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(12) Patent Application Publication (10) Pub. No.: US 2016/0347799 A1 BARRY Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2016/0347799 A1 BARRY Et Al US 2016.0347799A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2016/0347799 A1 BARRY et al. (43) Pub. Date: Dec. 1, 2016 (54) INSECTICIDAL PROTEINS FROM PLANTS Related U.S. Application Data AND METHODS FOR THEIR USE (60) Provisional application No. 62/051,720, filed on Sep. (71) Applicants:PIONEER HI-BRED 17, 2014, provisional application No. 61/937.295, ppl1canus: filed on Feb. 7, 2014. INTERNATIONAL, INC., Johnston, IA (US); E. I. DUPONT DE Publication Classification NEMOURS AND COMPANY., (51) Int. Cl Wilmington, DE (US) C07K I4/45 (2006.01) CI2N 5/82 2006.O1 LU LIU, PALO ALTO, CA (US); AMY (52) U.S. Cl. LUM, HAYWARD, CA (US); ERIC CPC ............. C07K 14/415 (2013.01); A0IN 37/18 SCHEPERS, PORT DEPOSIT, MD (2013.01); C12N 15/8286 (2013.01) (US); NASSERYALPANI, JOHNSTON, IA (US); GENHAI ZHU, (57) ABSTRACT SAN JOSE, CA (US) Compositions and methods for controlling pests are pro s vided. The methods involve transforming organisms with a (73) Assignees: PIONEER HI-BRED nucleic acid sequence encoding an insecticidal protein. In INTERNATIONAL INC particular, the nucleic acid sequences are useful for prepar JOHNSTON, IA (US). E i. DU PONT ing plants and microorganisms that possess insecticidal DE NEMOURS AND CoMPANY activity. Thus, transformed bacteria, plants, plant cells, plant s tissues and seeds are provided. Compositions are insecti WILMINGTON, DE (US) cidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors (21) Appl. No.: 15/116,740 for Subsequent transformation into organisms of interest (22) PCT Filed: Feb 6, 2015 including plants, as probes for the isolation of other homolo a vs. gous (or partially homologous) genes. The pesticidal pro (86). PCT No.: PCT/US 15/14824 teins find use in controlling, inhibiting growth or killing Lepidopteran, Coleopteran, Dipteran, fungal, Hemipteran S 371 (c)(1), and nematode pest populations and for producing composi 2) Date: Aug.g 4, 2016 tions with insecticidal activitv.y Patent Application Publication Dec. 1, 2016 Sheet 1 of 21 US 2016/0347799 A1 Fig. Polypodiaceae DaValiaceae Oleancraceae -C Tectariaceae Eupolypodsh.c LOmariopsicaceae mid Dryoptericaceae Eupolypods, OnOceaceae Blechnaceae Polypodiales Eupolypods|FFE Thelyptericaceae C Woodsiaceae