Methods of producing homoplasmic modified plants or parts thereof

Abstract

The present invention relates to a method of selecting modified plastids or mitochondria, or selecting plants or parts thereof comprising such modified plastids or mitochondria, and a method of producing a homoplasmic modified plant comprising modified plastids or mitochondria, and to expression constructs for performing said methods.

Description

[0001]METHODS OF PRODUCING HOMOPLASMIC MODIFIED PLANTS OR PARTS THEREOF FIELD OF THE INVENTION The present invention relates to a method of selecting modified plastids or mitochondria, or selecting plants or parts thereof comprising such modified plastids or mitochondria, and a method of producing a homoplasmic modified plant comprising modified plastids or mitochondria. The present invention also relates to an expression construct encoding a protein, specifically a TALEN, which is capable of cleavage of a target site in plastid or mitochondrial DNA under control of an inducible system, and its use to produce homoplasmic modified plants. BACKGROUND TO THE INVENTION Plastids and mitochondria in plant cells provide essential elements of the genome in plants, consequently their genomes provide important targets for genetic modification in order to alter the phenotypes of plants. In each plant cell, the genome copy numbers in plant plastids (the form before the differentiation of chloroplasts) and mitochondria are very high – up to 10,000 copies may exist for the chloroplasts and up to 500 copies for the mitochondria. The features of high copy number and maternal inheritance in plastids and mitochondria provide advantages for plastid and mitochondrial transformation applications for high expression levels of transgenes and prevention of transgene contamination through dissemination via pollen. In addition, some genes which control important commercial features which may be desirable to modify, such as cytoplasmic male sterility, are located in mitochondrial genomes. The tools available for genetic modification of plant plastid and mitochondrial genomes are limited, unlike those available for genetic modification of plant nuclear genomes. A reliable method of mitochondrial genomes and efficiently selecting the modified plants therefrom has not yet been achieved and methods for modifying plastid genomes are limited to those species amenable to stable plastid transformation. In part, this is due to the high copy numbers of the genome in each plant cell, which makes homogenous genetic modification of the plastid and mitochondrial genomes to create a homoplasmic plant difficult. Furthermore, the organelle genomes commonly exhibit very low spontaneous mutation rates, which are 5-20 fold lower than the nuclear genome (1). These low mutation rates prevent the variation in organelle gene sequences required for breeding improved crops. This includes single nucleotide polymorphisms (SNPs) in organelle genes that give rise to multiple alleles of a chloroplast or mitochondrial gene. This lack of variation is a barrier to understanding the functions of organelle genes and isolating new alleles for breeding improved crops. Current techniques for plastid transformation are very inefficient and limited to a few plant species because relatively few plant species are amenable to plastid transformation and because the commonly used marker gene (aadA) for selecting the homogenous status of mutated plastid genomes (homoplasmy) is not reliable in many plant species, especially cereals. For mitochondria, stable plant mitochondrial transformation has not yet been reported because there is no suitable selection method. Furthermore, while co-transformation of the gene of interest and aadA, which provides spectinomycin resistance, for isolating plants with mutations in plastid genes is commonly used in some plant species, this method has disadvantages. First, it has not been successful in cereals. Second, only a fraction of selected transformants contain the desired targeted organellar mutation because the marker gene can be inserted without insertion of the intended targeted mutation. Third, usually several rounds of selection are needed to ensure plastid homoplasmy of modified plants. Fourth, plants bearing the aadA marker will be regulated as genetically modified crops which is undesirable and will also contain unnecessary foreign genetic material as well as an unnecessary metabolic burden. Although it is possible to remove the aadA marker gene from the engineered plastid genome, prolonged vegetative growth and additional rounds of regeneration under selection or seed generations are needed which is time consuming and therefore undesirable. Fifth, plastid transformation with specific sequences is inherently limited to introducing single variants of a given sequence, thus limiting the genetic diversity that can be obtained. In summary, genetic modification of plastids and mitochondria has been hindered by the reasons mentioned above, especially the lack of efficient methods to select for plants having modified plastids or mitochondria, and therefore the lack of efficient methods to create genetic uniformity i.e. homoplasmy in the resulting plants. This has significantly limited the use of plastid and mitochondrial gene engineering in commercial crop farming, especially in cereals. One key example of the application of genetically modifying plant organelles is in the generation of hybrids. Hybrid F1 crop usually provide higher yield and resistance to biotic / abiotic stresses. F1 seeds are produced by sexual hybridisation between male line (pollen donor) and female line (pollen receiver). A key step to control the genetic uniformity of the F1 progeny requires preventing self-pollination of the female parent line during hybridisation. This can be done by de-tasselling the male flower of the female parent (for example in maize) or using a female parent that is cytoplasmic male sterile (for example in oil- seed rape and rice). Cytoplasmic male sterility (CMS) is therefore a desirable trait for commercial hybrid seed production from self-pollinated crops. Wheat has not benefitted from hybrid production, even though the predicted increase of yield in hybrid wheat is around 20%. A major factor limiting wheat hybrid development has been the lack of CMS systems in wheat breeding lines. CMS is a trait controlled by the mitochondrial genome, and thus reliable methods for selecting mutated mitochondrial genomes and producing homoplasmic modified plants in this respect would open the route for creating commercial cultivars of hybrid wheat. An efficient method for plastid and mitochondrial transformation would be highly advantageous for generation of CMS plants and therefore hybrid crops, amongst other applications. One or more aspects or embodiments of the present invention seek to address at least these problems, or one or more alternative problems in the art. SUMMARY OF THE INVENTION According to a first aspect of the present invention, there is provided a method of producing a homoplasmic modified plant or part thereof, the method comprising: (a) Modifying plastid or mitochondrial DNA of the plant or part thereof at one or more target sites to produce a modified plant or part thereof having plastid or mitochondrial DNA comprising one or more modified target sites; (b) Introducing into the plant or part thereof, optionally the modified plant or part thereof, an expression construct comprising: one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; (d) Regenerating the modified plant or part thereof; and (e) Optionally repeating steps (c) and (d). In one embodiment, there is provided a method of producing a homoplasmic modified plant or part thereof, the method comprising: (a) Modifying plastid DNA of the plant or part thereof at one or more target sites to produce a modified plant or part thereof having plastid DNA comprising one or more modified target sites; (b) Introducing into the plant or part thereof, optionally the modified plant or part thereof, an expression construct comprising: one or more nucleic acids which encode a plastid targeted protein capable of cleavage of any non-modified target sites in the plastid DNA under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; (d) Regenerating the modified plant or part thereof; and (e) Optionally repeating steps (c) and (d). In one embodiment, there is provided a method of producing a homoplasmic modified plant or part thereof, the method comprising: (a) Modifying mitochondrial DNA of the plant or part thereof at one or more target sites to produce a modified plant or part thereof having mitochondrial DNA comprising one or more modified target sites; (b) Introducing into the plant or part thereof, optionally the modified plant or part thereof, an expression construct comprising: one or more nucleic acids which encode a mitochondria targeted protein capable of cleavage of any non-modified target sites in the mitochondrial DNA under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; (d) Regenerating the modified plant or part thereof; and (e) Optionally repeating steps (c) and (d). In one embodiment, the method may be carried out on a plurality of plants or parts thereof. In one embodiment, the method comprises a further step of selecting the plant or part thereof if it maintains or has improved viability compared to the plant or part thereof of step (a). Suitably after inducing expression of the protein (step (c)) and before regeneration (step (d)). It will be appreciated that the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA under the control of an inducible system can be introduced prior to, during or after the step of modifying plastid or mitochondrial DNA of the plant or part thereof. Introduction prior to or during the step of modifying plastid or mitochondrial DNA of the plant or part thereof is useful where the targeted protein is used to cause or facilitate modification at the target site. In such cases, inducing expression of the protein from the expression construct is used to cause or facilitate modification of the target site through cleavage at the target site by the targeted protein. According to a second aspect of the present invention, there is provided a method of selecting modified plastids or mitochondria, the method comprising: (a) Providing one or more plastids or mitochondria that have undergone a modification process to modify one or more target sites in the plastid or mitochondrial DNA; (b) Introducing into the one or more plastids or mitochondria an expression construct comprising: one or more nucleic acids which encode a protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; and (d) Selecting the plastids or mitochondria which maintain or have improved viability compared to the plastids or mitochondria of step (a). In one embodiment, the modified plastids or mitochondria may be present in a cell, suitably in a plant cell. Therefore in one embodiment the method may comprise selecting cells comprising modified plastids or mitochondria, suitably comprising providing one or more cells which comprise one or more plastids or mitochondria that have undergone a modification process, introducing into the one or more cells an expression construct, and selecting the one or more cells which maintain or have improved viability. In such an embodiment, the one or more nucleic acids may encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites. In one embodiment, the method may comprise providing one or more cells which comprise one or more plastids or mitochondria, introducing into the one or more cells an expression construct, introducing into the one or more cells a heterologous nucleic acid, suitably a repair template, suitably a template adapted for homology directed repair (HDR), such as a DNA repair template, and selecting the one or more cells which maintain or have improved viability. In such an embodiment, the one or more nucleic acids may encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites. In one embodiment, the method may further comprise step (e) regenerating the selected plastids or mitochondria, suitably which may comprise regenerating one or more cells containing the selected plastids or mitochondria. In one embodiment, the method may further comprise repeating steps (c) to (e). In one embodiment, there is provided a method of selecting modified plastids, the method comprising: (a) Providing one or more plastids that have undergone a modification process to modify one or more target sites in the plastid DNA; (b) Introducing into the one or more plastids an expression construct comprising: one or more nucleic acids which encode a protein capable of cleavage of any non-modified target sites in the plastid DNA under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; and (d) Selecting the plastids which maintain or have improved viability compared to the plastids of step (a). In one embodiment, the method may further comprise step (e) regenerating the selected plastids, suitably which may comprise regenerating the cells containing the selected plastids. In one embodiment, the method may further comprise repeating steps (c) to (e). In one embodiment, there is provided a method of selecting modified mitochondria, the method comprising: (a) Providing one or more mitochondria that have undergone a modification process to modify one or more target sites in the mitochondrial DNA; (b) Introducing into the one or more mitochondria an expression construct comprising: one or more nucleic acids which encode a protein capable of cleavage of any non-modified target sites in the mitochondrial DNA under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; and (d) Selecting the mitochondria which maintain or have improved viability compared to the mitochondria of step (a). In one embodiment, the method may further comprise step (e) regenerating the selected mitochondria, suitably which may comprise regenerating the cells containing the selected mitochondria. In one embodiment, the method may further comprise repeating steps (c) to (e). In one embodiment, the methods of the second aspect are ex vivo methods. According to a third aspect of the present invention, there is provided a method of selecting plants or parts thereof comprising modified plastid or mitochondrial DNA, the method comprising: (a) Providing one or more plants or parts thereof that have undergone a modification process to modify one or more target sites in the plastid or mitochondrial DNA; (b) Introducing into the one or more plants or parts thereof an expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA, under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; (d) Selecting the plants or parts thereof which maintain or have improved viability compared to the plants of step (a). In one embodiment, the method may further comprise step (e) regenerating the selected plants or parts thereof. In one embodiment, the method may further comprise repeating steps (c) to (e). Suitably in such embodiments, the method produces homoplasmic modified plants or parts thereof. In one embodiment, there is provided a method of selecting plants or parts thereof comprising modified plastid DNA, the method comprising: (a) Providing one or more plants or parts thereof that have undergone a modification process to modify one or more target sites in the plastid DNA; (b) Introducing into the one or more plants or parts thereof an expression construct comprising one or more nucleic acids which encode a plastid targeted protein capable of cleavage of non-modified target sites in the plastid DNA, under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; (d) Selecting the plants or parts thereof which maintain or have improved viability compared to the plants of step (a). In one embodiment, the method may further comprise step (e) regenerating the selected plants or parts thereof. In one embodiment, the method may further comprise repeating steps (c) to (e). Suitably in such embodiments, the method produces homoplasmic modified plants or parts thereof. In one embodiment, there is provided a method of selecting plants or parts thereof comprising modified mitochondrial DNA, the method comprising: (a) Providing one or more plants or parts thereof that have undergone a modification process to modify one or more target sites in the mitochondrial DNA; (b) Introducing into the one or more plants or parts thereof an expression construct comprising one or more nucleic acids which encode a mitochondria targeted protein capable of cleavage of non-modified target sites in the mitochondrial DNA, under the control of an inducible system; (c) Inducing expression of the protein from the expression construct; (d) Selecting the plants or parts thereof which maintain or have improved viability compared to the plants of step (a). In one embodiment, the method may further comprise step (e) regenerating the selected plants or parts thereof. In one embodiment, the method may further comprise repeating steps (c) to (e). Suitably in such embodiments, the method produces homoplasmic modified plants or parts thereof. In one embodiment of any of the methods, the modification process used to modify the one or more target sites in the plastid or mitochondrial DNA, or modifying the one or more target sites in the plastid or mitochondrial DNA in step (a) of the methods is by using an error prone polymerase, suitably by exposing the plastid or mitochondrial DNA to an error prone polymerase. In one embodiment, the error prone polymerase is introduced into the plant or part thereof, the plastid or the mitochondria, suitably by transformation, to modify the plastid or mitochondrial DNA at one or more target sites. In some preferred embodiments of any of the methods set out herein, the modification process used to modify the one or more target sites in the plastid or mitochondrial DNA uses a heterologous nucleic acid, suitably a repair template, suitably a template adapted for homology directed repair (HDR), such as a DNA repair template. Accordingly, in some embodiments, the modifying of the one or more target sites in the plastid or mitochondrial DNA in step (a) of the methods is achieved by using a heterologous nucleic acid, suitably a repair template, suitably a template adapted for homology directed repair (HDR), such as a DNA repair template. In one embodiment, the heterologous nucleic acid is introduced into the plant or part thereof, or the plastid or the mitochondria, suitably by transformation, suitably by particle bombardment, to modify the plastid or mitochondrial DNA at one or more target sites. In some preferred embodiments of the methods set out herein, the modification process used to modify the one or more target sites in the plastid or mitochondrial DNA uses a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA. Accordingly, in some embodiments the modifying of the one or more target sites in the plastid or mitochondrial DNA in step (a) of the methods is achieved by using a mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA. In some embodiments, a nucleic acid encoding the targeted protein is introduced into the plant or part thereof, or the plastid or the mitochondria, suitably by transformation, suitably by particle bombardment, to modify the plastid or mitochondrial DNA at one or more target sites. Thus, the modification of the plastid or mitochondrial DNA of the plant or part thereof at one or more target sites to produce a modified plant or part thereof can be achieved through cleavage at the target site by a suitable targeted protein capable of cleavage (e.g., a programmable nuclease). Suitable proteins capable of cleavage (e.g., programmable nucleases) are discussed hereinbelow. In some preferred embodiments of the methods set out herein, the modification process used to modify the one or more target sites in the plastid or mitochondrial DNA uses a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA and a heterologous nucleic acid (suitably a repair template, suitably a template adapted for homology directed repair (HDR), such as a DNA repair template). Accordingly, in some embodiments the modifying of the one or more target sites in the plastid or mitochondrial DNA in step (a) of the methods is achieved by using a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA and a heterologous nucleic acid (suitably a repair template, suitably a template adapted for homology directed repair (HDR), such as a DNA repair template). As will be appreciated by the skilled person, cleavage at the target site will facilitate integration of a desired sequence present in the heterologous nucleic acid, e.g. through HDR. Thus, the modification of the plastid or mitochondrial DNA of the plant or part thereof at one or more target sites to produce a modified plant or part thereof can be achieved through gene editing using a suitable targeted protein capable of cleavage (e.g., a programmable nuclease) in combination with a suitable template nucleic acid. Suitable protein capable of cleavage (e.g., programmable nucleases) and heterologous nucleic acid are discussed hereinbelow. In one embodiment of any of the methods, the inducible system is an oestradiol inducible system, alternatively referred to as an ‘estradiol’ inducible system. According to a fourth aspect of the present invention, there is provided an expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA, under the control of an oestradiol inducible system. In one embodiment, there is provided an expression construct comprising one or more nucleic acids which encode a plastid targeted protein capable of cleavage of a target site in plastid DNA, under the control of an oestradiol inducible system. In one embodiment, there is provided an expression construct comprising one or more nucleic acids which encode a mitochondria targeted protein capable of cleavage of a target site in mitochondrial DNA, under the control of an oestradiol inducible system. In one embodiment, under the control of the inducible system means that the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA are operably linked to a binding site for an oestradiol inducible system. In one embodiment, the expression construct further comprises one or more nucleic acids which encode the components of the oestradiol inducible system. In an alternative fourth aspect of the present invention, there is provided a pair of expression constructs, the first expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA, under the control of an oestradiol inducible system (suitably operably linked to a binding site for an oestradiol inducible system), and the second expression construct comprising one or more nucleic acids which encode components of the oestradiol inducible system. In one embodiment, there is provided a pair of expression constructs, the first expression construct comprising one or more nucleic acids which encode a plastid targeted protein capable of cleavage of a target site in plastid DNA, under the control of an oestradiol inducible system (suitably operably linked to a binding site for an oestradiol inducible system), and the second expression construct comprising one or more nucleic acids which encode components of the oestradiol inducible system. In one embodiment, there is provided a pair of expression constructs, the first expression construct comprising one or more nucleic acids which encode a mitochondria targeted protein capable of cleavage of a target site in mitochondrial DNA, under the control of an oestradiol inducible system (suitably operably linked to a binding site for an oestradiol inducible system), and the second expression construct comprising one or more nucleic acids which encode components of the oestradiol inducible system. In one embodiment, the oestradiol inducible system is an XVE system. In one embodiment therefore the components of the oestradiol inducible system are a DNA-binding domain comprising the bacterial repressor LexA (X), an activator domain comprising the Herpes Simplex Virus transactivator VP16 (V), and an inducer binding domain comprising the regulatory region of the human oestrogen receptor (E). In one embodiment, the expression construct or constructs of the fourth aspect are used in any of the