PET I T I ON FO R TO L E R A NC E E XEMPTION Ge n et ica ll y E n gine e re d C itrus tr is t ez a l'i ru s ex 1 > r e ss in g S pina ch Defe n s in P r o t e in s 2 , 7 a nd 8 (G E C TY-SoD) S outhe rn Garde n s Cit r us N ur s e r y, LL C Octo b e r 20 1 5



### Ge n e ti ca ll y E n g in ee r e d 

### Citr u s t ris t ez a v iru s Ex pr ess in g S pin ac h D e fe n s in Prote in s 2, 7 a nd 

### 8 (GE C T V-S oD ) in C i t ru s F ruits (C rop G r o up 

### 10

- 10 ) : P e titi on for Es tab li s h m e n t o f a n Exe m p tio n fr o m th e R e q ui r e m e n t of 

### a T o le ra nce o n a T e m po rary Ba s is

Prepare d b y Vi c ki e A. F o rs t e r , Pr inc ip a l F ors te r & 

### A ss o c ia t es C o n s u lt in g, LL C Fo r , S o u th e rn G a rd e n s C itru s N u r se ry , LL C

Da t e Oct o b e r 28, 2 0 1 5

S ubmi tte d b y So uthern G a rd e n s C it ru s N ur se ry , LL C 1 82 0 C o u nty Ro a d 833 C lew i s to n , FL 33440

V olum e 1 

### of 2 1 

### A dmin ist r a ti ve Ma t e ri a l s


### P age 1 o f 1 3P ET ITI O:'\ FOR TOLERA NCE EXE**IPTJ O~**  Genetica ll y E n g in ee r e d Citrus tris teu 1 1

• im s express in g S pinach De f e n s in Prot e in s 2, 7 and 8 (GE CTV -SoD ) So uth e rn Ga rd e n s C itru s ' ursery, LLC October 20 I 5 

### Introduction Spinach 

### (Spina c ia oleracea) dcfcnsin genes 2 , 7 a nd 8 have bee n insert e d into 

### Citru s tristeza virus, i so late T 36 of stra in T36, a decl in e iso late na ti ve to F l orida. Th e genet i ca ll y engineered C T Y i s in corp ora ted int o c itru s trees v ia inocu lat i on: A full

- genome eDNA of CTV iso late T36 in binary pl as mid s was used to agroi nfiltr a t c 

### Nicotiana benthamiana leaves. When N. 

### benthamiana pl a nt s became in fected, as d e termin e d by No rth e rn blot ana lys is and ELISA, expressed sap o f the tobacco plant s was use d to bark fl ap o r ste m s lash in oc ul ate c itru s tree s. C itrus tree s initi al l y inoculate d as de sc ri bed above, and s ub se quent l y det ermined to have been s ucce ss fu ll y inf ec ted with GE CTY expres s in g o ne o r two s pinach defens in protein s (GE CTY-SoD), have se rved as th e so ur ce material for inoculating , via bud woo d g raf tin g, c itru s trees th a t are fie l d t es te d fo r res i s tance or to l e ran ce - b y reducin g o r e li minating di sease sy mpt oms- to c itru s g ree nin g (CG), a l so known as Hu a ngl ong b ing ( HLB ). Spinach has a long history of safe u se in the hum a n diet. Residues of s pina c h 6C;.f..:
. ts:n prote in s 2 and 7, So02 and So07, respectively , are exempted from the requircm er:t '),.a tolerance (on a temporary ba s is) w hen expressed in modified ( rONA) cit ru s tre es (40 CFR Part 174.535). Defcns in 8 (So 0 8) from s p in ach i s s imilar to So02 and So07, and i s likew i se expec t ed to b e rapidl y d eg raded in th e hum an d i gest ive tract (report l D SGC

- 042 s t..b1.1itt..:d w ith thi s app l ication). Add it iona l st udie s inc luded in support of the t emporary**t0lcr l'lCC**  

### exempt i on ( report ID s SGC-040 a nd SGC-04 1) o n the bioinformatics of the se**r..i.uc h**  

### dcfcn s in s s ho w no relations hi p to any known toxin or a ll ergen. Human exposure to a va ri e t y of natural pl ant v iruse s and plant v iral pro t e in s is commort in the diet. Plant v iru ses arc not pathogenic to hum an s. Human ex p osure to a va riety of**natun~** 

