loi cỏc lipit trung tớnh v cỏc protein tp, sau ú c phõn on tip thnh photpho lipit v glycolipit Do s cú mt ca gossypol liờn kt vi photpho lipit quỏ trỡnh tỏch du, photpho lipit t ht bụng ch c s dng cho cỏc mc ớch cụng nghip Photpho lipit bụng cú nhiu u im hn so vi photpho lipit t cỏc loi ht khỏc bi hu ht cỏc axit bộo cú mt u ch cú di liờn kt ụi vy chỳng ớt b oxy hoỏ v b ụi hn (Cherry and Leffler, 1984) T ging bụng khụng to gossypol (glandless cotton), cú th cú c sn phm photpho lipit t ht bụng dựng cho thc phm (Cherry v Leffler, 1984) Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt PHN III THễNG TIN LIấN QUAN N SINH VT CHO GEN Tờn ca sinh vt cho gen S kin ny cú hai sinh vt cho khỏc l: Streptomyces hygroscopicus v Bacillus thuringensis 1.1 Sinh vt cho gen chng chu thuc tr c - Tờn khoa hc: Streptomyces hygroscopicus Tờn thụng thng: Streptomyces V trớ phõn loi: Gii: Bacteria Lp: Actinobacteria H: Streptomycetaceae Chi: Streptomyces Loi: Streptomyces hygroscopicus Chng:ATCC21705 Sinh vt cho gen khỏng sõu gen cry2Ae 1.2 - Tờn khoa hc: Bacillus thuringiensis Tờn thụng thng: Bacillus V trớ phõn loi: Gii: Bacteria Lp: Bacilli H:Bacillaceae Chi: Bacillus Loi: B thuringiensis chng: dakota 1715 Thụng tin liờn quan Lch s s dng an ton - Bacillus thuringiensis (Bt) rt ph bin t nhiờn v cú c mt th k cỏc nghiờn cu ó chng minh lch s s dng an ton ca loi vi khun ny Cỏc chng Bt c phõn loi chung vo nhúm cỏc vi sinh vt khụng gõy bnh phõn loi vi sinh vt ca mt s quc gia (OECD, 2007) Mt s chng khỏc ca vi khun Bt c ng ký lm thuc tr sõu v c quan tõm nhiu thõn thin vi mụi trng bi cỏc c tớnh chuyờn bit ca chỳng (ch nh hng n nht cỏc cụn trựng gõy hi) v khụng tn ti dai dng mụi trng Mt lng cỏc ging cõy trng, c bit v cỏc loi cõy nh bụng, ngụ, khoai tõy, thuc lỏ, c chua, mớa ó c chuyn gen to cỏc loi protein endotoxin t Bt Bt l mt loi vi khun thụng dng cú kh nng tn ti mụi trng mt thi gian di nú to cỏc bo t cú kh nng chng chu tt vi cỏc iu kin bt li ca mụi trng Vi khun Bacillus u cú dng hỡnh que, gram dng ch to mt bo t vi Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt mi t bo Cỏc chng Bt c c trng bi s to thnh ca mt hoc nhiu tinh th parasporal protein cựng lỳc vi vic hỡnh thnh bo t Cỏc tinh th parasporal ny cha thnh phn cú tớnh khỏng sõu -endotoxins cựng vi cỏc protein kt ni v cỏc c t Cyt Cỏc c t ny to hot tớnh khỏng sõu cho cỏc sn phm thuc tr sõu thng mi x lý cỏc loi sõu b thuc h cỏnh vy lepidopteran, dipteran, v coleopteran Kt qu thu c t mt s lng ln nghiờn cu trờn ngi v ng vt qua nhiu nm cho thy tớnh tr sõu ca nú gn nh khụng cú ri ro cho ngi v ng vt, v khụng cú bt c bng chng no cho thy c tớnh tr sõu sinh hc ca vi khun Bt gõy bt k du hiu lõy nhim hay ng c ca thc phm (McClintock et al., 1995; Betz et al., 2000; Rosenquist et al., 2005) Túm li, c tớnh tr sõu sinh hc ca Bt ó cú mt lch s s dng an ton - S hygroscopicus ó c chng minh l an ton s dng qua bi bỏo khoa hc tng hp ca Herouet v cng s (2005) B mt bo t ca sinh vt ny thng nhn nheo khi bo t mu xỏm, cú mu en v nht trng thnh Cỏc vi sinh vt ny thuc nhúm nhng vi sinh vt tim nng cho vic to cỏc gen khỏng khỏng sinh cú chc nng vụ hot tớnh c ca chớnh chỳng (Freyssinet, 2007) õy l nhng vi sinh vt d dng, hiu khớ, cú kh nng trao i cht theo ng oxy hoỏ mnh m t l mụi trng t nhiờn thớch hp cho s tn ti, phỏt trin v sinh trng ca cỏc vi sinh vt ny Con ngi tip xỳc nhiu vi vi khun v vi khun ny phỏt tỏn rng t nhiờn (Kutzner, 1981) Do ú, vic s dng cỏc loi c v rau l ngun tip xỳc thụng thng ca ngi v ng vt vi loi vi khun ny Khụng cú bng chng no cho thy nh hng bt li tip xỳc vi cỏc vi khun ny Thụng tin v vic tỡm thy t nhiờn cỏc cht khỏng dinh dng, c t v cht gõy d ng Qua nhiu thp k s dng cỏc sn phm Bt vi sinh vt thng mi, kh nng gõy c vi ng vt cú vỳ ca chỳng c ỏnh giỏ Cỏc d liu c t hc ca cỏc Bt khụng ch nh hng xu no ca -endotoxins vi sc kho ca ng vt cú vỳ Mt kho sỏt trờn nhiu nghiờn cu v s nhim v gõy bnh ch mt kiu hỡnh loi b cỏc vi sinh vt Bt trờn loi gm nhm c thớ nghim cho n theo ng tiờu hoỏ, theo ng khớ th hay tiờm vo tnh mch Khụng cú nh hng bt li no (bao gm nh hng ti s tng trng lng c th, t l cht hay cỏc biu hin lõm sng, tỏc ng ti cỏc b phn bờn ng vt thớ nghim) c ch cỏc nghiờn cu ú nh hng ca vi sinh vt Bt C ch tỏc ng ca -endotoxins t Bt cụn trựnh mn cm ó c lm rừ (OECD, 2007) Nhiu nghiờn cu ó tỡm thy mt s chng Bt gõy c cho chut cú h thng dch b phỏ hy hay chut b nhim virus gõy cỳm Tuy nhiờn, cỏc tỏc gi ch rng ú l tỏc ng ca cht c khỏc t Bacillus ch khụng phi -endotoxins Cỏc chng Bt tng t khỏc khụng cha -endotoxins cú th gõy c cho ng vt cú vỳ bi cỏc c t gõy tan huyt (haemolytic toxins) (OECD, 2007) C quan chu trỏch nhim ỏnh giỏ an ton thc phm ca chõu õu (EFSA) ó ỏnh giỏ nguy c gõy d ng tim n ca -endotoxin biu hin ngụ, chng hn nh Cry1Ab Cỏc ỏnh giỏ v kh nng gõy di ng ca protein Cry c thc hin vi nhiu phng phỏp v cỏch tip cn khỏc nhau, a cỏc bng chng giỏn tip cho thy nguy c gõy di ng l Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt rt thp Cỏc bng chng ú bao gm: ngun cho protein Cry khụng gõy d ng, khụng cú s tng ng v trỡnh t ca Cry protein vi cỏc cht gõy d ng ó bit, s mn cm v b phõn gii nhanh chúng bi pepsin Cho ti nay, mc dự cú s rt nhiu nghiờn cu khoa hc, khụng cú phng phỏp no c tỡm ỏnh giỏ mt cỏch thuyt phc v kh nng gõy di ng ca mt cht b nghi ng sau cỏc kim tra sng lc s b (OECD, 2007) S hygroscopicus bn thõn khụng c bit n l vi sinh vt gõy bnh cho ngi hay liờn quan ti cỏc nh hng bt li cho sc kho ca ngi (Locci, 1995) Streptomyceteae phõn b ph bin t v mụi trng nc Hu ht chỳng l loi hot sinh bt buc Rt ớt thụng tin v vai trũ ca Streptomycetes mụi trng t nhiờn, nhiờn s cú mt ca chỳng v s lng ca chỳng mụi trng l rt ln t, cỏc phn phõn hu l nhng ngun cú cha nhiu Streptomycetes Tuy nhiờn, cng cú mt s ký sinh trờn cõy trng v ng vt Mc dự Streptomyceteae thng c xp vo nhúm vi sinh vt hiu khớ bt buc, nhng chỳng cú th phỏt trin c t vi lng oxy thp, tr lng CO2 quỏ 10% Trong t khụ, lng Streptomycete gim, nhng bo t ca chỳng cú th chu c iu kin khc nghit hn so vi t bo sinh dng cú th sng sút c Streptomyces hygroscopicus khụng c bit n l loi gõy d ng v gõy c Nhiu thnh viờn ca ging ny cú th to cỏc khỏng sinh dung cho y hc (Freyssinet, 2007) Mt s Streptomyces c phõn lp t ngi v ng vt, gõy bnh cho vt ch ca chỳng, nhng nhng chng ny l cỏ bit Tuy nhiờn, vi thc vt thỡ mt s loi Streptomyces spp c bit n l cỏc vi sinh vt gõy bnh cho cõy trng (Kutzner, 1981) S hygroscopicus v cỏc loi Streptomyces khỏc phõn b rng rói t nhiờn Chỳng l mt thnh phn ph bin sinh quyn th gii v rt ớt loi Streptomyces l tỏc nhõn gõy bnh cho ngi, ng vt v thc vt Mt s loi ca Streptomyces tng t vi S hygroscopicus v rt nhiu s chỳng cú cỏc thnh phn ng dng bar/pat Khụng mt thnh phn ng dng no c bỏo cỏo l c v gõy d ng cho ngi v ng vt (Kutzner, 1981 M-204308-01-1) Mt s loi khỏc thuc nhúm ny cũn sn sinh cỏc loi khỏng sinh cha bnh hu ớch (Freyssinet, 2007) Thụng tin v vic ó v ang s dng S tip xỳc c bn nht ca ngỡ vi -endotoxins cõy trng l tiờu th cỏc cõy trng thc phm S tip xỳc vi cỏc sol khớ to quỏ trỡnh ch bin nguyờn liu t cõy trng Bt (chng hn ht) cng l mt cỏch tip xỳc khỏc, mc dự nh Nhiu quc gia yờu cu cỏc nghiờn cu v d lng thuc tr sõu xỏc nh hm lng ln nht cỏc thuc tr sõu hoỏ hc hoc trờn nguyờn liu nụng nghip Do khụng cú tớnh c vi ng vt cú vỳ nờn -endotoxins c kim tra hm lng rt ln, cỏc nghiờn cu thụng thng v d lng vy khụng cn thit Thuc tr sõu cha Bt c ng ký cỏc nc yờu cu v d lng (a.k.a Maximum Residue Levels) c tr yờu cu v vic t mt d lng s hc D liu ch cõy trng chuyn gen khỏng sõu ang c s dng thng mi hin cú cha hm lng rt thp -endotoxins cỏc phn n c ca cõy trng Da trờn lch s s dng an ton ca Bt v cỏc kt qu thớ nghim gõy c cp tớnh cú th kt lun cỏc -endotoxins mang nguy c gõy c cho ngi v ng vt l khụng ỏng k (OECD, 2007) Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt PHN IV THễNG TIN V THC VT BIN I GEN Quỏ trỡnh bin i gen Cỏc ging bụng GHB119 thuc loi, Gossypium hirsutum L v c phõn bit vi cỏc ging bụng bi tớnh trng khỏng sõu v kh nng chng chu thuc tr c ammonium glufosinat Tớnh trng khỏng sõu bụng GHB119 cú c nh tinh th protein tr sõu, Cry2Ae, gen cry2Ae mó húa, cú ngun gc t loi vi khun t thụng dng Bacillus thuringiensis subsp dakota (B.t dakota) Protein Cry2Ae cú hiu qu kim soỏt u trựng cỏc loi cụn trựng b cỏnh vy nh u trựng sõu hi qu bụng (CBW, Helicoverpa zea), u trựng hi chi thuc lỏ(TBW, Heliothis virescens) v u trựng sõu khoang (FAW, Spodoptera frugiperda), l cỏc loi sõu hi trờn cõy bụng Tớnh trng chng chu thuc tr c (OECD, 1999) gen bar biu hin, gen khỏng thuc tr c t nhiờn c phõn lp t vi khun t Streptomyces hygroscopicus Gen bar biu hin cho phộp vic sn sinh enzyme Phosphinothricin-Acetyl-Transferase (PAT) cú kh nng acetyl húa L-glufosinate ammonium v ú giỳp cho cõy trng cú th chng chu thuc tr c cú gc ammonium glufosinat (Freyssinet, 2002) Phng phỏp chuyn gen Mụi trng vi khun Agrobacterium tumefaciens c s dng chuyn vector pTEM12 vo mụ ca bụng Gossypium hirsutum ging Cỏc mụ phụi c tip xỳc vi mụi trng nuụi cõy cú cha A tumefaciens, chng C58C1Rif (Van Larebeke et al., 1974 M147632-01-1) ó b lm mt c tớnh mang vector pTEM12 Sau nuụi cy, cỏc mụ bụng tip tc c tỏi sinh thnh cõy