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WO 2015/038734 A2 19 March 2015 (19.03.2015) P O P C T (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2015/038734 A2 19 March 2015 (19.03.2015) P O P C T (51) International Patent Classification: Not classified International, Inc., 7250 N.W. 62nd Avenue, Johnston, Iowa 5013 1-0552 (US). SCHELLENBERGER, Ute; c/o (21) International Application Number: Pioneer Hi-Bred International, Inc., 7250 N.W. 62nd Av PCT/US20 14/055 128 enue, Johnston, Iowa 5013 1-0552 (US). UDRANSZKY, (22) International Filing Date Ingrid; c/o Pioneer Hi-Bred International, Inc., 7250 N.W. 11 September 2014 ( 11.09.2014) 62nd Avenue, Johnston, Iowa 5013 1-0552 (US). WEI, Jun-zhi; c/o Pioneer Hi-Bred International, Inc., 7250 (25) Filing Language: English N.W. 62nd Avenue, Johnston, Iowa 5013 1-0552 (US). (26) Publication Language: English XIE, Weiping; c/o Pioneer Hi-Bred International, Inc., 7250 N.W. 62nd Avenue, Johnson, Iowa 5013 1-0552 (30) Priority Data: (US). ZHU, Genhai; c/o Pioneer Hi-Bred International, 61/877,625 13 September 2013 (13.09.2013) US Inc., 7250 N.W. 62nd Avenue, Johnston, Iowa 5013 1-0552 (71) Applicant: PIONEER HI-BRED INTERNATIONAL, (US). INC. [US/US]; 7100 N.W. 62nd Avenue, Johnston, Iowa (74) Agent: BAUER, S. Christopher; Pioneer Hi-Bred Interna 5013 1-1014 (US). tional, Inc., 7250 N.W. 62nd Avenue, Johnston, Iowa (72) Inventors: DIEHN, Scott; c/o Pioneer Hi-Bred Interna 5013 1-0552 (US). tional, Inc., 7250 N.W. 62nd Avenue, Johnston, Iowa (81) Designated States (unless otherwise indicated, for every 5013 1-0552 (US). ENGLISH, James; c/o Pioneer Hi-Bred kind of national protection available): AE, AG, AL, AM, International, Inc., 7250 N.W. 62nd Avenue, Johnston, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, Iowa 5013 1-0552 (US). LIU, Lu; c/o Pioneer Hi-Bred In BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, ternational, Inc., 7250 N.W. 62nd Avenue, Johnston, Iowa DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, 5013 1-0552 (US). ONG, Azalea; c/o Pioneer Hi-Bred In HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, ternational, Inc., 7250 N.W. 62nd Avenue, Johnston, Iowa KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, 5013 1-0552 (US). ORAL, Jarred; c/o Pioneer Hi-Bred In MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, ternational, Inc., 7250 N.W. 62nd Avenue, Johnston, Iowa PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, 5013 1-0552 (US). ROSEN, Barbara; c/o Pioneer Hi-Bred [Continued on nextpage] (54) Title: INSECTICIDAL PROTEINS AND METHODS FOR THEIR USE (57) Abstract: Compositions and methods for controlling pests are provided. The meth Fig. 6 ods involve transforming organisms with a nucleic acid sequence encoding an insecticid- al protein. In particular, the nucleic acid se quences are useful for preparing plants and microorganisms that possess insecticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are insecticidal nuc leic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transforma tion into organisms of interest including plants, as probes for the isolation of other ho mologous (or partially homologous) genes. The pesticidal proteins find use in con trolling, inhibiting growth or killing Lepidop- teran, Coleopteran, Dipteran, fungal, Hemi- < pteran and nematode pest populations and for producing compositions with insecticidal activity. 00 © o o w o 2015/038734 A2 1inn nil 1mil i mil il i 1ill inn i l i urn i imill 11 i i SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, Declarations under Rule 4.17: TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. — as to applicant's entitlement to apply for and be granted (84) Designated States (unless otherwise indicated, for every apatent (Rule 4.1 7(H)) kind of regional protection available): ARIPO (BW, GH, Published: GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, — without international search report and to be republished TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, upon receipt of that report (Rule 48.2(g)) DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, — with sequence listing part of description (Rule 5.2(a)) LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). INSECTICIDAL PROTEINS AND METHODS FOR THEIR USE REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY A sequence listing having the file name "5345PCT_sequence_listing.txt" created on August 28, 2014, and having a size of 576 kilobytes is filed in computer readable form concurrently with the specification. The sequence listing is part of the specification and is herein incorporated by reference in its entirety. FIELD OF THE INVENTION This disclosure relates to the field of molecular biology. Provided are novel genes that encode pesticidal proteins. These pesticidal proteins and the nucleic acid sequences that encode them are useful in preparing pesticidal formulations and in the production of transgenic