### phnt vi ru ses a nd pl a nt vira l prote in s i s co mm on in th e d i et. Pla nt v iru ses are not pathogen i c to hum a n s. The hum a n gas tro intest inal trac t harbors a wide var i ety of p lant v iruses w ithin the inte s t ines. There arc no report s of ad ve rse effects from CTV exposu res by any rout e of admi ni s trat ion (Government of Aus tra lia , De pt. Agricult ur e and Fi s her i es, 20 15 ). This petition, to exempt from the re quirement of a tole rance, genetica ll y engineered 

### Citrus tristeza virus express in g sp in ac h dcfe ns in proteins 2, 7 and 8 (G E CTV -SoD) in citrus fru its (4 0 CF R 1 80.4 l (c) ( 1 3)), dem o ns trate s th e sa f ety of these prote in s ba sed o n their hi s tory of safe u sc in common ly co n s um ed food s and v ia lack of tox i city or a ll erge n ic ity of s pinach dcfcn s in protein s to hum ans and li ves to ck anim a l s. Thi s petition conta in s data or information r ega rding th e e l ements se t fo rth in Section 40 8(a) of the FFDCA; however EPA ha s not fu ll y eva lu ated the s ufficiency of the s ubmitted data at thi s time or w h e th e r the data s upp ort s gra ntin g of the exem pti o n.

Page 2 of 13P ETIT I ON FOR TOL E R ANCE E X E MPTION Ge n et ic a ll y E n g i n ee r e d Citrus tris t ez a virus ex p r ess in g S p in ach D e f e n s i n Prot e i n s 2 , 7 and 8 (CE CTV-So D ) So uth er n Gardens C itru s Nurse r y, LLC October 20 1 5 

### Contents 

### Tit l e Page .................................................................. .. ......... . ... ........................ ....... ........... .... . .. . Introd uct i on ................................................................ .......................... ................. .. .. .. ............... 2 Sect ion A: Na m e , C he mi ca l Id e ntity and Co mp os iti o n of th e Active In gre di e nt: Sp inac h Defe n s in Pro t e in s ...... . ............. .. .... . . . . . .............. ............... ........... ..... . .... .................... 4 Sect i on 8: A mo unt , Fre qu e ncy, an d Timin g o f Ex press i o n of th e Pl a nt- I nc o rp o rat edProtec tant .......... .................................... ....................... ..................... ..... . ................. 6 Sect io n C: Report s of In ves ti ga ti o n s Made w ith Respec t to th e Safety of Sp in ac h Defe n s in Prote in s i. Matnmalian Toxicology
........ ........... .. ..... ...... ................... . ...... . . ......................... .. 7 Sec ti o n D: Report o n the A mo unt of Res idu e Found in Food a n d/or Feed Co mm odi ti es and a D escr ipt io n o f th e A na l y tic a l Method
..................................... ... ................. . .. 9 Sec ti o n E: Pract i ca l Met h od for Detect in g and Mea s urin g F oo d Res idu es and Method s tc r Removing Residues Which May Excee d Proposed T o le ra n ces ........ ..... . ..... . . ..... I 0 Sec t ion F: Proposed Exe mpti on from th e Re quire m e nt o f a Temporary Tolerance for Ge ne ti ca ll y E ngi n eered Ci trus tris t eza vi rus 

### expressing S pin ac h Defensin Prote in s 2, 7 a nd 8 

### in C itru s Fruits (Crop Group 10

- 1 0)
... . .... ....................... . ... .. . . I I Sec ti o n G: Reaso n ab l e Gro und s in S upp o rt of th e Peti ti o n ..................................................... 1 2

Page 3 of 13PETITION FOR TOLERANCE EXEM PTIO N Ge n et ica ll y E n g ineer ed Citru s tristeza virus ex pr ess in g S pin ac h D e rens i n Prot e in s 2 , 7 and 8 (GE C TV-SoD) So uth ern Gar d en s C itru s N ur se r y, LL C October 2015 Section A Na me , C hemical Id e ntity, and C omposition of the Active Ingred i e nt: Ge neticall y Engineered 

