# US 9,750,205 B2 · Quartet breeding > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,750,205 B2 - Document ID: US9750205B2 - Status: Granted - Kind: B2 - Application number: 13/774,705 - Filed: 2013-02-22 - Published: 2013-09-12 - Granted: 2017-09-05 - Earliest claimed: 2011-09-29 - Assignees: Rijk Zwaan Zaadteelt En Zaadhandel B.V. - Applicants: Rijk Zwaan Zaadteelt En Zaadhandel B.V. - Inventors: Cornelis Maria Petrus van Dun - Web page: https://patentgrep.com/patent/US9750205B2 ## CPC - C12N 15/8212: {Colour markers, e.g. beta-glucoronidase [GUS], green fluorescent protein [GFP], carotenoid} - A01H 1/04: Processes of selection {involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection} - A01H 1/08: Methods for producing changes in chromosome number - A01H 3/04: by treatment with chemicals - C12N 15/82: for plant cells {, e.g. plant artificial chromosomes (PACs)} - C12N 15/62: DNA sequences coding for fusion proteins - C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]} - C12N 15/8287: {for fertility modification, e.g. apomixis} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES ## Abstract The invention relates to a method for the production of a set of seeds which are genetically identical to the male gametes from which they arise, which may comprise placing a limited number of paternal gametes that have the form of tetrads or dyads on the stigma of a flower to fertilize maternal egg cells to obtain a number of zygotes; and inducing the loss of maternal chromosomes from the zygotes to obtain a seed set containing a limited number of seeds in which the maternal chromosomes are absent. In a preferred embodiment the father plant exhibits suppression of chromosome recombination or second division restitution (SDR) during meiosis. ## Claims ### Claim 1 A method for the production of a set of Arabidopsis seeds which are genetically identical to the male gametes from which they arise, comprising: a) placing two or four paternal gametes from Arabidopsis that have the form of tetrads or dyads on the stigma of an Arabidopsis flower to fertilize maternal egg cells, to obtain a number of zygotes; b) inducing the loss of maternal chromosomes from the zygotes to obtain a seed set containing a limited number of seeds in which the maternal chromosomes are absent, wherein the two or four paternal gametes is equal to or lower than the number of egg cells contained in the female reproductive organ attached to the stigma and wherein the loss of maternal chromosomes from the zygote is induced by using a haploid inducer line as the female, wherein the female is a transgenic plant comprising a heterologous transgene expression cassette comprising a promoter operably linked to a polynucleotide encoding a recombinantly altered CENH3 polypeptide, and having a corresponding inactivated endogenous CENH3 gene, wherein the paternal gametes that have the form of tetrads or dyads result from interference with microspore tetrad separation and wherein interference with microspore tetrad separation comprises interference with a QRT1 gene. ### Claim 2 (depends on 1) The method as claimed in claim 1, wherein the paternal gametes are produced by a father plant which exhibits second division restitution (SDR) during meiosis. ### Claim 3 (depends on 1) The method as claimed in claim 1, wherein the interfering with the QRT1 gene prevents transcription thereof. ### Claim 4 (depends on 1) The method as claimed in claim 1, wherein the interfering with the QRT1 gene consists of destabilizing the QRT1 gene mRNA or transcript. ### Claim 5 (depends on 1) The method as claimed in claim 1, wherein the interfering with the QRT1 gene consists of introducing one or more mutations into the QRT1 gene, leading to perturbation of its biological function, wherein the one or more mutations are introduced randomly by one or more chemical compounds, by physical means, or by insertion of a genetic element, or wherein the one or more mutations are introduced specifically by homologous recombination or oligonucleotide-based mutation induction. ### Claim 6 (depends on 4) The method as claimed in claim 4, wherein the destabilizing the QRT1 gene mRNA or transcript is by binding with antisense RNA, RNAi molecules, Virus-Induced Gene Silencing (VIGS) molecules, co-suppressor molecules, RNA oligonucleotides or DNA oligonucleotides. ### Claim 7 (depends on 5) The method as claimed in claim 5, wherein the one or more chemical compounds is ethyl methanesulphonate, nitrosomethylurea, hydroxylamine, proflavine, N-methyl-N-nitrosoguanidine, N-ethyl-N-nitrosourea, N-methyl-N-nitro-nitrosoguanidine, diethyl sulphate, ethylene imine, sodium azide, formaline, urethane, phenol or ethylene oxide, or wherein the physical means are UV-irradiation, fast-neutron exposure, X-rays, gamma irradiation, or wherein the genetic element is a transposon, T-DNA or retroviral element. ### Claim 8 (depends on 2) The method as claimed in claim 2, wherein second division restitution is achieved by subjecting the father plant to environmental stress wherein the environment stress is temperature stress, NO2, nitrous oxide (N2O), or a combination thereof.