## Pla n t Pest Ri s k A s s es s m e n t for D A S

- 40 2 78

- 9 C o rn

Dow A g roSci e n ce s L L C ( re f e r r e d to h e r ea ft e r a s D A S ) p e t i t i on e d AP H I S ( AP H I S num b e r 09

- 233

- 01p ) for a d e te r m i n a t i on that g e n e t ic a l l y e n g i n ee r e d ( G E ) c o r n ( Z e a ma y s ) e v e nt DA S

4 027 8

- 9 is un l ike l y to po s e a plant p e st risk a nd, t h e r e f o r e , should no lon g e r b e a re g ulat e d

a rt i c le und e r re g ulations a t 7 C ode of R e g ulations ( C F R ) p a rt 340. Th e earlier 2010 version of the petition was revised on April 12, 2011 to update the summary on page 4 and was received by
APHIS on J une 22, 2011. This version of the petition is referenced hereaft er as DAS 2011.
AP H I S a dm i nis t e rs 7 C F R p a rt 3 40 und e r the a uthori t y of the pl a nt pest provisions of the P lant

P rot e c t i on A c t of 200 0 1 . This p l a nt pest risk a ssessment w a s co n du c ted to d e te r m i ne wh e t h e r

e v e nt DAS

- 40278

- 9 c o r n is unl i k e l y to po s e a pla n t pest risk.



### H i s t o r y o f D e v e l o pm ent of DA S

- 40 27 8

- 9 Her b i c i d e
- T o l e r a n t 2 

### C o rn

DA S

- 40278

- 9 c o r n is a GE c o r n l i ne that h a s b e e n pr o vided inc r e a s e d to le r a n c e to t r ea t m e nt wi t h ph e no x y a u x in h e rbi c id e s a nd r e si s tan c e to a r y lo x y p h e no x y p ropion a te ( A OP P ) a ce t y l c o e n z y m e A ca rbo x y la s e ( AC C a s e ) inh i bi t or ( “ fop ” ) h e rbi c i d e s . The in t rodu c e d g e n e t i c mat e ri a l ( D N A)

r e sul t s in t he pro d u c t i on of a n a r y lo x y a l k a no a te d i o x y g e n a se ( A A D

- 1 ) e n z y m e that in a c t i v a tes
h e rbi c ides of the a r y lo x y a lkano a te f a m i l y , includ i ng ph e no x y a u x ins and A OPP AC C a se inh i bi t o r s

. I f a d e te r m i n a t i on of non r e g ulat e d st a t us is r eac h e d

, D A S

- 402 7 8

- 9 c o r n would be t h e fi r st comm e r c ial l y a v a i la ble c o r n v a ri e t y with both i m p r ov e d to l e r a n c e to ph e no x y a u x in

h e rbi c ides a nd r e si s tan c e to “ fop” h e rbi c ides.

The most w e l l

- known a n d wid e l y

- used ph e no x y a u x in he r bicide is 2, 4

- dichlo r oph e no x y a c e t i c

ac id (2,4

- D ) w hich h a s b ee n us e d f o r m a n y d e ca d e s a s a p r e

- plant or pos t

- e me r g e nt h e rbi c ide to

c ontrol bro a dl e a f (di c ot) w ee ds in c o r n fi e ld s . 2,4

- D is also l a b e led f or u s e f or ma n y other w ee d

c ontrol ap p l i ca t i ons including with other c rops, on p a stur e s and r a n g e land, a nd b y both

c om m e r c ial a nd r e sident i a l e nt i t i e s for w e e d c ont r ol i n turf a nd la w ns. The mode of ac t i on a nd
the r e lative bioc h e m i ca l s e le c t i vi t y of 2, 4

- D for b ro a dle a f pl a nts is un c le a r , but i t i s believ e d to
fun c t i on a s a pl a nt g ro w t h re g ulator ( a u x in hormon e ) mimic whi c h ca u s e s a bnorm a l c e ll div i sion

a nd g r owth le a di n g to p la nt i nju r y a nd d e a th ( D A S 201 1 , p a g e s 154

- 155; E P A 2005 ) . Thou g h it
is not a bro a dle a f p l a nt, c o r n ca n su f f

e r inju r y f r o m 2, 4

- D d e p e ndi n g on t h e gro w th s ta g e a nd
method of a ppl i ca t i on ( D AS 201 1 , p a g e s 103

- 105 )

.

1 Secti o n 40 3 (1 4 ) o f t h e P la n t P ro tecti o n A ct (7 USC Sec 7702(14 ) ) d e f i n es p la n t p est a s :

“ P la n t P est

- T h e te r m “ p l a n t p est” m e a n s a n y l i v i n g s t a g e o f a n y o f t h e f o ll o w i n g t h a t c an d ire c tly o r i n d ire c t l y i n

j u r e, c a us e d a m a g e t o , o r c a us e d i s e a s e in a n y p l a n t o r p la n t prod u ct: ( A ) A pro t o z o a n

. (B ) A n on h u m a n a n i m al. ( C ) A p a r asi t ic p l a n t. ( D) A b a cte r i u m

. ( E) A fu n g us

. ( F) A v ir u s o r v ir o i d

. ( G) A n i n f e c ti o u s a g e n t o r o t h er

p at h o g e n

. ( H) A n y a r ticle si mi lar to o r allied w i t h a n y o f t h e a r ticle s s p e c ified i n t h e pr e c e d i n g su bp a r a g r a ph s .”

2 T h e a pp lica n t h as d esc r i b ed DA S

- 40278

- 9 c or n as “ h e rb ic i d e t o le r a n t” a n d h i s t or ical l y A P H I S h as al s o r e f e rr ed to
GE p la n ts w i t h d i m i n is h ed h e rb ici d e s e ns it i v i t y as “ h e rb ici d e t o le r a n t ”

. H o w e v e r , t h e p h e n o t y p e w o u ld f all u n d er

t h e W e e d Scie n ce S o cie t y o f A m e r ic a ’ s d e f i n i t i o n o f “ h e rb ici d e r esi s ta n c e ” s i n ce D A S

- 40278

- 9 h as an i nh e r ited

a b ili t y to s u r v i v e a n d r e prod u ce f o ll o w i n g e x po s u r e to a do s e o f h e rb ici d e n o r m al l y le t h al t o t h e w i ld t y p e v a r ie t y (W S S A 1998)

. B y t h e W S S A d e f i n i t i o n , “ r esi s t a n ce [ to an h e rb ici d e] m a y b e n a t u r al l y o c c u rr i n g o r i n d u c e d b y

su c h tec h n i q u es as g e n e tic e ng i n e er i n g o r s ele c ti o n o f v a r i a n t s prod u c e d b y ti s su e c u l t u r e o r m u t a g e n es i s .”

He rb ici d e t o le r a n c e , b y t h e W S S A d e f i n iti o n , o n l y a pp lies t o p la n t s p e c ies w i t h an “ i nh e r e n t ab ili t y ” to su r v i v e a n d

r e prod u ce a f ter h e rb ici d e tre a t m e n t.

1A O P P AC C a se inh i bi t o r s , or “ fop” h e rbi c ides, a re pos t

- e me r g e nt g r a m i nicid e s , me a ni n g that
the h e rbi c ides s e l e c t i v e l y c o ntrol e me r g e d g r a ss (Po a ce a e ) w e e d s . The s e le c t i vi t y of the “ fops”
is b a s e d on a n in c r e a s e d s e nsi t iv i t y of g r a ss spe c i e s to AC C a s e s comp a r e d to other plant spe c ies ( D e vine 199 7 ; Konishi a nd S a s a ki 199 4 ). The c o nseq ue n c e o f t r e a t m e nt wi t h a n AC C a se inh i bi t or h e rbi c ide is to block p r odu c t i on of fa t t y a c id b i o s y nthesis i n se n si t ive pl a nts. C o r n, a plant in t he P o ace a e f am il y , a lo n g with o t h e r r e la t e d ce r ea l p l a nt v a ri e t i e s, is c onv e nt i on a l l y

s e nsi t ive to tr ea t m e nt wi t h “ fop” h e rbi c id e s, a nd t h e se h e rb i c ides h a ve not b ee n tr a di t ional l y lab e led for w ee d c ontrol in co r n fi e lds (W SS A 2007). W hi l e some ce re a l c rop plants h a ve d e v e loped some tole r a n c e to “ f op” t r ea t m e nt due t o i m p r ov e d h e rbi c ide d e t o x ific a t i on ( D A S

201 1 , p a g e s 155

- 156; D e vine 1997 )

, a nd c o rn v a r i e t i e s with re si s tan c e to “ f op” tr ea t m e nt ha v e b ee n iso l a ted usi n g mu t a t i on s c r e e ni n g ( M a rsh a ll e t al. 199 2 ), if a d e t e rmin a t i on of

non r e g ulat e d status i s r eac h e d, DA S

- 4 027 8

- 9 c o r n m a y b e the f irst co r n v a ri e t y with r e si s tan c e

to t h e se h e rbi c ides to b e c ome w i d e l y c om m e r c ial l y a v a i la ble. I n addition, DAS has indicated
that DAS

- 40278

- 9 may be stacked with glyphosate and other herbicide resistance traits to generate commercial hybrids with resistance to multiple herbicides (pg. 4, DAS 2011).

I f a d e t e rmin a t i on of no n re g ulat e d st a tus i s r e ac h e d, D A S

- 40278

- 9 would p r ovide c o r n g r ow e rs with a ddi t ional opt i ons for the pos t

- e me r g e nt c o n trol of both b r o a dle a f a nd g r a ss w e e d s . This

c o r n pr o du c t would a lso provide a nother tool to address t he inc r ea s e d incid e n c e of g l y phos a te

a nd ace to la c tate s y nthas e ( A L S ) inh i bi t or r e si s tant w e e d s . The p e t i t i on e r h a s s t udied v a rious

a g r onom i c a nd c omposi t i on a l c omponents of DA S

- 40278

- 9 c o r n sin c e 20 0 6 throu g h c onfin e d

fi e ld t e st s , a nd, in a cc o r d a n c e with 7 C F R p a rt 340.6( c ) , h a s p r e s e n t e d d a ta in t he submi t ted

p e t i t i on f o r a d e te r m i n a t i on that D A S

- 40278

- 9 c o r n should no lon g e r b e r e gulat e d und e r 7 C F R

p a rt 340.

Addit i on a l l y , t he p e t i t i on e r submitted D A S

- 40278

- 9 c o r n f or a Consultation of B ioe n g ine e r e d

F oods to the F DA on O c tob e r1, 2009 und e r the F D A ’s ‘ S tat e ment of P ol ic y : F o o ds D e riv e d

f rom Ne w P la nt V a ri e t i e s ’ ( FDA 1992). FDA evaluated D AS’ submission to determine
whether DAS

- 40278

- 9 corn raises any safety or regulatory issues with respect to the intended modification or with respect to the food itself. Based on the information provided by the company and other information available to the agency, FDA did not identify any issues under the Federal Food, Drug and Cosmetic Act that would require further evaluation at this time (BNF No. 000120, April 8, 2011: http://www.fda.gov/Food/FoodScienceResearch/Biotechnology/Submissions/ucm254647.htm

. )

The U.S. E nvironmental P rotection A gency (EPA) regulates the distribution, sale, use and testing of herbicides under the Federal Insecticide, Fungicide and Rodenticide Act (FIFRA) (7

U.S.C. 136 et seq) . EPA also sets tolerance limits for residues of herbicides on and in food and animal feed, or establishes an exemption from the requ irement for a tolerance, under the Federal Food, Drug and Cosmetic Act (FFDCA) (21 U.S.C. Chapter 9). Prior to registration for a new use for a new or previously registered h erbicide , EPA must determine through submitted test
data that the herbicide does n ot cause unreasonable adverse effects on the environment and non

target species when used in accordance with label instructions and will result in a reasonable certainty of no harm to humans . EPA must also approve the language used on the h erbicide label

i n accordance with 40 CFR part 158. DAS has s ubmitted information in support of label amendments to the EPA for the use of 2,4

- D and quizalofop herbicides for weed management in
fields with DAS

- 40278

- 9 corn (DAS 2011, pp 113

- 115) . These actions are curren tly under

2review by EPA.