methods of the invention. All features related to the expression construct or vector products may equally apply to the expression construct used in the methods. In one embodiment, the plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA targets a gene that controls plant growth, suitably a gene selected from the group consist of: accD, psbA, rbcL, atpB, 16S rDNA, and 23S rDNA. According to a fifth aspect of the present invention, there is provided a vector comprising the expression construct or constructs of the fourth aspect. According to a sixth aspect of the present invention, there is provided a plastid or mitochondria comprising the expression construct or constructs of the fourth aspect or the vector of the fifth aspect. In one embodiment, there is provided a plastid comprising the expression construct or constructs of the fourth aspect or the vector of the fifth aspect. In one embodiment, there is provided a mitochondria comprising the expression construct or constructs of the fourth aspect or the vector of the fifth aspect. According to a seventh aspect of the present invention, there is provided a plant or part thereof, such as a plant cell, comprising the expression construct or constructs of the fourth aspect or the vector of the fifth aspect. According to a seventh aspect of the present invention, there is provided use of the expression construct or constructs of the fourth aspect or the vector of the fifth aspect for selecting plants or parts thereof comprising modified plastid or mitochondrial DNA. In one embodiment, there is provided use of the expression construct or constructs of the fourth aspect or the vector of the fifth aspect for selecting plants or parts thereof comprising modified plastid DNA. In one embodiment, there is provided use of the expression construct or constructs of the fourth aspect or the vector of the fifth aspect for selecting plants or parts thereof comprising modified mitochondrial DNA. According to an eighth aspect of the present invention, there is provided use of the expression construct or constructs of the fourth aspect or the vector of the fifth aspect for producing homoplasmic modified plants or parts thereof. In one embodiment, the expression construct or constructs of the fourth aspect or the vector of the fifth aspect is used in combination with an error prone polymerase. In one embodiment, the expression construct or constructs of the fourth aspect or the vector of the fifth aspect is used in combination with a heterologous nucleic acid, suitably a repair template, suitably a template adapted for homology directed repair (HDR), such as a DNA repair template. In one embodiment, the expression construct or constructs of the fourth aspect or the vector of the fifth aspect is used for inducing expression of the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA in the plant or part thereof at an optimal time. In one embodiment, the expression construct or constructs of the fourth aspect or the vector of the fifth aspect is used for inducing expression of the one or more nucleic acids which encode a plastid targeted protein capable of cleavage of a target site in plastid DNA in the plant or part thereof at an optimal time. In one embodiment, the expression construct or constructs of the fourth aspect or the vector of the fifth aspect is used for inducing expression of the one or more nucleic acids which encode a mitochondria targeted protein capable of cleavage of a target site in mitochondrial DNA in the plant or part thereof at an optimal time. Suitably at an optimal time to produce homoplasmic modified plants or parts thereof. According to a ninth aspect of the present invention there is provided a homoplasmic modified plant or part thereof, such as a plant cell, produced by the method of the first aspect. The invention disclosed herein provides a novel selection method for ensuring the genetic uniformity (homoplasmy) of plant organelles having mutated genomes, particularly plastid or mitochondria having mutated genomes. Compared to the commonly used aadA selection marker method, the invention can provide higher efficiencies for isolating and recovering organelle-engineered plants, can be applied to any plant species, and can be used in combination with established methods of random mutagenesis to create genetic diversity. In some embodiments the invention leads to genetically improved plants that are not subject to strict GMO regulations. Advantages of the invention include: Use of a protein capable of cleavage of non-modified target sites in plastid or mitochondrial DNA for implementing selection of plastids or mitochondria that have been modified at target site in their DNA does not rely on any marker gene inserted into the plastid genome. This should have three major advantages: 1. Enabling plastid and mitochondria transformation in agronomically important crops such as cereals. This will expand the benefits of plastid transformation to cereals for reducing the crop loss due to pests and diseases. Mitochondrial transformation will enable artificial creation of the cytoplasmic male sterility trait, which will significant reduce the cost for production of hybrid seeds. Hybrid crops usually perform better than their parent lines as exemplified by the maize seed market, which is dominated by hybrid seeds. Additionally, use of mitochondrial transformation for artificial creation of the cytoplasmic male sterility trait will reduce the cost and time required to create CMS breeding lines. 2. Enabling saturation mutagenesis in a targeted sequence (30-35 bp) of the plastid / mitochondrial genome in 3 months. This could help to quickly discover new and advantageous mutations in the plastid and mitochondrial genomes, which can potentially improve gene functions in plastids and mitochondria, and provide desirable traits. 3. The engineered plastid / mitochondrial genomes are transgene-free, while the nuclear transgene expressing the protein capable of cleavage of a non-modified target site in plastid or mitochondrial DNA can be removed easily via crossing with the wild type plant. Transgene- free crops can be exempt from GM regulations in many countries, including the US, Canada, England and China. DETAILED DESCRIPTION OF THE INVENTION Herein is disclosed a novel method for selecting mutagenized plant organellar genomes, particularly mutagenized plastid and mitochondrial genomes. The invention relates to the use of targeted nucleases to reduce or completely remove the wild type plastid genome through DNA cleavage (double strand break). Targeted nucleases used in this invention may include restriction enzymes and programmable nucleases such as meganuclease, transcription activator-like effector nuclease (TALEN), zinc finger nucleases (ZFNs), and CRISPR-Cas systems etc. Such nucleases can recognise and cleave specific DNA target sites, if targeted to cleave a non-modified target site, any mutations (such as point mutation, insertions and deletions, for example mutations that are introduced using a DNA molecule) that are introduced within the target site will prevent further cleavage by the corresponding targeting nuclease. Such targeting nucleases can be expressed from the nucleus and targeted to organellar genomes, suitably plastid or mitochondrial genomes. Wild type organellar genomes cleaved by the sequence-targeting nuclease will become vulnerable to degradation while mutated organellar genomes will be inert to the same nuclease, and as such gain a replication advantage. As a result, through division of the organelles with modified genomes and the resulting cells or plants containing them, the copy number of the mutated organellar genome will increase and eventually become homoplasmic within the cell, and the plant. In the present invention, the nuclease is controlled by an inducible expression system, suitably the estradiol-inducible XVE system. In the presence of the inducer, the nuclease is expressed, and plant cells with a desired mutation in the target DNA can be selected and ‘fixed’ in a cell or a plant by the above mechanism. Cells or plants without mutated organelles will have difficulty or fail to regenerate into plants as substantially all of the non-modified DNA will be cleaved by the nuclease. Inducible expression of the nuclease can be fine-tuned to optimise the selection efficiency of homoplasmic mutant plants, and to induce expression of the selective nuclease at an optimal time in the development of the plant or in an optimal location in the plant, such as when and / or where the plant cell has the lowest organelle genome copy number. Further features of the invention will now be described under the following headed sections. Features in any section may be combined with any aspect or embodiment of the invention in any workable combination. Homoplasmic The present invention relates to processes of making modifications to plastid or mitochondrial genomes, selecting for said modified plastid or mitochondrial genomes, and subsequently creating homoplasmic modified plants therefrom. As used herein the term ‘homoplasmic’ or ‘homoplasmy’ means that substantially all copies of the plastid or mitochondrial genome are the same, with reference to a given organelle, plant cell, or to an entire plant. Suitably, homoplasmic as used herein in the context of the present invention means that substantially all copies of the plastid or mitochondrial genome in a given organelle, cell or plant contain an intended modification, introduced by the methods of the invention. Suitably therefore that substantially all copies of the plastid or mitochondrial genome in a given organelle, cell or plant contain a modification at the or each target site. Suitably therefore that substantially all copies of the plastid or mitochondrial genome in a given plant contain a modification at the or each target site. Suitably the term ‘homochondric’ or ‘homochondry’ or ‘homochondriomy’ may also be used herein interchangeably with the terms ‘homoplasmic’ or ‘homoplasmy’ but specifically to refer to substantially all copies of the mitochondrial genome being the same, with reference to a given organelle, plant cell, or to an entire plant. Suitably homoplasmy can be quantified by there being less than one point mutation per million nucleotides of each copy of the plastid or mitochondrial genomes in a given organelle, plant cell, or an entire plant (https: / / www.pnas.org / doi / full / 10.1073 / pnas.2001998117). Suitably therefore substantially all copies of the genome are the same. By the same it is meant genetically homogenous. By ‘substantially all’ as used above it is meant that at least 95% of the copies of the plastid or mitochondrial genome in a given organelle, plant cell, or an entire plant are the same. Suitably at least 96%, at least 97%, at least 98%, at least 99% or 100% of the copies of the plastid or mitochondrial genome in a given organelle, plant cell, or an entire plant are the same. Suitably therefore at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the copies of the plastid or mitochondrial genome in a given organelle, cell or plant contain a modification at the or each target site. In the art it has been difficult to create homoplasmic modified plants due to the high copy number of the plastid and mitochondrial genomes in each plant cell. Typical modification procedures will only modify some of the genome copies, not all, meaning the resulting plants are genetically heterogeneous. The present invention aims to provide a more efficient way to arrive at a homoplasmic plant where each copy of the plastid or mitochondrial genome contains a desired or intended modification at a target site. Suitably, so that the resulting plants are genetically homogeneous. Plastid or Mitochondria Suitably the present invention may relate to any plastid or mitochondria or any organelle derived therefrom. Suitably a plastid or mitochondria at any stage of development is encompassed in the present invention. Suitably a ‘plastid’ as referred to herein is any membrane bound organelle found in the cells of plants or algae. Suitable plastids may include any of the following organelles: Proplastids, Chloroplasts, Etioplasts, Leucoplasts, Amyloplasts, Elaioplast, Proteinoplasts, and Chromoplasts. Suitably the plastid is a chloroplast. Suitably a ‘mitochondrion’ as referred to herein is any membrane bound organelle found in eukaryotic cells which carries out respiration. Suitably the mitochondrion is found in plant, fungi, and algae cells. Suitably therefore the present invention relates primarily to plants and plant cells, but may also relate to fungi, algae, and fungal and algal cells. Suitably any reference herein in any of the methods or embodiments to a plant or a part thereof may be replaced with a fungus or a part thereof, or an alga or a part thereof, including a fungal or an algal cell. Suitable mitochondria may include any of the following organelles: aerobic or typical mitochondria, anaerobic mitochondria, hydrogenosomes, mitosomes. Suitably the mitochondrion is an aerobic or typical mitochondrion. Plant or part thereof Suitably the present invention primarily relates to selecting modified plants which comprise modified plastid or mitochondrial genomes, and the creation of homoplasmic modified plants therefrom. Suitably the invention may relate to any plant or part thereof. Suitably the plant or part thereof may be a dicot or monocot, or a part thereof. Suitably the plant or part thereof may be an agronomically important species. Suitable plants or parts thereof of the present invention include crops and plants of agricultural, horticultural, or economic significance. ‘Crop’ as used herein means a plant species or variety that is grown to be harvested as food, livestock fodder, fuel raw material, medical uses, or for any other economic purpose. As a non-limiting example, said crops can be maize, cereals, such as wheat, rye, barley and oats, sorghum, rice, sugar beet and fodder beet, fruit, such as pome fruit (e.g. apples and pears), citrus fruit (e.g. oranges, lemons, limes, grapefruit, or mandarins), stone fruit (e. g. peaches, nectarines or plums), nuts (e.g. almonds or walnuts), soft fruit (e.g. cherries, strawberries, blackberries or raspberries), the plantain family or grapevines, leguminous crops, such as beans, lentils, peas and soya, oil crops, such as sunflower, safflower, rapeseed, canola, castor or olives, cucurbits, such as cucumbers, melons or pumpkins, fibre plants, such as cotton, flax or hemp, fuel crops, such as sugarcane, miscanthus or switchgrass, vegetables, such as potatoes, tomatoes, peppers, lettuce, spinach, onions, carrots, egg-plants, asparagus or cabbage, ornamentals, such as flowers (e.g. petunias, pelargoniums, roses, tulips, lilies, or chrysanthemums), shrubs, broadleaved trees (e.g. poplars or willows) and evergreens (e.g. conifers), grasses, such as lawn, turf or forage grass or other useful plants, such as coffee, tea, tobacco, hops, pepper, rubber or latex plants. Suitable plants may include any of the following or parts thereof: Musa textilis, Medicago sativa, Prunus dulcis, Pimpinella anisum, Malus sylvestris, Prunus armeniaca, Areca catechu, Arracacia xanthorhiza, Maranta arundinacea, Cynara scolymus, Helianthus tuberosus, Asparagus officinalis, Persea americona, Pennisetum americanum, Vigna subterranean, Musa paradisiaca, Hordeum vulgare, Phaseolus vulgaris, Phaseolus vigna spp., Beta vulgaris, Citrus bergamia, Rubus spp., Piper nigrum, Acacia mearnsii, Vaccinium spp., Bertholletia excelsa, Artocarpus altilis, Vicia faba, Brassica oleracea botrytis, Sorghum bicolor, Brassica oleracea gemmifera, Fagopyrum esculentum, Brassica oleracea capitate, Brassica rapa, Brassica spp., Theobroma cacao, Cucumis melo, Carum carvi, Elettaria cardamomum, Cynara cardunculus, Ceratonia siliqua, Daucus carota, Anacardium occidentale, Manihot esculenta, Ricinus communis, Brassica oleracea botrytis, Apium graveolens, Sechium edule, Prunus spp., Castanea sativa, Cicer arietinum, Cichorium intybus, Cichorium intybus, Capsicum spp., Cinnamomum verum, Cymbopogon nardus, Citrus medica, Citrus veticulata, Trifolium spp., Syzygium aromaticum, Cocos nucifera, Colocasia spp.; Xanthosoma spp., Coffee spp., Cola spp., Brassica napus, Zea mays, Valerianella locusta, Gossypium spp., Vigna unguiculate, Vaccinium spp., Lepidium sativum, Cucumis sativus, Ribes spp., Annona reticulata, Colocasia esculenta, Phoenix dactylifera, Moringa oleifera, Phaseolus spp., Allium sativum, Allium cepa, Pisum sativum, Triticum durum, Xanthosoma spp.; Colocasia spp., Solanum melongena, Solanum tuberosum, Cichorium endivia, Lygeum spartum, Foeniculum vulgare, Trigonella foenumgraecum, Ficus carica, Corylus avellane, Furcraea macrophylla, Linum usitatissimum, Phormium tenax, Pelargonium spp.; Geranium spp., Zingiber officinalis, Langenaria spp; Cucurbita spp., Cicer arietinum, Citrus paradise, Vitis vinifera, Lygeum spartum, Dactylis glomerata, Arachis hypogaea, Psidium guajava, Corylus avellane, Cannabis sativa, Crotalaria juncea, Agave fourcroydes, Lawsonia inermis, Humulus lupulus, Armoracia Rusticana, Indigofera tinctorial, Jasminum spp., Corchorus spp., Brassica oleracea acephala, Ceiba pentandra, Hibiscus cannabinus, Brassica oleracea gongylodes, Lavandula spp., Allium ampeloprasum, Citrus limon, Cymbopogon citratus, Lens culinaris, Lespendeza spp., Lactuca sativa, Glycyrrhiza glabra, Citrus aurantifolia, Citrus limetta, Linum usitatissimum, Litchi chinensis, Eriobotrya japonica, Lupinus spp., Macadamia spp., Myristica fragrans, Agave atrovirens, Citrus reticulata, Mangifera indica, Manihot esculenta, Secale cereal, Mespilus germanica, Cucumis melo, Penicum miliaceum, Eleusine coracana, Setaria italica, Echinochloa crusgalli, Eleusine coracana; Mentha spp., Morus spp., Morus alba, Agaricus spp.; Pleurotus spp. Volvariella, Brassica nigra; Sinapis alba, Prunus persica, Phormium tenax, Guizotia abyssinica, Myristica fragrans, Avena spp., Elaeis guineensis, Abelmoschus esculentus, Olea europea, Papaver somniferum, Citrus sinensis, Citrus aurantium, Dactylis glomerate, Metroxylon spp., Borassus flabellifer, Carica papaya, Pastinaca sativa, Pyrus communis, Pisum sativum, Carya illinoensis, Capsicum annuum, Diospyros kaki; Diospyros virginiana, Cajanus cajan, Ananas comosus, Pistacia spp., Prunus domestica, Punica granatum, Citrus grandis, Solamum tuberosum, Ipomoea batatas, Cucurbita spp., Chrysanthemum cineraraiefolium, Aspidosperma spp., Cydonia oblonga, Cinchona spp., Chenopodium quinoa, Raphanus sativus (including Cochlearia armoracia), Boehmeria nivea, Agrostis spp., Boehmeria nivea, Rheum spp., Oryza sativa; Oryza glaberrima, Rose spp., Hevea brasiliensis, Secale cereal, Lolium spp., Carthamus tinctorius, Metroxylon spp., Onobrychis viciifolia, Valerianella locusta, Tragopogon porrifolius, Achras sapota, Citrus reticulata, Brassica ileracea capitate, Scorzonera hispanica, Sesamum indicum, Butyrospermum paradoxum, Agave sislana, Citrus aurantifolia, Glycine max, Triticum spelta, Spinacia oleracea, Secale cereal, Cucurbita spp., Fragaria spp., Sorghum bicolor Sudanense, Saccharum officinarum, Helianthus annuus, Crotalaria juncea, Citrus limetta, Iopmoea batatas, Citrus reticulata, Xanthosoma sagittifolium, Manihot esculenta, Colocasia esculenta, Camellia sinensis, Eragrostis abyssinica, Phleum pratense, Nicotiana tabacum, Lycopersicum esculentum, Lotus spp., Aleurites spp., Brassica rapa, Urena lobate, Vanilla planifolia, Vicia sativa, Juglans spp., Citrullus lanatus, Acacia mearnsii, Triticum spp., Hordeum spp., Dioscorea spp., and Ilex paraguariensis. As used herein unless clearly indicated otherwise, the term "plant" is intended to mean a plant at any developmental stage, as well as any part or parts of a plant that may be attached to or separate from a whole intact plant. Such parts of a plant include, but are not limited to, organs, tissues, and cells of a plant including, plant calli, plant clumps, plant protoplasts and plant cell tissue cultures or explants from which plants can be regenerated. Examples of particular plant parts include a stem, a leaf, a root, an inflorescence, a flower, a floret, a fruit, a pedicle, a peduncle, a stamen, an anther, a stigma, a style, an ovary, a petal, a sepal, a carpel, a root tip, a root cap, a root hair, a leaf hair, a seed hair, a pollen grain, a microspore, an embryos, an ovule, a cotyledon, a hypocotyl, an epicotyl, xylem, phloem, parenchyma, endosperm, a companion cell, a guard cell, and any other known organs, tissues, and cells of a plant. Furthermore, it is recognized that a seed is a plant part. A "plant cell" is a structural and physiological unit of a plant, comprising a protoplast and a cell wall. A plant cell is considered to be a plant part herein. The plant cell may be in the form of an isolated single cell or a cultured cell, or as a part of a higher organized unit such as, for example, plant tissue, a plant organ, or a whole plant. A "plant organ" is a distinct and visibly structured and differentiated part of a plant such as a root, stem, leaf, flower bud, or embryo, or any of those parts listed above. In one embodiment, the plant or part thereof is wheat. Suitably any species of wheat selected from: Triticum aestivum, Triticum spelta, Triticum durum, Triticum dicoccum, Triticum turanicum, and Triticum monococcum. Target Site The methods of the present invention encompass steps of modifying plastid or mitochondrial DNA at one or more target sites, or steps of providing plastids or mitochondria that have already been modified at one or more target sites in their DNA. Suitably the one or more target sites are target sites in the plastid or mitochondrial genome. Suitably in the plastid or mitochondrial DNA which makes up the genome. Suitably the one or more target sites may be one or more intended sites of modification in the plastid or mitochondrial DNA. Suitably the plastid or mitochondrial genome or DNA is located within a plastid or mitochondrion respectively. Suitably the plastid or mitochondrial genome or DNA is located within a plant cell. Suitably the plastid or mitochondrial genome or DNA is located within a plastid or mitochondrion respectively, within a plant cell. Suitably the or each target site may be anywhere in the plastid or mitochondrial genome. Suitably the one or more target sites may be the located in a gene, or outside of a gene. Suitably the one or more target sites may be the located in the coding region or non-coding region of a gene, for example in a regulatory region of a gene. Suitable regulatory regions which may comprise the or each target site include: promoters, silencers, enhancers, insulators, operators, terminators, 5’UTRs, 3’UTRs, etc. Suitably the gene may be any gene that is encoded by a plastid genome. Suitably the gene may be any gene that is encoded by the chloroplast genome, suitably any gene that is mentioned in Jansen et al.2012, incorporated herein by reference. Suitably the gene may be any gene that is encoded by the mitochondrial genome, suitably any gene that is mentioned in Knoop 2012, incorporated herein by reference. In one embodiment, the target site is located in the 5’UTR of a gene, suitably the 5’UTR of a protein-coding gene or the 5’UTR of a non protein-coding gene such as a regulatory RNA- encoding gene. A 5’UTR is also referred to in the art as a leader sequence or 5’ leader sequence. Suitably the target site may encompass the junction between the coding region and the non- coding region of a gene, such as the junction between the 5’UTR and the first exon of a gene, or the junction between an exon and an adjacent intron of a gene. Suitably, the target site may span across the start codon (ATG) of a gene. Suitably, the target site may comprise the start codon (ATG) of a gene. Suitably the one or more target sites may be located in a gene which controls an important agronomic trait, suitably in a coding or non-coding region of a gene which controls an important agronomic trait. Suitably the one or more target sites may be located in the coding region of a gene which controls an important agronomic trait. Suitably such as a gene which controls growth, yield, disease resistance pest resistance, sterility etc. Suitably, genes which control