hon chnh trờn mụi trng thớch hp cú b sung 500 mg/l loi b phn cũn li ca Agrobacterium, v c chn lc bng ammonium glufosinat Cõy ó phỏt trin c a vo trng nh li v tip tc c kim tra kh nng chng chu vi thuc tr c Liberty, cho hoa v to ht (Criel 2008 M-311228-01-1) Mụ t kiu hỡnh ca tớnh trng mi c chuyn Cỏc ging bụng GHB119 thuc loi Gossypium hirsutum L v cú c im nhn bit khỏc bit vi cỏc ging bụng khỏc hai trớnh trng: khỏng sõu v chng chu thuc tr c ammonium glufosinat Cỏc kho nghim trờn ng rung ó c tin hnh ỏnh giỏ cỏc thay i v mt sinh hc ca bụng GHB119 v kh nng tỏi sinh, phỏt tỏn, tn ti so sỏnh vi bụng khụng chuyn gen Khụng cú s thay i v kh nng sinh sn, hỡnh thỏi hoa hay s tn ti v phỏt tỏn ca ht phn ó c cỏc nh chn to ging quan sỏt chng trỡnh phỏt trin ging (vn ang tip tc t nm 2004) (Villagrỏn, 2008b M-300865-01-1) S khỏc bit nht ca bụng GHB119 l kh nng khỏng li u trựng ca cụn trựng b cỏnh vy v chng chu thuc tr c ammonium glufosinate Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt Lch s cp phộp ca bụng GHB119 trờn th gii GHB119 ó c xem xột v tớnh an ton lm thc phm, thc n chn nuụi ti mt s nc trờn th gii GHB119 ó c cp phộp ti quc gia l: Hoa K, AustraliaNew Zealand, Canada, Nht Bn, Trung Quc v Hn Quc Danh sỏch cỏc quc gia cp phộp cho s dng lm thc phm, thc n chn nuụi i vi bụng GHB119 Quc gia Australia/New Zealand Canada Trung Quc Nht Bn Hn Quc Hoa K Thc phm Thc n chn nuụi 2011 2012 2014 2013 2013 2012 2011 2012 2014 2013 2013 2012 Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt PHN V NH GI RI RO CA THC VT BIN I GEN N SC KHE CON NGI V NG VT Nghiờn cu c tin hnh trờn mu bụng GHB119 v i chng khụng chuyn gen so sỏnh thnh phn dinh dng ca ht bụng nguyờn v ht bụng tỏch si Cỏc mu thuc t 16 im nm 2007 v 2008 ti Tõy Ban nha c s dng phõn tớch 50 thnh phn D liu chi tit cho phộp ỏnh giỏ cỏc phõn tớch thng kờ, kt qu cỏc phõn tớch ny cho thy s tng ng v thnh phn ca bụng GHB119 vi i chng khụng chuyn gen Coker 312 Cỏc d liu nghiờn cu c phõn tớch bi Bayer CropScience Chõu u (Oberdoerfer, 2009a; Oberdoerfer 2009b) Ht bụng c cu to bi phn chớnh: ht vi v cú si di (lint) v t bụng (linter); lỏ mm giỳp cho vic hỡnh thnh phụi; v mm Sau si di ca ht c loi b ti cụng on tỏch si, ht bụng cũn li 12,71% t, 31,76% v v 55,53 % ht (gm lỏ mm v phụi) (Jones and King, 1993) Ti Hoa K, quỏ trỡnh ch bin cụng nghip thc t ca c ht bụng thu c cỏc sn phm khỏc nhau: 8,51% t; 25,15% v; 16,14% du; 46% khụ du bụng v tn tht 4,21% vic chuyn v ch bin (Jones and King, 1993) T t ht bụng cha 90-95% cellulose, khụng cha lignin v ch cú mt lng nh sỏp, pectin, axit hu c v cỏc cht vụ c to tro (Jones and King, 1993) Cỏc ht bụng ó tỏch si di cha 25-40% v V tru chim khong 38% ht, vi 1% du, 3% protein v thnh phn cht x cũn li ging cellulose v lignin (Jones and King, 1993) Cỏc thnh phn c chn phõn tớch l cỏc thnh phn dinh dng quan trng v c bn ca bụng ú l cỏc thnh phn proximate, cht x, cỏc thnh phn vi lng nh khoỏng, vitamin E, cht chng dinh dng, axit amin tng v tng cỏc axit bộo Nhỡn chung, tt c cỏc giỏ tr trung bỡnh ca ht bụng tỏch si tng ng vi cỏc giỏ tr thu c ca cỏc ging bụng thng mi Tt c cỏc giỏ tr tr giỏ tr trung bỡnh thu c ca Coker 312 v bụng GHB119 l nm khong cỏc giỏ tr tham kho cho cỏc ging bụng hin ang thng mi húa Ba giỏ tr khụng nm khong l protein thụ, cht x ty axit v Maggie, ú protein thụ v Maggie thc t l cao hn i chng khụng chuyn gen so sỏnh vi cỏc ging thng mi húa Vi cỏc axit bộo, mt s khỏc bit gia cỏc bin phỏp x lý ó c tỡm thy Cỏc ch s ca ging i chng khụng chuyn gen v cỏc ging bụng chuyn gen cho thy s tng ng cao vi cỏc ging ang thng mi húa (xem Bng 17) Túm li, tt c cỏc thnh phn ỏnh giỏ thu c bụng chuyn gen l tng t nh cỏc ging bụng thng mi húa Khụng cú s khỏc bit no liờn quan n sinh hc c xỏc nh gia i chng khụng chuyn gen v cỏc ging bụng chuyn gen ỏnh giỏ v an ton thc phm ca protein Cry2Ae, trỡnh t axit amin cho thy s tng ng rừ rt gia protein ny vi cỏc protein khỏc thuc nhúm protein Cry2 cú biu hin cỏc s kin bin i gen khỏc, v cú lch s s dng an ton Khụng cú bng chng no c tỡm thy v s tng ng ca nú vi cỏc cht gõy c, gõy d ng, cht chng dinh dng ó bit hay cỏc bin i sau dch mó Protein Cry2Ae cho thy bn phn no Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt vi nhit v dch gi rut ca ngi, nhng li b phõn hy nhanh dch gi d dy ca ngi Kim tra c cp tớnh cho thy khụng cú bng chng no v h thng gõy c, thng nht vi cỏc nghiờn cu khoa hc v c tớnh ca protein Cry Phõn tớch thnh phn ca protein mi cho thy kh nng nh lng ca protein Cry2Ae cỏc sn phm ch bin cú ngun gc t ht bụng GHB119 tr du ht bụng, l loi thc phm ch yu ca bụng m ngi s dng