I n 2009, the percent of c o r n p l a nted in the United S t a tes that poss e ssed either r e si s tan c e to an h e rbi c ide or stacked resistance to both herbicides and insects

, both of which were co n f e r r e d
thro u g h bio t ec hnol o gy , was approximately 22% and 46%, respectively, and 68% combined ( USDA N ASS 200 9 ). The prim a r y h e rbi c ide r e si s tan c e t r a it in use h a s be e n g l y phos a t e

r e si s tan c e, a nd the a d o pt i on of th i s tr a it in other major c rops, s u c h a s c ot t on a nd s o y b ea n, is

e v e n h i g h e r. W ee d r e si s t a n c e to he r bici d e s is a c on ce rn in a g ri c ul t u ra l p r odu c t i on a nd the

w id e

- s p r e a d a dopt i on of h e rbi c i d e r e si s tant c rops, e spe c ial l y G E

- d e riv e d g l y p hosat e

- r e si s tant

c rops, h a s dr a mati c a l l y c h a n g e d the a pproa c h that f a

rm e rs t a ke to avoid y ie l d los s e s f r om

w ee ds ( D u k e a nd P owl e s 200 9 ; Gi a n e ssi 2008 ) . G l y phos a t e

- r e si s tant c r o ps h a ve b ec o m e wid e l y a dopted since their in t ro d u c t i on in t he m i d

- late 1990s for sever a l r e a son s . G l y phos a t e

wo r ks no n

- s e le c t i v e l y on a wi d e ra n ge of pl a nt spe c i e s, is a r e l a t i v e l y low

- c ost h e r b ici de,

a l l ows ‘ no

- t i l l ’ f ar m i ng pra c t i ce s, a nd h a s m i ni m a l animal to x icolo g ic a l a nd e nviro n ment a l

i mpa c t. Ho we v e r, in c r e a s e d sel ec t i on p r e s s u r e re sul t i n g f rom the wid e

- spr e a d a dopt i on of

g l y phos a t e

- r e si s tant c rops, a long with t he r e d u c t i ons in the use of othe r h e rbi c i d e s a nd w ee d

man a g e ment p r ac t i ce s, h a s r e sul t e d in both w ee d population sh i fts a nd g r o wing num b e rs of

g l y p hosate r e si s tan t biotypes a mo n g some w ee d populations ( Duke a nd P owl e s 2009; Ow e n 2008). I n o r d e r to c ombat th i s tr e nd, a nd to a void de c r ea s e d c r op y ields th a t r e sult f r om w ee d

c ompeti t ion, g r o w e rs must c ont i nue to diversify and a d a pt their w e e d man a g e ment str a t e g ie s to adopt best management practices ( Norsworthy et al. 2012). If DA S

- 40278

- 9 c o r n b ec o mes a v a i l a ble to them, c o rn fa rm e rs who c hoo s e to g r o w it will h a ve a dd e d opt i ons for

w ee d c on t rol if requested herbicide label changes mentioned above are approved by the EPA

. The use of DA S

- 40278

- 9 c o r n c ould a l l ow f or i m p r oved c o r n c rop y i e lds wh e n it is g r o wn in
the vi c in i t y of w e e ds r e si s tant to g l y phos a t e , p a rticul a r l y with proposed lab e l i n g c h a n g e s to

a l l o w the e x p a nd e d a ppl i ca t i on of 2,4

- D a t a c ritic a l t i m e p e riod f or c ontroll i ng w ee ds t h a t

comp e te w i t h c o r n dur i ng the f i rst 3

- 5 w e e ks b e f o r e the c o rn r e ac h e s 6

- 8 inch e s in h e i g ht a nd proposed l a b e l ch a n g e s to a l l ow o v e r

- the

- top pos t

- e me r g e nt use of qui z a lofop ( D A S 201 1

, p a g e s 108 a nd 113). Th e p rac t ic e of using h e rb i c ides w i t h a l t e rn a t i ve modes of ac t i on c ould also pot e nt i a l l y di m in i sh the populations of those g l y phos a t e

- r e si s tant w ee ds a nd r e du c e t h e l i k e l i hood of the d e v e lop m e nt of n e w h e rbi c ide

- r e si s tant w e e d p opulation s if properly implemented within a diversified weed management cropping system ( D A S 201 1 , p a g e s 16

- 18 a nd A pp e ndix 6 ; Duke a nd P owl e s 2008, 2009; Di l l et a l. 2008; O we n 200 8 ; Norsworthy
et al. 2012 )

.

The h e rbi c i d e 2,4

- D w a s in t rodu c e d more than 60 y ea rs a g o a nd is used th r ou g hout t he w o rld

f

or the tr ea t m e nt of b r o a dle a f wee ds. I n 2002, 2,4

- D w a s r a n k e d a s the third most used h e rbi c ide
b y ac t i ve i n g r e dient in t he U

. S . for a ll pur p oses ( ~ 40 m i l l ion pound s ), b e hind g l y phos a te (

~ 102

m i l l ion pounds) a nd a tr a z i ne (~77 m i l l ion pound s ) ( G ian e ssi a nd R e i g n e r 2 0 0 6). Th a t same

r e port f ound that the u s e of 2, 4

- D r e main e d r e lative l y st e a d y fr om 1992 to 2002, wh e re a s

g l y phos a te u s a g e inc r ea s e d more than 5

- fold ov e r the s a me ti m e p e riod. T h is i n c r e a se in

g l y phos a te u s e is attribut a ble to the introdu c t i on a nd wid e

- spr e a d a dopt i on of GE g l y pho s a te

r e si s tant c rop sp e c ies ( e

. g ., s o y b e a ns, c o rn, c ot t on, ca nola) a nd rising a dop t ion of no

- or

re d u ce d

- t i ll fa rming pra c t i ce s that t y pic a l l y a cc ompani e s t h e use o f g l y phos a t e . T h e re port a lso

i ndic a t e d that ov e r a ll h e rbi c i d e use on c o r n d e c re a s e d du r i n g that s a me 10 y ea r time f r a me ( f r om ~213 m i l l ion pounds i n 1992 t o ~159 mil l ion pounds i n 2002 )

, a s f a rm e rs in c r e a sin g l y fa v o r e d
the use of GE c rops w h i c h a l l o w e d for f e w e r t y p e s and a small e r ov e r a ll a mount o f h e rbi c ide to

3b e use d

.

The EPA did a n a n a l y sis of h e rbi c ide u s a g e in c o r n fi e lds for 2004 that a l s o show e d that the use
of g l y phos a t e inc r e a s e d d r a matic a l l y f r om 1987 to 2001 wh e re a s 2, 4

- D us a ge r e main e d

e ssential l y u n c h a n g e d d u ring that t i me ( K i e l y 20 0 4 )

. I n 2005, h e rb i c ides w e re a ppl i e d to 97

p e r c e nt o f c o r n ac r e a ge p lant e d in 19 stat e s r e p r e s e n t ing 93 p e r c e nt o f a ll c o r n plant e d in t he
Uni t e d S tat e s. The h e rbi c ide most wid e l y used on c o r n w a s a tr a z ine (66 p er ce n t , ~57 mil l ion pounds a ppl i e d), g l y pho s a te wa s s e c ond ( 31 p e rc e n t , ~23 mil l ion pound s ), follow e d b y s

m e to l ac hlor a nd a ce toch l or (both at 23 p e r ce n t

, < 2 4 m i l l ion pounds and < 30 m i l l ion pounds,

r e spe c t i v e l y )

. B y c ompa r ison, 2, 4

- D wa s ap p l i e d o n less than 8 p e rc e nt of 2 0 05 c o r n a c re a g e ( ~2 m i l l ion pounds appli e d) ( USDA N ASS 2006 )

. According to the USDA National Agricultural Statistics Service Chemical Use Database, t he average percentage of all corn acreage treated with 2,4

- D r emained relatively stable from 1990 to 2003 t ypically ranging from
about 8 to 12 percent, with slightly higher peaks in 1990, 1994 and 1995

, and slightly lower usage in 2002 and 2003 for program states included in the survey ( data accessed via
ht t p: / /ww w .p e st m a n a g e ment.info/n a ss / inde x .ht m l on 12/19/2013 ). Th e se d a ta d e monstr a te t h a t

t h e re is a his t o r y of s u cce ssful a nd e f f

ec t i ve u se of 2,4

- D a s an h e rbi c id e , both g e n e r a l l y a nd

sp ec ifi c a l l y f or c o r n c rops, as one of many options to manag e w e e d spe c ies. B e c a use c o r n is not

a bro a dl e a f p l a nt, i t alr e a d y h a s some to le r a n c e to 2,4

- D ; a nd the a ddi t ion of a tr a it f or inc rea s e d
resistance to 2, 4

- D in DA S

- 40278

- 9 c o r n will g iv e g r o w e rs t h e opt i on to a pp l y 2,4

- D du r ing
dif f

e r e nt g r o wing windows, using dif f

e r e nt a p pl i ca t i on methods a nd h e r b icide m i x e s , if approved by EPA ( D A S 201 1 , p a g e s 11 3

- 115 ) . T h e e x p ec tation f r om t he p e t i t ioner is t h a t th i s

a dd e d tr a it will h e lp t o m i ni m i z e the e me r g e n c e of h e rbi c ide r e si s tant w ee d s b eca use b a s e d on

fi e ld e f fi c a c y stud i e s ( D AS 2 01 1 , p a g e s 103

- 105) the DA S

- 40278

- 9 c o rn p lants a re less

susc e pt i ble to in j u r y a ss o c iat e d with p r opos e d a p p l i ca t i on r a tes of 2, 4

- D a p pl i e d e i t h e r a t t he

p re

- e me r g e nt s ta ge or l a ter p ost em e r g e nt s ta g e s, a nd thus will a l l ow f or more robust w ee d

c on t rol opt i ons. As stated earlier the label change request submitted by DAS to EPA is under
review.