plant growth (including genes that are involved in plant growth) may include one or more chloroplast genome encoded genes, suitably accD, psbA, rbcL, ycf3, petN (ycf6), atpB, petA, petB, petD, psbJ, psbE, psbF, psbL, psbJ, ycf5 (ccsA), psaJ, petG, psbM, psaA, a gene encoding a ribosomal protein subunit (such as a small subunit or a large subunit), an NADH dehydrogenase subunit, a gene encoding ribosomal DNA (rDNA), a gene encoding transfer RNA (tRNA), and / or other chloroplast genome encoded genes. Suitably, genes which control plant growth (including genes that are involved in plant growth) may include one or more mitochondrial genome encoded genes, suitably a gene encoding an ATP synthase subunit, a cytochrome c oxidase subunit, an NADH dehydrogenase subunit, a tRNA, an rDNA, a ribosomal protein subunit (such as small subunit or a large subunit), an uncharacterised ORF gene, and / or other mitochondrial genome encoded genes. Suitably the one or more target sites may be located in more than one gene. Suitably any reference herein to a target site in a gene may encompass multiple target sites in multiple different genes. In one embodiment, the one or more target sites are located in a gene which when modified controls cytoplasmic male sterility. In one embodiment, the one or more target sites are located in the coding region of a gene which when modified controls cytoplasmic male sterility. Suitably such genes may be mitochondrial genes related to respiration. Non-limiting examples of such genes include mitochondrial genes encoding ATP synthase subunits and NADH dehydrogenase subunits. Suitably such genes may be atp9, atp6c or nad7. Suitably, modification of one or more target sites in such genes controls cytoplasmic male sterility. Suitably therefore, the modification of one or more target sites in such genes converts the genes into CMS controlling genes. Suitably the modification at each target site in the plastid or mitochondrial DNA may comprise any type of genetic modification. Suitably the modification at a given target site may be a mutation such as a deletion, insertion, substitution, translocation, frameshift, inversion, rearrangement, truncation, repeat, expanding repeat, SNP etc. Suitably a substitution mutation may comprise a point mutation, suitably which may be a missense mutation, or a nonsense mutation. Suitably the modification in each instance may be of one or more nucleotides in the plastid or mitochondrial DNA. In some cases, the modification at a target site may comprise the insertion of a transgene. Suitably each target site may comprise a different modification. However typically, each target site comprises the same type of modification. Suitably, the modification at each target site in the plastid or mitochondrial DNA is an insertion mutation. Suitably, the modification at each target site in the plastid or mitochondrial DNA is an insertion of a sequence, suitably of a heterologous nucleic acid sequence. Suitably, the modification is an insertion which introduces a transgene into the plastid or mitochondrial DNA. Suitably, the modification at each target site in the plastid or mitochondrial DNA is a substitution mutation. Suitably, the modification at each target site in the plastid or mitochondrial DNA is a single nucleotide substitution mutation, i.e. a point mutation. Suitably which introduces an SNP into the plastid or mitochondrial DNA. Suitably modification of the plastid or mitochondrial DNA may be achieved by any means known in the art. Suitably by any means of mutagenesis of DNA known in the art, as explained below. Suitably, the protein used in the methods of the invention is capable of cleavage of any non- modified target sites in the plastid or mitochondrial DNA. Suitably a non-modified target site, in contrast to a modified target site discussed above, is a target site which does not contain a modification. Suitably a non-modified target site is the same site in the plastid or mitochondrial DNA as a modified target site, but which was not modified by the modification process or step (a) of the methods or step (iii) of the methods. Suitably due to the number of copies of the plastid and mitochondrial genome in each cell, processes of modification will not modify every target site in every copy of the genome. Some target sites will remain non-modified after a process or step of modification of plastid or mitochondrial DNA. Suitably the or each non- modified target site therefore comprises the unmodified sequence of DNA at the target site. Suitably the non-modified target site therefore comprises the wild type sequence of DNA at the target site. Modifying plastid or mitochondrial DNA / Modification Processes As explained above, the methods of the present invention encompass steps of modifying plastid or mitochondrial DNA at one or more target sites, or steps of providing plastids or mitochondria that have already undergone a modification process to modify one or more target sites in their DNA. Suitably the plastid or mitochondrial genome may be modified by any known means. Suitably the plastid or mitochondrial genome may be modified by any technique or process which creates a modification in plastid or mitochondrial DNA. Suitable modifications are explained above. Any suitable method of modifying DNA may be used. Suitably random mutagenesis or site-directed / targeted mutagenesis may be used. Suitable methods of modifying DNA may include: exposure of the plastid or mitochondrial DNA to a chemical or physical mutagen, transformation of the plastid or mitochondrial DNA with a transgene, a heterologous nucleic acid (which may encode a transgene), exposure of the plastid or mitochondrial DNA to a DNA modifying enzyme. Suitable chemical mutagens may include alkylating agents, base analogues, intercalating agents, deaminating agents, hydroxylating agents, cross-linking agents, radiomimetic agents, aromatic amines, nitrosamines, aldehydes, alkyl sulfonates, and alfatoxins. Some specific examples include: ethyl methanesulfonate (EMS), methylnitrosourea (MNU), 1-ethyl-1- nitrosourea (ENU), N-ethyl-N-nitrosourea, N-methyl-N-nitrosourea, Ethyleneimine, Diethyl sulfate, N-methyl-N1-nitro-N-nitrosoguanidine, 1-ethyl-2-nitro-1-nitrosoguanidine, Dimethyl sulfate, Methyl methanesulfonate, N,N-diethylnitrous amide, N,N-dimethylnitrous amide, Diepoxybutane, sodium azide (SA), vinyl chloride, benzene, phenol, 5-bromo-uracil, 5-bromo- deoxyuridine, ethidium bromide, acridines, psoralens and formaldehyde, for example. Suitably the dose of chemical mutagen used is an effective dose. Suitably the dose is equivalent to the concentration of the chemical per the duration of the treatment (Spencer- Lopes et al., 2018). The chemical's concentration and time of exposure varies depending on the chemical itself and the plant material / plant species considered. The evaluation of LD50at M1 generation can be considered the best proxy to quantify the amount of mutagen to be used (Lee et al., 2014). Also, temperature (usually in the range 20–25°C) is an important parameter to consider in chemical mutagenesis (Mba et al., 2010; Spencer-Lopes et al., 2018). The pH of the solution used to dissolve the chemical mutagen should be neutral or slightly acidic (pH 6–7) to reduce the decomposition of the mutagen itself. Overall, the chemical concentration, the time of exposure and the pH is to be adjusted depending on the chemical and the plant tissue to be treated (Leitao, 2012). Suitable effective doses of a chemical mutagen will be known in the art. Suitable physical mutagens may include radiation such as ionising or non-ionising radiation. Suitable examples of ionising radiation are: X-rays, alpha radiation, beta radiation, and gamma rays. Suitable examples of non-ionising radiation are UV radiation suitably UV-A, UV-B, UV- C rays, Fast neutron bombardment (FNB), protons, microwaves, RF radiation, and Ion beams, for example. Other forms of physical mutagenesis may include high temperature, mechanical stress, high pressure, sonication, electric fields, oxidative stress, desiccation. Suitably the exposure to the physical mutagen can be chronic, acute or fractionated. Suitably in chronic exposure, the plant or part thereof, plastid or mitochondrion is exposed to a relatively low dose of radiation for a long period of time (suitably weeks or months). Conversely, in acute exposure, the plant or part thereof, plastid or mitochondrion is exposed to a high dose of radiation for a short period of time (suitably seconds or minutes) (Mba et al., 2010). The fractionated approach consists of irradiating a plant or part thereof, plastid or mitochondrion at discrete time intervals to allow the material to recover between treatments (Kodym et al., 2012; Bado et al., 2015). Suitably the dose of radiation used is an effective dose. Suitably it depends on the species and the genotype of plant considered (Lee et al., 2014). In physical mutagenesis, the dose is currently measured in gray (Gy) which corresponds to 1 Joule per kg of treated material (1 Gy = 1 J kg−1) (Spencer-Lopes et al., 2018). A general rule is to apply an irradiation dose which results in a lethal dose of 50% of treated material (LD50) or reduction of 30–60% in growth (GR30-60) of the treatment material in M1 (Mba, 2013; Bado et al., 2015). Suitable effective doses of a physical mutagen will be known in the art. In one embodiment, the modification is carried out by exposing the plastid or mitochondrial DNA to a heterologous nucleic acid (i.e. a heterologous nucleic acid sequence). Suitably the modification is achieved by transforming the plant, or part thereof, with the heterologous nucleic acid or otherwise delivering the heterologous nucleic acid sequence into the plastid or mitochondrion of the plant or part thereof. Suitably transformation of a plant, or part thereof, with a heterologous nucleic acid sequence, which may be a transgene, may be carried out by known methods of transformation with a heterologous nucleic acid such as Agrobacterium mediated gene transfer, calcium phosphate- DNA co-precipitation, DEAE-dextran-mediated transfection, polybrene-mediated transfection, electroporation, microinjection, liposome fusion, lipofection, protoplast fusion, retroviral infection, and biolistics (gene gun). Suitably transformation may be carried out by known methods such as nanocarrier delivery by infiltration or bombardment for example (as described for example in Law et al.2023, incorporated herein by reference) and polyethylene glycol (PEG) transformation (as described for example in Golds et al. 1993, incorporated herein by reference). Suitably such methods are further described hereinbelow. Suitable, methods of transformation described hereinbelow in relation to expression construct and vectors and apply equally to the transformation of a heterologous nucleic acid sequence. In one embodiment, transformation is by particle bombardment (i.e. biolistic or microprojectile bombardment), suitably using gold or tungsten particles, suitably performed using a gene gun. Suitably, it is understood that the term “heterologous nucleic acid” refers to a nucleic acid sequence which does not naturally occur in the plant or part thereof into which it is being introduced, or a nucleic acid molecule which originates from outside said plant or part thereof such as a synthetic nucleic acid molecule for example. Suitably, the heterologous nucleic acid may modify or be capable of modifying the target site. Suitably by serving as a repair template, suitably a repair template such as a DNA repair template for repairing the target site. Suitably therefore, the heterologous nucleic acid may be referred to as donor DNA. Suitably, modification of the target site by the heterologous nucleic acid may be facilitated by inducing a break at the target site, such as a single-stranded or a double-stranded break, suitably a single-stranded or double-stranded DNA break or any other type of DNA damage such as the creation of an abasic site. Suitably, such a break may be induced in the target site by any known means in the art. Suitably, a break may be induced by chemical or physical mutagens, i.e. chemical or physical mutagenesis, as described hereinabove. Suitably, a break may be induced by targeted mutagenesis methods such as by providing a programmable nuclease such as a TALEN (including a pair of TALENs), ZFN (including a pair of ZFNs), meganuclease, or a CRISPR endonuclease such as Cas9 or Cas12a targeted at the target site. Suitably, the programmable nuclease may be provided, suitably provided in combination with the heterologous nucleic acid, by delivering it into the plant or part thereof, suitably by any transformation method. Suitably for example, the programmable nuclease may be one or more purified proteins, such as a TALEN protein or a pair of TALEN proteins, a ZFN protein or a pair of ZFN proteins, meganuclease protein, or a Cas9-gRNA ribonucleoprotein complex, appropriately tagged with a suitable plastid or mitochondria targeting peptide, which may be delivered into the plant or part thereof (e.g. a plant protoplast) for example by PEG transformation or electroporation. Suitably, the programmable nuclease may be provided by being co-delivered (i.e. contemporaneously delivered; i.e. simultaneously delivered) alongside the heterologous nucleic acid into the plant or part thereof, or by being delivered into the plant or part thereof shortly before or shortly after the heterologous nucleic acid is delivered into said plant or part thereof. Suitably, “shortly” may be up to 72 hours, suitably up to 60 hours, 48 hours, suitably up to 36 hours, suitably up to 24 hours such as up to 12, 10, 8, 6, 4 or 2 hours. Suitably, the programmable nuclease may be delivered on the same day as the heterologous nucleic acid into the plant or part thereof. Suitably, the target site of the programmable nuclease may be the same or different to the target site of the targeted protein capable of cleavage of any non-modified target site of step (b). Suitably, the programmable nuclease may be the same protein as the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA of step (b). Suitably therefore, the protein of step (b) may be used for introducing the breaks to facilitate modification of the target site using the heterologous nucleic acid, or it could be a different programmable nuclease as described hereinabove. Suitably, the break may be induced by providing (e.g., expressing) in the plant or part thereof or a cell of the plant or part thereof the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA. Suitably, there may be introduced into the plant or part thereof an expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein (e.g., a programmable nuclease) capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA under the control of an inducible system, expressing said construct and contemporaneously, shortly before or shortly after delivering a heterologous nucleic acid into the plastid or mitochondria of the plant or part thereof. Suitably, delivery of the heterologous nucleic acid may be after, shortly after such as 12 h, 24 h, 36 h 48 h, 60 h or 72 h after the expression of the construct, less than 24 h, 48 h or 72 h after the expression of the construct, more 24 h, 48 h or 72 h after the expression of the construct, or together with (i.e. contemporaneously or simultaneously with) the expression of the construct. Suitably, delivery of the heterologous nucleic acid may be before, shortly before such 1 h, 2 h, 3 h, 4 h, 5 h or 6 h before the expression of the construct, less than 8 h before the expression of the construct, more 8 h before the expression of the construct. Suitably therefore, there is provided a method of producing a homoplasmic modified plant or part thereof, the method comprising: (i) Introducing into the plant or part thereof an expression construct comprising: one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA under the control of an inducible expression system; (ii) Inducing expression of the protein from the expression construct; (iii) Modifying the non-modified target site to produce a modified plant or part thereof having plastid or mitochondrial DNA comprising one or more modified target sites, suitably wherein modification is by delivering a heterologous nucleic acid into the plant or part thereof, suitably into the plastid or the mitochondria of the plant or part thereof, and optionally repeating step (ii); (iv) Regenerating the modified plant or part thereof; and (v) Optionally repeating steps (ii), (iii) and (iv). Suitably, the method may be carried out on a plurality of plants or parts thereof. Suitably, the method may comprise a further step of selecting the plant or part thereof if it maintains or has improved viability compared to the plant or part thereof of step (i). Suitably after modifying the plastid or mitochondrial DNA (step (iii)) and before regeneration (step (iv)). Suitably, the delivery of the heterologous nucleic acid into the plant or part thereof may occur while the plastid or mitochondria targeted protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is present, or while said protein is expressed. Suitably, while said protein is present in the plastid or mitochondria. In some embodiments, the heterologous nucleic acid may be delivered into the plant or part thereof at the same time as, or on the same day as, the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In some embodiments, the heterologous nucleic acid may be delivered into the plant or part thereof within 72 h, 48 h, 24 h, 12 h, 8 h, 6 h, 4 h or within 2 h of the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In some embodiments, the heterologous nucleic acid may be delivered into the plant or part thereof up to 72 h, 48 h, 24 h, 12 h, 8 h, 6 h, 4 h or up to 2 h after the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In one embodiment, the method is an ex vivo method. Suitably therefore, there is provided a method of selecting modified plastids or mitochondria, the method comprising: (i) Introducing into one or more plastids or mitochondria an expression construct comprising: one or more nucleic acids which encode a protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA under the control of an inducible system; (ii) Inducing expression of the protein from the expression construct; and (iii) Modifying the non-modified target site to produce one or more plastids or mitochondria that have undergone a modification process, suitably wherein modification is by delivering a heterologous nucleic acid into the one or more plastids or mitochondria, and optionally repeating step (ii); (iv) Selecting the plastids or mitochondria which maintain or have improved viability compared to the plastids or mitochondria of step (i). In one embodiment, the modified plastids or mitochondria may be present in a cell, suitably in a plant cell. Therefore in one embodiment the method may comprise selecting cells comprising modified plastids or mitochondria, suitably comprising providing one or more cells which comprise one or more plastids or mitochondria, introducing into the one or more cells an expression construct, expressing the construct, modifying the one or more plastids or mitochondria by introducing into the one or more cells a heterologous nucleic acid and optionally express the construct again, and selecting the one or more cells which maintain or have improved viability. In such an embodiment, the one or more nucleic acids may encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites. Suitably, the delivery of the heterologous nucleic acid into the one or more plastids or mitochondria may occur while the plastid or mitochondria targeted protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is present, or while said protein is expressed. Suitably, while said protein is present in the plastid or mitochondria. In some embodiments, the heterologous nucleic acid may be delivered into the one or more plastids or mitochondria at the same time as, or on the same day as, the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In some embodiments, the heterologous nucleic acid may be delivered into the one or more plastids or mitochondria within 72 h, 48 h, 24 h, 12 h, 8 h, 6 h, 4 h or within 2 h of the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In some embodiments, the heterologous nucleic acid may be delivered into the one or more plastids or mitochondria up to 72 h, 48 h, 24 h, 12 h, 8 h, 6 h, 4 h or up to 2 h after the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In one embodiment, the method is an ex vivo method. Suitably therefore, there is provided a method of selecting plants or parts thereof comprising modified plastid or mitochondrial DNA, the method comprising: (i) Introducing into one or more plants or parts thereof an expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA, under the control of an inducible system; (ii) Inducing expression of the protein from the expression construct; (iii) Modifying the non-modified target site in the plastid or mitochondrial DNA to produce one or more modified plants or parts thereof, suitably wherein modification is by delivering a heterologous nucleic acid into the plant or part thereof, suitably into the plastid or the mitochondria of the plant or part thereof, and optionally repeating step (ii); (iv) Selecting the plants or parts thereof which maintain or have improved viability compared to the plants of step (i). In one embodiment, the method may further comprise step (v) regenerating the selected plastids or mitochondria, suitably which may comprise regenerating one or more cells containing the selected plastids or mitochondria. In one embodiment, the method may further comprise repeating steps (ii) to (v). Suitably, the delivery of the heterologous nucleic acid into the plant or part thereof (suitably into the plastid or the mitochondria of the plant or part thereof) may occur while the plastid or mitochondria targeted protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is present, or while said protein is expressed. Suitably, while said protein is present in the plastid or mitochondria. In some embodiments, the heterologous nucleic acid may be delivered into the plant or part thereof (suitably into the plastid or the mitochondria of the plant or part thereof) at the same time as, or on the same day as, the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In some embodiments, the heterologous nucleic acid may be delivered into the plant or part thereof (suitably into the plastid or the mitochondria of the plant or part thereof) within 72 h, 48 h, 24 h, 12 h, 8 h, 6 h, 4 h or within 2 h of the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In some embodiments, the heterologous nucleic acid may be delivered into the plant or part thereof (suitably into the plastid or the mitochondria of the plant or part thereof) up to 72 h, 48 h, 24 h, 12 h, 8 h, 6 h, 4 h or up to 2 h after the protein capable of cleavage of a non-modified target site in the plastid or mitochondrial DNA is expressed. In one embodiment, the methods is an ex vivo method. Suitably, breaks in the target site may enhance the efficiency with which the target site is modified by the heterologous nucleic acid, suitably by inducing or upregulating DNA repair pathways such as non-homologous end-joining (NHEJ) which may insert a heterologous nucleic acid such as a double-stranded DNA into the break site, and / or DNA-templated repair pathways such as homologous recombination (HR), microhomology-mediated end-joining (MMEJ), polymerase theta-mediated end joining (TMEJ), single-strand annealing (SSA) which may insert or copy over the sequence of the heterologous nucleic acid into the target site, and / or other DNA repair pathways. Alternatively, breaks may not be necessary to include in the target site for the heterologous nucleic acid to modify the target site. Suitably, the target site may be modified without any breaks in the target site through natural cellular DNA repair pathways which may modify an unbroken target site using the heterologous nucleic acid (typically requiring the heterologous nucleic acid to have one or more regions of complementarity to the target site such as homology arms on either side