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WO 2016/016855 Al 4 February 2016 (04.02.2016) P O P C T

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WO 2016/016855 Al 4 February 2016 (04.02.2016) 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 2016/016855 Al 4 February 2016 (04.02.2016) P O P C T (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every C12N 15/113 (2010.01) A01H 5/08 (2006.01) kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, (21) International Application Number: BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, PCT/IB2015/055802 DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (22) International Filing Date: HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, 31 July 20 15 (31 .07.2015) KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, (25) Filing Language: English PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, (26) Publication Language: English SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (30) Priority Data: 628200 1 August 2014 (01.08.2014) (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (71) Applicant: THE NEW ZEALAND INSTITUTE FOR GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, PLANT AND FOOD RESEARCH LIMITED [NZ/NZ]; TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, a Crown Research Institute established under the Crown TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, Research Institutes Act 1992, 120 Mt Albert Road, Mt A l DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, bert, Auckland, 1025 (NZ). 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, (72) Inventors: IRELAND, Hilary Sara; 36 Lynden Avenue, GW, KM, ML, MR, NE, SN, TD, TG). Hillcrest, Auckland, 0627 (NZ). SCHAFFER, Robert James; 9 Mount Royal Avenue, Mount Albert, Auckland, Published: 1025 (NZ). YAO, Jia-long; 270 Muriwai Road, — with international search report (Art. 21(3)) Waimauku, Auckland (NZ). — with sequence listing part of description (Rule 5.2(a)) (74) Agents: AJ PARK et al; Level 22, State Insurance Tower, 1 Willis Street, Wellington (NZ). (54) Title: METHODS AND MATERIALS FOR PRODUCING CORELESS FRUIT (57) Abstract: The invention provides materials and methods for producing coreless fruit, or plants that produce co- FIGURE 7 reless fruit. The invention involves combining reduced expression of AGAMOUS (AG) with parthenocarpy. Parthenocarpy can be induced by hor mone treatment, or can be provided by reduced or eliminated expression of PISTILATA (PI) or APETALA3 (AP3). The invention provides methods and materials for producing the plants and coreless fruit by genetic modifica tion (GM) and non-GM means. The in vention also provides the plants and co reless fruit. METHODS AND MATERIALS FOR PRODUCING CORELESS FRUIT TECHNICAL FIELD The present invention relates to methods and materials for producing coreless fruit. BACKGROUND ART Cores present in many fruit carry seed which, under suitable conditions, can germinate to ultimately produce a new fruit bearing plants. Fruit are typically attractive to animals, and seed ingested with the fruit, may be deposited by animals at distant locations from the original fruit bearing plant, resulting in spread of the fruit plant species. While a seed bearing core is clearly an evolutionary advantage for many of plants, presence of the core can be an inconvenience to humans. The cores of many fruits are fibrous and tough, and are therefore unpleasant for humans to eat and may be difficult to digest. For these reasons cores are often discarded by those eating fruit, or removed before fruit are further processed and/or incorporated into other food products. Such disposal or removal of cores represents a significant waste of the biomass of the fruit, and adds significantly to the cost of fruit processing. It is therefore an object of the invention to provide novel methods and compositions for producing coreless fruit, or at least to provide the public with a useful choice. SUMMARY OF THE INVENTION METHODS Reducing or eliminating AG In one aspect the invention provides a method for producing a coreless fruit, the method comprising reducing, or eliminating, expression of at least one AGAMOUS (AG) protein in a plant. In a further aspect the invention provides a method for producing a plant that produces at least one coreless fruit, the method comprising reducing or eliminating expression of an AGAMOUS (AG) protein in the plant. Method including the step of inducing parthenocarpy In one embodiment the method includes the additional step of inducing parthenocarpy in the plant. Therefore in one aspect the invention provides a method for producing a coreless fruit, the method comprising the steps: a) reducing, or eliminating, expression of at least one AGAMOUS (AG) protein in a plant, and b) inducing parthenocarpy in