T he “ fo p ” h e rbi c i d e s ( A A O P AC C a se inh i bi t o r s) h a ve b e e n re g is t e r e d for c rop u se f o r ov e r 20

y ea rs. Th e y h a ve not tra di t ional l y b ee n us e d to c ontrol w ee d s p e c ies in c o rn f ields b e c a use, a s a

g r a ss (Poac ea e f a m i l y ) s p ec ies, c o rn is d a m a g e d b y A O P P AC C a se inh i bi t or ac t i vi t y

. T h e use

o f “f op” h e rbi c ides h a s b e e n p r i m a ri l y on b r o a dle a f c rops, su c h a s s o y b ea n, t o c ontrol gr a ss

w e e d spe c ies, a ltho u g h c e rt a in ce re a l p l a nt v a ri e t i e s h a v e a lev e l of to le r a n c e to s ome “ fops

. ”

A cc o r di n g to t he USDA N a t i on a l A g ri c ul t u ra l S t a t i st i c s S e rvi c e ( N A SS ) A g ri c ul t u r a l C h e m i ca l

Use D a ta b a s e , “ fop” t y pe h e rbi c ides w e re u s e d for w ee d c ontrol on a t l ea st 23 f o od c rop sp e c ies
b e tw ee n 1990 a nd 2006, to t a l i ng ov e r 16 mill i on pounds of ac t i ve i n g r e d i e n t ( ht t p: / /ww w .p e st m a n a g e ment.info/n a ss / inde x .ht m l , A cce ssed 3/201 0 ). I f a d e te r m i n a t i on of

nonre g ulat e d st a tus i s r ea c h e d, DA S

- 4027 8

- 9 c o r n would be the f irst GE c o rn v a ri e t y with

nonre g ulat e d st a tus t h a t a lso has r e si s tan c e to t he “ fop” c l a ss of h e rbi c ides. The p e t i t i on e r h a s ind i ca ted th a t “ f o p ” h e rb i c ides c ould be us e d to m a in t a in se e d pu r i t y in D A S

- 40278

- 9 c o r n b ree di n g nur s e ri e s , h y b r i d pr o du c t i on fi e ld s , a nd g e n e r a l l y f o r the c ontrol of gr a ss w e e ds in

c o r n

. Qui z a lofop is cu r r e nt l y und e r r e g is tr a t i on r e view ( ht t p: / /ww w

. e p a

. g ov/oppsrr d1/r e g is t r a t ion_r e v i e w / ) b y th e E P A (72 FR 71893

- 71896).

As r e quir e d un d e r F I F R A, met a bol i sm and re si d ue d a ta, a lo n g with prop o s e d lab e l i ng c h a n g e s will be reviewed by t he EPA, for the use of 2,4

- D and “ fop ”

- t y pe h e rbi c i d es (sp ec i fi c a l l y

4qui z a lofop) in DA S

- 40278

- 9 c o r n fi e lds ( D A S 20 1 1 , p a g e 18 )

. U nd e r F I F RA, it i s unl a w f ul t o

use a n h e rbi c ide “ in a m a nn e r i n c onsistent with i t s lab e l i n g ” witho u t an e x p e rime n tal use p e r m i t
i ssued ( 7 U.S. C

. 136j).

When t he petition was submitted, r e lative l y f e w w ee d sp e c ies h a d d e v e loped r e si s tan c e to

s y nthetic a u x ins or AC C a se inh i bi t or h e rbi c id e s c om p a r e d to two o t h e r c las s e s of h e rbi c ides.
The I n t e rn a t i o n a l S u r v e y of H e rbi c ide Resist a nt W e e ds ( I S HR W ) re g u l a r l y u p d a tes a w e bsi t e with l is t s of “ fi e ld sel e c ted, g e n e t i ca l l y inhe r i t e d r e s i stant w ee d bio t y p e s that survive a ra t e of
herbi c i d e to which the ind i g e nous popul a t i on w a s co n trolled” that a re di s c ov e r e d a nd v e ri f ied

g l o b a l l y

. A cc o r di n g to the I S HR W ( ht t p: / /ww w .w ee dsci e n c e .or g , ac c e ssed 04/2010 ) , the r e we re 28 b r o a dle a f w e e ds (8 in the U.S.) with sel e c ted r e si s tan c e to s y n t h e t i c a u x ins (in c lud i ng ph e no x y a u x i n s such a s 2,4

- D )

, a nd 38 g r a ss w e e ds (15 in the U

. S .) with r e si s tan c e to AC C a se inh i bi t o r s (in c lud i ng “ fop s ” ). At that time, w i t hin t he U.S., non e of the b r o a d l e a f w e e ds

r e si s tant to s y nthetic a u x i ns h a d b e e n fo u nd in co r n fi e lds, a nd g r a ss w e e ds r e si s tant to “ f ops”

h a ve a lso not be e n a pro b l e m f or c o r n g r o w e rs a s AC C a se inh i bi t o r s ha v e not t radi t ional l y b ee n

us e d to control w ee ds in c o r n fi e lds a f t e r p l a nt i n g

. B y c ompa r ison, the r e were 1 0 8 w ee ds found

w o rl d

- wide that h a ve d e v e loped re si s tan c e to A L S inh i bi t o r

- t y p e h e rb i c ides, 68 th a t h a ve d e v e lo p e d r e si s tan c e to P hoto s y st e m I I inh i bi t o r s ( such a s a tr a z ine ) , 18 that h a ve b e e n fo u nd to
be re si s tant to g l y phos a te, a nd, a c c o r ding t o the I S HR W

, many of the s e re si s tant w ee ds w e re found in U.S. c o r n fi e ld s . The p e t i t i on e r h a s also ind i ca t e d that the h e rbi c ide r e si s tan c e tr a i t s of

D A S

- 40278

- 9 c o r n m i g ht be c omb i n e d (or “ sta c k e d ” ) with o t h e r h e rbi c ide r e s i s tan c e t r a i t s (i. e

. g l y pho s a te a nd g lufosinat e ) ( D A S 201 1

, p a g e s 4, 18, 114

- 115, a nd 151) t o use a s pa r t of a w e e d

c ontrol s t r a te g y for g r o w e rs. When the petition was submitted, w i t hin t he U. S . the r e were not

a n y w ee ds th a t ha d shown r e si s tan c e to e i t h e r g l yphos a te o r a tr a z ine a n d e i t h e r a s y nthetic a u x in

or a “ fo p ” h e rb i c ide , nor were there any r e si s t a nt t o both 2, 4

- D a nd a nother c lass of h e rbi c ide,
thou g h ins t a n ce s h a d b e e n identifi e d in o t h e r c o u ntri e s

. The I S HRW h a d ind i ca ted how e v e r, that the r e were f ive inst a nc e s of w e e ds within t he U

. S . showing re si s tan c e to both ACC a se inh i bi t o r s and a nother h e rbi c ide mode of ac t i on c l a ss, thou g h none o f th e se w e re lo c a ted w i t hin

c o r n fi e ld s . The u s e of D A S

- 40278

- 9 c o r n, re g a r d less of the sta c ki n g with other h e rb i c i d e

r e si s tan c e t r a i t s that m a y or m a y n ot o cc u r , could pro v ide g r o w e rs with anoth e r w e e d c ontrol
opt i on, a nd should e n a ble the c ont i nu e d use o f n o

- or re d u ce d

- t i l la g e in c o r n c rops in a re a s wh e re g l y phos a t e h e rbi c i de re si s ta n t w e e ds h a ve a pp ea r e d. Herbi cide resistant weeds continue
to evolve

, and herbicide use patterns and the impacts of DAS

- 40278

- 9 corn on herbicide resistant weed management are addressed in the E nvironmental I mpact S tatement (EIS) prepared for this petition ( see http://www.aphis.usda.gov/biotechnology/petitions_table_pending.shtml )

.



### D e s c r i pt i o n o f I n s e r ted Gene t i c M a te r i a l

DA S

- 40278

- 9 c o r n w a s p r odu ce d b y t r a nsfo r mat i on of e mb r y o g e nic sus p e nsion c ul t u re s of the

r e c ip i e nt c o rn line Hi

- II using si l icon ca rbi d e wh i sker f ibe r s (Petolino e t al. 2003; P e to l ino and

A r nold 2009) to introdu c e a 6236 b a se p a ir r e stri c t i on f r a g ment e x p re ss i on ca ssette c on t a i n in g the g e n e t i c mat e ri a l that e n a bles the p r od u c t i on of the A A D

- 1 e n z y me ( D A S 201 1 , p a g e 2 1 )

. The p e t i t i on e r h a s es t a bl i shed that D A S

- 40278

- 9 c o r n c ontains on l y th i s e x p re ss i on ca ssette

a s d e s c rib e d in the p e t i t i on ( D A S 201 1 , p a g e s 26

- 57) a nd that th i s in s e rt e d g e n e t i c mat e ri a l

r e sul t s in t he pro d u c t i on of the A A D

- 1 pr o tein with i n t h e D A S

- 40278

- 9 c o rn pl a nts (D A S

201 1 p a g e s 58

- 63 and 132

- 140 ).

5Co r n plants c ontaining t h e inse r ted g e n e t i c mat e r i a l w e re in i t i a l l y iso l a ted b y g r o wing

tr a nsfo r med pl a nt t is s ue on medium cont a in i ng R

- h a lo x y f

op , a n A O P P A C C a se inh i bi t or ( D A S 201 1 , p a ge 21). W hole p lants w e re g e n e r a ted f r o m t he surviving t i ssue a n d s e le c ted plants w e re b re d with d e sir a ble c o r n l ines to g e n e r a t e e l i te c o r n v a ri e t i e s co n taini n g the g e n e t i c inse r t i on. D A S c onfi r med via Sou t h e rn blot an a l y s e s that t h e f inal c o r n p rodu c t ( DA S

- 40278

- 9) do e s not c ontain a n y plasmid se q u e n ce s outside of the inte n d e d 6236 b a se p a irs a nd

c ontains on l y a si n g le in t ac t i ns e rt in the c o r n g e nome with no evid e n c e of dupl i ca ted or

m i ss i ng D N A f rom that inse r t i on. I n h e ritance p a t te r ns of the inse r ted g e n e t i c mat e ri a l also
d e mons t r a te the in t ac t n e ss a nd f u n c t i on a l s t a bi l i t y o f the inse r ted D NA ov e r s e v e r a l b r ee di n g

g e n e r a t i on s

. ( D A S 201 1 , p a g e s 30

- 57 ).

A d e tailed d e s c ription of the i nse r ted g e n e t i c mat e ri a l i s p r ovided in the p e t i t ion (D A S

201 1 , p a g e s 26

- 27 )

, a nd is sum m a ri z e d b e low:

The in t rodu c e d g e ne r e s p onsible f or c on f e r r i n g the h e rbi c ide r e si s tan c e e n c od e s an

a r y lo x y a lkano a t e dio x y g e n a se ( AAD

- 1 ) e n z y m e. The g e n e , aa d

- 1, w a s or i g inal l y obtain e d f rom the soil b ac te r ium Sphingob i um h e rbicido v oran s , a nd w a s subs e q u e nt l y a l t e r e d to be pl a n t

opt i m iz e d ( the o r i g inal nu c leic a c id sequ e n c e w a s c h a n g e d b eca u s e plants f a v or dif f

e r e nt
codons for opt i mal D N A

- p r o t e in t r a nslatio n ) . The f inal p r otein p r odu c t ( A A D

- 1) p rodu c e d in

DA S

- 40278

- 9 c o r n is n e a r l y identic a l (99. 3 % identi t y ) t o the p r otein p r odu c e d b y S.
h e rb i c idovorans dif fe ri n g on l y b y the a dd i t i on of one a m i no ac i d

, a l a nine, a t pos i t i on 2 ( D A S

2011 , p a ge 58; W ri g ht et a l. 2009). D A S pro v id ed e viden c e t h a t t his s in g le a m i no ac id ch a n ge d id n ot i mpa c t t he f un c t i on of the e n z y me n o r w a s t h e re a n y e viden c e that the p r otein

c ontribut e d to a n y c h a n g e s to t he a g r onom i c pro p e rties, dise a se sus c e pt i bi l i t y , o r c o mposi t ion
of D A S

- 40278

- 9 c o r n c om p a r e d to other c om m e r c ial l y a v a i l a ble c o r n (s e e “ P otential I mp a c ts of

G e n e t i c Mod i fi ca t i ons on Alt e r e d Dise a se a nd P e st S usc e p t ib i l i t i e s” b e low).