### Citru s trist ez a viru s Expressing Sp i nach Defensin Proteins 2 , 7 and 8
C hem ica l Na m e: N / A CAS N umber s: 103220

- 14-0 (P l a nt D efe n s in s (as a cla ss) Formu l a: A min o Ac i d Seq u e n ces of Sp in ac h D e f e n s in Prote in s 2, 7 and 8 are prov id e d wit h thi s s ubmi ss io n i n Report ID SGC -0 39 Mo lecular W eig h t: 4227 (SoD7) Da lton s to 7 099 (So D S) Dalton s Com m o n Name : Genetically E ngin ee red 

### Citrus triste za virus Ex p• · essing S pin ac h Def e n s in Prote in s 2, 7 and 8 Co de N umb er: N/ A Trade Na me: Genetically E ngin ee r e d 

### Citrus tristeza virus Ex pre ss ing S pinach Defensin Proteins 2, 7 and 8 S pina c h 

### (Spi na c i a oleracea) d efe n s in ge n es 2 , 7 a nd 8 h a ve be e n in serted into 

### Citrus tris teza viru s, i so late T36 of s tr a in T 36, a dec l in e i so la te n a ti ve to Florida. Th e ge n e tic a ll y e ng in ee r e d CTV is in co rp o rated in to c itru s tree s v ia i n oc ul a tion: A fu ll- ge nom e eDNA ofCTV i so l a te T

36 in b in ary pl as mid s was u se d to agro i nfiltra te 

### Nicot iana b en tham iana leaves. When N. 

### b e nthamiana plant s b ec a m e in fected , as d e termin e d b y No rth e rn b lo t analys is a nd E LI SA, ex pressed s ap of th e tob acco pl a nt s was u se d to bark flap o r s te m s l as h in ocu late c itru s trees . C itru s trees initially in ocu l ated as d escribed above, a nd s ub se quentl y determ in e d to h ave be e n s u cce ss fully inf ec te d w ith GE C TV expressi n g o n e o r two s pina c h d efe n s in prot e in s (GE CTV -So D) , serve as th e so urc e ma te ri a l f or ino c ul a tin g, v i a budwood g ra ftin g, citr u s trees that a re fie ld teste d for res i s tance o r to le rance - by reducin g o r e l imin at in g di s ease sy mptom s -to c itru s g re e nin g (CG), a l s o known a s Hu a n g lo ng bin g (HLB)
. A l i s t of th e d a ta a nd i nfonnation o n th e product c h a r ac teri za tion s ubmitt e d w ith thi s p e tition to s upport th e re qu es te d tol e ra n ce exe mpti o n are pro v ided b y re fer e n ce on the fo ll ow in g p age.

Page 4 of 13P ETITI ON F O R TOL E RA NCE. E X E MPT I O N Ge n et ic a ll y E n g in e er e d C itrus t ri s tez a v i ru s e xpr ess in g S pin ac h De f e n s in Prote in s 2 , 7 a nd 8 ( GE C T V

- S oD ) So uth e rn Ga rd e n s C i tru s N ur se r y, LL C O c t o b e r 20 1 5 

### PROD UCT AN ALYSIS METHOD OF S U PPORT G uid e lin e / OCS PP

885. 1 10 0 Produ c t Id e n tity
885. 12 0 0 Ma nuf ac turin g Process
885. 1 3 0 0 Fo rm a ti o n o f U nint e n t i o n a l In g re di e nt s 885
. 1400 A n a lys is of Sa m p les
885. 1 5 0 0 Ce rtifi ca ti o n of L imit s S tud y Ti t le

" P ro du c t C h arac te r iza ti o n of Ge ne tica ll y E n g in ee red 

### C itrus tristeza vi r us Ex press in g S pin ac h D efe n s in P r o te in s 2, 7 a nd 8 (GE CTV

- So D ), SGC- 0 39". Vol u me 2

" Produ c t C h aracte ri zat io n of Ge ne ti ca ll y E n g in ee red 

### C itrus tris teza vi rus Ex press in g S pin ac h D efe n s in P rote in s 2, 7 a nd 8 (GE C TV -So D ) , SGC- 0 39". Vo l um e 2