for facilitating homologous recombination), or equally by DNA breaks that may occur naturally as part of the normal life and metabolism of a cell. Suitably therefore, as mentioned hereinabove, modification may be carried out by in a single step of transforming the plant, or part thereof, with a heterologous nucleic acid. Suitably, the heterologous nucleic acid may be capable of modifying the target site or a region of DNA encompassing the target site by targeting (e.g. binding to) said target site or region, suitably by comprising one or more regions of identity to the target site or the region of DNA encompassing the target site, suitably one or more regions or partial or complete identity to the target site or said region. Suitably, partial identity may be at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or higher levels of identity. In one embodiment, the one or more regions of identity are completely identical to the target site or a region of DNA encompassing the target site. Suitably, the one or more regions of identity may be located on either end (i.e. on both ends, i.e. on both the 5’ and 3’ end) of the heterologous nucleic acid, suitably flanking a central region. Suitably, the central region may be non-identical, suitably less than 20% or 50% identical, suitably different to, the target site. Suitably, the central region may encode a gene such as a transgene or cis-gene. Suitably, the central region may be at least partially identical, such as more than 50% or 70% identical, or completely identical, to the target site. Suitably, the one or more regions of identity may be referred to as homology arms, suitably flanking homology arms on either end of the heterologous nucleic acid. Suitably, homology arms may be short homology arms, suitably for example 5, 10, 20, 30, 40 or 50 nucleotides or base pairs in length. Suitably, homology arms may be long homology arms, suitably for example 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 or more nucleotides or base pairs in length. In one embodiment, the heterologous nucleic acid is adapted for homology directed repair (HDR), suitably having homology arms (i.e. regions of identity) on both ends, suitably the 5’ and 3’ end, of the heterologous nucleic acid. Suitably the homology arms are 100% identical to a region of DNA that encompasses the target site. HDR repair templates and how to design effective HDR templates are known in the art. Suitably, the one or more regions of identity (such as the homology arms) may enable the heterologous nucleic acid to target (e.g. bind to) the target site through base pairing complementarity. Suitably, whereby the targeting or binding of the heterologous nucleic acid to the target site modifies or is capable of modifying the target site. Suitably by templated DNA repair processes such as for example homology directed repair (HDR), homologous recombination (HR), microhomology-mediated end-joining (MMEJ), polymerase theta- mediated end joining (TMEJ), single-strand annealing (SSA) or other DNA repair pathways. Targeting of the heterologous nucleic acid to a target site may be considered a form of site- directed / targeted mutagenesis. Suitably, the heterologous nucleic acid may be capable of modifying the target site without comprising any region of identity to the target site. Suitably, the heterologous nucleic acid may be inserted into the target site by end-joining such as by NHEJ, suitably thereby modifying the target site. Suitably, as mentioned hereinabove, modification by a heterologous nucleic acid molecule does not necessarily require inducing breaks in the target site since for example breaks in the target site may naturally occur as part of the normal life and metabolism of a cell. Alternatively, breaks may be induced in the target site to facilitate the insertion of the heterologous nucleic acid as described hereinabove. Suitably, where no breaks are induced in the target site, this may be considered a form of random mutagenesis. Suitably, the heterologous nucleic acid may be a DNA nucleic acid molecule (e.g., double- or single-stranded), an RNA nucleic acid molecule, or a DNA-RNA hybrid nucleic acid molecule. Suitably, the heterologous nucleic acid may be a circular nucleic acid molecule, suitably a vector, suitably a DNA vector or RNA vector. Suitably, the heterologous nucleic acid may be linear nucleic acid molecule, suitably a linear DNA or linear RNA. Suitably, the heterologous nucleic acid may be single-stranded, double-stranded or a partially double-stranded nucleic acid molecule, suitably a single-stranded DNA or RNA molecule, suitably a double-stranded DNA or RNA molecule, suitably a single-stranded linear DNA or RNA molecule, suitably a double-stranded linear DNA or RNA molecule. In one embodiment, the heterologous nucleic acid is a DNA vector, suitably a double-stranded DNA vector. In one embodiment, the heterologous nucleic acid is a linear DNA molecule, suitably a linear double-stranded DNA molecule. Suitably, the heterologous nucleic acid may be at least 30, 50, 70, 100, 500 or 1000 nucleotides or base pairs long. Suitably, the heterologous nucleic acid may be up to 10,000, 5000, 3000, 1500, 1000, 500, 120, 100, 70, 50 or 30 nucleotides or base pairs long. Suitably, the heterologous nucleic acid may encode one or more transgenes, transgene cassettes or expression cassettes for example. Suitably, the heterologous nucleic acid may not necessarily comprise a selection marker such as an antibiotic selection marker, suitably the heterologous nucleic acid does not comprise a selection marker. Suitably, it is possible to provide a homoplasmic modified plant or part thereof with the methods of the invention whereby the modifications are introducing using a heterologous nucleic acid which does not comprise a selection marker. Suitably, the heterologous nucleic acid may also encode or comprise a sequence that is partially identical to that of the target site, comprising one or more modifications, suitably one or more modifications with respect to the target site, such as one or more nucleotide or trinucleotide substitutions, insertions and / or deletions in the sequence of the target site, i.e. the heterologous nucleic acids may encode certain modifications in the target site. Suitably, the heterologous nucleic acid may be at least partially identical to a region of DNA that encompasses the target site, comprising one or more modifications, suitably one or more modifications with respect to the region of DNA or the target site. Suitably, transgenes may encode genes involved in regulating plant growth, drought tolerance, resistance to biotic or abiotic stresses, production of recombinant proteins or peptides, production of bio-compounds, cytoplasmic male sterility, and more. Suitably, transgenes may encode genes including herbicide resistance genes (e.g. bar gene), pest resistance gene (e.g. Cry toxin gene), CMS causal genes (sugar beet orf129, Brassica juncea orf288, maize atp6c), any gene that confers cytoplasmic male sterility, and more. Suitably, a plurality nucleic acid sequences may be employed to modify a plurality of target sites in the plastid or mitochondria of a plant or part thereof. Suitably, each of the plurality of nucleic acid sequence may or may not comprise homology arms that are homologous to their respective target sites. Suitably therefore in embodiments of the methods of the invention, step (a) of modification of the plastid or mitochondrial genome may comprise: i) providing one or more plastids or mitochondria that have undergone modification using a heterologous nucleic acid to modify one or more target sites in the plastid or mitochondrial DNA, or: ii) modifying plastid or mitochondrial DNA of the plant, or part thereof, at one or more target sites using one or more heterologous nucleic acids, such as by transforming the plant or part thereof with one or more heterologous nucleic acids, suitably wherein the or each heterologous nucleic acid may comprise homology arms that are homologous to its respective target site, to produce a modified plant or part thereof having plastid or mitochondrial DNA comprising one or more modified target sites. Suitably, modification of the plastid or mitochondrial DNA using a heterologous nucleic acid is achieved by transforming the plant or part thereof, or the plastid or mitochondria, with the heterologous nucleic acid. In further aspects of the invention there may be provided a kit of parts comprising a heterologous nucleic acid, and an expression construct comprising one or more nucleic acids encoding a plastid or mitochondria targeted protein capable of cleavage of a non-modified target sites in plastid or mitochondrial DNA under the control of an inducible expression system. Optionally further comprising a second expression construct comprising one or more nucleic acids encoding the components of an inducible system. Optionally further comprising instructions for use of the heterologous nucleic acid and one or more expression constructs to modify plastid or mitochondrial DNA, suitably in a plant or part thereof, and to select plants or parts thereof, plastids or mitochondria, comprising modified plastid or mitochondrial DNA. Optionally further comprising suitable reagents for using the expression constructs, such as reagents for transformation of plants or parts thereof, plastids or mitochondria. In some preferred embodiments of the various aspects and embodiments described above, the heterologous nucleic acid is a template nucleic acid suitable for homology directed repair (HDR) comprising homology arms flanking a desired sequence to be inserted at the target site. Suitably, the heterologous nucleic acid may be a synthetic organelle genome such as a synthetic plastid genome (e.g. a synthetic chloroplast genome) or a synthetic mitochondrial genome. Suitably, a synthetic organelle genome is one which has been synthesised outside of the plant or part thereof, suitably synthesised in vitro or ex vivo. Suitably, a synthetic organelle genome may comprise one or more modifications with respect to the corresponding native (i.e. natural, i.e. endogenous) genome. Suitably therefore, modifying a non-modified target site in the plastid or mitochondrial DNA of a plant or part thereof may include introducing into the plastid or mitochondria of the plant or part thereof a synthetic plastid or mitochondrial genome, respectively. Suitably the synthetic plastid or mitochondrial genome may comprise one or more modifications with respect to the native plastid or mitochondrial genome, respectively. Suitably therefore, the one or more plastids or mitochondria that have undergone a modification process to modify one or more target sites may include one or more plastids or mitochondria comprising a synthetic plastid or mitochondrial genome, respectively. Suitably therefore, modified plastid or mitochondrial DNA may include plastid or mitochondrial DNA comprised on a synthetic plastid or mitochondrial genome, respectively. Suitably, a synthetic organelle genome may replace or be a replacement of the native (i.e. natural, i.e. endogenous) genome of the respective organelle of the plant or part thereof. Suitably, a synthetic plastid genome may replace or be a replacement of the native genome of the corresponding plastid of the plant or part thereof, e.g. a synthetic chloroplast genome may replace or be a replacement of the native genome of the chloroplast. Suitably, a synthetic mitochondrial genome may replace or be a replacement of the native mitochondrial genome. Suitably, a synthetic organelle genome may become homoplasmic within the plant or part thereof. Suitably therefore, a synthetic organelle genome may, in some cases entirely, replace the natural genome of the respective organelle of the plant or part thereof. Suitably, a synthetic organelle genome may be up to 150,000 bp or 200,000 bp long, or longer. Suitably therefore, the methods described herein may produce a plant or part thereof that is homoplasmic for a synthetic plastid or mitochondrial genome. Suitably therefore, the methods described herein may be for selecting synthetic plastids or mitochondria. Suitably therefore, the methods described herein may be for selecting a plant or part thereof comprising a synthetic plastid or mitochondrial genome. Suitable DNA modifying enzymes may include a nuclease, a meganuclease, an endonuclease, a base editor such as a cytosine deaminase, and / or a polymerase for example. In one embodiment, the modification is carried out by exposing the plastid or mitochondrial DNA to a polymerase enzyme. Suitably by transforming the plant, or part thereof with a nucleic acid encoding a polymerase enzyme. Suitably using the methods of transformation mentioned above and further described hereinbelow. Suitably the polymerase enzyme is a DNA polymerase. Suitably the polymerase is an error prone polymerase. Suitably the polymerase is an error prone DNA polymerase. Suitably such that the polymerase introduces errors, i.e. modifications into the plastid or mitochondrial DNA when replicating the DNA. Suitably the error prone polymerase has a high error rate, and suitably a low fidelity. Suitably the error prone polymerase has a probability of introducing nucleotide substitutions or frameshift mutations in the plastid or mitochondrial DNA, suitably at an error rate of 10−3to 7.5x10−1. Suitably the error prone polymerase may lack proofreading exonuclease activity. Suitable error prone polymerases may be selected from polymerase families A, B, X and Y. Suitable error prone polymerases may be selected from a terminal deoxynucleotidyl transferase (TdT), a DNA polymerase (including any of β, ζ, κ, η, ι, λ, μ, γ DNA polymerases), Rev1 and a plant organellar polymerase (POP). In one embodiment, the error prone polymerase is a plant organellar polymerase (POP). Suitably the plant organellar polymerase may be derived from any species of plant, algae or protozoan. Suitably the organellar polymerase may be derived from the following species of plant, for example: Arabidopsis thaliana, Brassica rapa, Nicotiana tomentosiformis, Oryza sativa, Physcomitrella patens, Solanum lycopersiucm, Zea mays, Petunia axillaris, Nicotiana tabacum. In some embodiments, the organellar polymerase may be derived from a species of moss, for example from Physcomitrella patens. In one embodiment, the plant organellar polymerase is derived from Nicotiana tabacum. In one preferred embodiment, the error prone polymerase is a modified plant organellar polymerase, suitably which has been modified to increase its probability of introducing errors into plastid or mitochondrial DNA i.e. to increase its error rate. Suitable such modified POPs are described in for example Ji and Day. Nucleic Acids Research, 2020, Vol.48 No.21, or WO2023 / 073333, incorporated herein by reference. Suitably therefore, the modification step of the methods of the invention is carried out by exposing the plastid or mitochondrial DNA to a plant organellar polymerase (POP). Suitably wherein the plant organellar polymerase (POP) enzyme may comprise an amino acid sequence according to SEQ ID NO: 22 or an amino acid sequence having at least 35% identity thereto, or a functional fragment thereof. Suitably the plant organellar polymerase (POP) enzyme may comprise an amino acid sequence having at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity to SEQ ID NO: 22. Suitably, the plant organellar polymerase may further comprise one or more modifications. In one embodiment, the plant organellar polymerase (POP) comprises an amino acid sequence which is a variant of SEQ ID NO:22, or an amino acid sequence having at least 35% identity thereto, or a functional fragment thereof. By ‘variant’ it is meant that the reference sequence, such as SEQ ID NO:22, contains one or more modifications. Suitably the plant organellar polymerase (POP) comprises an amino acid sequence according to SEQ ID NO:22, or an amino acid sequence having at least 35% identity thereto, or a functional fragment thereof, comprising a modification at or corresponding to position L903, and optionally one or more further modifications at the following positions: D390, E392, R862, E904, and N1065 of SEQ ID NO:22, or positions corresponding thereto. Suitably wherein the amino acid sequence may have at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity to SEQ ID NO: 22, and comprise one or more modifications at the positions listed above. “Identity” or “percent identity” refers to the degree of sequence variation between two given nucleic acid or amino acid sequences. For sequence comparison, typically one sequence acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of (Smith and Waterman, 1981), by the homology alignment algorithm of (Needleman and Wunsch, 1970), by the search for similarity method of (Pearson and Lipman, 1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection. One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, which is described in (Altschul et al., 1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (on the world wide web at ncbi.nlm.nih.gov / ). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighbourhood word score threshold (Altschul et al., 1990) These initial neighbourhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when the cumulative alignment score falls off by the quantity X from its maximum achieved value, the cumulative score goes to zero or below due to the accumulation of one or more negative-scoring residue alignments, or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, a cutoff of 100, M=5, N=- 4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix ((Henikoff and Henikoff, 1992). In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (Karlin and Altschul, 1990). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a test nucleic acid sequence is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid sequence to the reference nucleic acid sequence is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001. A "functional fragment" refers to a protein fragment that retains the function of the full length protein. As such, a functional fragment of an plant organellar polymerase enzyme is a fragment, portion or part of such a protein that is capable of catalysing the replication of organellar DNA. In one embodiment, the plant organellar polymerase (POP) is an error prone organellar (DNA) polymerase. In one embodiment the plant organellar polymerase (POP) is a modified plant organellar (DNA) polymerase. In one embodiment the plant organellar polymerase (POP) is a mutated plant organellar (DNA) polymerase. Suitably therefore, the modification step of the methods of the invention is carried out by exposing the plastid or mitochondrial DNA to an error prone plant organellar polymerase, wherein in one embodiment, the plant organellar polymerase (POP) enzyme comprises an amino acid sequence according to SEQ ID NO:22 wherein the amino acid sequence comprises a modification at position L903, and optionally one or more further modifications at the following positions: D390, E392, R862, E904, and N1065 of SEQ ID NO:22. Suitably wherein the POP is error prone. Suitably therefore, the modification step of the methods of the invention is carried out by exposing the plastid or mitochondrial DNA to an error prone plant organellar polymerase, wherein in one embodiment, the plant organellar polymerase (POP) enzyme comprises an amino acid sequence according to SEQ ID NO:22 wherein the amino acid sequence comprises a modification at position L903, and further modifications at the following positions: D390 and E392 of SEQ ID NO:22. Suitably wherein the POP is error prone. Suitably the one or more modifications comprise one or more amino acid substitutions. Suitably any amino acid may be used for the substitution. Suitably any proteinogenic amino acid may be used for the substitution. Suitably the substitution is a conservative substitution. By ‘conservative’ it is meant that an amino acid with similar characteristics may be used for the substitution. Conservative amino acid substitutions” refer to the interchangeability of residues having similar side chains, and thus typically involves substitution of an amino acid in a polypeptide with amino acids within the same or similar defined class of amino acids. By way of example, an amino acid with an aliphatic side chain may be substituted with another aliphatic amino acid, e.g., alanine, valine, leucine, and isoleucine; an amino acid with hydroxyl side chain may be substituted with another amino acid with a hydroxyl side chain, e.g., serine and threonine; an amino acids having aromatic side chains may be substituted with another amino acid having an aromatic side chain, e.g., phenylalanine, tyrosine, tryptophan, and histidine; an amino acid with a basic side chain may be substituted with another amino acid with a basic side chain, e.g., lysine and arginine; an amino acid with an acidic side chain may be substituted with another amino acid with an acidic side chain, e.g., aspartic acid or glutamic acid; and a hydrophobic or hydrophilic amino acid may be substituted with another hydrophobic or hydrophilic amino acid, respectively. Suitably position L903 or a position corresponding thereto is substituted with an amino acid selected from Methionine (M), Asparagine, Phenylalanine (F) and Alanine (A) . In one embodiment, L903 or a position corresponding thereto is substituted with phenylalanine (F). Therefore the plant organellar polymerase enzyme comprises the modification L903F, or the same modification at a corresponding position. Suitably positions D390 and E392 or a position corresponding thereto are substituted with an amino acid selected from alanine (A), valine (V), Leucine (L), Isoleucine (I).In one embodiment, D390 or a position corresponding thereto is substituted with alanine (A). Therefore the plant organellar polymerase enzyme comprises the modification D390A or the same modification at a corresponding position. In one embodiment, E392 or a position corresponding thereto is substituted with alanine (A). Therefore the plant organellar polymerase enzyme comprises the modification E392A or the same modification at a corresponding position. Suitably ‘corresponding position’ as used herein means the same amino acid position in a different reference sequence, suitably in a different reference sequence to that of SEQ ID NO:22, suitably in a different organellar polymerase sequence. Therefore whilst the statements herein refer to SEQ ID NO:22, the invention is not restricted to the plant organellar polymerase of SEQ ID NO:22, each modification may be located at a position corresponding to an amino acid position denoted above in another plant organellar polymerase enzyme sequence. Therefore the invention equally refers to other organellar DNA polymerase enzymes having different amino acid sequences with the same modifications. It is possible to compare organellar DNA polymerase polypeptides by sequence comparison and locate conserved regions that correspond to the amino acid positions listed above. Sequence comparison to find corresponding positions may be carried out by aligning the amino acid sequences of two or more proteins, using an alignment program such as BLAST®. Methods for the alignment of sequences for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA and TFASTA. GAP uses the algorithm of Needleman and Wunsch ((1970) J Mol Biol 48: 443-453) to find the global (i.e. spanning the complete sequences) alignment of two sequences that maximizes the number of matches and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of the similarity between the two sequences. The software for performing BLAST analysis is publicly available through the National Centre for Biotechnology Information (NCBI). Homologues may readily be identified using, for example, the ClustalW multiple sequence alignment algorithm (version 1.83), with the default pairwise alignment parameters, and a scoring method in percentage. Global percentages of similarity and identity may also be determined using one of the methods available in the MatGAT software package (Campanella et al., BMC Bioinformatics.2003 Jul 10;4:29. MatGAT: an application that generates similarity / identity matrices