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United States Patent (10) Patent No.: US 9,556.448 B2 Olivier Et Al USOO9556448B2 (12) United States Patent (10) Patent No.: US 9,556.448 B2 Olivier et al. (45) Date of Patent: Jan. 31, 2017 (54) IDENTIFICATION AND THE USE OF KRP 7,807,872 B2 * 10/2010 Frankard .............. CO7K 14,415 MUTANTS IN PLANTS 800,286 8,431,775 B2 4, 2013 Hegstad et al. (75) Inventors: Jean Paul Olivier, Seattle, WA (US); E. E: 38: y s al Dayna L. Loeffler, Seattle, WA (US) 2004/0019926 Al I/2004 Frankard et al. 2007/00441.71 A1 2/2007 Kovalic et al. (73) Assignee: Targeted Growth, Inc., Seattle, WA 2007.0056.058 A1* 3, 2007 Olivier ................. CO7K 14,415 (US) 800,290 2008/0134355 A1 6/2008 Van Camp (*)c Notice:- r Subject to any disclaimer, the term of this 2008/03075462008/0216193 A1 12/20089/2008 VeylderSavidge etet al. patent is extended or adjusted under 35 2009/0070894 A1 3/2009 Frankard et al. ............. 800,278 U.S.C. 154(b) by 190 days. 2009 OO87878 A9 4, 2009 La Rosa et al. 2009, O144863 A1 6/2009 Song et al. (21) Appl. No.: 14/111.292 2011/O135647 A1 6, 2011 Nakamura et al. 2012/0131698 A1 5, 2012 Olivier et al. 1-1. 2012/0284813 A1 11/2012 Olivier et al. (22) PCT Filed: Apr. 11, 2012 2014/0331362 A1 11/2014 Olivier et al. (86). PCT NO. PCT/US2012/033047 2016,0002656 A1 1/2016 Olivier et al. S 371 (c)(1), FOREIGN PATENT DOCUMENTS (2), (4) Date: Dec. 18, 2013 WO WO-99.64599 A 12, 1999 WO WOOOf 60087 A2 10, 2000 WO WO-2005/007829 A2 1, 2005 (87) PCT Pub. No.: WO2012/142106 WO WO-2005/024029 A2 3, 2005 WO WO 2006/081029 A2 8, 2006 PCT Pub. Date: Oct. 18, 2012 WO WO 2007/O16319 A2 2/2007 WO WO 2009/092009 A2 T 2009 (65) Prior Publication Data WO WO-2010/0990.83 A1 9, 2010 WO WO 2012/142116 A2 10, 2012 US 2014/O143900 A1 May 22, 2014 OTHER PUBLICATIONS Cheng et al., 2013, Plant J., 75:642-655.* Related U.S. Application Data Extended European Search Report in corresponding European Pat (60) Provisional application No. 61/474,201, filed on Apr. ent Application No. 127716777, mailed Sep 1, 2014. 11, 2011. AZZi et al., “Interaction Between the Cell-Cycle-Control Proteins p34* and p9''''', Evidence for Two Cooperative Binding (51) Int. Cl. Domains in p9'.” Eur: J. Biochem. 203:353–360 (1992). CI2N IS/00 (2006.01) Coats et al., “Requirement of p27 for Restriction Point Control AOIH 5/00 (2006.01) of the Fibroblast Cell Cycle.” Science 272:877-880 (1996). CI2N 5/82 (2006.01) Elmore et al., “Glyphosate-Resistant Soybean Cultivar Response to C07K I4/45 (2006.01) Glyphosate.” Agron. J. 93:404–407 (2001). AOIH I/04 2006.O1 AOIH 5/10 3:08: (Continued) (52) U.S. Cl. Primary Examiner — Jason Deveau Rosen CPC ............. C12N 15/8261 (2013.01); A0IH I/04 (74) Attorney, Agent, or Firm — Cooley LLP (2013.01); A0IH 5/10 (2013.01); C07K (57) ABSTRACT 14/415 (2013.01) The invention provides a plant cell, part, tissue culture or (58) Field of Classification Search whole plant comprising at least one disrupted KRP gene of None the present invention. The present invention also provides See application file for complete search history. methods of increasing weight, size, and/or number of one or more organs, and/or yield of a plant by utilizing the dis (56) References Cited