" P ro du c t C ha racte ri za ti o n of Ge ne ti ca ll y E n g in ee r e d 

### C itrus tris teza virus Ex press in g S pin ac h D efe n s in Pro t e in s 2 , 7 a nd 8 (GE C T V-So D ), SGC

- 039". Volume 2

" Pro du c t C h a ra c t e ri za ti o n of Ge n et i ca ll y En g in ee re d 

### Cit rus tris t eza virus Ex press ing S pin ac h D efens in Pro t e in s 2, 7 a nd 8 (GE C T Y-So D ), SGC

- 039". Vo l ume 2 C onfid e nti a l S tatement s of Form ul a , E P A Form 8570-4 V o l um e 1



### Page 5 of 13P ET I T I ON FOR TOLERANCE EXEM PTIO N Ge n eti ca ll y E ngin ee red Ci tru s tris tez CI virus express in g Spi n ac h Defen s in Pr o t ein s 2, 7 a nd 8 (GE C T V-So D) So uth er n Gar d e n s C itru s Nurse r y, LL C Oc tob e r 20 I S 

### Sec tion B Amount, Frequency, a nd Timing of Ex press ion of the Ge n etic a ll y Engi n eered Micro bi a l Pesticide Express ing S pinach Defensin Prote in s 2 , 7 and 8 Draft Prod uc t la b e lin g is in c lud e d in Volum e I of thi s s ubmi ss ion.

Page 6 of 13P ETITIO N FOR TOLERAN C E EX E MPTION Ge n e ticall y E ngin ee r e d Citru s tri s t e: a vims e x pr ess in g S pin ac h Def e n s in Prot e in s 2 , 7 and 8 ( GE C TV

- SoD) So uth e rn G ardens C itru s ' urse r y, LL C Oc t o b e r 20 I 5 Section C R e port s of the Investigations Made with R e spect to the Safety of Spinach Defensin Proteins 2 , 7 and 8 

### i. Mammalian Toxicolo S pin ac h h as a lo n g hi s tory o f sa f e u sc in th e hum a n di e t. R e s idu e s o f s pinach d e f e n s in pro t e in s 2 a nd 7 , So D 2 a nd So D 7 , re s p ec ti ve ly , a rc exe mpt e d f ro m th e re quire m e nt of a to l e ra n ce ( on a te mp o ra ry b as i s ) w h e n ex presse d in modifi e d (rO N A ) c itru s tr ees ( 40 C FR Pa rt 1 74.53 5 ). D efe n s in 8 (So D 8) fr o m s pin ac h is s im il ar to So D 2 a nd So D 7, a nd i s li k e wi se ex p e ct e d to b e ra pidl y d eg ra d e d in th e hum a n di ge s ti ve trac t (re port 

### JD S G C -04 2 s ubmitte d w ith t hi s a pp li cat i o n )
. A dditi o nal s tudi es in c lud e d in s upp o rt o f th e te mp o rary to le ra n ce exe mpti o n (re p o rt ID s S GC -040 and SGC- 041 ) o n th e bi o informati cs o f th e s e s pin ac h de f c n s in s s h ow n o re la ti o n s hip to a n y kn ow n tox in o r a ll e rge n . Hum a n ex p o s ure to a var i ety o f n a tur a l pl a nt v iru s es a nd p l a nt v ira l pro te in s i s co mm o n in th e di e t . Pl a n t v iru ses a r c n o t p a th oge ni c t o human s . Hum a n e x p o s ure t o a va ri ety of n a tur a l pl a nt vi ru s e s a nd pl a nt v ir a l prote in s i s co mm o n in th e di e t. Pl a nt v iru s es a re no t p a th oge ni c to hum a n s . Th e hum a n gas tr o inte s t in a l trac t h a rb o r s a w id e va ri ety of pl a nt v iru ses w it hin th e i nt es tin es. Th e re a re n o re p o rt s o f a d ve r s e e ff ects f ro m C TV ex p o s ure s b y a n y ro ute of ad mini s t ra ti o n (Gove rnm e nt o f A u s tra l ia , D e pt. Ag ri c ulture a nd Fi s h e ri e s, 20 15)
. A li s t o f th e d a t a a n d inf orm a t io n s ubmitt e d w ith thi s p e t i ti o n t o s upp o rt thi s t o le ra n ce exe mpti o n i s provi d e d b y refe re n c e b e low. 