Vit Nam Gen cry2Ae cú ngun gc t vi khun t thụng thng Bacillus thuringiensis chng dakota, mó húa protein Cry2Ae Protein ny l mt loi Bt-endotoxin ca lp Cry2, cú kh nng kim soỏt u trựng b cỏnh vy, l loi sõu hi ph bin trờn bụng, gm cú sõu hi qu (Heilcoverpa zea) v sõu hi chi thuc lỏ (Helioihis virenscens) Gen bar cng cú ngun gc t mt loi vi khun t thụng dng l Streptomyces hygroscopicus Gen ny mó húa cho protein phosphinothricin acetyl transferase (PAT) protein l enzyme cú tớnh c hiu cao Protein PAT protein thuc v nhúm cỏc enzyme acetyltransferase thụng dng vi cõy trng v ng vt PAT cú cu trỳc bc 2, phn ng dch, lng phõn t v chc nng tng t vi cỏc enzyme acetyltransferase khỏc cú mt ch dinh dng ca ngi v ng vt Khụng cú bỏo cỏo no v s lien quan n c tớnh hay gõy d ng ca nhúm cỏc enzyme acetyltransferase S cú mt ca protein PAT chui thc phm v thc n chn nuụi khụng phi l mi xy Streptomyces hygroscopicus l loi vi khun hoi sinh t thụng thng v c bit l khụng gõy bnh cho ngi v ng vt Protein PAT giỳp kh nng chng chu vi vic s dng thuc tr c cú gc ammonium glufosinat l hot cht chớnh Protein PAT c biu hin khỏ nhiu cõy trng bin i gen, l cỏc cõy trng ó c ỏnh giỏ an ton mt cỏch cht ch bi c quan chc nng ca nhiu quc gia trờn th gii Vic tớch hp v biu hin ca locus chuyn gen bụng GHB119 h gen ca bụng G hirsutum ó c phõn tớch c im thụng qua cỏc tiờu chun quc t v ỏnh giỏ an ton cỏc sn phm cụng ngh sinh hc Thụng tin ny c a vo h s ny h tr cho vic ỏnh giỏ an ton thc phm ca protein Cry2Ae v PAT/bar Bng di õy trỡnh by tng hp cỏc ỏnh giỏ an ton v kt lun i vi bụng GHB119 Vn quan tõm Kiu d liu Xỏc nh nguy c c im ca sinh vt cho v sinh vt nhn Gen bar cú ngun gc t Streptomyces sp, cú mt rng rói t nhiờn Khụng cú Gen cry2Ae cú ngun gc t vi khun Bacillus thuringensis, cú mt rng rói t nhiờn Khụng cú Mt on chốn n c phõn tớch chi tit cú mt bụng GHB119 m khụng cú mt ca gen khỏng khỏng sinh Cỏc nghiờn cu cho Khụng cú Bin i di truyn v cỏc chc nng liờn quan Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt thy tớnh n nh ca gen biu hin v di truyn tớnh trng bụng GHB119 Kh nng nh hng ti mụi Kh nng tng tỏc ca cỏc sn phm bụng trng GHB119 ti mụi trng l cc k hn ch, khụng cú khỏc bit no gia nhp khu sn phm ny v cỏc sn phm t bụng thụng thng khỏc Khụng cú c tớnh nụng hc Khụng liờn quan Khụng cú Kh nng gõy c v d ng ca sn phm gen mi chuyn, cht chuyn húa v ca ton cõy Protein PAT cú mt rng rói t nhiờn v cỏc sn phm thng mi húa ca cỏc loi cõy trng bin i gen khỏc Tng hp ỏnh giỏ an ton chi tit ca protein PAT khụng tỡm thy bt k quan ngi no Bụng GHB119 khụng khỏc so vi cỏc ging bụng ang thng mi Khụng cú Tng hp ỏnh giỏ an ton chi tit ca protein Cry2Ae khụng tỡm thy bt k quan ngi no Bụng GHB119 khụng khỏc bit so vi cỏc ging bụng ang thng mi húa Cỏc c im thnh phn, dinh dng ó chng minh c s tng ng sinh hc v thnh phn, dinh dng v cỏc ch tiờu an ton Khụng cú nh hng ca quỏ trỡnh ch bin n c im ca thc phm, thc n chn nuụi Khụng cú s thay i v thnh phn dinh dng hay cú mt ca cỏc nhõn t dinh dng chớnh cỏc sn phm ch bin Khụng cú Kh nng thay i Khụng cú cỏc c im c hin cú th dn vic hp th qua ch dinh n vic tng lờn hp th qua ch dinh dng dng s vi bụng truyn thng Khụng cú Kh nng nh hng dinh dng lõu di Khụng cú s thay i thnh phn dinh dng hay cú mt ca cỏc cht dinh dng chớnh Khụng cú Cỏc nh hng ch ớch v khụng ch ớch vic chuyn gen Tớnh tng ng c bn vi bụng khụng chuyn gen tr nhng tớnh trng cú ch ớch a vo Khụng cú Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt PHN VI XUT CC BIN PHP QUN Lí RI RO CA THC VT BIN I GEN I VI SC KHE CON NGI V VT NUễI Bayer CropScience ó tin hnh ỏnh giỏ an ton ca thc vt bin i gen v thc phm, thc n chn nuụi cú ngun gc t bụng GHB119 Cỏc nghiờn cu v bỏo cỏo cho n khụng xỏc nh c bt k nguy c tim n no liờn quan ti bụng chuyn gen mang s kin GHB119 Kt lun v ỏnh giỏ ri ro i vi bụng mang s kin GHB119 l an ton v cú thnh phn tng ng vi i chng truyn thng v cỏc ging bụng thng mi K hoch giỏm sỏt xut a vic giỏm sỏt chung cỏc nh hng bt li cú th cú, lp tc hay chm tr, trc tip hay giỏn tip ca bụng chuyn gen GHB119 n sc kho ca ngi v mụi trng PHN VII KT LUN V KIN NGH Khụng cú ri ro liờn quan n bụng chuyn gen GHB119 no c tỡm thy sau phõn tớch v ỏnh giỏ i chng truyn thng c s dng phõn tớch so sỏnh Vic thay i di truyn cú ch ớch l tng li ớch v mt nụng hc, khụng lm thay i cỏc thnh phn dinh dng hay giỏ tr ca chỳng Khụng cú quan ngi cú ch ớch no liờn quan n sc kho ca ngi Thc phm v thc n chn nuụi cú ngun gc t GHB119 s khụng thay th hay lm thay i thc phm v thc n chn nuụi truyn thng GHB119 khụng mang nhng c im c bit no lm tng ch dinh dng so sỏnh vi bụng truyn