The aad

- 1 g e n e is flank e d on e i t h e r side b y re g u l a to r y (no n

- c odi n g ) s e qu e n ce s that have b ee n d e riv e d f r om t he c o r n g e nome. I n f r ont of aa d

- 1 ( on the 5’ e nd) is the Z

. m a y s ubiqu i t i n 1 ( Z mUbi 1 ) p r omo t e r w hi c h is a s e qu e n c e of D N A th a t promot e s the c onsti t ut i ve e x p re ss i on of

g e n e t i c mat e r i a l , p a rticu l a r l y in m ono c ots (Christ e nsen e t al. 199 2 , Christ i a nsen a nd Qu a il
199 6 ) . This pa r t i c ular promot e r s e qu e n c e w a s p r e vious l y used in a pl a nt produ c t t h a t has b ee n

p re vious l y d e te r m i n e d to h a ve non r e g u l a ted status ( A P H I S B RS P e t i t i on 03

- 181

- 01p) (see http://www.aphis.usda.gov/biotechnology/petitions_table_pending.shtml ) a nd t h e use o f th a t

sequ e n c e in D A S

- 40278

- 9 c o r n d o e s not r a ise a n y n e w issues. On the downstr ea m side of the
aad

- 1 g e n e is t he Z . ma y s p e ro x idase 3 ’ u ntr a nslat e d r e g ion ( Zm P e r5 3’ UT R ) , whi c h is a

r e g ulato r y seq u e n c e of c o r n D N A th a t promo t e s the e f f i c ient tr a n s c ription a nd tr a nslation of a

c o r n g e n e ( Ainl e y e t al. 2002 )

. B oth t he Z mUbi 1 promoter a nd the Z m P e r 5 3’ U TR gen e t i c

s e qu e n ce s s e rve on l y a s a r e g ulato r y r ole, a nd a re not t r a nslat e d with t he A A D

- 1 pr o tein ( D A S

201 1 , App e ndix 2, p a g e s 1 3 2

- 140 )

.

On e i t h e r side of the a b o ve g e n e t i c mat e ri a l, DAS h a s also includ e d the R B 7 M a trix a t t ac hment

re g ion (MAR). T his g e n e t i c mat e ri a l , d e riv e d f r om tob acc o ( Nicotiana t ab a c um ) h a s b ee n d e monstr a ted to i m p r ove the stabili t y o f e x p re ss i on of inse r ted g e n e t i c ma t e ri a l b y bind i ng to

nu c le a r m a t ri ce s within a plant ce ll ( H a ll e t al. 19 9 1; Al l e n e t a l. 2000; Ma n kin et a l. 200 3 )

. MAR s e qu e n ce s do not e n c ode f or a p r otein p r od u c t a nd a r e a lso consid e red sol e l y a s

6re g ulato r y e lem e nts. Also p a rt of t h e 6236 b a se p a ir i ntrodu c e d f r a g m e nt a r e sh o rt in t e rv e ni n g D N A s e qu e n ce s t h a t w e r e p a rt of the o r i g inal c lon i n g plasmid. Th e se seq ue n ce s do not c ontain

a n y c oding s e qu e n c e but r e p r e s e nt t he l i nking portions of the plasmid that l i e b e tw ee n the D N A

s e qu e n c e in s e rtion points f or the r e g u l a to r y r e g io n s and c odi n g r e g ion of t h e e x p re ss i on

ca ssett e

.

I n summ a r y , t h e on l y p o r t i on of the introdu ce d D NA f ra g m e nt t h a t i s tr a nslat e d in t o a p r otein

p r odu c t or that c ould po t e nt i a l l y a l te r the nutritional or f und a men t a l comp o n e nt c h a r ac te rist i c s of the c o r n plants is the aa d

- 1 g e n e . An a n a l y sis of t he i m p ac ts of that ins e rt i on with i n DA S

40278

- 9 c o r n a s it p e rt a ins t o p l a nt pest risk is pr e s e nted throu g h the r e st of th i s do c ument.



## Plant Pest Ri s k A s s es s m e n t

AP H I S is r e spons i ble for r e g ulating GE o r g a nis m s and plants und e r the pl a nt pest p r ovis i ons in
the P l a nt P rot ec t i on A c t (PPA ) of 2000, a s am e nd e d (7 U S C § 7701 e t seq .) to ensu r e that th e y do not pose a plant pest r isk t o the e nvironme n t. AP H I S re g ulations a t 7 C ode of F e d e r a l

R e g ulations ( C F R ) Pa rt 3 40, whi c h w e r e promu l g a ted pu r suant to a u thori t y g r a nted b y the

P P A, re g ulate the introd u c t i on (impor t a t i on, in t e rst a te mov e ment, or re l ea se in t o t he

e nvironme n t) of ce rt a in GE o r g a nis m s a n d pr o du c t s . A G E o r g a nism is no lon g e r subj ec t t o the pl a nt pest p r ovis i ons of the PP A or to t he r e g ulato r y r e quir e ments of Pa r t 340 w h e n AP H I S
d e t e rmin e s that it is u n l i k e l y to pose a plant p e st risk. A G E o r g a nism is conside re d a r e g u lat e d

a rticle un d e r P a rt 340 if the donor o r g a ni s m, r ec ip i e nt o r g a nis m , v e c tor, or v e c tor a g e nt u s e d in

en g in e e ri n g the o r g a nism b e lon g s to one of the t a x a l i sted in the re g ulation (7 C F R 340.2) a nd
is a lso conside re d a plant p e s t . A G E o r g a n ism is also r e g ulat e d un d e r P a rt 340 wh e n AP H I S

h a s r ea son to b e l i e ve that the G E o r g a nism m a y be a plant p e st or AP H IS do e s not h a ve info r ma t ion t o d e te r m i ne if the G E o r g a nism is unl i k e l y to pose a plant p e st r i sk.

A p e rson m a y p e t i t i on the a g e n c y that a p a rticul a r r e g ulat e d a rticle is unl i k e l y to pose a plant p e st risk, a nd the r e fo re , i s no lon g e r re g ulat e d un d e r the pl a nt pest provisi o ns of the PP A or the

re g ulati ons of P a rt 340. The p e t i t i on e r is r e qui r e d to provide info r mation under §340.6 ( c ) ( 4)to
d e monstr a te th a t t he re g u lat e d a rticle that is the s u bje c t of the p e t i t i on do e s not pose a plant p e st
risk a nd that it should no lon g e r b e subj ec t t o the r e g u l a t i ons of 7 C F R p a rt 340. A G E o r g a nism
is no lon g e r subj e c t t o the r e g ulato r y r e qui r e men t s of P a rt 340 or the pl a nt p e st provisions of the

PP A wh e n AP H I S d e t e r m i n e s that it i s unlike l y t o pose a plant p e st risk.

As d e s c rib e d ea rli e r in t his do c ument, the in t rodu c e d g e netic mat e ri a l i n D A S

- 40278

- 9 c o r n

c onf e rs e nh a n c e d to l e r a n c e a nd r e si s tan c e to tr ea t ment with sel ec t he r bici d e s to the G E pl a nts. No plant p e st or plant p e s t

- d e riv e d mat e ri a l w a s u s e d to g e n e r a te the DA S

- 40278

- 9 c o r n plants. DAS h a s pl a nted DA S

- 4 027 8

- 9 c o r n a s a r e g u l a t e d a rticle und e r A P H I S BRS
Notifi ca t i on of Environm e ntal R e le a se s i n c e 2006 ( D AS 201 1 , p a g e 14 4 )

.

P otential i m p ac ts t o be a ddr e ssed in th i s plant p e st risk a ssessment a re t ho s e that p e rt a in t o the

use o f DA S

- 40278

- 9 c o r n a nd i t s prog e n y in t he a bsen c e of c onfin e ment a s othe r wise a f fo r d e d
und e r c ondi t ions of 7 C F R p a rt 340 for the in t ro d u c t i on of g e n e t i ca l l y e n g i n ee r e d o r g a nis m s. Of the in f o r mation r e qu e s t e d b y AP H I S f o r submission of a p e t i t i on f o r no n re g ulat e d st a t us, AP H I S e x a m i n e d info r mation subm i t t e d b y the p e t i t i on e r ( a s per 7 C F R p a rt 340.6 ( c ) ( 4 ) ) r e la t e d
to p l a nt p e st risk c h a r ac te r is t ics, dise a se a nd p e st s usc e pt i bi l i t ies, e x p re ss i on of the g e n e

7pro d u c t, n e w e n z y mes or c h a n g e s to p l a nt m e tabolism, w ee din e ss (in c lud i ng c ontrol opt i ons) of
the r e g u l a ted a rticl e , the possib i l i t y of e f f

e c ts of the re g ulat e d ar t i c le on no n

- ta r g e t o r g a n i sm s ,

a nd a n y i m p a c ts on the wee diness o f a n y other plant with which it c a n in t e rb r ee d. Ot h e r issues

re lat e d to a g r icultur a l or c ul t ivation pr ac t i ce s (in c lud i ng but not l i m i t e d to t hose r e lat e d to
ch a n g e s in h e rbi c id e s used to c on t r o l w ee ds in c o r n ) , non

- ta r g e t o r g a nis m s, be ne f i c ial

org a nis m s, thr ea te n e d a nd e nd a n g e r e d s p ec ies a nd other e nvironme n t a l i mpa c ts h a ve b e e n

a d d r e s s e d in an Environm e ntal Ass e ss me nt ( E A) and subsequent EIS that h a v e b e e n prep a r e d to

a n a l y z e potenti a l i mpa c ts of a d e te r m i n a t i on of non r e g u lat e d st a tus of DA S

- 402 7 8

- 9 c o r n b y AP H I S ( see http://www.aphis.usda.gov/biotechnology/petitions_table_pending.shtml

. )



### P o ten t i a l I m p a c t s of Genet i c M o d i f i c a tio ns on Al tered D is e as e and P e s t
Su s cep t i b i l i t i es

AP H I S a s s e ssed w h e t h e r DA S

- 40 278

- 9 c o r n is l i k e l y to ha v e si g n i fi ca nt l y inc rea s e d dise a s e

a nd p e st s usc e pt i bi l i t y c omp a r e d to c onv e nt i on a l c o rn. This ass e ss m e nt en c o m p a sses a thoro u g h c onsid e r a t i on of in t roduc e d tr a i t s and in t e ra c t i ons with p e sts a nd dis ea s e

.