### TO X ICOLOGY M E THOD OF S U PPORT G uid e lin e / OCS PP

885
. 3 0 50 Ac u te ora l tox i c ity
- wa i v er re qu e s t

885.3 1 0 0 Ac ut e d e m1al tox i c ity
- wa ive r re qu es t
885.3 1 50 Ac ut e pulm o n ary tox ic ity
- wa i ve r req u e s t St ud y Ti t le

" Ge n e ti ca ll y E n g in ee re d 

### C it r us tris tez a virus Ex pres s in g S pin ac h D cfc n s in Prot ei n s 2 , 7 a nd 8 (GE C TV -So D ) : R a ti o n a le fo r No t Co ndu c tin g a G uid e lin e S tud y - Ac u te O ra l T ox i c ity / P a th ogen ic i ty " , SGC

- 043
. Volum e 3

" Ge n e ti c all y E n g in ee r e d 

### C itr us tr i s t ez a v i rus Ex pre s s in g S pin ac h D e f e n s i n Pro te in s 2 , 7 a nd 8 (GE C T V -So D ): R a ti o n a l e for No t Co ndu c tin g a G uideline Stud y- Ac ut e D e rm a l T ox i c ity / P a th o l ogy" , SGC- 0 4 4
. Volume 4

" Ge n e ti ca ll y E ng in e e r e d 

### Ci trus tr i s tez a virus Ex pre ss in g S p i nac h D c f c n s in Pro te in s 2 , 7 and 8 ( GE C T V-So D ): R a ti o na le f o r No t Co ndu c tin g a G uid e lin e S tud y - Ac ut e Pulm o n ary T o x i c ity/ Pa th oge ni c i ty ", SGC -0 4 5
. Volume 5

Page 7 of 13P ETITI ON FOR TOL E RAN CE EXEMPT I ON Ge n etic a ll y E n g in ee r e d Citru s t ris t eza 1 • irus e xpre ss ing Spinach De f e n s in Prot ein s 2 , 7 and 8 (G E C TV- S oD ) So uth er n G arden s Citru s N ursery, LL C Octob e r 20 I S 

### TOXICOLOGY METHOD OF SUPPORT, CON TI N UED G uid e lin e / OCS PP 88 5. 32 00 Ac ut e i nj ec ti o n tox i c ity

- w ai ve r re qu es t
885.340 0 H y p e r se n s iti v ity in c id e nt s - wa i ve r re qu es t
885.35 00 Ce ll C ulturewa i ve r re qu est

Non- S t a bili ty o f So D 8 in G uid e lin e di ges ti ve fluid s - wa i ve r re qu est
No n - Bi o in fo rm at ics G uid e lin e Sea rc h o f A ll e rge n s
No n- B io inf o rm a ti cs G uid e lin e Sea rc h o f Tox in s
No n- Eva lu at i o n of G uid e lin e S t a bi li ty of So D 8 Stud y Ti t le

" Gen e ti ca ll y E n g in ee re d 

### C itrus tris teza v iru s Ex press in g S pin ac h D e f e n s in Pro te in s 2 , 7 a nd 8 ( GE CT V

- S o D ) : Rati o n a le f or No t Co ndu c tin g a G uid e l in e S tud y - Ac ut e Inj ec ti o n T ox ici ty /P a tho ge ni c i ty", SGC -04 6. Volume 6 "Ge n e ti ca ll y E n g in ee re d 

### C itr us tr is teza v iru s Ex press in g S pin ac h D efc n s in P ro te in s 2, 7 and 8 (GE C TV

- S o D ) : Ra ti o n a le fo r No t Co ndu c tin g a G uid e lin e S tud y - H y p e rs en s iti v ity In c id e nt s", SGC- 04 7. Volum e 7