Aspleniaceae Polypod Ptericaceae DennStaectaceae Polypodiopsida SaCColomataceae -----C Lindsaeaceae Metaxyaceae DickSOniaceae C Cyatheaceae Cootaceae Cyatheales Core leptospOrangiates Plagiogyriaceae Culcitaceae LOXOmataceae Thyrsopteridaceae Salviniaceae Marsleaceae Salviniaceales Schizaeaceae AnemiaCeae Schizaeaceales Lygodiaceae Matoniaceae Leptosporangiate Diptericaceae Gleicheniales Gleicheniaceae Hymenophyllaceae Hymenophyllales OSMUndaceae Osmundales Fern EquisetaceaeMarataceae EquisetalesMarataes -J EquisetopsidaMaratopsida OphiogloSSaceaePsilotaceae Psilotalesge. PsilotooscaO Patent Application Publication Dec. 1, 2016 Sheet 3 of 21 US 2016/0347799 A1 Fig.2b - - - - MOTIF 20 ----- 120 140 Pt.IP-83Aa PTDGRQS---DTELHLTFMKL AI-QREE-NRWEITAADG----MNWGVY : 132 PtP-83Ca PIDGSQ- - - -ATTLRLPFMQL AR-VIEQNVKSEITATDG----MNWGIY : 129 PIP-83C ETDGSE- - - -AAWLELPEMEL T--LIEQ-YRSEIKAADD----AKWGTY : 128 Pt.IP-83CC PIDGSQ- - - -ATTLRLPFMQL. AR-VIEQNVKSEITATDG----MNWGIY : 129 Pt.IP-83Cd PIDGSQ----ATTLRLPFMQL AR-VIEQNVKSEITATDG----MNWGIY : 129 Pt.IP-83Ce PIDGSQ- - - -ATTLRLPFMOL AR-VIEONVKSEITATDG----MNWGIY : 129 PEIP-83CfE PTGGSR----AATLCLPFMKV AT-LIER-FRSEIAAADG----MNWGTY : 128 Pt.IP-83Fa PEVDQRPHMAQCSLDFPFMRL VVGSWHENVGGVMQAFSSDATPSNIGIY : 139 gi-598378568 SSGNGWP--QVALLQGPLVQYNGN---------- YSPWTG- - - - - NWSAT : 127 PtIP-83Aa SSE : CCCCCCC---CCHE HH 132 PtIP-83Ca SSE : CCCCCC- - - -C 129 PtIP-83Cb SSE : CCCCCC----HHHH 128 PtIP-83Cc SSE : CCCCCC----CCCCCCC 129 PtIP-83Cd SSE : CCCCCC----CCCCCCC 129 PtIP-83Ce SSE : CCCCCC----CCCCCCCH 129 PtIP-83Cf SSE : CCCCCC----CEECCCCC 128 Pt.IP-83Fa SSE : 139 gi-598378568 127 160 180 200 Pt.IP-83Aa IHA EVOVGVLTMSWSSVLRVSALRSVITSGFRA------------- WSW 169 Pt.IP-83Ca : IYG KVERSPLLPSNA-ILAWWADRCISAR- - - - - - - - - - - - - - - HNH : 163 Pt.IP-83Cb : WHA EVOLSPLFNGWP-YLVVEAQRCIITAAM--------------- HNT : 162 Pt.IP-83CC YG KWERSPLPSNA-AWWADRCTTSAR- - - - - - - - - - - - - - - HNH : 163 Pt.IP-83Cd YG EVERSPLLPSNA-LAWWADRCTTSAR- - - - - - - - - - - - - - - ENE : 163 Pt.IP-83Ce YG KVERSPLLPSNA-ILAWWADRCISAR- - - - - - - - - - - - - - - HNH : 163 Pt.IP-83Cf IHG EVOVSPLYPNNS-ILGVWADRSIITSAF--------------- HNW : 162 Pt.IP-83Fa : LHADRFIYROATSPASNFVLPLDVRVSFGSSTYSGPTIRPDWONLNVSNI : 189 gi-598378568 : PLL QDTYR ----FFLHGQQLLILSGT------------------ 150 PtIP-83Aa SSE : CCC HCCCEE------------- se 169 PtIP-83Ca SSE : 163 PtIP-83Cb SSE : 162 PtIP-83Cc SSE : 163 PtIP-83Cd SSE : 163 PtIP-83Ce