thng Khụng cú bng chng no cho thy nh hng lõu di v dinh dng v sc kho vi nhng sn phm thc phm cú ngun gc t GHB119 Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt REFERENCES Arnaut, G., Boets A., Vannest S., Van Rie J., and Van Houdt S 2002 Novel Bacillus thuringiensis insecticidal proteins Patent Application: WO 02/057664 A2 2005 68 pages #M-392710-01-1 Betz, F.S, Hammond, B.G., Fuchs, R.L., 2000 Safety and advantages of Bacillus thuringensis protected plants to control insect pests Regulatory Toxicology and Pharmacology 32 Bolivar F; Rodriguez R L; Greene P J; Betlach M C; Heyneker H L; Boyer H W; Crosa J H; Falkow S, 1977 Construction and characterization of new cloning vehicles II A multipurpose cloning system Gene, , M-147993-01-2 Berger L.A., Vaissiốre B.E., Moffett J.O., Merritt, S.J 1988 Bombus spp (Hymenoptera: Apidae) as pollinators of male-sterile Upland cotton on the Texas High Plains Environ Entomol 17 789-794 M-208075-01-1 Bressani, R., 1997 High protein quality vegetable mixtures for human feeding Insituto de Nutricion de Centro America y Panama (INCAP) 1-10 M-207987-01-1 Cherry, J P.; Leffler, H R Seed (Chapter 13) ARS-USDA, Pennsylvania, USA; M-208069-01-1Cornelissen M; Vandewiele M, 1989 Nuclear transcriptional activity of the tobacco plasmid psbA promoter, Nucleic Acids Research, , 11 pages, M-147994-01-2 Capt A 2010c Heat stability study with the Cry2Ae protein Bayer CropScience Internal report pages #M-363360-01-1 Capt A 2011c In silico analysis of putative Open Reading Frame (ORF) sequences for identifying potential homologies to known toxins and allergens Bayer CropScience Internal report 36 pages # M-354512-03-1 Capt A 2011d In silico analysis of putative Open Reading Frame (ORF) sequences for identifying potential homologies to known toxins and allergens Bayer CropScience Internal report 412 pages # M-411784-01-1 Cornelissen M., Vandewiele, M 1989 Nuclear transcriptional activity of the tobacco plastid psbA promoter Nucleic Acids Research 17 19-29 11 pages #M-14799401-2 Criel I 2008 Description of the GHB119 transformation methodology Bayer CropScience Internal report 161 pages #M-311228-01-1 Currier T.C 2005 Analysis to Determine if the GEM2 Protein from Cotton Leaves is Glycosylated Bayer CropScience Internal report 30 pages #M-255733-01-1 Currier T.C 2008 Protein Expression Analysis of Cotton Event GHB119, Expressing Cry2Ae and PAT/bar Proteins, USA, 2007 Bayer CropScience Internal report 161 pages #M-312367-03-1 Damgaard, P.H., Granum, P.E., Bresciani, J., Torregrossa, M.V., Eilenberg, J., Valentino, L (1997) Characterization of Bacillus thuringiensis isolated from infections in burn wounds FEMS Immunology and Medical Microbiology 18,47-53 M-301514-01-1 De Almeida R.P.E., Gossele V., Muller C.G., Dockx J., Reynaerts A., Botterman J., Krebbers E and Timko M.P 1989 Transgenic expression of two marker genes under the control of an Arabidopsis rbcS promoter: Sequences encoding the Rubisco transit Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt peptide increase expression levels Mol Gen Genet (1989) 218:78-86 #M-180194-01-1 De Beuckeleer, 2003 Description of the amino acid sequence of the PAT protein encoded from the bar gene Bayer CropScience Internal report pages #M-084188-01-2 Depicker A., Stachel S., Dhaese P., Zambryski P and Goodman H.M 1982 Nopaline Synthase: Transcript Mapping and DNA Sequence Journal of Molecular and Applied Genetics Vol 1, No 6561-573 #M-131630-01-2 Deblaere R; Bytebier B; De Greve H; Deboeck F; Schell J; van Montagu M; Leemans J, 1985 Efficient octopine Ti plasmid-derived vectors for Agrobacteriummediated gene transfer to plants, Vrije Universiteit Brussel;Laboratorium voor Genetische Virologie, pages, M-147995-01-1 Ensminger A.H., Ensminger M.E Kondale J.E., Robson J.R.K 1994 Foods and Nuitrition Encylcopedia 2nd ed Ann Harbor, MI M-208031-01-1 FEDIOL, 2008 EU-27 consumption of vegetable oils and fats M-366266-01-1 Fling M.E., Kopf J., Richards C 1985 Nucleotide sequence of the transposon Tn7 gene encoding an aminoglycoside-modifying enzyme, 3(9)-O-nucleotidyltransferase Nucleic Acids Research 13 (19): 7095-7106 12 pages #M-231609-01-1 Franck, A.W (1989) Food uses of cottonseed protein Developments in Food proteins - New York, p 31-80 M 208064-01-1 Fryxell P.A 1979 The natural history of the cotton tribe (Malvaceae, tribe Gossypieae),Texas A&M University Press College Station and London M-208062-01-1 Fryxell PA 1992 A revised taxonomic interpretation of Gossypium L (Malvaceae) Rheedea 2: 