" Ge n et i ca ll y E n g in ee re d 

### C itrus tris teza v i rus Ex press in g Spin ac h D e f e n s in Pro te in s 2 , 7 a nd 8 ( G E C TV -So D ): Ra ti o n a le f o r No t Co ndu ct in g a G uid e lin e Stud y - Ce ll C ulture", SGC -04 8
. Volume 8 "Ge n e tic a ll y E n g in ee re d 

### Citrus tr is teza v i ru s Ex press in g S pin ac h D efe n s in Pro te in 8 ( GE C TV

- SoD 8): R a ti o na le fo r No t Co ndu c tin g a Di ges t ib ili ty S tu dy in S imu la te d Gas tr i c F luid ", SGC -0 49
. V o lum e 9

" Bi oi nf o rm a tic A n a lys i s o f Po te nti a l A ll c rge ni c ity o f Dc f e n s in Pro t e in s 2, 7 a nd 8 fr o m Spin ac h f o r u sc in C itru s F rui ts". No n- G uid e li n e. SGC -0 4 0
. Volume 10

" Bi o inf o rm a ti c A na lys i s o f P o t e nti a l T ox i c ity o f D e f e n s in P r

o te in s 2, 7 a nd 8 from S pin a ch f o r u sc in C it r us Fruit s". SGC-04 1. Volume 11 "Eva lu a ti o n o f th e S imil a riti es of Spin ac h D e fe n s i n So D S Co mp a re d to Th ose of So D 2 a nd So D 7 Prot e in s to Es tim a t e th e Di ges tibi li ty of S o D 8 f o r th e Sa f ety Assess m e nt o f Ge n e ti ca ll y E n g in ee r e d C ro p s", SGC- 04 2. Volume 12

P age 8 o f 13PET I T I ON FOR TOLERA NCE EXE I PT I O:\' Ge n eti ca ll y E n g in eere d Ci tru s t riste:a vi ru s ex pre ss ing S pina c h Defen s in Prot ein s 2 , 7 a nd 8 (GE CTV-So D) So uth e rn Ga rd e n s C itru s N urse r y , LLC Oc t ober 20 I S 

### Sec tion 0 Report o n the A mount of Re s idue Fo und in Food a nd / or Fee d Co mmoditi es and a 

### De s cription of th e A nal y tical Met hod 

### Pre limin a r y researc h c itru s sa mpl e s (f ruit and leaves) fr o m trees in fec t e d w ith GE CTY-So D 2 o r GE CTY

- So D7 we r e s hipp e d from So ut h e rn Ga rd e n s C itru s N urse ry in C l ew is ton , FL , to LifeSen s ors, In c., Ma lve rn , PA , to measu re a ny d et ec tabl e re s i due s of SoD2 or So D 7 v ia we s t e rn blot. Wes t ern b l ot method is the c urre nt Analyt ica l E nf

orceme nt Met hod o n fil e w ith the E PA fo r s pinach defen s in protein (MRID 49409303). Addi ti ona l information on the s e analyse s i s prov ided in report I D SGC-039. T he a n a l y ti ca l e nf o rce me nt meth od (we s te rn b lo t) h as be e n rev iewe d prev i ous l y and det ermin ed to b e "Acce pt ab le" by th e Agency, MR ID 49409303. RESID UE METHOD OF SUPPO RT 

### Gu id eline/ OCSPP St ud y T itle 885.2300 ; Express i on in "Ex pres s io n of Sp in ac h De fen s in 2 a nd 7 Pro t e in s in Leaf 885 .2 500 C itr u s L eaf T i ss ue Ti ss u e of Ci tru s Events SGC- 11 208-5, SGC-4 11 08-7 , SGC41332-6 a n d SGC

- 11 319 -8", SGC-008. MR ID 

### 49409303

Pa ge 9 of 1 3PETIT I O:\' FOR TOLERAi\CE EXEMPT I ON Ge n etica ll y E n g ine e red Citm s**riste~a** vim s ex pre ss in g Sp in ac h D e fen s in Pr ote in s 2 , 7 a n d 8 (GE CTV- oD)

o uth er n Gar d e n s C itru s l\' ur se r y, LL C Oc t ober 20 I 5 

### Section E A Practical Met hod for Detecting and Measuring Food Residues and Met hods for Removing Re s idues Which May Excee d Proposed Tolerance s


### I t i s no t prac ti ca l , a nd th ere i s n o nee d f o r re mova l of resid ues of ge ne ti ca ll y e n g ine e r e d Ci tru s tri s teza virus 

### o r res idu es of s pin ac h defensin prote in s 2, 7 a nd 8, from c itru s ti ss ues a nd commodi ti es, a s an exe mp t i o n from th e requirement of a t olera nce i req u ested f or the se proteins w he n ex pressed in c it ru s. A to l e ra nce is not nee ded to protect the public hea lth
.