the plant. In a further aspect the invention provides a method for producing a plant that produces at least one coreless fruit, the method comprising the steps: a) reducing, or eliminating, expression of at least one AGAMOUS (AG) protein in a plant, and b) inducing parthenocarpy in the plant. Reducing or eliminating AG in parthenocarpic plant In one embodiment the plant in which expression of at least one AGAMOUS (AG) protein is reduced or eliminated is a parthenocarpic plant. Methods for inducing parthenocarpy Pathenocarpy be induced by any means. In one embodiment parthenocarpy is induced by application of plant hormones to flowers of the plant. In a further embodiment parthenocarpy is induced manipulating expression of genes controlling fruit set. In one embodiment parthenocarpy is induced manipulating the expression of at least one PISTILSTA (PI) gene or protein. In one embodiment parthenocarpy is induced reducing or eliminating expression of at least one PISTILSTA (PI) gene or protein. In one embodiment parthenocarpy is induced manipulating the expression of at least one APETALA3 (AP3) gene or protein. In one embodiment parthenocarpy is induced reducing or eliminating expression of at least one APETALA3 (AP3) gene or protein. Mutant parthenocarpic plants In one embodiment the parthenocarpic plant is a mutant plant with reduced or eliminated expression of at least one PISTILSTA (PI) gene or protein. In a further embodiment the parthenocarpic plant is a mutant plant with reduced, or eliminated, expression of at least one APETALA3 (AP3) gene or protein. The mutant plant may be a naturally occurring mutant plant. Alternatively the mutant may be an induced mutant. Reducing or eliminating AG and PI In one aspect the invention provides a method for producing a coreless fruit, the method comprising reducing, or eliminating, expression of at least one AGAMOUS (AG) protein and at least one PISTILATA (PI) protein in a plant. In a further aspect the invention provides a method for producing a plant that produces at least one coreless fruit, the method comprising reducing or eliminating expression of an AGAMOUS (AG) protein and at least one PISTILATA (PI) protein in the plant. In one embodiment the reducing or eliminating expression of the at least one PISTILATA (PI) protein induces parthenocarpy. Reducing or eliminating AG and AP3 In one aspect the invention provides a method for producing a coreless fruit, the method comprising reducing, or eliminating, expression of at least one AGAMOUS (AG) protein and at least one APETALA3 (AP3) protein in a plant. In a further aspect the invention provides a method for producing a plant that produces at least one coreless fruit, the method comprising reducing or eliminating expression of an AGAMOUS (AG) protein and at least one APETALA3 (AP3) protein in the plant. In one embodiment the reducing or eliminating expression of the at least one APETALA3 (AP3) protein induces parthenocarpy. Non-GM selection method for reduced or eliminated AGAMOUS (AG) In a further aspect the invention provides a method for identifying a plant with a genotype indicative of producing, or being useful for producing, at least one coreless fruit, the method comprising testing a plant for at least one of: a) reduced, or eliminated, expression of at least one AGAMOUS (AG) protein, b) reduced , or eliminated, expression of at least one polynucleotide encoding an AGAMOUS (AG) protein, c) presence of a marker associated with reduced expression of at least one AGAMOUS (AG) protein, and d) presence of a marker associated with reduced expression of at least one polynucleotide encoding an AGAMOUS (AG) protein. In one embodiment presence of any of a) to d) indicates that the plant will produce, or be useful for producing, at least one coreless fruit. In a further embodiment the plant identified is a mutant plant with reduced or eliminated expression of an AGAMOUS (AG) gene or protein. The mutant plant may be a naturally occurring mutant plant. Alternatively the mutant may be an induced mutant. Non-GM selection method for reduced or eliminated PISTILATA (PI) In a further aspect the invention provides a method for identifying a plant with a genotype indicative of producing, or being useful for producing, at least one coreless fruit, the method comprising testing a plant for at least one of: a) reduced, or eliminated, expression of at least one PISTILATA (PI) protein, b) reduced , or eliminated, expression of at least one polynucleotide encoding an PISTILATA (PI) protein, c) presence of a marker associated with reduced expression of at least one PISTILATA (PI) protein, and d) presence of a marker associated with reduced expression of at least one polynucleotide encoding a PISTILATA (PI) protein.
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