rupted KRP genes of the present invention. Furthermore, methods of breeding plants to produce new plants having U.S. PATENT DOCUMENTS increased weight, size, and/or number of one or more 5,958,769 A 9, 1999 Roberts et al. organs, and/or yield are provided. The present invention E. A 63. She et al provides isolated Kinase Inhibitor Protein (KIP) Related 6.5 593 58 B1 5/2003 Murray Protein (KRP) polynucleotide sequences and isolated KRP SE R 1858 Irisha et al polypeptide sequences and methods of their use. Exemplary 7329799 B23. 3/2008 Savidge. C07K 14/415 plants include wheat, rice and soybean. 426,531 7,803,990 B2 9, 2010 Abbitt 5 Claims, 14 Drawing Sheets US 9,556.448 B2 Page 2 (56) References Cited Wang et al., “Genome-Wide Analysis of the Cyclin Family in Arabidopsis and Comparative Phylogenetic Analysis of Plant Cyclin-Like Proteins.” Plant Physiol. 135: 1084-1099 (2004). OTHER PUBLICATIONS Zhou et al., “Plant CDK Inhibitors: Studies of Interactions With Cell Elmore et at, “Glyphosate-Resistant Soybean Cultivar Yields Com Cycle Regulators in the Yeast Two-Hybrid System and Functional pared with Sister Lines,” Agron. J. 93:408-412 (2001). Comparisons in Transgenic Arabidopsis Plants.” Plant Cell Rep. Feig and Cooper, "Inhibition of NIH 3T3 Cell Proliferation by a 20:967-975 (2002). Mutant ras Protein with Preferential Affinity for GDP.” Mol. Cell. Zhou et al., “The Plant Cyclin-Dependent Kinase Inhibitor ICK1 Biol. 8:3235-3243 (1988). has Distinct Functional Domains for in vivo Kinase Inhibition, Fernandez-Cornejo, Agriculture Information Bulletin No. (AIB786) Protein Instability and Nuclear Localization.” Plant J. 35:476-489 81 pp, Feb. 2004. (2003). Fero et al., “A Syndrome of Multiorgan Hyperplasia with Features International Search Report based on International Patent Applica of Gigantism, Tumorigenesis, and Female Sterility in p27Kipl.- tion No. PCT/US2012/033047, mailed on Jun. 28, 2012. Deficient Mice,” Cell 85:733-744 (1996). Written Opinion by International Search Authority based on Inter Firpo et al., “Inactivation of a Cdk2 Inhibitor during Interleukin national Patent Application No. PCT/US2012/033047, mailed on 2-Induced Proliferation of Human T Lymphocytes.” Mol. 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Annual Review in Genetics, 40: 77-105 (2005). Nakayama et al., “Mice lacking p27' Display Increased Body Leenhardt et al. “Wheat lipoxygenase activity induces greater loss Size, Multiple Organ Hyperplasia, Retinal Dysplasia, and Pituitary of carotenoids than vitamin E during breadmaking'. J Agric Food Tumors,” Cell35:707-720 (1996). Chem, 54(5):1710-1715 (2006). Polyak et al., “Cloning of p27, a Cyclin-Dependent Kinase McKibbin et al., “Transcripts of Vp-1 homeologues are misspliced Inhibitor and a Potential Mediator of Extracellular Antimitogenic in modern wheat and ancestral species'. PNAS, 99(15): 10203 Signals.” Cell78:59-66 (1994). 10208 (2002). Russo et al., “Crystal Structure of the p27 Cyclin-Dependent NCBI BLAST alignment of SEQ ID No. 11 and SEQ ID No. 7 Kinase Inhibitor Bound to the Cyclin A-Cdk2 Complex,” Nature obtained from http://blast.ncbi.nlm.nih.gov/Biast.cgi on Nov. 11, 382:325-331 (1996). 2013. 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