N one of the inse r ted g e n e t i c s e qu e n ce s contained in DA S

- 40278

- 9 c o r n a re d e riv e d f rom

a n y plant p e sts l i sted in 7 C F R p a rt 340.2 or p e sts l i sted e lse w h e re in 7 C F R p a rts 3 0 0

- 399.
The d e s c ript i on of the g e n e t i c mod i fi ca t i on, including t he in t rodu c e d g e n e t i c e l e ments a nd
the re su l t ing e x p re ss i on a nd fun c t i on of the AA D

- 1 p r otein p r odu c t in DA S

- 4 0 278

- 9 c o r n, h a s b ee n s u m m a ri z e d a bov e

. D A S routin e l y mon i tor e d their c o r n fi e ld t ri a ls f o r c o r n dise a s e s such a s ea r rots, le a f bli g hts, rusts, a nd le a f spots a lo n g with inse c t dam a g e f r o m

co r n p e sts such a s le a f ho p p e r s, m i tes, thrips, a phids, b ee t l e s, g r a ssho p p e rs, a nd c o rn

e a r w o r m s. D A S c ondu c ted a g r on o m i c f ield tri a ls during the 2008 g r owi n g se a son ac ross 27
lo ca t i ons r e p r e s e ntative of the m a jor c o r n

- g r owi n g a r ea s of the U

. S

. ( D A S 201 1 , p a g e s 6 4

- 75). Th e re w e re n o statis t ic a l l y si g nifi c a nt dif fe r e n ce s in a g r onom i c c h a ra c te r is t i c s such a s vi g o r, lod g i n g o r y ield that m i g ht be ind i ca t i ve of a l te red s e nsi t iv i t y to pests and d i s ea s e s b e t w ee n D A S

- 40278

- 9 c o r n, d e riv e d h y b r i d s and their c ont rol c ounte rp a rts und e r st a n d a rd

c o r n m a n a g e m e nt pr a c t i ce s ( D A S 201 1 , p a ge 70) or in r e sponse to spe c ific h e rbi c ide

t r e a t m e nts ( D A S 201 1 , p a g e

. 6 7). I n one e x p e rime n t, d i s ea se a nd ins e c t d a m a g e r a t i n g s

w e re t a k e n for tr ea t m e nts t h a t i n c l ud e d D A S

- 40278

- 9 h y b r i d c o r n spr a y e d with 2,4

- D, qui z a lofop, 2,4

- D a nd qui z a lofop, a nd n e i t h e r h e rb i c ide, a s w e ll a s the n ea r

- iso l ine c ontrol

h y b r id p l a nted a t s i x ( 6) fi e ld s i tes; R ichl a nd, I A; C a r l y le, I L ; W y om i n g , I L ; R o c kvi l le, I N;

Yo r k, N E ; a nd B r a n c ht o n, Ont a rio, Can a d a ( D A S 201 1 , p a g e 7 1 ). The d a ta submit t e d b y DAS c onfi r med th a t t he ins e rt e d g e n e t i c mat e ri a l d i d n ot con fe r a n y a l t e r e d dis e a se or p e st

susc e pt i b i l i t y to D A S

- 4 0 27 8

- 9 c o r n. S p ec i fic dise a se a nd in s ec t s t r e ssor o bse r v a t i ons w e r e

c ol l ec ted f rom 2006 to 20 08 a c ross lo c a t i ons r e p r e s e nt i ng a bro a d r a n g e o f U.S. co r n

g ro w i n g re g ions ( D A S 201 1

, p a g e s 6 4

- 75 )

.

The d a ta p r e s e nted in the p e t i t i on ( D A S 201 1 , p a g e s 76

- 101) ind i ca te no m a te r ial dif f

e r e n c e in

c omposi t ional a nd nutrit i on a l quali t y of D A S

- 40 2 78

- 9 c o r n c om p a r e d to o t h e r c om m e r c ial l y

a v a i l a ble c o rn a p a rt f rom the p re s e n c e of the A A D

- 1 pr o tein. A f e w v a r i a bles did show
stati s t i ca l l y si g nifi c a nt dif fe r e n ce s b e tw e e n the c o mposi t ional c h a r a c t e rist i c s of DA S

- 40278

- 9

c o r n a nd c on t rol c o r n g r o wn a t t he same lo c a t i on s . Ho we v e r, these dif f

e r e n ce s w e re r e lative l y small a nd none of the v a l u e s for the f o ra g e a nd g r a in composit i on c h a ra c t e rist i c s w e re outs i d e the ra n g e o f n a tur a l v a ri a bi l i t y of c onv e nt i on a l c o r n r e port e d b y the I nte r n a t i on a l L i f

e S c ien ce s

8I nst i tu t e Crop Compos i t i on D a tab a se (Rid le y e t a l . 2004; IL S I 2006) o r in the OECD c onse n sus do c ument on c o r n c omp o si t ion ( O ECD 20 0 2). T h e r e fo r e , with t he e x ce pt i on of the A AD

- 1

p r otein whi c h is a t m ost 0.02% of c o r n t i ssue d r y w e i g ht ( ra n g e : 0.42

- 210 n g / m g d e p e ndi n g on the tissu e , see D A S 201 1 , p a ge 60 ), the c omposi t ion of DA S

- 40278

- 9 c o rn is not b i olo g ic a l l y dif fe r e nt t h a n c onv e nt i o n a l co r n. D a t a in t he p e t i t i on (D A S 201 1 , p a g e s 68

- 70) sho we d no
si g nifi ca nt d if f

e r e n c e in g r a in y i e ld betw e e n a D A S

- 40278

- 9 c o r n h y b r id a nd a n e a r is o g e nic

c ontrol line ac ross 21 lo c a t i ons (6 stat e s). Th e se d a ta c ol l ec t i v e l y support t he p e t i t i on e r ’ s cl a im
that D A S

- 40278

- 9 c o r n d o e s not h a ve altered s u s ce pt i bi l i t y to a n y inse c t s , plant p a thogens, or
other plant p e sts ov e r its no n

- tr a n s g e nic c ontrol.

The AA D

- 1 p rot e in i s d e riv e d f r om a g ra m

- n e g a t i ve soil b ac te r ium Sphin g obium

h e rbicido v orans that is not asso c iat e d with plant d ise a se ( B a lkwi l l et a l. 2 0 06; Kohler 1999;

Z ipp e r e t al. 1996; Ho r v a th et a l. 1990). S. h e rbic i do v orans w a s or i g inal l y iso l a ted using a soi l

- c olu m n e n r ichm e nt foll o w e d b y c ul t u r e g r owth w i t h ph e no x y a u x in he r bi c ide a s the sole

c a rbon a nd e n e r g y so u r c e s ( H o r v a th et a l. 1990). The A AD

- 1 pr o tein e x p re ssed b y the D A S

40278

- 9 c o r n, with t he e x ce pt i on of the s ec ond a m i no ac i d is i d e nt i ca l t o pr o teins found in

o t h e r soil b ac te r ia ( N on

- r e dund a nt P rot e in B a sic L o ca l Al i gnment S ea r c h Tool ( B L A S T )

, ht t p: / /ww w .n c bi.n l m.n i h .g ov /

, a cce s s e d 4/2010), in c lud i ng Rhodofer a x sp P 230 a nd D e l f

t i a

a c idovorans , whi c h h a v e a lso be e n iso l a t e d f rom h e rbi c ide

- e n r ic h e d subs tr a te ( M ül l e r e t a l. 2001) a nd a r e not consi d e r e d pl a nt pests (W e n e t al. 1999). Using B L AS T

, the A A D

- 1 pr o t e in

w a s c lassifi e d a s a n R

- 2

, 4

- dichlo r oph e no x y p r opi o n a te dio x y g e n a s e , a nd it h a s also b ee n n a med

a s a n (R )

- dichl o rp r op/α

- k e to g lu t a r a te dio x y g e n a s e ( RdpA) ( Mülle r , e t al. 200 4 ; W ri g ht e t al.
2009 ) . The e n z y m e h a s o n l y b e e n a ssoci a t e d with t he met a bol i sm of h e rbi c ides a nd the r e is no
ind i ca t i on that i t h a s a n y other f un c t i on.

I n summ a r y , all e viden c e r e vie we d b y AP H I S in d i ca te th a t DA S

- 4027 8

- 9 c o r n is just a s

susc e pt i ble to plant p a tho g e ns a nd other plant p e st s p ec ies a s co n v e nt i on a l c o r n a nd that the r e is no a dd e d plant p e st risk t h a t r e sul t s f r om t he inse r t ion and e x p re ss i on of the g e n e t i c mat e r i a l
d e s c rib e d in t he p e t i t i on

.



### P o ten t i a l I m p a c t s B as e d on the Re l a t i v e Weed i n e s s of DA S

- 4 02 78

- 9 Co rn

Co r n ( Z e a ma y s ) is not a plant p e st i n the U ni t e d S tat e s ( U S D A

- AP H I S 20 0 0 )

. C o r n is not

t y

pi c a l l y c o n side re d a s a w ee d (Cro c k e tt 1977; Holm e t al. 1979; Mu e nsch e r 19 8 0) a nd is n ot
l i sted on the Fe d e r a l no x ious w ee d l i st (7 C F R p a rt 36 0 ) . Co r n possess e s f e w o f the

c h a ra c te r is t ics of notab l y suc c e ssful w ee ds a nd c o rn is g r o wn a s a c rop thr o u g hout t he w o rld
without a n y r e port that it is a s e rious w ee d o r th a t i t fo r m s pe r si s tent fe r a l p opulations ( B a k e r 1965; K ee ler 198 9 ) . H o w e v e r, c o r n s e e d ca n g e r m i n a te in undesir e d loc a t ions a nd would
then be c onsid e r e d a w ee d. A c om m on e x a mp l e of this i s the a pp ea r a n c e of c o rn se e dl i n g s in

s o y b e a n fi e lds fol l owing a c o r n c r op whi c h h a s b e e n a ss oc i a ted w i t h r e du c e d s o y b ea n y ields ( B e c k e tt a nd S to l ler 1988 ) . Compa re d to o t h e r c o r n v a ri e t i e s, D A S

- 40278

- 9 h a s i m p r ov e d
fitn e ss ( re d u ce d in j u r y ) in t he p r e s e n c e of ce rt a in h e rbi c ides, whi c h tr a nslat e s in t o f e w e r

o p t i ons for the re moval of volun t ee r pl a nt s . No n e thel e ss, t h e re a r e ma n y a v a i l a ble options for
the c o ntrol of D A S

- 40278

- 9 plants so t h a t g r ow e rs c a n a void l osses in f ields wh e re unw a nted

D A S

- 4027 8

- 9 m i g ht be gro w i n g

.

AP H I S a s s e ssed w h e t h e r DA S

- 40278

- 9 c o r n is a n y more l i k e l y to b e c ome a w ee d t h a n a n y

9other c o r n v a r i e t i e s cu rr e nt l y u nd e r c ul t ivation. T h is a s s e ss m e nt en c ompass e s a thorou g h

c onsid e r a t i on of the b a sic biolo g y of c o r n a nd a n e v a luation of the unique c h a ra c te r is t ics of

DA S

- 40278

- 9 c o r n e v a l u a ted und e r fi e ld condit i ons. DAS c ondu c t ed a g r o nom i c f ield tri a ls during the 2008 g r owi n g s ea son a c ro s s 27 loc a t i o n s r e p re s e ntative of the m a jor c o r n

- g r owi n g

a r e a s o f the U. S

. ( D A S 2 01 1 , p a g e s 64

- 75 ) . T h e r e w e re f e w stati s t i ca l l y sign i fi ca nt d i f f

e r e n c e s b e tw ee n D A S

- 40278

- 9 c o r n , d e riv e d h y b rids a nd their c ontrol co u nte rp a rt s , r e g a rd l e ss of

h e rbi c ide t r ea t m e nt. The si g nifi ca nt d if f

e r e n ce s t h a t w e re s ee n ( D A S 201 1

, p a ge 67), in e a r h e i g ht a nd a visual e st i mate of pl a nt h e a l t h ( st a y gr e e n )

, r e f l ec ted v e r y sma l l d i f fe r e n ce s in morpholo g ic a l f e a tu r e s a nd w e re not a cc ompan i e d b y s i g nifi ca nt o v e r a ll t r ea t m e nt e f f

e c ts ( D A S

201 1 , p a ge 66). No di f f

e r e n ce s in ph e no t y pic c h a r ac te r is t ics th a t would c o n tribute to
enh a n ce d w ee diness w e re ob s e rv e d b e tw ee n D A S

- 40278

- 9 c o r n a nd c ontrol lines f or t he

r a n g e o f ph e no t y pic e ndpoin t s ass e ssed.