using protein or DNA sequences.). Minor manual editing may be performed to optimise alignment between conserved motifs, as would be apparent to a person skilled in the art. Furthermore, instead of using full-length sequences for the identification of homologues, specific domains may also be used. The sequence identity values may be determined over the entire nucleic acid or amino acid sequence or over selected domains or conserved motif(s), using the programs mentioned above using the default parameters. For local alignments, the Smith-Waterman algorithm is particularly useful (Smith TF, Waterman MS (1981 ) J. Mol. Biol 147(1 );195-7). In the present case, a corresponding position in a different organellar DNA polymerase sequence may be found by aligning the amino acid sequence of said other organellar DNA polymerase with SEQ ID NO:22 and locating the same amino acid position as those listed. Suitably therefore, the modification step of the methods of the invention is carried out by exposing the plastid or mitochondrial DNA to an error prone plant organellar polymerase, wherein in one embodiment, the plant organellar (DNA) Polymerase enzyme comprises or consists of an amino acid sequence according to SEQ ID NO:23. Suitably therefore in preferred embodiments of the methods of the invention, step (a) of modification of the plastid or mitochondrial genome may comprise: providing one or more plastids or mitochondria that have undergone modification with a plant organellar polymerase suitably an error prone plant organellar polymerase to modify one or more target sites in the plastid or mitochondrial DNA, or: modifying plastid or mitochondrial DNA of the plant or part thereof at one or more target sites using a plant organellar polymerase suitably an error prone plant organellar polymerase to produce a modified plant or part thereof having plastid or mitochondrial DNA comprising one or more modified target sites. Suitably modification of the plastid or mitochondrial DNA using the plant organellar polymerase is achieved by transforming the plant or part thereof, or the plastid or mitochondria, with a nucleic acid encoding the plant organellar polymerase and optionally a suitable plastid or mitochondrial targeting peptide. Suitably, further comprising inducing expression of the nucleic acid in the plant or part thereof, in order modify the plastid or mitochondrial DNA. Suitable means of transforming a plant or part thereof with a plant organellar polymerase to effect modifications in plastid or mitochondrial DNA are explained hereinbelow and also in application WO2023 / 073333, incorporated herein by reference. In further aspects of the invention there may be provided a kit of parts comprising a first expression construct comprising one or more nucleic acids encoding a plant organellar polymerase suitably an error prone plant organellar polymerase , and a second expression construct comprising one or more nucleic acids encoding a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in plastid or mitochondrial DNA under the control of an inducible system. Optionally further comprising a third expression construct comprising one or more nucleic acids encoding the components of an inducible system. Optionally further comprising instructions for use of the expression constructs to modify plastid or mitochondrial DNA, suitably in a plant or part thereof, and to select plants or parts thereof, plastids or mitochondria, comprising modified plastid or mitochondrial DNA. Optionally further comprising suitable reagents for using the expression constructs, such as reagents for transformation of plants or parts thereof, plastids or mitochondria. Protein capable of cleavage of a non-modified target site in DNA The present invention, in some aspects and embodiments, makes use of a protein which is capable of cleavage of DNA, suitably which is capable of cleavage of non-modified target sites in DNA, i.e. cleavage of target sites having a wild type sequence. Suitably as explained elsewhere herein, the protein is used as a selective cleavage tool to cleave any plastid or mitochondrial DNA which is not modified at a desired target site, i.e. remains unmodified after the initial step of modification in the methods described herein. Thereby leaving only desired plastid or mitochondrial DNA which is modified at the target site. With several rounds of regeneration the protein can selectively cleave all plastid or mitochondrial DNA which is not modified at a desired target site to leave plants having only modified plastid or mitochondrial DNA i.e. homoplasmic plants. Suitably the protein may be any protein capable of site specific DNA cleavage. Suitably the protein is a site directed nuclease. For example nucleases, meganucleases, Zinc finger nucleases, CRISPR-Cas systems, transcription activator-like effector nucleases (TALEN)s, and / or restriction enzymes. Suitably such proteins may be nicking enzymes, such as Cas9 nickase variants, S1 nuclease and / or nicking restriction enzymes. The skilled person can readily target known site directed nucleases (such as CRISPR / Cas9 or other CRISPR nucleases, TALENs or ZFNs) to any desired target sited in the plastid or mitochondrial DNA discussed above. In the case of CRISPR / Cas9 or other CRISPR nucleases the method suitably comprises providing a guide RNA to direct the Cas9 protein to the desired target site. Accordingly, in any aspect or embodiment in which the targeted protein capable of cleavage is CRISPR / Cas9 or other CRISPR nuclease system, a suitable guide RNA is provided which targets the targeted protein to the target site. The guide RNA can suitably be expressed (transcribed) from any suitable nucleic acid construct or the guide RNA it can be delivered to the cell, plastid or mitochondrion directly through other techniques described herein. Suitable systems express a guide RNA are well known in the art, as discussed below. In the case of TALEN or ZFN it is the protein code of the site-specific nuclease that determines the binding site of the site-specific nuclease. Suitably, the site-directed nuclease comprises at least one zinc finger nuclease (ZFN), Transcription Activator-Like Effector Nuclease (TALEN), RNA-guided DNA nuclease (e.g. CRISPR / Cas9 or other CRISPR nuclease, such as CRISPR / Cpf1, transposon associated nucleases, or HEARO nucleases), or a meganuclease. Suitably the cleavage of the DNA may be single stranded or double stranded cleavage. Suitably the cleavage of the DNA is double stranded cleavage. Suitably the protein is a nuclease. It should be noted that the term ‘nuclease’ is intended to cover any biological enzyme which creates a single or double stranded cut of a target nucleic acid. Accordingly, the term includes nickases and recombinases, as well as more conventional nucleases which cause single or double stranded breaks. Suitably in one embodiment, the nuclease introduces double stranded breaks into the plastid or mitochondrial DNA. Suitably the protein is a transposon associated nuclease, suitably derived from the IS200 / IS605 transposon family. Suitably such technology is described in https: / / www.nature.com / articles / s41586-023-05826-x. Suitably the protein is a TnpB nuclease. Suitably the protein is a HEARO nuclease. Suitably such technology is described in https: / / www.nature.com / articles / s41467-022-35257-7. Suitably the protein is a Zinc finger nuclease. ZFN technology is described extensively in the literature and, inter alia, in the following patent documents: US 6,479,626, 6,534,261, 6,607,882, 6,746,838, 6,794,136, 6,824,978, 6,866,997, 6,933,113, 6,979,539, 7,013,219, 7,030,215, 7,220,719, 7,241,573, 7,241,574, 7,585,849, 7,595,376, 6,903,185, 6,479,626, 8,106,255, 20030232410, and 20090203140, all of which are incorporated by reference. ZFNs can be obtained commercially from Sigma-Aldrich (St. Louis, MO, US) under the CompoZr® Zinc Finger Nuclease Technology branded products and services. Suitably the protein is a CRISPR-Cas system. CRISPR / Cas technology is described extensively in the literature (e.g. Cong et al. ‘Multiplex Genome Engineering Using CRISPR / Cas Systems’, Science, 15 February 2013: Vol.339 no.6121 pp.819-823) and, inter alia, in the following patent documents: US 8,697,359, US2010076057, WO2013 / 176772, US8,771,945, US2010076057, US2014186843, US2014179770, US2014179006, WO2014093712, WO2014093701, WO2014093635, WO2014093694, WO2014093655, WO2014093709, WO2013 / 188638, WO2013 / 142578, WO2013 / 141680, WO2013 / 188522, US8546553, WO2014 / 089290, and WO2014 / 093479, all of which are incorporated by reference. CRISPR / Cas systems can be obtained commercially from Sigma-Aldrich (St. Louis, MO, US) under the CRISPR / Cas Nuclease RNA-guided Genome Editing suite of products and services, or from Thermo Fisher Scientific, Inc. (Waltham, MA, US) under the GeneArt® CRISPR branded products and services. CRISPR / Cpf has also been widely described in the literature. Of course, in this rapidly developing field other techniques for genetic editing are likely to become available. Such techniques could, in many cases, be readily adapted for use in the present invention. In one preferred embodiment, the protein is a TALEN (Feng et al.2013 Cell Res.23, 1229- 1232, Sander & Joung Nat. Biotechnol.32, 347-3552014). TALEN technology is described extensively in the literature and, inter alia, in the following patent documents: US8420782, US8470973, US8440431, US8440432, US8450471, US8586363, US8697853, EP2510096, US8586526, US8623618, EP2464750, US2011041195, US2011247089, US2013198878, WO2012 / 116274, WO2014110552, WO2014070887, WO2014022120, WO2013192316, and WO2010008562, all of which are incorporated by reference. TALENs can be obtained commercially from Thermo Fisher Scientific, Inc. (Waltham, MA, US) under the GeneArt® TALs branded products and services (formerly marketed under the Life Technologies brand). Transcription activator-like effectors (TALEs) can be quickly engineered to bind practically any DNA sequence. The term TALEN, as used herein, is broad and includes a monomeric TALEN that can cleave double stranded DNA without assistance from another TALEN. The term TALEN is also used to refer to one or both members of a pair of TALENs that are engineered to work together to cleave DNA at the same site. TALENs that work together may be referred to as a left-TALEN and a right- TALEN, which references the handedness of DNA. Transcription activator-like effector (TALE) DNA binding domains contain a highly conserved 33-34 amino acid sequence with the exception of the 12th and 13th amino acids. These two locations are highly variable (Repeat Variable Diresidue (RVD)) and show a strong correlation with specific nucleotide recognition. This simple relationship between amino acid sequence and DNA recognition has allowed for the engineering of specific DNA binding domains by selecting a combination of repeat segments containing the appropriate RVDs. Fusion of a TALE DNA binding domain with effector domains such as an endonuclease can create a TALE nucleases (TALENs). Suitably in the present invention the TALEN is formed of a pair of TALE domains, suitably a first (left) TALE domain and a second (right) TALE domain, fused to one or more endonuclease domains, termed the first (left) TALEN domain and the second (right) TALEN domain. Suitably the first (left) TALEN domain and the second (right) TALEN domain may comprise a plurality of TALE repeat sequences and an endonuclease domain. Suitably each of the plurality of TALE repeat sequences comprises an RVD capable of recognising the target site in the plastid or mitochondrial DNA. Suitably the first (left) TALE domain and the second (right) TALE domain may each comprise between 1.5–33.5 TALE repeats. Suitably each TALE repeat is around 33 to 34 amino acids in length. Suitably the sequence of each TALE domain may be designed to recognise and bind to a chosen target site in plastid or mitochondrial DNA, suitably to a chosen non-modified target site in plastid or mitochondrial DNA. Suitable programs and software are available in the art to design TALE domains to form a TALEN which will recognise and bind to a chosen target site in DNA. Suitably each of the TALEN domains may comprise a spacer. Suitably the spacer sequence may be located between the plurality of TALE repeat sequences and the endonuclease domain. The spacer sequence may be encoded by a sequence of 12 to 30 nucleotides. Suitably each TALE domain is targeted to bind to a non-modified target site in plastid or mitochondrial DNA. Suitably wherein each TALE domain is complementary to a half of a non- modified target site in plastid or mitochondrial DNA. Suitably the first (left) TALE domain comprises the ability to bind to a first half of the non-modified target site in plastid or mitochondrial DNA, and the second (right) TALE comprises the ability to bind to a second half of the non-modified target site in plastid or mitochondrial DNA. Suitably the first and second TALEN domains cooperate to effect double stranded cleavage of the plastid or mitochondrial DNA. Suitably the TALEN comprises TALE domains having a standard scaffold known in the art. Suitably such TALE scaffolds may be developed from TALE domains, for example by truncating the N or C terminus of known wild type TALE domains. Many different TALE domains and scaffolds are therefore known in the art. Suitable TALE domains may be derived from the gene AvrBs4 or AvrBs3. Suitable TALE domains may be selected from A4-NC or TALE13, for example, or derivatives thereof. In one embodiment such TALE scaffolds are selected from: (N-terminus / C-terminus) +153 / +47, which may suitably be derived from an A4- NC TALE domain, and +136 / +63, which may suitably be derived from a TALE13 domain. Suitably therefore each TALE domain comprises a +153 N-terminus (SEQ ID NO:24) and a +47 C-terminus (SEQ ID NO:25) (+153 / +47 architecture, Mussolino et al.2011 Nucleic Acids Res.39(21), 9283-9293). Alternatively each TALE domain comprises a +136 N-terminus (SEQ ID NO:26) and a +63 C-terminus (SEQ ID NO:27) (+136 / +63 architecture, Miller et al.2010 Nature Biotechnology 29, 143-148). Suitably wherein the numbers indicate the number of remaining amino acids in the N / C termini of each TALE domain. Suitably the first TALE domain comprises a sequence of SEQ ID NO: 6 or 4. Suitably the second TALE domain comprises a sequence of SEQ ID NO:7 or 5. Suitably wherein the RVDs highlighted in bold may be changed to allow the TALEN to bind to a desired target site in the plastid or mitochondrial DNA. Suitably therefore, the first TALE domain comprises a sequence of SEQ ID NO: 6 or 4 and the second TALE domain comprises a sequence of SEQ ID NO:7 or 5 wherein the RVDs highlighted in bold are modified to bind to a target site in the plastid or mitochondrial DNA In one embodiment, the protein used in the methods of the invention is a TALEN, wherein the TALEN comprises a first TALE domain having a sequence according to ID NO:6 or 4, and a second TALE domain having a sequence according to ID NO: 7 or 5. Suitably wherein the RVDs highlighted in bold are modified to allow the TALEN to bind to a desired target site in the plastid or mitochondrial DNA. Suitably the one or more endonucleases are selected from FokI, FokI cleavage domain, or a modified FokI cleavage domain. Suitably the one or more endonucleases are selected from I- TevI, I-TevI nuclease and linker domains, I-TevI nuclease domain, or a modified I-TevI nuclease domain. Suitably the one or more endonucleases are selected from I-SceI, I-SceI cleavage domain, or a modified I-SceI cleavage domain. Suitably the one or more endonucleases are selected MutH, MutH cleavage domain, or a modified MutH cleavage domain. Suitably the one or more endonucleases are selected from mito-nickase, mito- nickase cleavage domain, or a modified mito-nickase. The person skilled in the art will be able to select one or more suitable endonuclease for use in the various embodiments of the invention. Suitably the protein is targeted to the plastid or mitochondria. Suitably therefore the protein comprises a targeting peptide fused to the N terminus or C terminus thereof. Suitably the targeting peptide is a plant plastid or a plant mitochondrial targeting peptide. Non-limiting examples of targeting peptides include those set forth in WO2017198859, US10745712 and US10563220, each of which in incorporated by reference in their entirety. In one embodiment, the targeting peptide for targeting to plastids is the presequence of rubisco small subunit. In one embodiment, the targeting peptide for targeting to mitochondria is the presequence of alternative oxidase 1 (AOX1). Suitably the presequence of the rubisco small subunit comprises the following sequence: MASSVISSAAVATRTNVAQASMVAPFNGLKSAVSFPVSSKQNLDITSIASNGGRVQCM (SEQ ID NO: 11). Suitably therefore the targeting peptide comprises a sequence according to SEQ ID NO:11. Suitably the presequence of AOX1 comprises the following sequence: MMMMMSRSGGNRVANTAMFVAKGLSGEVGGLRALYGGGVRSES (SEQ ID NO:28). Suitably therefore the targeting peptide comprises a sequence according to SEQ ID NO:28. Suitably the TALEN comprises a targeting peptide fused to the N or C terminus of each TALE domain. In one embodiment, the TALEN comprises the presequence of rubisco small subunit fused to the N terminus of each TALE domain. In one embodiment, the TALEN comprises the presequence of AOX1 fused to the N terminus of each TALE domain. Suitably each TALE domain having a targeting peptide may comprise the +153 N-terminus (SEQ ID NO: 24) and +47 C-terminus (SEQ ID NO: 25). Suitably each TALE domain having a targeting peptide may comprise the +136 N-terminus (SEQ ID NO: 26) and +63 C-terminus (SEQ ID NO: 27). Suitably the first TALE domain having a targeting peptide comprises or consists of a sequence of SEQ ID NO: 4. Suitably the second TALE domain having a targeting peptide comprises or consists of a sequence of SEQ ID NO:5. Suitably wherein the RVDs highlighted in bold may be changed to allow the TALEN to bind to a desired target site in the plastid or mitochondrial DNA. In one embodiment, the protein used in the methods of the invention is a TALEN, wherein the TALEN comprises a first TALE domain having a sequence according to ID NO:4, and a second TALE domain having a sequence according to ID NO: 5. Suitably wherein the RVDs highlighted in bold may be changed to allow the TALEN to bind to a desired target site in the plastid or mitochondrial DNA. Inducible System The protein which is capable of cleavage of DNA, suitably which is capable of cleavage of non-modified target sites in DNA in the present invention, is suitably controlled by an inducible system which can be used to express the protein when desired in the plant or part thereof, and to the level of expression desired in the plant or part thereof. Suitably the protein capable of site specific DNA cleavage is under the control of an inducible system. Suitably, expression of the protein capable of site specific DNA cleavage in the plant or part thereof is under the control of an inducible system. Suitably any inducible system known in the art may be used to control the expression of the protein. Suitably inducible systems which may be used in the methods of the invention include systems induced by the presence of an inducer, the absence of an repressor, or any other suitable physical or chemical change. By way of non-limiting example, an inducible system for use in embodiments of the invention may be a forskolin-inducible system, a hypoxia-inducible system, a tetracycline-regulatable (e.g. inducible or repressible) system, an alcohol-inducible system, a steroid-inducible system, a mifepristone (RU486)-inducible system, an ecdysone-inducible system, a rapamycin- inducible system, a metallothionein-inducible system, a hormone-inducible system, a plant- hormone or analogue inducible system, a cumate-inducible system, a temperature-inducible system, a pH-inducible system and a metal-inducible system. As will be discussed further below, various suitable inducible system have been described in the art and others are discussed herein. The person skilled in the art will be able to select one or more suitable inducible system for use in the various embodiments of the invention. Suitably the inducible system is a chemically inducible proximity system (CIP). Suitable such small molecule–based systems for controlling protein abundance or activities is described in Liang et al 10.1126 / scisignal.2001449 which is incorporated herein by reference. Gene expression may be induced chemically by induced proximity caused by a chemical molecule combining two protein binding surfaces as shown in (Belshaw et al., 1996) which is incorporated herein by reference. Transcriptional activation of a gene of interest by chemically induced proximity by a molecule combining two protein binding surfaces is shown in Fig.3 of (Belshaw et al., 1996). Suitably a CIP system comprises a DNA binding domain, an activator domain, and an inducer binding domain. In some cases, the DNA binding domain is linked to a first inducer binding domain and the activator domain is linked to a second inducer binding domain. In some cases, a first DNA binding domain is linked to a first inducer binding domain and a first activator domain, and a second DNA binding domain is linked to a second inducer binding domain and a second activator domain. Suitably any activator domain may be used. Suitably the activator domain is selected from Gal4, Oaf1, Leu3, Rtg3, Pho4, Gln3, Gcn4 in yeast, and p53, NFAT, NF-κB, VP16 and VP34. Suitably any DNA binding domain may be used. Suitably the DNA binding domain is selected from: LexA, dI-SceI and Gal4. Suitably such CIP systems operate by binding to an inducer, suitably to a chemical inducer. Suitable inducers may be selected from any small molecule chemical. The person skilled in the art will be able to select a suitable CIP system for use in the various embodiments of the invention. For example, the CIP system may utilise a non-steroidal ecdysone agonist such as tebufenozide. Suitably therefore the inducible system is a tebufenozide inducible system, suitably as described in Martinez et al.1999 The Plant Journal 19(1). Suitably the CIP is a tebufenozide inducible system. Suitably the chemical inducer is tebufenozide. Suitably therefore the one or more inducer binding domains are tebufenozide binding domains. For example, the CIP system may be a dexamethasone inducible system. Suitably the dexamethasone inducible system comprises a TGV system. Suitably the TGV system comprises a Tet repressor, the rat glucocorticoid receptor hormone binding domain and the transcriptional activation domain of Herpes simplex virion protein VP16 which can mediate dexamethasone-inducible transcription of the synthetic promoter PTop10 (Böhner et al., 1999. The Plant Journal 19(1), 87-95). Suitably therefore the inducible system is a dexamethasone inducible system. Suitably the CIP is a dexamethasone inducible system. Suitably the chemical inducer is dexamethasone. Suitably therefore the one or more inducer binding domains are dexamethasone binding domains. Suitably the inducible system is an oestradiol inducible system. Suitably the CIP system is an oestradiol CIP system. Suitably wherein the chemical inducer is oestradiol. Suitably therefore the one or more inducer binding domains are oestradiol binding domains, suitably each inducer binding domain comprises the regulatory region of the human oestrogen receptor. Suitably the oestradiol inducible system comprises an XVE system (LexA-VP16-ER). Suitably the XVE system is described in Zuo et al. 2000 The Plant Journal 24(2), 265-273 and comprises a DNA-binding domain comprising the bacterial repressor LexA (X), an activator domain comprising the yeast transactivator VP16 (V), and an inducer binding domain comprising the regulatory region of the human oestrogen receptor (E). Suitably the XVE system operates with two XVE proteins each comprising said domains; suitably a first XVE protein and a second XVE protein which are operable to dimerise in the presence of the inducer oestradiol. Suitably the oestradiol inducible system comprises a sequence according to SEQ ID NO: 9. Suitably the DNA binding domain of the CIP system is capable of binding to a binding site upstream of the nucleic acid encoding the protein. Suitably the binding site is located upstream of the promotor which is operably linked to the nucleic acid encoding the protein. Suitably, when using the XVE inducible system, the binding site is a LexA operator sequence, otherwise known as the ‘OlexA’ operator sequence or site. Suitably therefore the bacterial repressor LexA is capable of binding to a LexA operator sequence upstream of the nucleic acid encoding the protein. Suitably the binding site may comprise more than one LexA operator sequence. Suitably the binding site may comprise between one and ten copies of the LexA operator sequence. In one embodiment, the binding site may comprise eight copies of the LexA operator sequence. Suitably the LexA operator sequence comprises one or more copies of a sequence according to SEQ ID NO:29. In the presence of the oestradiol inducer, suitably the first and second inducer binding domains of a first and second XVE protein comprising the regulatory region of the human oestrogen receptor (E) bind to a molecule of oestradiol. Suitably, upon binding of the inducer binding domains to the oestradiol inducer, the XVE proteins dimerise and move to the nucleus . Suitably, once inside the nucleus, the LexA DNA binding domains can bind the LexA operator sequence, bringing the VP16 activator domains into proximity with the RNA polymerase activating complex, and thereby stimulate transcription of the downstream nucleic acid encoding the protein. Suitably therefore in the presence of the inducer the inducible system is activated and causes the nucleic acid encoding the protein to be transcribed and the protein to be produced in the plant or part thereof. Suitably in the presence of oestradiol the XVE inducible system is activated and causes the protein to be transcribed and produced in the plant or part thereof. Suitably the inducible system is inducible by an effective concentration of an inducer. Suitably by exposing the plant or part thereof to an effective concentration of an inducer. Suitably therefore, inducing expression of the protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA from the expression construct comprises exposing the plastids, mitochondria, plants or parts thereof to an effective concentration of an inducer. Suitably, to an inducer capable of inducing the inducible system i.e. oestradiol when using an oestradiol inducible system. Suitably exposing may comprise applying the inducer to the plant or part thereof, for example by spraying, fumigating, or by adding it to soil, during irrigation, or by adding to water provided to the plant. Suitably the effective concentration of inducer may comprise a range of suitable concentrations of inducer. Suitably the concentration of the inducer may range between about 0.5uM to 50uM, suitably between about 1uM and 40uM, suitably between about 1.5uM and 30uM, suitably between about 2uM and 20uM. The inventors have discovered that the concentration of the inducer affects the efficiency of generating homoplasmic plants in the methods described herein. Suitably such that the concentration of the inducer can be tuneable to create the desired effect in the plant or part thereof. Suitably a concentration of inducer of about 2uM gives high plant survival rates, and rapid regeneration, but produces a mixture of homoplasmic and heteroplasmic plants. In some embodiments, the concentration of inducer, suitably oestradiol, is about 2uM. Suitably a concentration of inducer of about 20uM gives lower plant survival rates, and slower regeneration, but produces homogeneous homoplasmic plants. In some embodiments, the concentration of inducer, suitably oestradiol, is about 20uM. Suitably the expression construct described herein may be used for inducing expression of the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA at an optimal time and / or in an optimal location. The inventors have further discovered that there are optimal times at which the protein should be expressed in order to most efficiently generate homoplasmic modified plants or parts thereof. Suitably therefore exposing the plant or part thereof to an effective concentration of the inducer may take place at an optimal time for inducing expression of the protein in the plant or part thereof. Suitably the plant or part thereof may be exposed to an effective concentration of the inducer at any time during the methods described herein. Suitably the plant or part thereof may be exposed to an effective concentration of the inducer at any time after the modification step (a) is completed. Suitably the plant or part thereof may be exposed to an effective concentration of the inducer at any time before the modification step (a) is completed. Suitably the plant or part thereof may be exposed to an effective concentration of the inducer when the copy number of the plastid or mitochondrial genome is at a minimum. Suitably this may be at the stage of egg production, or de-differentiation in the plant. Suitably, egg production takes place in the ovule, at the stage of sexual maturity of the plant, after flowers have developed. Suitably de-differentiation takes place in callus formation. Suitably the exact time of such developmental stages may vary with the species of plant, but will be known to the skilled person. Suitably the plant or part thereof may be exposed to an effective concentration of the inducer at an optimal time for inducing expression of the protein in a given plant organ, such as calli, embryos, meristems, root tips, etc. Suitably exposing the plant or part thereof to an effective concentration of the inducer may take place when the plant has reached sexual maturity. Suitably exposing the plant or part thereof to an effective concentration of the inducer may take place on the day of pollination. Suitably exposing the plant or part thereof to an effective concentration of the inducer may take place after flowering. Suitably 1 days, 2 days, 3 days, 4 days, 5 days, 6, days, 1 week, 2 weeks, 3 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or 2 months after flowering. Suitably exposing the plant or part thereof to an effective concentration of the inducer may take place during callus formation, suitably which may be during regeneration of the plant or part thereof. Suitably regeneration may take place in tissue culture, or by growing a plant from seed. Suitably therefore exposing the plant or part thereof to an effective concentration of the inducer may take place during tissue culture, suitably during regeneration of the plant or part thereof. Suitably exposing the plant or part thereof to an effective concentration of the inducer may take place on day 1, day 2, day 3, day 4, day 5, day 6, day 7, after 1 week, 2 weeks, 3 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or 2 months of tissue culture. Suitably exposing the plant or part thereof to an effective concentration of the inducer may take place on day 1 of tissue culture. Suitably therefore exposing the plant or part thereof to an effective concentration of the inducer may take place during seed germination, suitably during regeneration of the plant or part thereof. Suitably exposing the plant or part thereof to an effective concentration of the inducer may take place when seeds are planted, during vernalization of the seed, or on day 1, day 2, day 3, day 4, day 5, day 6, day 7, after 1 week, 2 weeks, 3 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or 2 months of seed germination. The inventors have further discovered that there may be optimal locations at which the protein should be expressed in order to most efficiently generate homoplasmic modified plants or parts thereof. Suitably therefore exposing the plant or part thereof to an effective concentration of the inducer may take place at an optimal location for inducing expression of the protein in the plant or part thereof. Suitably the plant or part thereof may be exposed to an effective concentration of the inducer at any location and at any time during the methods described herein. Suitably the plant or part thereof may be exposed to an effective concentration of the inducer at any location after the modification step (a) is completed. Suitably the plant or part thereof may be exposed to an effective concentration of the inducer at any location before the modification step (a) is completed. Suitably expression of the protein which is capable of cleavage of DNA at an optimal location in the plant may be achieved by using a tissue specific promoter, such that the protein may be under the control of a tissue specific promoter. Suitable tissue specific promoters include AtRPS5Apro which is specific to egg cells, the early embryo and shoot meristem. Further suitable desirable tissues in which to induce expression of the protein, and tissue specific promoters are described hereinbelow in relation to the expression construct used to express the protein which is capable of cleavage of DNA. Additional suitable tissue specific promoters will be known by the person skilled in the art. Suitably tissue specific promoters may be used alone, or in combination with an inducible system to allow induction of expression of the protein capable of cleavage DNA at optimal times and also in optimal locations. Expression construct Suitably the protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA is under the control of an inducible system as described above. Suitably therefore, the invention further provides an expression construct which may be used to express the protein in a plant or part thereof. Suitably the expression construct is inserted into the plant or part thereof to control expression of the protein therein, by typical methods in the art described below. "Expression construct" as used herein means a nucleic acid sequence capable of directing expression of a particular nucleic acid sequence in an appropriate host cell, comprising a promoter operably linked to the nucleic acid of interest which is operably linked to termination signal sequences. It also typically comprises sequences required for proper translation of the nucleic acid sequence. The expression construct comprising the nucleic acid of interest may be chimeric, meaning that at least one of its components is heterologous with respect to at least one of its other components. The expression construct may also be one that is naturally occurring but has been obtained in a recombinant form useful for heterologous expression. Typically, however, the expression construct is heterologous with respect to the host, i.e., the particular nucleic acid of the expression cassette does not occur naturally in the host cell and must have been introduced into the host cell or an ancestor of the host cell by a transformation event. The expression of the nucleic acid sequence in the expression construct may be under the control of, for example, a constitutive promoter or of an inducible promoter that initiates transcription only when the host cell is exposed to some particular external stimulus. In the case of a multicellular organism, such as a plant, the promoter can also be a tissue-specific promoter, which is specific to a particular tissue, or organ, or stage of development. The expression of the nucleic acid sequence in the expression construct may be under the control of, for example, an organ or cell type specific promoter that initiates transcription when present in a host cell in a particular organ or in a host cell of a particular cell type. For instance, the promoter may be a pollen, anther, egg cell, tapetum cell, root cell or callus specific promoter. Suitably, the expression of the nucleic acid sequence in the expression construct may be under the control of a developmentally regulated promoter. The expression construct may be under the control of a tissue specific promoter. Specific expression of the nucleic acid sequence in the expression construct in egg cells, embryo cells and shoot meristem may be particularly advantageous, because plastid DNA copy numbers may be significantly lower in such cells than in mesophyll cells. Therefore in one embodiment, the expression construct comprises an egg cell, or an embryo, or a callus, or a shoot meristem specific promoter. Suitably the tissue specific promoter is selected from AtRPS5Apro (specific to egg cells, early embryo and shoot meristem), Wuschel promoter (specific to shoot meristems), Clavata3 promoter (specific to shoot meristems), STM promoter (specific to shoot meristem), DD45 (specific to egg cell and early embryo), YAO promoter (specific to shoot apical meristem) for example. Suitably egg, embryo or shoot meristem cells comprising the expression construct comprising such a tissue specific promoter may be used to regenerate a whole plant or plants. Specific expression of the nucleic acid sequence in the expression construct in root cells may be particularly advantageous, because plastid DNA copy numbers may be significantly lower in root cells than in mesophyll cells. Therefore in one embodiment, the expression construct comprises a root specific promoter. Suitably the root specific promoter initiates expression of the expression construct when present in a root cell. Suitably root cells comprising the expression construct comprising a root specific promoter may be used to regenerate a whole plant or plants. Suitably the root specific promoter is selected from: Patatin promoter, or root cortex PEP promoter, YAO promoter (specific to root meristem), ANAC033 / SOMBRERO (SMB) promoter, for example. Specific expression of the nucleic acid sequence in the expression construct in pollen may be particularly advantageous, because organelle DNA copy numbers, including plastid and / or mitochondrial DNA copy numbers, may be significantly lower in pollen compared to other plant cells such as mesophyll cells. Furthermore, typically only paternal DNA is present in the mitochondria of pollen, suitably therefore expression of the nucleic acid sequence in the expression construct in pollen may be a way to modify only the paternal organelle DNA. Therefore in one embodiment, the expression construct comprises an anther specific promoter, or a pollen mother cell specific promoter, or a pollen specific promoter. Suitably the pollen specific promoter initiates expression of the expression construct when present in a pollen cell. Suitably the anther specific promoter initiates expression of the expression construct when present in an anther cell. Suitably the pollen mother cell specific promoter initiates expression of the expression construct when present in a pollen mother cell. Suitably pollen cells, or pollen mother cells comprising the expression construct comprising a pollen specific promoter may be germinated and developed to a whole plant or plants. A suitable pollen specific promoter may be the LAT52 promoter. Suitably a pollen specific promoter may be an anther specific promoter. Suitable such promoters include the TA29 promoter, or the A9 promoter, for example. Suitably the expression construct comprises one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA, under the control of an inducible system. Suitably, the nucleic acid encoding the protein is operably linked to an inducible system. Suitably the nucleic acid encoding the protein is operably linked to a binding site for an inducible system. Suitably the nucleic acid encoding the protein is operably linked to a LexA operator sequence. Suitably the binding site, such as the LexA operator sequence, is located upstream of the or each nucleic acid encoding the protein. Suitably the binding site, such as the LexA operator sequence, is located upstream of a promoter and the or each nucleic acid encoding the protein. Suitably therefore, the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA are operably linked to a binding site for an inducible system and a promoter, suitably upstream of the one or more nucleic acids. Suitably any promoter may be used for expression of the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA. Suitable promoters are constitutive promoters. Suitably these promoters will drive strong expression of the protein once the inducible system is activated by the relevant inducer. Suitable promoters may be selected from: the core promoter of the Rsyn7 promoter and other constitutive promoters disclosed in WO 99 / 43838 and U.S. Patent No.6,072,050; the core CaMV 35S promoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroy et al. (1990) Plant Cell 2:163-171); ubiquitin (Christensen et al. (1989) Plant MoI. Biol.12:619-632 and Christensen et al. (1992) Plant MoI. Biol.18:675-689); pEMU (Last et al. (1991) Theor. Appl. Genet.81 :581-588); MAS (Velten et al. (1984) EMBO J.3:2723-2730); ALS promoter (U.S. Patent No.5,659,026), and the like. Other constitutive promoters include, for example, those described in U.S. Patent Nos.5,608,149; 5,608,144; 5,604,121 ; 5,569,597; 5,466,785; 5,399,680; 5,268,463; 5,608,142; and 6,177,611. In one embodiment, the promoter is CaMV 35S promoter. Suitably however the promoter used in the expression construct encoding the plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA is a minimal promoter. Suitably a minimal promoter which is operable function in combination with a LexA operator sequence. Suitably therefore, the expression construct comprises one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA, operably linked to an LexA operator sequence and a minimal 35S promoter, suitably wherein the LexA operator sequence and minimal 35S promoter are upstream of the or each nucleic acid sequence. Suitably the minimal 35S promoter comprises a sequence according to SEQ ID NO:30. Alternatively, the expression construct comprises one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA, operably linked to an LexA operator sequence and a minimal tissue-specific promoter, suitably wherein the LexA operator sequence and minimal tissue specific promoter are upstream of the or each nucleic acid sequence. Suitably wherein the tissue specific promoters are selected from those listed above. Suitably the protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA is described hereinabove. Suitably the inducible system is described hereinabove. Suitably the expression construct may also comprise one or more nucleic acids encoding the components of the inducible system. Suitably the expression construct may comprise one or more nucleic acids encoding the components of a chemically inducible proximity (CIP) system. Suitably the expression construct may also comprise one or more nucleic acids encoding the one or more DNA binding domains, the one or more activator domains and the one or more inducer binding domains of the inducible system. Alternately, a second expression construct may comprise the one or more nucleic acids encoding the components of the inducible system. Suitably therefore in a further aspect of the invention there is provided a pair of expression constructs, the first expression construct comprising one or more nucleic acids encoding the protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA under the control of an inducible system, (suitably by being operably linked to a binding site for an inducible system), and the second expression construct comprising one or more nucleic acids encoding the components of the inducible system. Suitably the one or more nucleic acids encoding the components of the inducible system comprise a promoter. Suitably operably linked thereto, suitably upstream of the or each nucleic acid. Suitably this is a constitutive promoter. Suitably the promoter will drive strong expression of the components of the inducible system in the cell, however the components will not associate to induce expression of the protein unless an inducer is also present. Suitable promoters may be selected from: the core promoter of the Rsyn7 promoter and other constitutive promoters disclosed in WO 99 / 43838 and U.S. Patent No.6,072,050; the core CaMV 35S promoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroy et al. (1990) Plant Cell 2:163-171); ubiquitin (Christensen et al. (1989) Plant MoI. Biol.12:619-632 and Christensen et al. (1992) Plant MoI. Biol.18:675-689); pEMU (Last et al. (1991) Theor. Appl. Genet.81 :581-588); MAS (Velten et al. (1984) EMBO J.3:2723-2730); ALS promoter (U.S. Patent No.5,659,026), G10-90 promoter (Ishige et al., 1999 ), and the like. Other constitutive promoters include, for example, those described in U.S. Patent Nos.5,608,149; 5,608,144; 5,604,121 ; 5,569,597; 5,466,785; 5,399,680; 5,268,463; 5,608,142; and 6,177,611. In one embodiment, the promoter is CaMV 35S promoter. Suitably the nucleic acids referred to herein may be one nucleic acid or more than one nucleic acid. Suitably at least one nucleic acid encodes the protein used in the invention, and the binding site of the inducible system, operably linked thereto. Suitably at least one nucleic acid encodes the components of the inducible system, such as the DNA binding domain, the activator domain and the first and second inducer binding domains. In some cases, the protein and the components of the inducible system may be encoded on the same nucleic acid. In some cases, the protein and the binding site of the DNA binding domain operably linked thereto may be encoded on one nucleic acid, and the components of the inducible system may be encoded on another nucleic acid. Suitably any combination of the nucleic acids may be present in one or more expression constructs. By “operably linked” or “operably associated” as used herein, it is meant that the indicated elements are functionally related to each other, and are also generally physically related. Thus, the term “operably linked” or “operably associated” as used herein, refers to polynucleotides on a single nucleic acid molecule that are functionally associated. Thus, a first polynucleotide sequence or nucleic acid molecule that is operably linked to a second polynucleotide sequence or nucleic acid molecule, means a situation when the first polynucleotide sequence or nucleic acid molecule is placed in a functional relationship with the second polynucleotide sequence or nucleic acid molecule. For instance, a promoter is operably associated with a polynucleotide sequence or nucleic acid molecule if the promoter effects the transcription or expression of said polynucleotide sequence or nucleic acid molecule. Similarly, an inducible system is operably linked to a polynucleotide sequence or nucleic acid molecule if the inducible system affects the transcription or expression of said polynucleotide sequence or nucleic acid molecule. Those skilled in the art will appreciate that the control sequences (e.g., promoter or nucleic acids encoding the components of the inducible system) need not be contiguous with the polynucleotide sequence or nucleic acid molecule to which it is operably associated, as long as the control sequences function to direct the expression thereof. Thus, for example, intervening untranslated, yet transcribed, sequences can be present between a promoter and a polynucleotide sequence or nucleic acid molecule, and the promoter can still be considered “operably linked” to or “operatively associated” with the polynucleotide sequence or nucleic acid molecule. In some cases, the term ‘operably linked’ may be considered the same as being ‘under the control of’, and such terms may be used interchangeably. Suitably the expression construct may further comprise other regulatory elements. The term "regulatory element" or “regulatory sequence” as used herein refers to a nucleic acid that is capable of regulating the transcription and / or translation of an operably linked polynucleotide. Regulatory elements include, but are not limited to, promoters, enhancers, introns, 5' UTRs, and 3' UTRs. Suitable promoters are described above. Expression cassettes may include in the 5 '- 3' direction of transcription, a transcriptional and translational initiation region (e.g., a promoter), a nucleic acid sequence encoding the protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA, and a transcriptional and translational termination region (e.g., termination region) functional in plants. The expression constructs may also comprise transcription termination regions. Where transcription terminations regions are used, any termination region may be used in the preparation of the expression cassettes. For example, the termination region may be native to the transcriptional initiation region, may be native to the operably linked nucleic acid of interest, may be native to the plant host, or may be derived from another source (i.e., foreign or heterologous to the promoter, the nucleic acid of interest, the plant host, or any combination thereof). Examples of termination regions that are available for use in the expression constructs and vectors of the present invention include those from the Ti-plasmid of A. tumefaciens, such as the octopine synthase and nopaline synthase termination regions. See also Guerineau et al. (1991) MoI. Gen. Genet.262: 141-144; Sanfacon et al. (1991) Genes Dev.5:141-149; Mogen et al. (1990) Plant Cell 2:1261-1272; Munroe et al. (1990) Gene 91:151-158; Ballas et al. (1989) Nucleic Acids Res.17:7891-7903; and Joshi et al. (1987) Nucleic Acid Res.15:9627-9639. Expression constructs may additionally contain 5' leader sequences. Such leader sequences can act to enhance translation. Translation leaders are known in the art and include: picornavirus leaders, for example, EMCV leader (Encephalomyocarditis 5' noncoding region) (Elroy-Stein et al. (1989) Proc. Natl. Acad. ScL USA 86:6126-6130); potyvirus leaders, for example, TEV leader (Tobacco Etch Virus) (Gallie et al. (1995) Gene 165(2):233-238), MDMV leader (Maize Dwarf Mosaic Virus) (Virology 154:9-20), and human immunoglobulin heavy-chain binding protein (BiP) (Macejak et al. (1991) Nature 353: 90-94); untranslated leader from the coat protein mRNA of alfalfa mosaic virus (AMV RNA 4) (Jobling et al. (1987) Nature 325: 622-625); tobacco mosaic virus leader (TMV) (Gallie et al. (1989) in Molecular Biology of RNA, ed. Cech (Liss, New York), pp.237-256); and maize chlorotic mottle virus leader (MCMV) (Lommel et al. (1991) Virology 81 :382-385). See also, Della- Cioppa et al. (1987) Plant Physiol.84:965-968. The expression construct may be optimized for increased expression in a transformed plant. Suitably, the nucleic acid encoding the protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA can be synthesized using plant-preferred codons for improved expression, or optionally using mitochondria or plastid preferred codons for improved expression therein. See, for example, Campbell and Gowri (1990) Plant Physiol. 