SSE : EccoccCCCCCEEEEEEE : 163 162 PtIP-83Cf SSE : CCCCCCCE-coccaccCCCCCCCc--------------- CCC : PtIP-83Fa SSE : EECCCH CCCCCCCCCCCCCCCCCCCCCC : 189 gi-598378568 : CCCCCCCCE--------- CCCCCEEECCC------------------ 150 Patent Application Publication Dec. 1, 2016 Sheet 10 of 21 US 2016/0347799 A1 Fig. 2. MOTIF 5 ------------------ (-- 82O 840 PtP-83Aa : TTOTGTVYILL GSTIFHDRRRDQVLPF AAAPLNYHYAYRLDTGDSTLT : BOO PtP-83Ca : STRSGIWYILL, GSTIEHDRRRDEWMTF AADPLNF YAYRLDTGEATLT : 786 PtP-83Cb : ATQTGTIYILL GSTVFHDRRREEVMTF AAVPLNYHYAYRLDTGEATLT : 785 PtP-83CC : STRSGIWYILL, GSTIEHDRRRDEVMT. AADPLNF YAYRLDTGEATLT : 784 PtP-83CC : STRSGIWYILL GSTIEHDRRRDEVMT. AADPLNE YAYRLDTGEATLT : 784 PtP-83Ce : STRSGIWYILL, GSTIEHDRRRDEWMTF AADPLNF YAYRLDTGEATLT : 784 PtIP-83Cf : ATQTGTVYILL, GSTIFHDRRRDEVMTF AAAPLNY YAYRLDTGETTLT : 785 PtP-83Ea : STNTGLVYMLL, GSRFLHDRKHEEVMDY ASILGPWYAYAYDLNTGATTLN : 817 gi-598378568 ATVPAYHYAYNLASETTED : 749 PtIP-83Aa SSE : EEEEECCCCCCCC : BOO PtIP-83Ca SSE : M CCCCCCCC : 786 PtIP-83Cb SSE : , , as CCCCCCCC : 785 PtIP-83cc SSE : CCCCCCCC : 784 PtIP-83Cd SSE : CCCCCCCC : 784 PtIP-83Ce SSE : CCCCCCCC : 784 PtIP-83Cf SSE : CCCCCCCC : 785 PtP-83Fa SSE : CCCCCCCC : 817 gi-598378568 CCCCCCCC : 749 MOTIF 23 ------------------- MOTIF 3 - - - - - - - - - - - - - - 860 880 900 PtF-83Aa : NEPSEQFANKFMQMTPFTRWRLRLSASAKENAGLAFPTATALDSTT IVI : 850 PtP-83Ca : NEPSEDFANTFMQMTPFTRWRLRLSASASENAELAFPTATAPDSTT 836 PtP-83Cb : NEPSEQEANTFMQMTPFTHWRLRLSASAAENKGLAFPTATAPDSTT 835 PtP-83CC : NEPSEEFANTFMQMTPFTRWRLRLSASASENAELAFPTATAPDSTT 834 PtP-83CC : NEPSEEFANTFMQMTPFTRWRLRLSASASENAELAFPTATAPDSTT 834 PtP-83Ce : NEPSEEFANTFMQMTPFTRWRLRLSASASENAELAFPTATAPDSTT 834 Pt.IP-83Cf : NOPSEHFANTFMQMTPFTRWRLRLSASAPENAGLAFPTATALDSTT 835 PtP-83Fa : NIPSQQYANTFMQMTPFNAWRLRLSASAVENQGLVFPTATSPDNTT 867 gi-598378568 : NRPDEEFARVFMRMTPFTRWKLRVSTSARENQGLAFPTAITPDATT : 799 PtIP-83Aa SSE : VV V. CCCCCCCCCCCCCCCEEVVVV VVV EE 850 PtIP-83Ca SSE : CCCCCCCCCCCCCCCCCCEEEEEECCCCCCCCCCCCCCCCCCCCEE 836 PtIP-83Cb SSE : CCCCCCCCCCCCCCCCCC CCCCCCCCCCCCCCCCCC 835 PtIP-83Cc SSE : CCCCCCCCCCCCCCCCCC CCCCCCCCCCCCCCCCCCC 834 PtIP-83Cd SSE : CCCCCCCCCCCCCCCCCC CCCCCCCCCCCCCCCCCCC 834 PtIP-83Ce SSE : CCCCCCCCCCCCCCCCCC CCCCCCCCCCCCCCCCCCC 834 PtIP-83Cf SSE : cCCCCCCCCCCCCCCCCC CCCCCCCCCCCCCCCCEE 835 Pt.IP-83Fa SSE : CCCCCCCCCCCCCCCCCCC CCCCCCCCCCCCCCCCCC 867 gi-598378568 : CCCCCCCCCCCCCCCCCCEEEE CCCCCCCCCCCCCCCCCCCEE : 799 Patent Application Publication