B a s e d on t h e a g ronomic fi e ld data a nd l i te r a ture s u r v e y a bout c o rn w e e diness potential, DA S

40278

- 9 c o r n l a c ks the a bi l i t y to p e rsist a s a trou b lesome or invasive w e e d. S t i l l , b eca use DA S

40278

- 9 c o r n m i g ht gr o w in a r e a s wh e r e it is und e sir e d, the G E c o r n , si m i l a r to other c o rn

v a ri e t i e s, would be c onsid e r e d a w ee d (B ec k e tt a n d S to l ler 19 8 8 )

. B e c a use the g e n e t i c e le m e nts
inse r ted into DA S

- 4027 8

- 9 c o r n c o n f e rs g rea t e r t o le ra n c e or r e si s tan c e to certain h e rbi c ides t h e

me a sur e s that c a n b e u s e d to r e move volunte e r D A S

- 40278

- 9 c o r n a re m o re l i m i ted c ompa re d
to other c o r n v a ri e t i e s . No n e thel e s s , a s discuss e d in t he n e x t pa r a g r a p h s, most of the c u r r e nt l y

u s e d c ontrol m ea sur e s for r e mov i ng volun t ee r c o r n a re a lso av a i l a ble f o r the r e moval of D A S

40278

- 9 c o r n.

C o r n is common l y g r o w n in rot a t i on with o t h e r crop va ri e t i e s includi n g , b ut not n ece ssa r i l y l i m i ted to, o a ts, fo ra ge c r ops, p ea nut, wh e a t, r y e , c ot t on, a nd s o y b e a n. C o r n se e d le f t i n a f i e ld

a ft e r h a rv e st i s common, a nd that s ee d c a n g e r min a te, or “ volun te e r,” d u ring the g r o wth of the following r otational c ro p

. I f this co r n is not dest ro y e d o r r e mo v e d, the su b s e qu e nt c r op planti n g m a y h a ve low e r y i e lds a n d r e du ce d v a lue due to c ompeti t ion a nd m i x ing with t he
volun t ee r c o r n ( B e c kie a nd O w e n 2007; B ec k e tt a nd S to l ler 1 9 8 8 ) . The a d v e nt and w ide

a dopt i on of h e rbi c ide r e si s tant c rops h a s a lr ea d y c h a n g e d the a pproa c h e s g ro w e rs c a n a dopt to

r e du c e c rop y i e ld l osses r e sul t ing f rom volunt ee r c o r n. F o r e x a mp l e , g l y p hosat e

- r e si s tant

s o y b e a n plants, g ro w n a ft e r g l y phos a t e

- r e si s tant c o r n , ca n h a v e re d u ct i ons in s o y b e a n y ield,
due to co m p e t i t i on with volun t ee r c o r n, if g l y ph o s a te is the on l y w ee d c o n trol m e thod us e d (Soltani e t al. 2006 )

. S i m i l a r stud i e s ha v e a lso be e n d o ne f or c o r n

- c ot t on r o tati o ns (C l e wis e t al. 2008; Thom a s et a l. 2007) a nd c o r n

- wh ea t ro t a t i ons (M a rtin 2008 )

.

One s u g g e sted c h e m i ca l method f or the c ontrol of volun t ee r c o r n a mo n g b r o a dle a f c rops is
the use o f g r a m i ni c ide A CC a se inh i bi t o r s whi c h include “ fops” ( D e e n e t al. 2 0 06; S ol t a n i et

a l. 2006 ) . Ho we v e r, the use of “ fop” AC C a se inh i bi t or h e rbi c ides would not be e f f

ec t i ve f o r the c ontrol of volunte e r D A S

- 4027 8

- 9 c o r n d u e to t he r e si s tan c e that h a s be e n c onf e r r e d b y

g e n e t i c e n g i n ee ri n g . No n e thel e s s, other AC C a se inh i bi t or g r a m i ni c i des, such a s

c y c lohe x a n e dione ( “ di m ” ) herbi c ides ( e

. g

. c lethodim or s e tho x y di m ) a re a lso e f f

ec t i ve f o r the

c ontrol of volunte e r c o r n in bro a dl e a f r o t a t i on a l c rops ( Cle w is et a l. 2 0 08; Thom a s et a l. 2007;

D ee n e t a l. 2006; S ol t a ni et a l. 2006; W SS A 2007 )

, a l t ho u g h the l e v e l of c ontrol with

s e th o x y dim h a s b e e n r e port e d to be l e ss e f f

ec t i ve a t m a nufa c tur e r r e c o m m e nd e d dos e s than
the “ fop” h e rbi c ides test e d (Soltani e t al, 2006 ) . A n other la r g e c lass of h e rb i c ides that c a n b e used f o r the e li m ination of volun t e e r c o r n i n c ludes the a c e to l ac tate s y nthesis ( A L S ) inhibitors,
such a s i m a z a mo x , i m a z a quin, and ima z e tha p y r (W SS A 20 0 7). The p e t i t i on e r h a s co n fi r m e d
10the sus ce pt i bi l i t y of DA S

- 40278

- 9 c o r n to b o th “dim” a nd A L S inh i bi t or h e rbi c ides a t
lab e le d

- u se lev e ls (D A S 201 1 , p a ge 115 a nd A p ril 29, 2010 c o rr e spon d e n c e with A P H I S)

. A L S inh i bi t or h e rbi c ides a s w e ll a s p a r a qu a t ca n a lso b e used to cont r ol volun t ee r c o r n in co r n to wh ea t ro t a t i ons (M a rtin 2008 )

.

Oth e r c om m on l y u s e d m e thods f or the c ontrol of volun t ee r c o r n a re a lso s t i ll a v a i l a ble f or use

o n DA S

- 40278

- 9. F o r e x a mp l e , n on

- s e le c t i ve h erbi c i d e s ( e

. g

. g l y phos a te a nd g lufosinat e ) a r e opt i ons for e l i m i n a t i ng D A S

- 40278

- 9 c o r n a nd i t s prog e n y , a ssu m i n g that the r e h a s b ee n no b ree di n g ( or s t ac ki n g ) or c ros s

- pol l i nation with o t h e r c o r n l i n e s that h a ve r e si s tan c e tr a i t s to
those h e rbi c ides. U s e of a n y h e rbi c ide for the c o n trol of volunte e r D A S

- 4 0 278

- 9 c o r n is subj ec t

to a n y la b e l i ng r e stricti o ns est a bl i shed b y the EPA, a s per F I F R A. Addit i on a l l y , the r e a re a lso

ma n y no n

- c h e m ic a l methods of wee d c ontrol (ti l l i n g , m e c h a ni c a l r e moval, e tc.) that a re

e f

f

e c t i ve for re mov i n g a n y u n w a n t e d D A S

- 40278

- 9 c o rn.

The poss i bi l i t y o f h e rbi c i de

- r e si s tant volun t e e r c o r n c ompeting with o t h e r c rops is not c onfin e d

to GE v a ri e t i e s. Ot h e r c u l t ivar

- d e v e lop m e nt m e t h ods h a ve g e n e r a ted c o r n v a ri e t i e s that have

r e du ce d s e nsi t iv i t y to he r bicid e s that could othe r w ise c ontrol for volun t e e r c o r n in c r ops such

a s s o y b e a n. F o r e x a mp l e , the r e a re c om m e r c ial l y a v a i l a ble c o r n h y b rid v a ri e t ies w i t h

r e si s tan c e to s e tho x y di m , a c y c lohe x a n e dione A CC a se inh i bi t or ( V a n g e ssel e t al. 1997 )

. S i m i l a r to D A S

- 40278

- 9, ma n y opt i ons for c ontrol of the h e rbi c i d e re si s tant c o r n a re a v a i la ble ( You n g a nd H a rt, 1997 )

. W hi l e fa rm e rs m a y h a ve to c h a n g e their man a g e men t str a t e g ies d ue to
DA S

- 4027 8

- 9, these c h a n g e s will not ne ce ss i tate a major d e p a rt u re f rom w e l l

- e stablished a nd
b r o a d l y used a g r icultur a l protocols.



### P o ten t i a l I m p a c t s f r o m Outcros s i ng ( G e ne F l o w ) t o Se x u a l l y
- c o m p a t i b l e W i l d

R e la t i v es

G e ne f low is a n a tur a l b i olo g ic a l pr o ce ss wi t h s i gnifi ca nt evolut i on a r y i m port a n ce

. A num b e r of

a n g iosp e rm t a x a a re b e l i e v e d to be d e riv e d f r om h y b r id iz a t i on or in t r o g re s sion b e tw ee n c lose l y

r e lat e d ta x a ( G r a nt 1981; S ol t is and So l t i s 1993; R ies e b e r g 1997; H e g de e t a l. 2006 ) . Ev e n in

e x is t ing f lor a s, the o cc u r r e n c e of h y b ridi z a t i on or in t ro g r e ss i on is r e port e d t o be wid e spr e a d ( K nobloch 1972; S ta c e 1 987; R ies e b e r g a nd W e n d e l 1993; P e te r son e t al. 2002 )

. I t h a s b e e n a

c om m on p rac t i c e b y p l a n t br ee d e rs to a rtifi c ial l y i ntro g r e ss t r a i t s f r om wi l d re latives into c rop plants to d e v e lop new c u l t i v a rs. H ow e v e r, g e n e f l ow f rom c rops to wild r e latives is also thou g ht
of a s h a vi n g a potential t o e nh a n c e the w e e diness of wild r e latives, a s obs e rv e d in ri ce , s o r g hum,
sunflow e r a nd ot h e r cro p s ( E l l str a nd e t al. 1999 )

.

AP H I S e v a luat e d the po t e nt i a l for g e n e in t rog re s s ion t o o cc ur f rom D A S

- 40278

- 9 c o r n to

s e x u a l l y c ompatib l e wild r e latives a nd c onsid e r e d wh e ther such in t r o g re ss i on would re sult in
inc rea s e d w e e diness. Cul t ivat e d c o r n, or mai z e , Z e a ma y s L . subsp. ma y s , i s se x u a l l y

c ompatib l e with o t h e r m e mbe r s of t h e g e nus Z e a , a nd to a m u c h less e r deg r e e with me mbe r s of
the g e nus T rip s a c um ( O ECD 2003 )

. W i l d dip l o id and t e tr a plo i d memb e rs of Z e a, c o l le c t i v e l y

r e f e r r e d to as teosin t e, are n o rmal l y confined to t he tropi c al a nd subtropical r e g ions of Me x ico, Guatemala, and Nica ra g ua (Wi l kes 1967; Fukun a g a et al. 20 0 5). As previously analyzed by

USDA APHIS ( 2013)

, despite accounts of some taxa of teosinte as having been cultivated in the

U ni ted States as fodder (Taba 1995) or as research specimens, it does not appear to be present in
the U nited S tates other than as an occasional botanical garden or research specimen . There are

11no known naturalized or native populations of teosinte currently growing in the United States.