108-165 M-420817-01-1 Freyssinet M 2007 General description of the bacterial qene bar and its gene product PAT as used for producing plants with a genetically-based tolerance to Libertyđ herbicide Bayer CropScience Internal report 24 pages #M-085622-01-2 Freyssinet, M 2007 Cotton crop biology and general information, Bayer CropScience, 50 pages, M-215504-03-2 Jones L.A and King C.C 1993 Cottonseed oil National Cottonseed Products Association Inc and The Cotton Foundation Memphis, Tennesse M-22804401-1 Gouffon 2011 Binding Sites for Bacillus thuringiensis Cry2Ae Toxin on Heliothine Brush Border Membrane Vesicles Are Not Shared with Cry1A, Cry1F, or Vip3A Toxin APPLIED AND ENVIRONMENTAL MICROBIOLOGY, May 2011, p 31823188 # M409337-01-1 Habex V 2005 Description of the amino acid sequence of the GEM2 protein Bayer CropScience Internal report pages #M-254766-01-2 Habex V 2011 Detailed insert characterization of Gossypium hirsutum transformation event GHB119 by Southern blot analysis Bayer CropScience Internal report 49 pages # M-308404-03-1 Hajdukiewicz, P., Svab, Z., Maliga, P (1994) The small, versatile pPZP family of Agrobacteriurn binary vectors for plant transformation Plant Molecular Biology 25, 989994 M 289172-01-1 Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt Harpster M.H., Townsend J.A., Jones J.D.G., Bedbrook J., Durismuir P (1988) Relative strengths of the 35S Cauliflower Mosaic Virus, 1', 2' and nopaline synthase promoters in transformed tobacco, sugarbeet and oilseed rape callus tissue Molecular and Genera! Genetics 212,182-190 M-135146-01-1 Herouet, C, Esdaiie, D.J., Mallyon, B.A., Debruyne, E., Schulz, A., Currier, T., Hendrickx, K., van der Klis, R.J., Rouan, D (2005) Safety evaluation of the phosphinothricin acetyltransferase proteins encoded by the pat and bar sequences that confer tolerance to glufosinate-ammonium herbicide in transgenic plants Regulatory Toxicology and Pharmacology 41:134-149 M-247779-01-1 Kỹtzner H.J (1981) The family Streptomycetaceae In The Prokaryotes: A Handbook on Habitats, Isolation and Identification of Bacteria, Starr M.P., Stolp H., Trtiper H.G., Ballows A., Schlegel H.G (Eds), p 2028-2090 Kowite W.J 2007 Production of RAC (Fuzzy Seed) Samples of GEM2 Cotton and the Non-transgenic Counterpart, USA, 2006 Bayer CropScience Internal report 29 pages #M-312403-01-1 Kowite, 2009 Production of Processed Commodities of Transgenic Insect-Resistant Cottonseed (Event GHB119) and the Non-transgenic Counterpart, USA, 2009 Bayer CropScience Internal report 19 pages #M-358440-01-1 Kowite, 2010 Composition of processed Fractions of Transgenic Event GHB119 Cottonseed and the Non-Transgenic Counterpart, USA, 2009 Bayer CropScience Internal report 60 pages #M-399271-01-1 Locci, R 1995 Streptomycetes and related genera In: Bergeys Manual of Systematic Bacteriology Vol S.T Williams, M.E Sharpe and J.G Holt, eds Williams & Wilkins, Baltimore, pp 2451-2492, M-135197-01-1 Mackie S.J.W 2008a Composition of RAC (Ginned Seed) of Cry2Ae GHB119 Cotton and the Non-transgenic Counterpart, USA, 2007 Bayer CropScience Internal report 229 pages #M-311235-02-1 Mackie S 2009 Analysis of processed commodities from transgenic event GHB119 cotton for Cry2Ae and Pat/bar proteins USA 2009 Bayer CropScience Internal report 47 pages #M-359968-02-1 Martone A 2009a Analyses of Raw Agricultural Commodity (Fuzzy Seed) of GHB119 Cotton from Catalonia for PAT/bar and Cry2Ae Bayer CropScience Internal report 99 pages #M-344919-02-1 MartoneA 2009b Analyses of Raw Agricultural Commodity (Fuzzy Seed) of GHB119 Cotton from Andalusia for Cry2Ae and PAT/bar Proteins, USA 2009 Bayer CropScience Internal report 120 pages #M-349875-01-1 Martone A 2011 Analyses of raw agricultural commodity (fuzzy seed) of Cry2Ae cotton event GHB119 for PAT/bar and Cry2Ae and its non-transgenic counterpart for PAT/bar and Cry2Ae proteins Bayer CropScience Internal report 126 pages #M-30165503-1 McClintock, J T., Schaffer, C R., Sjoblad, R.D (1995) A comparative review of the mammalian toxicity of Bacillus ihuringiensis-based pesticides Pesticide Science 45, 95105 Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt Moens, S (2008) Confirmation of the absence of vector backbone sequences in Gossypium hirsutum transformation event GHB119 Unpublished Bayer CropScience Report; October 22, 2008 M-311127-01-1 Moens S 2009 Determination of additional flanking sequences and the corresponding pre-insertion locus of Gossypium hirsutum transformation event GHB119 Bayer CropScience Internal