pest-resistant plants. BACKGROUND OF THE INVENTION Biological control of insect pests of agricultural significance using a microbial agent, such as fungi, bacteria or another species of insect affords an environmentally friendly and commercially attractive alternative to synthetic chemical pesticides. Generally speaking, the use of biopesticides presents a lower risk of pollution and environmental hazards and biopesticides provide greater target specificity than is characteristic of traditional broad-spectrum chemical insecticides. In addition, biopesticides often cost less to produce and thus improve economic yield for a wide variety of crops. Certain species of microorganisms of the genus Bacillus are known to possess pesticidal activity against a range of insect pests including Lepidoptera, Diptera, Coleoptera, Hemiptera and others. Bacillus thuringiensis (Bt) and Bacillus popilliae are among the most successful biocontrol agents discovered to date. Insect pathogenicity has also been attributed to strains of B. larvae, B. lentimorbus, B. sphaericus and B. cereus. Microbial insecticides, particularly those obtained from Bacillus strains, have played an important role in agriculture as alternatives to chemical pest control. Crop plants have been developed with enhanced insect resistance by genetically engineering crop plants to produce pesticidal proteins from Bacillus. For example, corn and cotton plants have been genetically engineered to produce pesticidal proteins isolated from strains of Bt. These genetically engineered crops are now widely used in agriculture and have provided the farmer with an environmentally friendly alternative to traditional insect-control methods. While they have proven to be very successful commercially, these genetically engineered, insect-resistant crop plants provide resistance to only a narrow range of the economically important insect pests. In some cases, insects can develop resistance to different insecticidal compounds, which raises the need to identify alternative biological control agents for pest control. Accordingly, there remains a need for new pesticidal proteins with different ranges of insecticidal activity against insect pests, e.g., insecticidal proteins which are active against a variety of insects in the order Lepidoptera and the order Coleoptera including but not limited to insect pests that have developed resistance to existing insecticides. SUMMARY OF THE INVENTION Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions include nucleic acid molecules encoding sequences for pesticidal and insecticidal polypeptides, vectors comprising those nucleic acid molecules, and host cells comprising the vectors. Compositions also include the pesticidal polypeptide sequences and antibodies to those polypeptides. The nucleic acid sequences can be used in DNA constructs or expression cassettes for transformation and expression in organisms, including microorganisms and plants. The nucleotide or amino acid sequences may be synthetic sequences that have been designed for expression in an organism including, but not limited to, a microorganism or a plant. Compositions also comprise transformed bacteria, plants, plant cells, tissues and seeds. In particular, isolated or recombinant nucleic acid molecules are provided encoding Pseudomonas Insecticidal Protein-72 (PIP-72) polypeptides including amino acid substitutions, deletions, insertions, and fragments thereof, and combinations thereof. Additionally, amino acid sequences corresponding to the PIP-72 polypeptides are encompassed. Provided are isolated or recombinant nucleic acid molecules capable of encoding a PIP-72 polypeptide of SEQ ID NO: 849 as well as amino acid substitutions, deletions, insertions, fragments thereof and combinations thereof. Nucleic acid sequences that are complementary to a nucleic acid sequence of the embodiments or that hybridize to a sequence of the embodiments are also encompassed. Also provided are isolated or recombinant PIP-72 polypeptides of SEQ ID NO: 849 as well as amino acid substitutions, deletions, insertions, fragments thereof and combinations thereof. Methods are provided for producing the polypeptides and for using those polypeptides for controlling or killing a Lepidopteran, Coleopteran, nematode, fungi, and/or Dipteran pests. The transgenic plants of the embodiments express one or more of the pesticidal sequences disclosed herein. In various embodiments, the transgenic plant further comprises one or more additional genes for insect resistance, for example, one or more additional genes for controlling Coleopteran, Lepidopteran, Hemipteran or nematode pests. It will be understood by one of skill in the art that the transgenic plant may comprise any gene imparting an agronomic trait of interest. Methods for detecting the nucleic acids and polypeptides of the embodiments in a sample are also included.
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