Syngenta Participations Ag

Syngenta Crop Protection Ag

Leland Stanford Junior University

Timothy Kelliher

Brent Delzer

Michael L Nuccio

Todd Lee Warner

William Paul Bullock

Suresh Babu Kadaru

Robert Arthur Dietrich

Satya P Chintamanani

Qiudeng Que

Kun Yu

Zhongying Chen

Jiang Li

Satya Chintamanani

Virginia Walbot

Julie Leonard Green

Wan Shi

Guozhu Tang

Jian Lv

Hongju Zhou

Juan Wei

Chunxia Liu

The presently disclosed subject matter relates to using a haploid inducing line (whether existing or created) and transforming the haploid line so that it encodes cellular machinery capable of editing genes. The transformed haploid inducing line is used as a parent in a cross between two plants. During pollination, the parental gametes fuse to form an embryo; and the gene editing machinery is also delivered to the embryo at this time. During embryonic development, one set of parental chromosomes are lost, and the gene editing machinery operates on the remaining set of chromosomes. Thus, at least one haploid progeny with edited genes is produced from the cross. The disclosure is also directed to methods of testing an edited haploid plant progeny for the presence of a first plant's genomic material.

Simultaneous gene editing and haploid induction

Provided here are methods of using a mutated patatin-like phospholipase IIα ('pPLAIIα,' renamed here MATRILINEAL) to induce haploid induction in plants, cloning a pPLAIIα to induce haploid induction in plants, and genetically engineering a plant to contain a mutated pPLAIIα. Also provided are methods of applying topical and spray chemicals, lipids, and RNAi molecules to plants during pollination in order to induce haploid production. Further provided are methods of chemically treating plants during pollination to induce haploids while also reducing embryo abortion and increasing seed set.

Haploid induction compositions and methods for use therefor

Systems and methods for producing a plurality of unique edits in a plant's T1 seed. In one example, a method comprises transforming at least one expression cassette into a plant cell or a plant tissue. The at least one expression cassette may comprise a nucleic acid that encodes a DNA modification enzyme; optionally, a nucleic acid that encodes at least one guide RNA (gRNA); and a floral mosaic (FMOS) regulatory sequence, wherein the FMOS regulatory sequence (i) mediates expression of the DNA modification enzyme in at least one of a floral primordia cell and a floral reproductive organ, and (ii) mediates a plurality of edits in the at least one of the floral primordia and the floral reproductive organ. The method may also include regenerating the plant cell or plant tissue into a T0 plant having a plurality of T1 seed, wherein the T1 seed contain a plurality of unique edits.

Compositions and methods for driving T1 event diversity

The presently disclosed subject matter relates to using a haploid inducing line (whether existing or created) and transforming the haploid line so that it encodes cellular machinery capable of editing genes. The transformed haploid inducing line is used as a parent in a cross between two plants. During pollination, the parental gametes fuse to form an embryo; and the gene editing machinery is also delivered to the embryo at this time. During embryonic development, one set of parental chromosomes are lost, and the gene editing machinery operates on the remaining set of chromosomes. Thus, at least one haploid progeny with edited genes is produced from the cross.

The present invention relates to wheat plants comprising a mutation causing an alteration of the amino acid sequence in centromere histone H3 (CENH3), which have the biological activity of a haploid inducer. Further, the present invention provides methods of generating the wheat plants of the present invention and haploid and doubled haploid wheat plants obtainable by crossing the wheat plants of the present invention with wildtype wheat plants.

Wheat CENH3 alleles

Provided are isolated cDNAs comprising a nucleotide sequence having at least 90% identity to SEQ ID NO: 33, SEQ ID NO: 37, SEQ ID NO: 52 or SEQ ID NO: 53. Also provided are expression cassettes; vectors; transgenic plant cells; plants, plant parts, and seeds; isolated polypeptides; amplicons and informative fragments of the presently disclosed nucleic acids; compositions that include amplification primer pairs; methods for producing plants that exhibit HI; methods for identifying the presence or absence of an allele associated with HI in a plant; methods for introgressing Haploid-inducing nucleotide sequences into plants; and methods for selecting parental plants predicted to produce progeny generations with plants that exhibit Haploid Induction trait.

Provided here are methods of using a mutated patatin-like phospholipase II&#x3b1; ('pPLAII&#x3b1;,' renamed here MATRILINEAL) to induce haploid induction in plants, cloning a pPLAII&#x3b1; to induce haploid induction in plants, and genetically engineering a plant to contain a mutated pPLAII&#x3b1;. Also provided are methods of applying topical and spray chemicals, lipids, and RNAi molecules to plants during pollination in order to induce haploid production. Further provided are methods of chemically treating plants during pollination to induce haploids while also reducing embryo abortion and increasing seed set.

Provided are isolated cDNAs comprising a nucleotide sequence having at least 90% identity to SEQ ID NO: 33, SEQ ID NO: 37, SEQ ID NO: 52 or SEQ ID NO: 53. Also provided are expression cassettes; vectors; transgenic plant cells; plants, plant parts, and seeds; isolated polypeptides; amplicons and informative fragments of the presently disclosed nucleic acids; compositions that include amplification primer pairs; methods for producing plants that exhibit HI; methods for identifying the presence or absence of an allele associated with HI in a plant; methods for introgressing Haploid&#x2014;inducing nucleotide sequences into plants; and methods for selecting parental plants predicted to produce progeny generations with plants that exhibit Haploid Induction trait.

Provided are expression cassettes; vectors; transgenic plant cells; plants, plant parts, and seeds; isolated polypeptides; amplicons and informative fragments of the presently disclosed nucleic acids; compositions that include amplification primer pairs; methods for producing plants that exhibit haploid induction (HI); methods for identifying the presence or absence of an allele associated with HI in a plant; methods for introgressing Haploid-inducing nucleotide sequences into plants; and methods for selecting parental plants predicted to produce progeny generations with plants that exhibit Haploid Induction trait.

Certain embodiments provide a method of altering the number of archesporial cells in a developing anther of a plant in certain embodiments, the method comprises exposing the anther to redox-modulatory conditions prior to differentiation of germline cells in the anther, thereby changing the redox potential of cells in the anther and altering the number of archesporial cells in the anther. This method may be employed to increase or decrease the number of archesporial cells in a developing anther, and may be employed to produce male sterile plants.

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Research Triangle Park, NC, United States of America

Timothy Kelliher