report 40 pages #M-356284-01-1 Moens S 2010 Full expression analysis of the trait genes and newly created ORFs of cotton event GHB119 Bayer CropScience Internal report 38 pages #M-356285-02-1 Moens S 2011a Bioinformatic analysis of Gossypium hirsutum transformation event GHB119 Bayer CropScience Internal report 206 pages #M-356287-03-1 Moens S 2011b Characterization of Cry2Ae and PAT/dar proteins expressed in Gossypium hirsutum transformation event GHB119 Bayer CropScience Internal report 39 pages # M-406615-03-1 Murakami, T., Anzai, H., imai, S., Satoh, A., Nagaoka, K., Thompson, C J (1986) The bialaphos biosynthetic genes of Streptomyces hygroscopicus: Molecular cloning and characterization of the gene cluster Molecular and General Genetics 205,42-50 M131622-01-1 National Cottonseed Products Association (NCPA) (2000) Cottonseed Oil CSO Bulletin National Cottonseed Products Association, Inc., Memphis, TN National Cottonseed Products Association (NCPA) (1999) Cottonseed and its Products CSIP, 10th Edition, 1999 National Cottonseed Products Association, Ihc, Memphis, TN Oosterhuis and Jernstedt, 1999 Morphology and anatomy of the cotton plant Origin, history technology and production M-291007-01-1 OECD, 1999 Series on harmonization of regulatory oversight in Biotechnology no.10 Consensus document on the general information concerning the gene and their enzymes that confer tolerance to glyphosate herbicide, Organisation for Economic CoOperation and Development, Paris, France, 26 pages, M-406417-01-1 OECD, 2008 Series on harmonization of regulatory oversight in Biotechnology no 45 Consensus document on the biology of cotton (Gossypium Spp), OECD Environmental Directorate, Paris, France, 64 pages, M-327025-01-1 OECD, 2009 Consensus document on compositional considerations for new varieties of cotton (gossypium hirsutum and gossypium barbadense): key food and feed nutrients and anti-nutrients, OECD Environmental Directorate, Paris, France, M-263195-021 Oka A; Sugisaki H; Takanami M, 1980 Nucleotide sequence of the kanamycin resistance transposon Tn903, Kyoto University, Chemical Research, Japan, 10 pages, M148000-01-2 Oberdoerfer R 2009a Composition of raw agricultural commodity (ginned cottonseed) of the insect-tolerant cotton (event GHB119) and the non-transgenic counterpart (Coker 312) grown in Spain in 2007 Bayer CropScience Internal report 78 pages #M-311121-02-1 Oberdoerfer R 2009b Composition of raw agricultural commodity (ginned Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt cottonseed) of the insect-resistant cotton (event GHB119) and the non-transgenic counterpart (Coker 312) grown in Spain in 2008 Bayer CropScience Internal report 59 pages #M-348855-01-1 OberdoerferR 2009c Nutritional impact assessment report on insect-resistant cotton (event GHB119) Bayer CropScience Internal report 50 pages #M-358325-01-1 Oberdoerfer R 2010 General Information for the Nutritional Impact Assessment of GM Cotton Traits Bayer CropScience Internal report 39 pages #M-328532-03-1 Oberdoerfer 2011a Nutritional Impact Assessment Report for the Insect-resistant Cotton (Transformation Event GHB119) Bayer CropScience Internal report 43 pages #M355725-04-1 Odell J.T., Nagy F and Chua N.H 1985 Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter Nature Vol 313 28 February 1985 #M-147628-01-1 Pecoraro-Mercier, C 2014a Cry2Ae protein - Amino acid sequence homology search with known allergens and known toxins Bayer CropScience, M-309268-05-2 Pecoraro-Mercier, C 2014b PAT/pat protein - Amino acid sequence homology search with known allergens and known toxins, Bayer CropScience, M-266573-06-1 Rattemeyer-Matschurat V 2008 Analysis of substantial equivalence of transgenic and non-transgenic cotton by means of t-test for differences Bt cotton (Cry 2Ae) vs nontransgenic counterpart Bayer CropScience Internal report 55 pages #M-303119-01-1 Rattemeyer-Matschurat V 2009 Analysis of substantial equivalence of transgenic and non-transgenic cotton by means of t-test for differences Bt cotton (Cry 2Ae) vs nontransgenic counterpart Bayer CropScience Internal report 116 pages #M-354833-02-1 Rouquie D 2006 Gem2 protein acute toxicity by oral gavage in mice Bayer CropScience Internal report 62 pages #M-280141-01-1 Rouquie D 2008a Cry2Ae protein heat stability study Bayer CropScience Internal report 49 pages #M-308763-01-1 Rouquie D 2008b Cry2Ae protein in vitro digestibility study in human simulated gastric fluid Bayer CropScience Internal report 55 pages #M-308906-01-1 Rouquie D 