The g e nus T rip s a c um c o ntains up to 16 rec o g ni z e d sp ec ies, most of w hich a re n a t i ve to M e x ico,

C e ntr a l and Sou t h Am e r i ca , but t h re e ( T . da c tylo i d e s, T . f

l oridatu m , a nd T

. lanc e ola t u m ) e x ist

a s wild and/or c ul t ivat e d s p ec ies i n the c ont i n e ntal U

. S . ( O ECD 2003 ) ; and t w o ta x a ( T

. fasci c ula t um a nd T . la t i f

o l i u m ) a lso oc c ur in P u e r t o R i c o ( U S D A , NR C , 2010). Thou g h m a n y of these spe c ies o cc ur wh e r e c o r n m i g ht be c ul t ivat e d, g e n e in t r o g r e ss i on f r o m DA S

- 4027 8

- 9

c o r n und e r n a tu r a l cond it i ons is hi g h l y unl i k e l y

. H y b rids of T rip s a c um spe c ies with Z e a a re dif f icult to ob t a in ou t side of a la b o ra t o r y a nd a re o ft e n st e r i l e or h a ve g r ea t l y re d u c e d f e rtil i t y ,

a nd no n e of them ca n withstand e v e n the mi l d e st winte r s. F u r the r mor e , no n e of the s e x u a l l y

c ompatib l e r e latives of c o rn in the U

. S

. a re c onsid e r e d to be w ee ds in the U

. S

. ( Holm e t

. a l. 1979).

I ntr o g re ss i on of g e n e s f r om co r n in t o teosinte or T rip s a c um spe c ies in t he U.S. h a s not b ee n d e s c rib e d to o c c ur in na t u re

. W hi l e some teosinte m a y b e c onsid e r e d w e e d s in c e rt a in i nstan ce s, th e y a re a lso u s e d b y s o me fa rm e rs f or b r ee di n g i mprov e d c o r n (S á n c h e z a nd Rui z , 1997, a nd

r e fer e n c e s the r e in). T e o s in t e is des c rib e d a s b e ing sus ce pt i ble to ma n y o f the same p e sts a nd
dise a s e s that atta c k c ul t iv at e d c o r n. I n the w i l d, in t ro g r e ss i ve h y b r id iz a t i on f r om co r n to

t e osin t e is cu rr e nt l y l i m i ted b y s e v e r a l f ac tors includi n g g e o g r a phic isol a t i on, dif fe r i ng d e g ree s

o f g e n e t i c incomp a t i bi l i t y , dif f

e r e n c e s in flow e ri n g t i me in some ca s e s, d e v e lop m e ntal

morp holo g y a nd t i m i ng of the re p rodu c t i v e stru c tur e s, disseminati o n, a nd do r man c y ( D o e bl e y 1990a a nd 1990b; G a l i n a t 1988; Ellstr a nd 2007 )

. F irst

- g e n e r a t i on h y b r ids a r e g e n e r a l l y less f i t
for surviv a l and dis s e m i n a t i on in t he wil d

, a nd show subst a nt i a l l y r e du c e d r e p r od u c t i ve

ca p a c i t y , whi c h s e rv e s a s a si g nifi c a nt constr a int to in t ro g r e ss i on. Gu a d a g n u olo et a l. (20 0 6) did

r e p o rt th a t h y b r ids b e tw ee n a g l y pho s a te

- r e s is t a nt m a i z e c ul t ivar a nd “c h a lco t e osin t e ” ( Z . ma y s

ssp. m ex icana ) show e d g re a ter v i g or a nd pro d u ce d mo r e se e ds th a n the w i l d p a r e nt.
Non e t h e less, in t he a bsence of sel ec t i ve p r e ssure f r o m g l y phos a te th er e w a s no dir e c t pos i t i ve or

ne g a t i ve i m p ac t of the tr a nsge n e on the fitness or vi g or o f e i t h e r t h e h y b r i ds or the pure mai z e

pr o g e n y

.

D a ta in c luded in the p e t i t i on d e monstr a ted th a t t h e re w e re no s i g nifi ca nt d if f

e r e n ce s in viabil i t y

a nd morpholo g y o f poll e n c ol l ec ted f rom D A S

- 4 0 278

- 9 c o r n a nd c ontrol c o r n (D A S 201 1

, p a g e s 73

- 74) a nd th e r e f o re the ou t

- c rossing r a te o f D A S

- 40278

- 9 c o r n is not e x p ec ted to be
di f f

e r e nt c ompa re d to o t h e r c o r n v a ri e t i e s. B a s e d on the d a t a pr e s e n t e d in t he p e t i t i on, D A S

4027 8

- 9 c o r n do e s not e x hib i t ch a r a c t e rist i c s that c a use it to be a n y w ee d i e r th a n oth e r cul t ivat e d c o r n a nd the e x tr e me l y l i m i ted pot e nt i a l for g e n e in t rog r e ss i on in t o teosinte or T

rip s a c um spe c ies is not e x p ec ted to be a n y dif f

e r e nt t h a n that of other c ul t ivat e d c o r n v a ri e t i e s. E v e n i n the unlike l y e v e nt t h a t g e ne flow w er e to o cc ur b e tw ee n D A S

- 40278

- 9 c o r n a nd wild

r e latives, the r e is no r e a son to e x p ec t t h a t t he poss i ble e x p re ss i on of aad

- 1 would tr a nsfo r m c o r n wild r e latives into more w ee d y s p ec ies, e s p ec ial l y in t he a bse n c e of h e rbi c ide s e l ec t i on p re s s u re

. H y b r ids, i f th e y o cc u r r e d, c ould possib l y h a ve di m in i shed susc e pt i bi l i t y to some h e rbi c ides but

wo uld s t i l l be expected to be susc e pt i ble to ma n y ot h e rs, a s is D A S 4027 8

- 9.

I n summ a r y , D A S

- 4027 8

- 9 c o r n will not adv e r s e l y i m p ac t se x u a l l y c omp a t i ble w i l d r e latives
or the wee diness a t t ribut e s of those wild r e latives.

12

### P o ten t i a l I m p a c t s of D A S

- 4 0 2 7 8

- 9 on O r ga n i s m s C o n s i dered Benefi c i a l t o A g r i c u l t u re

The p e t i t i on e r a lso ass e s s e d the pot e nt i a l for D A S

- 40278

- 9 c o r n to i mpa c t “ non

- ta r g e t”

o r g a nis m s, including th o se c onsid e r e d b e n e fi c ial t o a g r icultur e , a nd d e te r m ined th a t t h e re would
be no e f f

ec t ( D AS 201 1 , p a ge 1 1 6). D A S

- 40278

- 9 is a g ronomi c a l l y a nd c omposi t ional l y si m i l a r to o t h e r c o r n v a ri e t i e s a n d will not adv e r s e l y a f f

e c t o t h e r o r g a nis m s comp a r e d to o t h e r

c o r n v a ri e t i e s. F u rth e rmo r e , AP H I S h a s found no e v i d e n c e or re a son to b e l i e v e that the

p re s e n c e o f the aad

- 1 g e ne o r the e x p re ssed A A D

- 1 pr o tein in D A S

- 40278

- 9 c o r n would h a ve

a n y i m p a c t on other o r g a nis m s, includ i ng o r g a nis m s be ne fi c i a l t o a g r iculture ( s u c h a s

e a r t hwo r ms, hon e y b ee s, p re d a tors or p a r a si t e s o f c o r n p e st s ). T h e A A D

- 1 p r otein is e x p re ssed
in a v a ri e t y of pl a nt t is s u e s in DAS

- 40278

- 9 c o r n w i th av e rage e x p re ss i on v a l u e s r a n g i n g f rom 2.87 n g/ m g d r y w e i g ht i n R1 sta g e ro o t t o 127 n g / m g i n pol l e n t i ssue ( D A S 201 1 , p a ge 60) a n d

e x p re ss i on v a lues we r e si m i l a r for spr a y e d tr ea t m e nts a s w e ll a s for plo t s spr a y e d a nd unspr a y e d with 2, 4

- D a nd qui z a lofop h e rbi c ides. The in s e rt e d g e n e t i c mat e r i a l i s not s ec r e ted, is not to x ic,

a nd do e s not pro d u c e a n y subs t a n c e that is se c r e ted or th a t would be c onsid e r e d to x ic. The

A A D

- 1 p r otein do e s not sha r e me a ningful a m i no ac i d s e qu e n c e si m i l a rities with known to x ins
bas e d on i n f o r mation r e por t e d b y D A S ( D A S 201 1 , p a ge 62). Amino a c id ho m o lo g ies w e re

e v a luat e d us i ng a g lo b a l sequ e n c e si m i l a ri t y s ea r c h a g a inst the Ge nBank no n

- r e dun d a nt protein
d a tas e t a nd the on l y s i g nifi c a nt ho mo l o g ies id e nt i fi e d w e re with o t h e r a lp h a

- k e t o glu t a r a te
d e p e nd e nt d i o x yg e n a s e s, the s a me c lass of e n z y m e s as A A D

- 1. D A S re p o rt e d that none of the
si m i l a r p r ote i ns r e turn e d b y the s ea r c h i d e nt i fi e d a n y s a fe t y c o n ce rns th a t m i g ht a r i se f rom the

e x p re ss i on o f A A D

- 1 pr o tein in plants. The aad

- 1 g e n e , iso l a ted f rom a soil ba c t e riu m , a nd the

A A D

- 1 p r otein a re both p r e s e nt in natu re . The soil ba c t e r iu m , Sphingob i um h e rbicido v oran s , is

n e i t h e r a p l a nt pest nor a known p e st of o r g a nis m s be n e fi c ial to a g ri c ul t u r e , a n d the AAD

- 1

pr o tein is si m i l a r to p r oteins found in other soil b ac te r ia t h a t a r e a lso n ot c onsid e r e d p e s ts ( W ri g ht et a l. 2009; B a lkwi l l et a l. 200 6 ).

A n y e f f

ec ts on no n

- t a r g e t or g a nis m s that could p o tential l y re sult f rom p r o p osed c h a n g e s in h e rbi c ide l a b e ls and/or any new herbicide metabolites produced as a result of the genetic modification will be e v a luat e d b y the E P A. This is addressed in the EIS prepared for this DAS petition.



### P o ten t i a l t o Tr a n s f e r Gene t i c In f o r m a t i o n f r o m DA S

- 40 2 7 8

- 9 C o rn t o Or g a n i s m s wi th w h i ch i t c a nn o t In t erb r eed

AP H I S e x a m i n e d the po t e nt i a l for the g e n e t i c ma t e ri a l i nse r ted into DA S

- 4 0278

- 9 c o r n to be
hori z ontal l y t r a nsf e r re d t o other o r g a nis m s without se x u a l r e p r odu c t i on a nd wh e ther such a n

e v e nt could le a d di r ec t l y or ind i r e c t l y to d i s ea s e , d a m a g e , in j u r y or h a rm to pl a nts, i n c lud i ng the

c r e a t i on of mo r e virulent p lant p e st s . The hori z on t a l gene tr a n s f e r ( H GT) b e tw ee n un r e lat e d

o r g a nis m s is one of the most in t e nsive l y stud i e d fi e lds i n the bi o

- s c ien c e s s i n c e 1940, a nd the
is s ue g a i n e d e x tra a t t e nt i on with t he re le a s e of t r a ns g e nic p l a nts i nto t he e n vironment ( D r ö g e e t

a l. 1998). P otential risks f r om s t a ble H GT f r om g e n e t i ca l l y e n g in e e r e d o r g a nis m s to anoth e r

o r g a nism without re p r o d u c t i on or human inte r v e n t i on w a s r e c e nt l y r e vie w e d ( K e e se 2008 )

. Me c h a nis m s of H GT i n c lude c onj u g a t i on, tr a nsf o r mation and tr a nsdu c t i on, a nd other dive r s e

me c h a nis m s of D NA a nd RNA uptake, re c omb i n a t i o n , a nd r e a r r a n g e m e nt, that o cc ur most

13notab l y thro u g h virus e s a nd mob i le g e n e t i c e lem e nts. H G T h a s be e n i m pl i ca ted a s a

c ontributor to major tr a nsi t ions i n e volu t ion, i n c lud i ng the sp r ea d of a nt i bio t ic r e si s tan c e

a mo n g st p a thogenic b a c te r ia a nd the e me r g e n c e o f in c r ea s e d virulen c e in b a c te r ia, e uk a r y ote s ,

a nd virus e s.