P

age 10 o f 13**PETITIO~'\** FOR TOLERA I'\CE EXEM PTION Ge n e t ica ll y E n g in ee r ed Ci tru s**triste ~a**  1 > iru s express in g S pin ac h Defen s in Proteins 2, 7 and 8 (GE CTV- oD) o u t h e rn Garde n s Citrus Nursery, LLC October 20 1 5 Sect ion F Proposed Exemption from the Requir e m e nt of a Time-limited Tolerance for S pinach Def e n s in Proteins in C itru s Fruits (Crop Group 1 0-10) 

### Southern Garde n s Cit ru s Nursery, LL C, is requ es tin g that the Age ncy g ra nt , o n a t e mp orary bas i s, a n exe mpti o n from th e requirem e nt of a to l era nce for res idu es of ge neticall y e n g in ee red Citrus tristeza virus 

### expressing sp in ac h d cfc n s in protein s 2, 7 and 8 de ri ve d f rom s pinach (Spina c ia ole r acea)

- 

### GE CTY-SoD

- i n C itru s Frui ts, Cro p Group I 0- 1 0 (40 

### CFR§ 1 80.4 1 

### (c)( 1 3)).



### Pa ge It 

### of l3PET I T I ON FOR TOLERANCE EXEMPTION Genetica ll y E n gi n eer e d Citrus tri s te:a l'irus expressing Spi n ac h De f e n s in Prote i n s 2, 7 a n d 8 (GE C TV-SoD) Southern Garden s Citr u s N u r se r y, LL C October 20 I S 

### Sectio n G Reasonable Grou n ds i n S u p p ort of the Petition S pin ac h d c f e n s in protein s a rc deri ve d from a food pl a nt ( s pina c h) th a t h as a l o ng hi s tory of sa f e u se i n t h e hum a n d i et. EPA ha s co nc luded (Memorandum , March 1 9 , 2 015 , from 

### J. Facey to K
. Ha y m es) that ne ith e r So D2 nor SoD7 ha ve am i no acid se quence s imi lariti es to known a ll e rg e ni c or to x ic prot e i n s, and t h at SoD 2 a n d SoD7 are d eg rad e d i n s i mu l ated gas tri c flu id
. These prote in s h ave no known de l eterious h uman h ea lth effects or any evidence of to x ic ity to p eo p l e, w ildlif e, or b ene fici a l in se ct s. 