Dec. 1, 2016 Sheet 11 of 21 US 2016/0347799 A1 Fig. 2 920 PtP-83Aa : TFHVTAIR IDWRHDEE--- 867 PtP-83Ca : TFHVTAIR WDWRQEEE--- 853 Pt.IP-83Cb : TFHVTAIR IDWRQEEE--- 852 PtP-83CC : TFHVTAIR VDWRQEEE--- 851 PtP-83CC : TFHVTAIR VDWRQEEE--- 851 Pt.IP-83Ce : TFHVTAIR WDWRQEEKEEE : 854 PtP-83Cf : TFHVTAIR IDWRHEEE--- 852 PtP-83Fa : TFYVTAIRRIDHRQEGDVE 886 gi-598378568 : RFYISAIRDFDWNIR- - - - - - 813 PtIP-83Aa SSE : ECCCCCC--- 867 PtIP-83Ca SSE : CCCCCCCC- - - 853 PtIP-83Cb SSE : ECCCCCCC--- 852 PtIP-83Cc SSE : CCCCCCCC- - - 851 PtIP-83Cd SSE : CCCCCCCC- - - 851 PtIP-83Ce SSE : 854 PtIP-83Cf SSE : 852 PtIP-83Fa SSE : 886 gi-598378568 813 Patent Application Publication Dec. 1, 2016 Sheet 13 of 21 US 2016/0347799 A1 Fig. 3b 6O1 W l 650 Pt(P-50Aa (547) QAD-RLSSIKPADLTNYLE MVSEMDMRTTRMLLELIRVLYIONAALOYEYLE. H PtIP-83Aa (597). OADRLAAICPADLTNYLEMATOMDMRTTRMLLGLLNILRIONAALRYEYL 651 7 OO PtIP-50Aa (597) OTPAPLNAWPVTMQTVWGLLVOOETAAINGLLOMGAPSDYTOEYAVRDVP PtIP-83Aa (647) LMPTELTTWPLGMDTVGDLLIAOENAALIGLMOLGPSSDFTSRHVVKDIP PtIP-50Aa (647) VRLLLGGGDWEFELPVRN--ADFPLTWCRVRIRYVDMRFDAAAE------ PtIP-83Aa (697) VNLLLDGEDWEFEIPVOAGMSSFPSSWSRVRIRHLEMHFVKEASGIGGE 751. 8 OO PtIP-50Aa (689) -HLPVTSTGEVYMLLOSSRFFEDRAKRENEFISYEGGMGLOYOYAYRLAT PtIP-83Aa (747) IHOPTTOTGIVYILLOGSTIFHDRRR--DOVLPFOAAAPLNYHYAYRLDT 8O1 85 O PtIP-50Aa (738). GDATVTNVPSEEYANTFMRLAPFTRWRLRLSASAPENKGLAFPTATLADA PtIP-83Aa (795) GDSTILTNEPSEOFANKFMOMTPETRWRLRLSASAKENAGLAFPTATALDS 851 873 PtIP-50Aa (788) TTRIKITFHVSAIRRISTRVAV PtIP-83Aa (845) TTOIVITFHVTAIROIDWRHDEEH Patent Application Publication Dec. 1, 2016 Sheet 21 of 21 US 2016/0347799 A1 Fig. 5e 801 850 PtIP-83Aa (793) DTGDSTLTNEPSEQFANKFMQMTPFTRWRLRLSASAKENAGLAFPTATAT, PtIP-83ca (779) DTGEATLTNEPSEDFANTFMQMTPFTRWRLRLSASASENAELAFPTATAP PtIP-83Cb (778) DTGEATLTNEPSEQFANTFMQMTPFTHWRLRLSASAAENKGLAFPTATAP PtIP-83cc (777) DTGEATLTNEPSEEFANTFMQMTPFTRWRLRLSASASENAELAFPTATAP PtIP-83Cd (777) DTGEATLTNEPSEEFANTFMQMTPFTRWRLRLSASASENAELAFPTATAP PtIP-83ce (777) DTGEATLTNEPSEEFANTFMQMTPFTRWRLRLSASASENAELAFPTATAP PtIP-83Cg (778) DTGEATLTNEPSEQFANTFMQMTPFTHWRLRLSASAAENEGLAFPTATAP Pt(P-83Cf (778) TTLTNPSEHFANTFMQMTPFTRWRLRTSASAPENAGLAFPTATAT, PtIP-83Da (777) f:------------------------------ -------------- PtIP-83Aa (843) Pt.IP-83Ca (829) PtIP-83Ch (828) PtIP-83Cc (827) Pt.IP-83Cd (827) Pt.IP-83Ce wVITFHVTAIRQVDWRQEEKEEE PtIP-83Cg (828) DSTTEIAITFHVMAIRQIDWRQEEE--- PtIP-83Cf
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