P o t e n tial f or Hori z o n tal G e n e T r a n s f e r t o Ba c te r ia or F un gi

HGT f r om a p l a nt spe c i e s to other b ac te r i a l spe c i e s is unl i k e l y to o cc ur b a s e d on the

f

ol l owing obse r v a t i ons. Althou g h t h e re a re ma n y opp o rtunities f or plants to dir ec t l y i nte rac t
wi t h fu n g i a nd b ac t e ria ( e

. g

. a s co m mens a ls, s y mb i onts, p a rasi t e s, patho g e ns, d ec om p ose r s, or in the g uts of h e rbivo re s), t h e re a re a l most no e volu t iona r y e x a mp l e s of H GT to b ac t e ria

f rom e uk a r y o t e s or f r om p l a nts t o fu n g i ( a s r e vie we d in K ee se 200 8 ). The on l y g e n e s l i k e l y

to be tr a nsf e rre d suc c e ssfu l l y f rom g e n e t ic a l l y e n g i n ee r e d plants to b ac te r ia a r e other b a c te r i a l

gen e s. Ho r i z ontal tr a nsf e r o f the aad

- 1 g e n e c onstru c t f rom the n uc l ea r g e no m e of DA S

40 2 78

- 9 c o r n a nd subsequ e nt e x p re ss i on in b ac te r ia is unlike l y to o c c u r

. F irst, m a n y

g e nom e s (or p a rts the r e o f ) h a ve b e e n se q u e n ce d f r o m ba c te r ia th a t a re c lose l y a ss o c iat e d with

p l a nts i n c lud i ng Agroba c terium a nd Rhi z obium ( Ka n e ko e t al. 20 0 0; W ood e t al. 2001;

K a n e ko e t al. 2002 )

.

Th e re is no e vide n c e that these o r g a nis m s c o ntain g e n e s de r i v e d f r om p l a n t s. S ec ond, in c a s e s wh e re r e view o f s e qu e n c e d a ta imp l ied th a t HGT o cc u r r e d, these e v e nts a r e inf e r r e d to oc c u r on

a n e volu t iona r y t i me s ca l e on the o r d e r of m i l l ions of y e a rs ( K oonin et a l. 2 001; B ro w n 200 3 ). Third, the 6236 b a se p a i r inse r t i on made to con f e r the h e rbi c ide tol e r a n c e to DA S

- 40 2 78

- 9 c o r n is a small f rac t i on of a p e r c e nt ( a ppro x i m a te l y 0. 0 0025 % ) of the total D N A (a ppro x i m a te l y 2500 m i l l ion base p a irs) f ound in t he c o r n g e nome. F o r t h , tr a nsge n e D N A p r o m ote r s and

c odi n g s e qu e n ce s a re opt i m iz e d for plant e x p re ss i on, not prok a r y ot i c b a c te r ial e x p re ss i on so

e v e n if HGT o cc u r re d, p rot e ins c o rr e spondi n g to t he tr a n s g e n e s a r e not l ike l y to b e pro d u ce d.

F inal l y , the F DA e v a luat e d HGT f r om t he use o f a nt i bio t ic re si s ta n c e m a rk e r g e n e s a nd

c on c lu ded t h a t the likelihood of tr a nsf e r of a nt i bio t ic re si s tan c e g e n e s f r om p l a nt g e no m e s to

m ic r oorg a nis m s in the g a stroin t e st i n a l t r a c t of humans or a ni m a ls, or in t he e nvironme n t, i s

r e mo t e ( F DA 199 8 ).

P o t e n tial f or Hori z o n tal G e n e T r a n s f e r t o V i r u s e s

AP H I S a lso consi d e r e d wh e ther hori z ontal tr a ns f e r of D N A f rom D A S

- 40278

- 9 c o r n to p l a nt
virus e s w a s l i k e l y to oc c ur a nd would le a d to t he c r e a t i on or s e le c t i on of a more virulent plant

p a thogen th r o u g h re c o m bination wi t h other plant virus e s. This i ssue h a s be e n c onsid e r e d b e fo r e b y other s c ien c e re v i e w p a n e ls and g ov e rnm e nt r e g ulato r y bodies ( f or a g e n e r a l r e view of the
issue s e e K e e se 2008 )

. B eca use t h e re a re no v i rus s e qu e n ce s u s e d a s p a rt of the ins e rt e d g e n e

ca ssette a nd no s e qu e n ce s that have b ee n i m pl i ca t e d in v i r a l r e c omb i n a t i on or p a th o g e nici t y , the r e is no r e a son to t hink t h a t v i r a l r ec omb i n a t i o n of the h e rbi c i d e

- r e si s tan c e c on f e rr i n g g e n e t i c

mat e ri a l wi l l oc c u r , o r th a t such the o r e t ic a l r e c omb i n a t i on would h a ve a n y i m p ac t on p l a nt pest
risk.

P o t e n tial f or Hori z o n tal G e n e T r a n s f e r t o P a r a sitic P la n ts

R ece nt l y , Yoshida e t al. ( 2010) throu g h a c omp a rat i ve g e nom i c s a n a l y sis i m pl i ca ted H G T for

14t h e inco rpor a t i on of a s p ec if i c g e n e t i c seq u e n c e in the p a r a si t ic pl a nt purple wi t c hw ee d ( Str i ga h e rmonthi c a )

, w hich in fe sts ce r e a l fi e lds (mono c o t s ) including c o r n a nd so r g hum ( Sorghum
bicolor ) . A cc o rding to t h is s tu d y , t h e inco rpo r a t i on of the spe c ific g e n e t i c s e qu e n c e ( w i t h a n unknown f un c t i on) o c c u r r e d b e tw e e n so r g hum a n d pu rple w i t c hw e e d b e f o re spe c i a t i on of pur p le
witch wee d ( S. h e rmonth i c a ) a nd r e lat e d c owp e a w i t c hw ee d ( S. g e snerioid e s ) , a p a r a si t ic pl a nt of dicots, f rom their c om m o n a n ce sto r

. I n other w o r ds, H G T b e tw e e n a p a r a s i t ic pl a nt and its host
is an e x tr e me l y r a re e v e nt

, a nd f u rth e rmo r e , S. h e r month ica is not f ound in t he U.S. and S.

asia t ica

, a nother re la t e d p a r a si t e of ce r ea l c rops, is on l y pr e s e nt i n No r th C a rolina a nd

S outh C a rolina ( USDA, N RC, 2010 )

.

F or a ll the r ea sons l is t e d a bov e , AP H I S c o n c ludes that HGT is un l ike l y to o cc ur a nd thus

po s e s no plant p e st risk.



## C o nclusi o n

AP H I S h a s r e vie w e d a nd c on d u c ted a p l a nt pest r i sk ass e ss m e nt on DA S

- 4 0278

- 9 c o r n. A P H IS

c o n c ludes that DA S

- 40278

- 9 c o r n is un l ike l y to pose a p l a nt p e st risk for the following reasons:

• Th ere is no p lant p e st risk f rom the ins e rt e d g e n e t i c mat e ri a l

.

• DA S

- 40278

- 9 c o r n w as not observed to disp lay atypical responses to d i s ea se or pl a nt

p e sts in t he f ield

.

• DA S

- 40278

- 9 c o rn does not h ave phenotypic characteristics that would c ontribute to
enhanced weediness ; and despite its greater herbicide resi stance, volunteers of DAS

40278

- 9 can still be controlled with some other currently ava ilable herbicides or other control methods in situations where they are considered weeds

. As with other herbicide resistant corn varieties available on the market, m anagement strategies c an be used to minimize the potential for s tacked herbicide resistant corn volunteers to become a weed management problem.

• DAS

- 40278

- 9 corn is not expected to increase weediness of other sexually

- compatible
wild relatives with which it can interbreed

.

• DAS

- 40278

- 9 corn i s not expected to have adverse e f f

e c ts on no n

- ta r g e ts or b e n e fi c i a l

org a nis m s in t he a g r o

- e c o s y stem

.

• Horizontal gene transfer from DAS

- 40278

- 9 to other species with which it cannot
interbreed is unlikely to occur and thus poses no plant pest risk.



### R eferenc e s

A i n l e y , M., K

. A r m s tr on g , S. B e l m a r , O . Fol k e r t s, N. H op k i ns, M

. A

. M e n k e, D. Pa r edd y , J . F. P e t o l i no, K . S m it h, a nd A . W o os l e y . 2002. R e g u l a t o r y Seq u enc e s f r om T r a n s g en i c P l a n t s ( U .S. P a t e n t
U S6384207)

A ll en, G. C

., S. S p i k e r , and W. F. T ho m pson. 20 0 0. U s e of m a t r i x a t t ach m ent r e g i ons ( MA R s) t o

m

i n i m i z e tr a n s g ene s il e nc i n g . Pl a nt Mo l ec u l ar B i o l o g y 43 : 361.

B a k e r , H

. B . 1965. C h a r a c t e r i s t i cs and m odes o f o r i g i n of w ee d s

. i n H

. G

. B a k er a n d G

. L. S t eb b i ns ( e ds )

. T he g ene t i cs o f c o lo n i z i ng sp e c i e s

. A cade m i c P r e s s, L ondon, U

. K

.

15Balkwill, D., J. Fredrickson, and M. Romine. 2006. Chapter 6.10: Sphingomonas and related genera. The Prokaryotes 7:605

- 629.

B ec k e tt

, T

. H

. and E. W. S t o ll e r . 1 9 88. V o l un t eer co r n (Zea mays ) i n t e rf e r en c e i n s o y beans (Glycine max)

. W eed S c ie nce 36: 1 5 9

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B ec k i e, H. J

. a nd M. D. K

. Ow en. 2007. H e r b i c i d e

- r e s i s t a n t c r ops as w e eds i n N o r t h A m e ri ca. CA B

r e v i ews: P A V S NN R 2 : 1

- 2 2.

B r o w n, J

. R . 2 0 03. A n c i e n t ho ri z o n t al g ene t r an s f e r

. N a t u r e Re v i ew s G en e t i cs 4 : 121

- 132.

C h ri s t e n s e n, A

. H

., R

. A

. S ha r r oc k , and P. H

. Q u a il

. 1 992. M a i z e p o l y ub i qu i t i n g ene s : s tr u c t u r e, t h e r m al

pe r t u r b a t i on o f ex p r e s s i on and t r a n s c r i p t s p li c i n g , and p r o m o t er a c t i v it y f o l l o wi ng tr a ns f er t o p r o t o p l a s t s by e l ec t r op o r a ti o n. P l ant M o l ec u l a r B i o l o g y 18 : 675

- 689.

C h ri s t e n s e n, A

. H

., Q u a i l

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