### C itrus tristeza virus i s endemic throu g hout the F l orida c itru s grow in g region s. Th e eng in eere d i so l a te, T 36, i s a ubiquitou s d ec l ine i so l ate nati ve to Florida. Hum a n ex po s ure to a va ri ety of n atur a l p la nt v iru ses a nd p l a nt v ir a l prote in s is co mmon in t h e di e t. Plant v iru ses a re n o t pathogenic to hum a n s. I nd ee d , t h e human gastrointe s tina l tra c t ha rb ors a w i de variety of plant v iru ses w ithin the intes tine s . Th e re arc no re port s of adver se ef f ec ts from C TV ex p os u res b y a n y ro ute o f admini s tration
. The ge n et ic material n ecessary f o r th e pro duct i on of the GE C TV -SoD prote i n s a re nucl e ic ac id s (DNA) which are co mmon to a ll form s of plant a nd a n im a l l if e. Th e r e are no kno w n in s t a n ces in wh i ch nu c l e i c acids ha ve caused to x ic effects as a re s ult of dietary expo s ure. S i g nific a nt d ie tary e x p os ur e to GE C TV expre ss i n g s pin ac h d e fen s in pro tein s 2, 7 a nd 8 i s un l ikc l y to occ ur du e to th e a nticip a ted l ow ex press ion of th e d e fcn s in prote in s from th e vec tor. Di e tary ex pos ur e to GE CTV i s not ex pect e d to b e g rea te r than ex p os ur e to w il d type C T V in c itru s produ cts. Di e ta ry ex po s u re to s pin ac h d e fen s in s, to the d eg ree th ey may occ ur v ia ingestion of fre s h and pro cesse d citru s co mmoditie s, wi ll be at leve ls far b e lo w ex i s tin g dietary ex po s ure to th ese pro te in s from regu la r con s umpt ion of s pina c h
. (Recent U.S. co n s umption d a ta i ndic ates a n ave ra ge of 2 lb s. of s pina c h are co n s um ed p e r p e r so n per yea r in the United St a tes ("Spinach Profile ", Agr icu ltura l Market i n g Reso urc e Ce nt e r ( Jun e 20 1 3)). Moreover, s uch ex po s ure s are u n like l y to be prob l e m a ti c b eca u se o f th e l ow tox ic ity a nd the high degree of di gcst ibi l ity of th e protein s. No n -occ upationa l expo s ure to GE CTV -SoD v ia a rout e other than co n s umption o f fres h c itru s and / o r pro cessed citrus commodit ies , s uch as drinking wate r , w i ll be minim a l to nonex is tent s in ce the se protein s ar e expressed on l y with i n p lant t i ss u es. Ba s ed on d ata a nd information s ubmitte d w i t h th i s p e tition , we ha ve co nc lud e d th a t th e re i s a reaso n ab l e ce rta in ty that no h a rm w ill res u lt to th e United State s popu lation, inc ludin g infant s a nd c hi l dre n , from agg regate ex po s ur e to residues of th ese prot e in s . Exe mptin g ge netica ll y e n g in ee red 

### C itru s tristeza vi rus ex press in g s pinach de fen s in pro te in s 2 , 7 and 8 (GE CTV

SoD) fr o m th e re quir eme nt of a to l e rance s hould po se no s i g nific a nt r i s k to hum a n s o r the environment.

P age 12 

### of l3P ETIT I ON FOR TOL E RAN CE EXEM PTI ON Ge n e ti ca ll y E n g ine e r ed C itrus tri s t ez a virus e xpre ss in g Sp in ac h Defen s in Prot e in s 2 , 7 and 8 (GE CTV-So D) Sout h ern G a rd en s C it rus N ur se r y, LL C October 20 I S Res idu es of th e p lant i n co rpor a ted -protec ta nt s s pin ac h d e f e n s i n s 2 a nd 7, So D 2 and So D 7 , res p ect i ve ly, a re exempted from th e re qu irement of a to le ran ce a t 40 CF R 

### § 1 74.535 : 

### § 174 . 535 S pinach Defen s in proteins ; te mporary exe mp tio n from the requirement of a tolera n ce. (a) Re s idu e s of th e d e fen s in prote in SoD2 d e ri ve d from s pin ac h 

### (Spina c ia o l eracea L.) in o r on citru s food co mm o dit i es a rc temporar il y exempt f rom th e re qu ire m e nt of a to le rance w h e n u se d as a p l a nt-in co rp o ra ted protec tant in c itru s p l ant s in accorda n ce w it h th e te m1 s of Ex p e rim e nt a l Use Permit No. 88232

- EU P

- 1
. Thi s te mp o rary exe mpti o n from the re quire m e n t of a to le ra nce ex pire s o n Apri l 1 8, 20 1 8
. (b) Re s idu es o f th e d efe n s in pro te in SoD7 d e ri ve d from s pin ac h 

### (Spina c ia oleracea L.) in o r on c itru s food co mm od iti es are te mporari ly exe mpt fr o m th e re quire m e nt of a to le ra nce w h e n u se d as a p l a nt-in corpo rat e d protec tant in c itru s pl a nt s in acco rd a nc e wit h the term s of Ex p er im e nt a l Usc Pe rmit No. 88232

- EU P

- 1
. Th i s te mp orary exe mpti o n from th e r e quire m e nt of a t o le ra n ce exp ires o n Ap ri I 1 8, 20 1 8.

Pa ge 13 

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