Methods for producing transgenic plants
US-2018273959-A1 · Sep 27, 2018 · US
US9750205B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9750205-B2 |
| Application number | US-201313774705-A |
| Country | US |
| Kind code | B2 |
| Filing date | Feb 22, 2013 |
| Priority date | Sep 29, 2011 |
| Publication date | Sep 5, 2017 |
| Grant date | Sep 5, 2017 |
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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.
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What is claimed is: 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. 2. 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. 3. The method as claimed in claim 1 , wherein the interfering with the QRT1 gene prevents transcription thereof. 4. The method as claimed in claim 1 , wherein the interfering with the QRT1 gene consists of destabilizing the QRT1 gene mRNA or transcript. 5. 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. 6. 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. 7. 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. 8. 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, NO 2 , nitrous oxide (N 2 O), or a combination thereof.
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