2008c Cry2Ae protein in vitro digestibility study in simulated intestinal fluid Bayer CropScience Internal report 52 pages #M-308911-01-1 Sanfaỗon H., Brodman P and Hohn T 1991 A dissection of the cauliflower mosaic virus polyadenylation signal Genes & Development 5:141-149 #M-135204-01-2 Schnepf, H E., Whiteley, H R (1981) Cloning and expression of the Bacillus thuringiensis crystal protein gene in Escherichia coli Proceedings of the National Academy of Sciences USA 78, 2893-7 United States Environment Protection Agency (US-EPA) (2007) Biopesticide active ingredients and products containing them 39 pages 104 United States Food and Drug Administration (US-FDA) (2006) Guidance for industry Recommendations for the early food safety evaluations of new non-pesticidal proteins produced by new plant varieties intended for food use pages Van der Klis R.J., De Pestel K., Van Rie J and Hendrickx K 2008 Cry2Ae description and characterization Bayer CropScience Internal report 14 pages #M-391944- Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt 01-1 Van Larebeke N., Engler G., Holsters M., Van den Elsacker S., Zaenen I., Schilperoort R.A., Schell, J 1974 Large plasmid in Agrobacterium tumefaciens essential for crown gall inducing ability Nature 252 169-170 pages #M-147632-01-1 Verhaeghe, S and Habex, V (2008) Full DNA sequence of event insert and integration site of Gossypium hirsỹtum transformation event GHB119 Unpublished Bayer CropScience Report; July 11, 2008 M-311234-01-1 Verhaeghe, S and De Pestel, K (2008a) Bioinformatics analysis of newly created ORFs from Gossypium hirsutum transformation event GHB119 Unpublished Bayer CropScience Report; October 20, 2008 M-311122-01-1 (Contains confidential commercial information) Verhaeghe, S and De Pestel, K (2008b) Bioinformatics analysis of the preinsertion locus of Gossypium hirsutum transformation event GHB119 Unpublished Bayer CropScience Report; October 20, 2008 M-311230-01-1 (Contains confidential commercial information) Verhaeghe, S and Criei, i (2008) Structural stability analysis of Gossypium hirsutum transformation event GHB119 in different generations, in different backgrounds and when grown in different environments Unpublished Bayer CropScience Report; October 20, 2008 M-311131-01-1 (Contains confidential commercial information) Villagrỏn C.A 2008a Production of Raw Agricultural Commodity (fuzzy seed) Samples, Agronomic Performance , soil microbial activity and invertebrate fauna and Biosafety Studies of Transgenic Cotton plants containing Glufosinate ammonium Tolerant (Cry2Ae) and Insect Resistance Cotton events, their Non-transgenic Counterparts and Commercial Cotton Varieties Catalonia, Spain 2007 Bayer CropScience Internal report 122 pages #M-391806-01-1 Villagrỏn C.A 2008b Production of Raw Agricultural Commodity (fuzzy seed) Samples, Agronomic Performance , soil microbial activity and invertebrate fauna and Biosafety Studies of Transgenic Cotton plants containing Glufosinate ammonium Tolerant (Cry2Ae) and Insect Resistance Cotton events, their Non-transgenic Counterparts and Commercial Cotton Varieties Andalusia, Spain 2007 Bayer CropScience Internal report 83 pages #M-391810-01-1 Villagrỏn C.A 2009a Production of Raw Agricultural Commodity (fuzzy seed) Samples, Agronomic Performance and Invertebrate Fauna and Biosafety Studies of Insect Resistant and Herbicide Tolerant GM Cotton GHB119 and its Non-transgenic Counterpart and Commercial Cotton Varieties Andalusia - Spain 2008 Bayer CropScience Internal report 225 pages #M-347131-01-1 Villagrỏn C.A 2009b Production of Raw Agricultural Commodity (fuzzy seed) Samples, Agronomic Performance , and Invertebrate Fauna and Biosafety Studies of Insect Resistant and Herbicide Tolerant GM Cotton GHB119, its Non-transgenic Counterpart and Commercial Cotton Varieties Catalonia, Spain 2008 Bayer CropScience Internal report 226 pages #M-349636-01-1 Wehrmann A., Van Vliet A., Opsomer C., Botterman J and Schulz A 1996 The similarities of bar and pat gene products make them equally applicable for plant engineers Túm tt bỏo cỏo ỏnh giỏ ri ro ca s kin bụng GHB119 ti sc khe ngi v ng vt Nature Biotechnology Vol 14 october 1996 #M-141685-01-1 Zambryski P 1988 Basic processes underlying Agrobacterium-mediated DNA transfer to plant cells Ann Rev Genet 22: 1-30 30 pages #M-234499-01-1 Zhu J., Oger P.M., Schrammeijer B., Hooykaas P.J.J., Farrand S.K., Winans S.C 2000 The bases of crown gall tumorigenesis Journal of Bacteriology 182 (14): 38853895 11 pages #M-290743-01-1