92:1-11 for a discussion of host-preferred codon usage. Methods are available in the art for synthesizing plant-preferred genes. See, for example, U.S. Patent Nos.5,380,831, and 5,436,391, and Murray et al. (1989) Nucleic Acids Res.17:477-498, herein incorporated by reference. In one embodiment, the expression construct comprises at least one nucleic acid encoding the components of an XVE inducible system operably linked to a constitutive promoter, and at least one nucleic acid encoding the lexA operator sequence upstream of a minimal constitutive promoter or a minimal tissue-specific promoter operably linked to a nucleic acid encoding a protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA. In another embodiment, there is provided a pair of expression constructs, comprising a first expression construct which comprises at least one nucleic acid encoding the components of an XVE inducible system operably linked to a constitutive promoter, and a second expression construct which comprises least one nucleic acid encoding the lexA operator sequence upstream of a minimal constitutive promoter or a minimal tissue-specific promoter operably linked to a nucleic acid encoding a protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA. Suitably in either embodiment, the construct comprises between one and ten copies of the LexA operator sequence. In one embodiment, the construct may comprise eight copies of the LexA operator sequence. Vectors The invention further provides a vector comprising the or each expression construct described above. Suitably, if more than one expression construct is used, then the vector may comprise all of the expression constructs, or one vector may comprise one expression construct. In embodiments where only one expression construct is used, there is provided a vector comprising the expression construct. In embodiments where a first expression construct and a second expression construct are used as described above, then there is provided a first vector comprising the first expression construct and a second vector comprising the second expression construct. Suitable vectors may be those used for plant transformations as known in the art. The vector can be either RNA or DNA, either prokaryotic or eukaryotic, and typically the vector is a virus or a plasmid. A number of vectors suitable for stable transformation of plant cells or for the establishment of transgenic plants have been described in, e.g., Pouwels et al., Cloning Vectors: A Laboratory Manual, 1985, supp.1987; Weissbach and Weissbach, Methods for Plant Molecular Biology, Academic Press, 1989; and Gelvin et al., Plant Molecular Biology Manual, Kluwer Academic Publishers, 1990. Typically, plant expression vectors include, for example, one or more nucleic acids under the transcriptional control of 5' and 3' regulatory sequences and a dominant selectable marker. For example, the vector may be pBIN 19 (Bevan, Nucl. Acids Res. (1984)). Suitably the vector may be pCB3300. The expression vector of the invention may include one or more regulatory sequences. For example, the expression vectors can contain a promoter regulatory region (e.g., a regulatory region controlling inducible or constitutive, environmentally- or developmentally- regulated, or cell- or tissue-specific expression), a transcription initiation start site, a ribosome binding site, an RNA processing signal, a transcription termination site, and / or a polyadenylation signal. Such a portion of an expression vector may be referred to as an expression construct. The expression construct may include one or more regulatory sequences that are functional in plants, as explained above. Suitably the vector may further comprise a selectable marker gene, suitably for determining if the transformation of the plant cell, or plant is successful. Suitable selectable marker genes for use in plants are well known, and may be selected from: herbicide resistance genes such as those conferring resistance against phosphinothricin, butafenacil, or glyphosate, antibiotic resistance genes such as those conferring resistance to kanamycin, hygromycin, alternatively, selection may be based upon expression of a visualisable marker gene such as GUS, that can be used to aid recovery of stably transformed plants. Suitably, the vector comprises the bar marker gene encoding the enzyme phosphinotricin acetyltransferase which confers phosphinotricin (PPT) resistance to the plant or part thereof. Suitably, the vector comprises the Hpt gene encoding the enzyme hygromycin phosphotransferase II which confers hygromycin resistance to the plant or part thereof. Suitably the methods of the invention may comprise a step after step (b) or step (i) of selecting the plants or parts thereof that contain the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA. Suitably such a selecting step may comprise exposing the plant or part thereof to a selective agent, such as phosphinotricin (PPT), and suitably selecting those plants or parts thereof which survive. Suitably only those plants successfully comprising the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA will have resistance to the selective agent due to expression of the marker gene. Introducing expression constructs / vectors into Plastids, Mitochondria or Plants In the methods of the invention, the or each expression construct, which may be comprised on a vector as described above, is introduced into the plastids, mitochondria or plant cells. Suitably references herein to introducing the expression construct may equally refer to introducing a vector comprising the expression construct. Suitably introducing an expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA comprises transforming the plant, part thereof, plastids, or mitochondria with said expression construct. “Transformation” refers to a process of introducing an exogenous nucleic acid molecule (for example, an expression construct or vector as described herein) into a plant cell or protoplast, or organelle and that exogenous nucleic acid molecule is incorporated into a host plant cell genome or an organelle genome (for example, chloroplast or mitochondria) or is capable of autonomous replication. Transformed” or “transgenic” refers to a cell, tissue, organ, or organism into which a foreign nucleic acid, such as an expression vector or recombinant nucleic acid molecule has been introduced. The nucleic acid molecule can be stably integrated into the genome of the host or the nucleic acid molecule can also be present as an extrachromosomal molecule. Such an extrachromosomal molecule can be auto-replicating. The nucleic acid molecule can also be introduced into the genome of the chloroplast or the mitochondria of a plant cell. Suitable methods of transforming plant cells are known in the art. For example, plant cells may be transformed by: Agrobacterium mediated transformation method and the Biolistics or particle-gun mediated transformation method. Suitable plant transformation vectors for the purpose of Agrobacterium mediated transformation include-those elements derived from a tumor inducing (Ti) plasmid of Agrobacterium tumefaciens, for example, right border (RB) regions and left border (LB) regions, and others disclosed by Herrera-Estrella et al., Nature 303:209 (1983); Bevan, Nucleic Acids Res.12:8711-8721 (1984); Klee et al., Bio- Technology 3(7):637-642 (1985). In addition to plant transformation vectors derived from the Ti or root-inducing (Ri) plasmids of Agrobacterium, alternative methods can be used to insert the nucleic acid molecules of this invention into plant cells. Such methods may involve, but are not limited to, for example, the use of liposomes, electroporation, chemicals that increase free DNA uptake, free DNA delivery via microprojectile bombardment, and transformation using viruses or pollen. Plastids and mitochondria and may be modified by the use of targeting peptides attached to the protein of interest, such that when the nucleus of the plant cell is transformed with the expression cassette encoding the protein by any of the methods above, the protein is expressed and then targeted to the plastid or mitochondria. In one such embodiment, the nucleus of a plant cell is transformed with the or each expression construct, or vector, by any of the methods above, and the proteins expressed therefrom comprise a targeting peptide which targets the proteins to the plastid or mitochondria. Suitable targeting peptides are described elsewhere herein. In some cases, however, direct transformation of plastids or mitochondria may take place, suitably this may carried out by any known technique such as those listed above, including: Biolistics or particle-gun mediated transformation methods, electroporation, free DNA delivery via microprojectile bombardment, peptide carrier mediated transformation (https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC8351593 / ) , and transformation using viruses. Although techniques for direct transformation of plastids and mitochondria are limited some examples have been achieved in the art, as described in https: / / www.annualreviews.org / doi / 10.1146 / annurev.arplant.55.031903.141633. For example using microprojectile transformation (Yoo et al. PeerJ.2020; 8: e8362, https: / / star- protocols.cell.com / protocols / 1675, ), or biolistics. In one such embodiment, the plastid or mitochondrion is transformed with the or each expression construct, or vector, by any of the methods above. Selecting Plastids, Mitochondria or Plants In the methods of the invention, once the protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA is expressed, suitably non-modified target sites in the plastid or mitochondrial DNA are preferentially cleaved by the protein, whilst plastid or mitochondrial DNA with modified target sites is resistant to cleavage Suitably the plants, parts thereof, plastids or mitochondria which have non-modified plastid or mitochondrial DNA at the target site have diminished viability after inducing expression of the protein. Suitably the plants, parts thereof, plastids or mitochondria which have diminished viability have plastid or mitochondrial DNA which has been cleaved by the protein. Suitably the plants, parts thereof, plastids or mitochondria which have plastid or mitochondrial DNA with modified target sites have maintained or improved viability after inducing expression of the protein. Suitably the plants, parts thereof, plastids or mitochondria which have maintained or improved viability have plastid or mitochondrial DNA has not been cleaved by the protein. Suitably these plants, parts thereof, plastids or mitochondria are selected. Suitably any of the methods of the invention may comprise a step of selecting the plants, parts thereof, plastids or mitochondria which maintain or have improved viability compared to the plastids or mitochondria of step (a) or step (i). Suitably maintained or improved viability means that the plant or part thereof does not have any necrotic regions, wilting, spotting, or cell death. Suitably maintained or improved viability means that the plant or part thereof may have maintained or increased growth, maintained or increased leaf number, maintained or increased root mass, maintained or increased budding or flowers, maintained or increased fruit number or weight, maintained or increased seed numbers or weight, etc. In some embodiments, maintained or improved viability may be survival, such that the plant or plant parts that are selected are those that survive. In the context of maintained or improved viability of mitochondria and plastids, this may mean that the mitochondria or plastids maintain or increase growth. In some embodiments, maintained or improved viability with respect to the mitochondria or plastids may be survival, such that the mitochondria or plastids that are selected are those that survive. Suitably diminished viability means that the plant or part thereof may have necrotic regions, wilting, spotting, cell death, decreased growth, decreased leaf number, maintained or increased root mass, decreased budding or flowers, decreased fruit number or weight, decreased seed numbers or weight, etc. In some embodiment, diminished viability may be lower survival, such that the plant or plant parts that are not selected are those that do not survive. Suitably diminished viability with regard to mitochondria and plastids means that the mitochondria or plastids have decreased growth. In some embodiment, diminished viability may be lower survival, such that the mitochondria and plastids that are not selected are those that do not survive. Suitably selection may therefore take place by visual inspection of the plant, part thereof, plastids or mitochondria. Alternatively, or additionally, selection may take place by sequencing the plastid or mitochondrial DNA to check for modified target sites. Suitably sequencing of the plastid or mitochondrial DNA may be carried out by methods known in the art. ‘Sequencing’ refers to determining the nucleic acid sequence of a piece of DNA, e.g. of. Plastid or mitochondrial DNA. Standard methods and commercial services are known in the art. Basic methods for DNA sequencing include the Maxam-Gilbert method and the chain termination method. High-throughput techniques have also been developed and may be used in the method of the present invention. These high-throughput techniques include, but are not limited to, Massively parallel signature sequencing (MPSS), Polony sequencing, 454 pyrosequencing, Illumina Hi-Seq / Mi-Seq (Solexa) sequencing, Combinatorial probe anchor synthesis (cPAS), SOLiD sequencing, Ion Torrent semiconductor sequencing, DNA nanoball sequencing, Heliscope single molecule sequencing, Single molecule real time (SMRT) sequencing. PacBio sequencing and Nanopore DNA sequencing. In some embodiments, the sequencing may be carried out using primers that are capable of binding to a desired modified target site in the plastid or mitochondrial DNA. For example, primers that are complimentary to at least a portion of a desired modified target site. Suitably therefore the sequencing will detect if said desired modification is present in the plastid or mitochondrial DNA. Suitably the primers are operable to amplify a fragment covering the desired modified target site i.e. the site in the plastid or mitochondrial DNA targeted by the protein. Suitably the fragment is less than around 1000bp. Suitably the fragment is between 50-1000bp, suitably 100-500bp, suitably 200-400bp, suitably around 300bp. Suitably the primers are operable to amplify said fragment by PCR. Suitably the PCR product can then be sent for sequencing to confirm if there is mutation present from the modification step of the methods. As used herein, the term “primer" refers to an oligonucleotide which is capable of annealing to a nucleic acid target and serving as a point of initiation of DNA synthesis when placed under conditions in which synthesis of a primer extension product is induced (e.g., in the presence of nucleotides and an agent for polymerization such as DNA polymerase and at a suitable temperature and pH). A primer (in some examples an extension primer and in some examples an amplification primer) may be single stranded for maximum efficiency in extension and / or amplification. The primer may be an oligodeoxyribonucleotide. A primer is typically sufficiently long to prime the synthesis of extension and / or amplification products in the presence of the agent for polymerization. The minimum length of the primer can depend on many factors, including, but not limited to temperature and composition (A / T vs. G / C content) of the primer. In the context of amplification primers, these are typically provided as a pair of bi-directional primers consisting of one forward and one reverse primer or provided as a pair of forward primers as commonly used in the art of DNA amplification such as in PCR amplification. Suitably once selected, the plants or parts thereof may then be regenerated as explained below, to reach a homoplasmic state. Regenerating Plants or parts thereof In the methods of the invention, once the protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA is induced and expressed, the protein cleaves non-modified target sites in the plastid or mitochondrial DNA. Suitably plastid or mitochondrial DNA with modified target sites remains intact and is not cleaved. Suitably through one or more rounds of regeneration of the modified plants, eventually only plastid or mitochondrial DNA with modified target sites remains, and all plastid or mitochondrial DNA with non-modified target sites is cleaved. Suitably therefore the methods of the invention may comprise a step of regenerating the plant or part thereof. Suitably this step is performed after the step of inducing expression of the protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA, and suitably after the step of selecting plants, parts thereof plastids or mitochondria if present in the method. “Regeneration” refers to the process of growing a plant from a plant part or from a plant cell (for example, plant protoplast or explant). Suitable methods of regeneration are known in the art. For example, the regeneration step may be carried out by taking an explant of the plant, or the plant part, and culturing it into a new plant, suitably into a shoot of a new plant. Such regeneration techniques rely on manipulation of certain phytohormones in a tissue culture growth medium. A suitable protocol for regenerating a plant is described in, for example, Ammirato et al., Handbook of Plant Cell Culture — Crop Species. Macmillan Publ. Co. (1984); Shimamoto et al., Nature 338:274-276 (1989); Fromm, UCLA Symposium on Molecular Strategies for Crop Improvement, Apr.16-22, 1990. Keystone, Colo. (1990); Vasil et al., Bio / Technology 8:429- 434 (1990); Vasil et al., Bio / Technology 10:667-674 (1992); Hayashimoto, Plant Physiol. 93:857-863 (1990); and Datta et al., Bio-technology 8:736-740 (1990). Such regeneration techniques are described generally in Klee et al., Ann. Rev. Plant Phys.38:467-486 (1987). The skilled person will be aware of which plant species are amenable to regeneration via tissue culture and which tissue culture conditions are suitable for a given plant species. This information is well known in the art. Suitably in some cases, if the step of regeneration is carried out by taking an explant and culturing using tissue culture, then the plant or plant part is amenable to tissue culture. Some plants are recalcitrant to tissue culture, however such tissue culture may still be achieved using the methods described in https: / / www.frontiersin.org / articles / 10.3389 / fpls.2023.1202235 / full. Suitably, for example, regeneration of cereals, soybean and cotton via tissue culture may take place using embryos excised from the seeds thereof, otherwise referred to as an embryo explant. Suitably for example, regeneration of maize via tissue culture may take place using leaves from said plants, otherwise referred to as a leaf explant. Alternatively, the step of regeneration may alternatively be carried out by collecting seed of the plant and germinating said seed into a new plant. Suitably the regenerated plants may then undergo further rounds of inducing expression of the protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA, optional selection, and regeneration. Optionally steps (c) and (d) of the methods of the invention maybe repeated. Suitably these steps of the methods may be repeated multiple times. Suitably, these steps may be repeated two times, three times, four times, five times or as many times as required to achieve the desired result. Suitably the steps of inducing the expression of the protein capable of cleavage of any non- modified target sites in the plastid or mitochondrial DNA, and regeneration, are repeated until a homoplasmic plant is generated. Suitable homoplasmic plants are defined hereinabove. Suitably DNA sequencing may be used to check if the resulting plants are indeed homoplasmic. Suitably the methods may comprise a step of sequencing the plastid or mitochondrial DNA of the plant or plant part, suitably sequencing the region of the plastid or mitochondrial DNA of the plant or plant part containing the target site. Suitably such a step may take place after the steps of the methods of the invention, suitably after any rounds of regeneration have taken place. Suitably such sequencing may take place after a step of amplifying the region of DNA containing the target site, suitably by PCR techniques known in the art. Suitably the sequencing step may be carried out by Sanger sequencing. Suitably Sanger sequencing will show a single new peak at the target site if homoplasmic modification at said target site has been achieved. Suitably the sequencing step may be carried out by next generation sequencing (NGS). Suitably NGS will show a high frequency (e.g. >95%) for the coverage of the modification out of the total coverage at the target site if homoplasmic modification at said target site has been achieved. Suitably in methods which include a step of selecting plants or plant parts, then the step of regeneration may take place after any selecting step in the methods. Suitably therefore, only the selected plants or parts thereof are regenerated. Suitably in such methods, the steps of inducing the expression of the protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA, selection, and regeneration may be repeated. Suitably once a homoplasmic plant is generated by the methods of the invention, the expression construct comprising: one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA under the control of an inducible system is removed from the plant or part thereof. Suitably therefore the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA are removed from the plant or part thereof, and the one or more nucleic acids encoding the inducible system are removed from the plant or part thereof. Optionally if a nucleic acid encoding an error prone polymerase is also present in the plant or part thereof, this may also be removed. Suitably therefore the method comprises a step of removing any heterologous nucleic acid sequences from the plant or part thereof. Suitably the step of removing may take place after the regenerating steps, and after any sequencing steps. Advantageously removing such sequences means that the homoplasmic plant contains the intended modification at target site in the plastid or mitochondrial DNA, but is not transgenic, and therefore may be sold and propagated more easily in many countries, and may be safer for human or animal consumption. Suitably removing these heterologous nucleic acid sequences, expression constructs, may be achieved by backcrossing of the plant with a plant of an interfertile species which does not contain any such nucleic acid sequences. In some embodiments, the plant may be backcrossed with a wild type plant, suitably a wild type plant of an interfertile species, which does not contain any such nucleic acid sequences. In other embodiments the plant may be backcrossed with a non-transgenic mutant plant, suitably a non-transgenic mutant plant of an interfertile species, which does not contain any such nucleic acid sequences, but which contains one or more other mutations. Suitably an interfertile species may be the same species of plant or another species which it is possible to breed with the plant, suitably there is no reproductive isolation between the two plants. Suitably the one or more other mutations have been introduced into said plant using non-transgenic techniques such as CRISPR. Suitably the one or more other mutations may be advantageous mutations, for example one or more mutations which improve performance of the plant, suitably which improve agronomic performance of the plant. Suitably the one or more mutations may increase drought resistance, pest resistance, or herbicide tolerance for example. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1. Design of the construct for estradiol inducible accD TALENs. (A) Binding regions of TALENs targeting the accD gene. Nucleotide sequence shows bases 730 to 783 (SEQ ID NO:1), where the adenine in the ATG start codon is the first base of accD coding sequence. Amino acid translations of the coding sequence are shown between the direct and complementary strands. N-Term / C-Term, N-terminus or C-terminus of the accD TALENs; LA, left arm of accD TALENs, binding to 14 nucleotides with 13.5 repeat variable diresidues (RVDs); RA, right arm of accD TALENs, binding to 16 nucleotides with 15.5 RVDs. Boxes with dashed line indicate the targeted base pairs by two arms. Dashed line in the middle of the sequence shows the predicted cleavage by the dimeric FokI endonuclease domains. (B) Schematic map showing the gene construct transformed into tobacco via Agrobacterium mediated transformation. White arrow boxes indicate promoters and transcription directions; grey boxes indicate transcription termination signals; LB and RB, T-DNA left and right borders; CaM35S Poly(A), cauliflower mosaic virus polyadenylation signal; Bar (PPTR), Streptomyces hygroscopicus bar gene coding sequence for phosphinotricin acetyltransferase conferring phosphinotricin (PPT) resistance; CaM35S, cauliflower mosaic virus promoter; G10-90, a synthetic constitutive promoter consisting of G-box 10 motif and CaMV -90 / 35S promoter (24); XVE CDS, chimeric transcription activator assembled with the DNA-binding domain of the bacterial repressor LexA (X), coding sequence for the acidic transactivating domain of VP16 (V) and the regulatory region of the human estrogen receptor (E); rbcS Term, pea rubisco small subunit terminator; LexA, LexA operator; m35S (arrow), -35 to +12 minimal regulatory of CaM35S promoter; CP Pre, petunia rbcS8 chloroplast targeting presequence; TALE (LA) and TALE (RA), coding sequences for the left and right arms of TALE targeting domains; FokI, DNA cleavage domains of endonuclease FokI; HSP Term (grey box), Arabidopsis heat shock protein 18.2 terminator; T3A Term (grey box), pea rbcsS3A poly(A) terminator. Black arrows indicate primers used for cDNA PCR in (C). (C) Schematic map of the housekeeping gene EF1-α. A pair of primers were designed to include the 74 bp intron between exon2 and exon3, which would produce a 290 bp product if any DNA contamination presented in the RT-PCR in (D). If using cDNA as the template, the primers pair would produce a shorter PCR product of 216 bp, as the 74 bp intron was omitted. (D) RT-PCR to detect the transcription of TALENs with / without estradiol induction. cDNA samples were transcribed from total DNA extracted from plant samples. WT, wild type; WAT-4, accD TALENs transgenic line 4; MAT-3 and MAT- 11, accD TALENs transgenic line 3 and 11 harbouring chloroplast-targeted mutator DNA polymerase (MuPOP). The absence of 290 bp bands but only the 216 bp bands from EF-α1 indicate no DNA contamination in the RT-PCR reaction. Figure 2. (A) Leaf disks from wild type and accD TALENs transgenic lines were regenerated for four weeks with or without 20 µM estradiol. Scale bar indicates 1 cm. On the estradiol regeneration medium (ERM), expression of accD TALENs strongly inhibited shoot regeneration of accD TALEN transgenic lines WAT-4, MAT-3, MAT-11. Regeneration of shoots from wild type (WT) leaves was not affected by 20 µM estradiol. (B) accD TALEN selection allows isolation of shoots containing homoplasmic point mutations in the TALEN binding region of the accD gene. Sanger sequencing chromatograms are shown for the region of the accD gene bound by the left arm (LA) of the accD TALEN. Base substitution mutations are highlighted with shading. Clear single thymidine (T) peaks indicate homoplasmic C to T and A to T base substitutions in isolated shoots MAT-3-1 and MAT-3-2, respectively. (C) T1 seedlings germinated from the seeds collected from MAT-3-20E1 / 2 / 3 T0 plants. MS medium used to grow the seeds was supplemented with 20 µM estradiol. Red boxes indicate the randomly selected seedlings used for Sanger sequencing. Sanger sequencing data shows the stable inheritance of the homoplasmic thymidine point mutations. Figure 3. Point mutations identified using the estradiol inducible accD TALEN system. Shown are boxes with dashed line indicate the sequences bound by accD TALENs; scissors show the predicted cleavage position; boxed amino acids indicate the amino acids located in the CxxC motif, which are critical for accD function; homoplasmic (HM) and Heteroplasmic (HR) base substitutions and a rearrangement (REA) substitution are located and numbered to indicate the number of independently isolated identical mutations. Base substitutions and amino acid changes are shown. Syn, synonymous mutation. Figure 4. Using seeds as the starting material for selecting accD mutants with the estradiol inducible accD TALEN system. Figure 5. Transgenic lines isolated from estradiol accD TALEN selection system. Altered bases added via point mutations are indicated for each of the mutants. Heteroplasmic point mutations are shown in white boxes and homoplasmic point mutations are shown in grey boxes. HM, homoplasmic mutant; HT, heteroplasmic mutant; ESC, wild type plants escaped from the selection; REA, mutant containing rearrangement mutation.; LOST, plant exhibited weak growth preventing its propagation in vitro. Figure 6. Germination of the seeds from MAT-3 T0 plants on the MS medium containing 20 µM estradiol. At 20 µM estradiol, TALEN selection is effective in preventing the growth and development of most seedlings. The only surviving seedling has no mutation in the target accD sequence, confirmed by Sanger sequencing. Figure 7. The homoplasmy level of induced point mutations in MAT3-20E-1 and MAT3-20E- 3 plants. Next generation sequencing reads from each sample were mapped to the reference sequence (accD, NCBI Z00044.2). The x axis indicates the position of the nucleotide in the accD coding sequence. The percentages given to each mutation indicate the ratio of the number of sequence reads containing the point mutation out of the total number of sequence reads for that nucleotide position. (A) Only a C to T mutation at position 741 was identified in MAT3-20E-1 with a homoplasmy level of 97.7%. (B) Only a A to T mutation at position 738 was identified in MAT3-20E-3 with a homoplasmy level of 98.4%. Figure 8. Read coverage of Next generation sequencing data across the accD gene of MAT lines MAT3-20E-1 and MAT3-20E-3. The x axis indicates the Fnucleotides 59798 – 61336 from the reference genome, NCBI Acc. Z00044.2. (A) The coverage of MAT3-20E-1 ranged from 298 to 5013, of which the coverage at the position revealing the C to T mutation was 433. (B) The coverage of MAT3-20E-3 ranged from 126 to 3324, of which the coverage at the position revealing the A to T mutation was 313. Figure 9. The binding positions of accD-TALEN (described in Example 1) and accD-TALEN- UPSTREAM (Example 2). (A) A schematic map of accD gene is shown including 5’UTR, coding sequence and 3’UTR regions. ATG indicates the location of start codon. Bars under the map indicate the binding position of the two arms of each of accD-TALEN-UPSTREAM and accD-TALEN. (B) Two boxes indicate the target sequences recognised by accD-TALEN- UPSTREAM LA and RA, respectively. Arrow indicates the translation direction starting from ATG. Figure 10. A regeneration assay to verify the inducibility of accD-TALEN-UPSTREAM in the WAT-UP (wild-type accD-TALEN-UPSTREAM) transgenic lines. Leaf explants for each transgenic line were incubated on the regeneration medium containing 8 mg / L PPT with or without 17-β-estradiol (inducer). Organelle genomes from WAT-UP lines were not mutagenised by any method. No shoots could therefore regenerate from explants on the estradiol medium, as expected. This indicates a strong selection pressure against the wild type cells by accD-TALEN-UPSTREAM. An ideal WAT-UP transgenic plant should meet two criteria: 1) good regeneration on the medium without inducer and 2) no regeneration on the medium with inducer. Line 5 could not regenerate on the medium without inducer, implying it is a false positive transgenic shoot survived from PPT selection during transformation of accD- TALEN-UPSTREAM construct. Lines 2, 3 and 6 were determined as good host candidates to receive the donor DNA (accD variant DNA sequence) via particle bombardment, which allowed efficient creation of accD mutant plants. Size expansion and calli growth (on the rim) were observed in the leaf explants from lines 13 and 15, while the growth of the leaf explants on the induction medium was strongly inhibited. Figure 11. RT-PCR of WAT-UP transgenic lines. LA, TALEN left arm; RA, TALEN right arm. Leaf materials from selected WAT-UP lines were sampled after 24 hours incubation on the regeneration medium with or without estradiol induction. Line 5 was used as a negative control as it is a PPT sensitive plant (false positive transformant). Primers for RT-PCR were SEQ ID NO:17-21. Figure 12. (A) The binding positions of atp1-TALEN. A schematic map of atp1 gene is shown including 5’UTR, coding sequence and 3’UTR regions. ATG indicates the location of start codon. Bars under the map indicate the binding position of the two atp1-TALEN arms. (B) Two boxes indicate the target sequences recognised by atp1-TALEN LA and RA, respectively. Figure 13. A regeneration assay to verify the inducibility of atp1-TALEN in the Watp1T transgenic lines. Allocations of leaf explants from each line are indicated by the right panel. Leaf explants from each transgenic line were incubated on the regeneration medium (RM) containing 8 mg / L PPT with or without 17-β-estradiol (inducer). Images were taken when the explants were incubated for 2 and 5 weeks, respectively. Organelle genomes from Watp1T lines were not mutagenised by any method. Leaf explants from line 4 and 5 showed size expansion and calli / shoots growth on RM, but no signs of calli / shoots growth were observed in these two lines on the RM with 20 µM inducer. Figure 14. RT-PCR of Watp1T transgenic lines. LA, TALEN left arm; RA, TALEN right arm. Leaf materials from Watp1T-4 and Watp1T-5 were sampled after 24 hours incubation on the regeneration medium with or without estradiol induction. Primers for RT-PCR were SEQ ID NO:17-21. Figure 15. Healthy Watp1T transgenic seedlings growing in the MS medium containing 8 mg / L PPT. Figure 16. Nucleotide substitutions in the target region of accD-TALEN. The positions of substituted bases in the donor construct has been pointed out by triangles. Figure 17. Map of pATR. EXAMPLES Example 1 Introduction Essential DNA genomes are present in the chloroplasts and mitochondria of plants. These organelle genomes commonly exhibit very low spontaneous mutation rates, which are 5-20 fold lower than the nuclear genome (1). These low mutation rates prevent the variation in organelle gene sequences required for breeding improved crops. This includes single nucleotide polymorphisms (SNPs) in organelle genes that give rise to multiple alleles of a chloroplast or mitochondrial gene. This lack of variation is a barrier to understanding the functions of organelle genes and isolating new alleles for breeding improved crops. It has previously been shown that organelle genes with SNPs encode beneficial crop traits such as herbicide resistance (2–4) and chilling tolerance (5). A method to introduce SNPs into organelle genes to generate multiple alleles would allow the identification of specific alleles with beneficial traits for crop breeding, and it would also allow the targeted creation of such alleles in varieties of interest such as elite cultivars used in plant breeding. Chloroplast transformation can introduce nucleotide substitutions into chloroplast genes (6). Chloroplast transformation has become a routine procedure for changing and studying chloroplast genes in tobacco (7). Applications of chloroplast transformation to isolate new alleles of chloroplast genes are limited by the relatively small number of plant species amenable to the technology. Species recalcitrant to chloroplast transformation include monocots such as the cereal crops wheat, maize, rice and barley. Other disadvantages of chloroplast transformation to introduce point mutations are the requirements for marker genes to select transgenic chloroplasts, and the fact that only a fraction of transformants will contain the desired nucleotide substitution in a chloroplast gene because the marker gene can be integrated without concomitant insertion of the nucleotide substitution. Usually several rounds of selection are needed to ensure a homogeneous population of mutant chloroplast genomes carrying the mutation with the marker gene are required. The presence of a marker gene, which can often confer antibiotic resistance, is undesirable due to regulatory concerns related to the presence of an unnecessary foreign gene in a crop and metabolic load. Therefore for applications in crops the chloroplast transformation marker genes will need to be removed once the desirable SNP has been introduced into a chloroplast genes. Several methods can be used to remove marker genes but they can leave DNA scars such as the target sites for site specific recombinases (6,8). These disadvantages limit the applications of chloroplast transformation to isolate new alleles of chloroplast genes that increase the yield and environmental resilience of major crops. A previous study isolated nine transplastomic tobacco plants containing point mutations in the rbcL gene of tobacco plants by chloroplast transformation (9). The results are limited by the relatively small number of bases changed compared to the number available in the target rbcL gene. This limits the potential to isolate a large number of rbcL alleles containing base substitutions located throughout the rbcL gene. Currently, mitochondrial transformation is not available in higher plants ruling out the isolation of new alleles of mitochondrial genes by transformation. CRISPR-Cas has been widely used for targeted mutagenesis in the plant nuclear genome (10). The type of mutations introduced by the initial version of CRISPR-Cas was limited to introduce deletions and insertions resulting from non-homologous recombination repair after DNA cleavage at the targeted location. Later, more derivatives overcame this limitation by combining the DNA recognition property of CRISPR-Cas system with cytosine or adenine deaminases (CD or AD). These fusion enzymes are known as cytosine or adenine base editors (CBE or ADE), allowing efficient base substitutions at specific sites, without cutting the DNA (11, 12). Nonetheless, plant organelle genomes are recalcitrant to CRISPR-Cas based gene-editing systems, as the DNA targeting property of CRISPR-Cas relies on complementary base-paring mediated by guide RNA (gRNA). This seems to rule out the use of CRISPR-Cas nucleases encoded by the nucleus to cleave plant organelle DNA because there seems to be no available method to target a gRNA expressed from a nuclear gene to the chloroplast or mitochondrion in plants (13). This makes transcription activator-like effector nucleases (TALENs) an attractive genome-editing tool for plant organelles, as TALENs only require the TALE protein domain to target specific DNA sequences. TALENs fused with mitochondrial targeting N-terminal presequences have been used to introduce deletions in the mitochondria genomes of Arabidopsis (14), tomato (15), Brassica napus (16) and rice (16, 17). More recently, TALE-CD fusions have been shown to be capable of introducing C:G to A:T substitutions at targeted locations in both chloroplasts and mitochondria (18–20). However, applications of TALE-CD for mutagenesis of plant organelle genes is significantly limited because TALE-CD is restricted to a single type of base substitution preventing changes at many nucleotide position within an organelle gene or regulatory sequence, or any organellar target site. A further limitation of base editors is the limited number of bases that can be changed. This means multiple base editor constructs are required to change different bases in an organelle gene, which is labour intensive and time consuming. Here is exemplified a method that enables targeted introduction of stable point mutations in plant organelles by a controllable selection method. This example uses a TALEN as the targeting nuclease and the chloroplast as the target organelle. By taking the advantage of the site-specific binding property of TALENs, chloroplast genomes carrying point mutations in the TALE recognition site will become resistant to cleavage and survive, while the wild type genomes will be cleaved and degraded. Eventually, TALEN-mediated counter-selection of wild-type chloroplast genomes will fix point mutations and enable isolation of homoplasmic chloroplast mutant plants. By applying this selection method, implemented in this example using TALENs, to a chloroplast mutator line, the inventors targeted the essential chloroplast accD gene in Nicotiana tabacum. The inventors chose an essential chloroplast gene because essential genes are recalcitrant to base substitution changes using chloroplast transformation (21). Most point mutations in essential genes are reversed to wild type by chloroplast DNA repair pathways. Here is shown that combining a chloroplast mutator with TALEN selection allows the facile isolation of stable mutant lines containing homoplasmic and inheritable point mutations in the chloroplast accD gene of Nicotiana tabacum with greater efficiency. This example demonstrates the high efficiency of the selection method for isolating alleles of difficult-to-change chloroplast genes and this method is applicable to a wide variety of crops, including monocots. Results Design of the estradiol inducible accD TALENs expression system DNA cleavage mediated by TALENs has been shown to be capable of targeting DNA sequences in nuclei, chloroplasts and mitochondria. The transcription activator-like effector (TALE) domain is responsible for sequence recognition. It contains an N-terminus recognising T and a programmable central array of amino acid repeats plus a C-terminus. Each repeat contains 34 amino acids with the 12thand 13thas repeat variable diresidues (RVDs) critical to specific base pairs recognition (22). With the endonuclease domain of FokI nuclease fused to the C-terminus of TALE, a pair of TALENs were designed to bind to opposite DNA strands with their C-termini juxtaposed, allowing cleavage at a specific position by the dimeric FokI endonuclease domain (23). To test the TALEN in the tobacco chloroplast genome, a pair of TALENs was designed to target the accD gene (SEQ ID NO:2–7), with the monomeric TALEN binding to the direct and complementary strands referred to as the left and right arms (LA and RA), respectively (Fig 1A). LA and RA were engineered to target the accD coding sequence to recognise 14 and 16 bases, respectively. To ensure chloroplast targeting of accD TALENs, the presequence from the petunia rubisco small subunit 8 (SEQ ID NO:10, 11) was added to the N-terminus of both TALEN arms. Chloroplast DNA is a multicopy sequence. This means any new mutations in chloroplast genes will be relatively rare compared to the vast excess of wild type chloroplast sequences present. Chloroplast DNA repair mechanisms including homology dependent repair act to remove mutations using the wild type sequences as templates. Strong expression of TALENs designed to cleave wild type sequences is necessary to selectively eliminate wild type chloroplast genomes. This provides strong negative selection against the wild type genome copies enabling preferential replication and amplification of mutant sequences that are not cleaved by the TALENs. The choice of promoter to express the TALEN is important. The use of a promoter that is active throughout development such as the CaMV 35S promoter results in relatively high and constant expression of the TALEN during growth and development. This may lead to cell death following transformation of the CaMV 35S TALEN construct into plants and cleavage of wild type chloroplast genomes. Cleavage would be prevented by the presence of mutated chloroplast genomes and mutations can be facilitated by concurrently mutagenizing the chloroplast, in this example this is performed by using a plant line expressing a high error-rate chloroplast polymerase (MuPOP, WO2023073383). To address this concern, an estradiol inducible system was used to control the expression of TALENs. The inducible control by estradiol is mediated by the expression of a fusion transactivator enzyme containing the DNA- binding domain (DBD) of the bacterial repressor LexA (X), the acidic transactivating domain of VP16 (V) and the regulatory region of the human estrogen receptor (E), so called the XVE system (24). XVE enzyme is constantly expressed, and its transactivator function is regulated by the estrogen receptor regulatory region (ER). Estradiol supplemented to the transgenic plants binds to ER, activating the LexA DBD and VP16 transactivator domain of XVE enzymes. The activated XVE enzymes bind to the LexA operator sequences upstream to the minimal CaM35S promoter (m35S) to enable TALENs expression and start the positive selection for the mutant chloroplast genomes. All transgenes including XVE (SEQ ID NO:8) and accD TALENs were cloned into a binary vector pCB3300 with a Bar marker gene ready for selecting transgenic plants (Fig 1B). The construct was transformed into the wild type line and a transgenic line expressing MuPOP. Fifteen T0 transgenic lines were isolated from each host background, namely wild type accD TALEN (WAT) lines and mutator accD TALEN (MAT) lines, respectively. To confirm the inducibility of XVE system, reverse transcription (RT)-PCR was performed with primers targeting to the cDNA regions specific to LA and RA, respectively. Among all tested transgenic lines, the tight control of the XVE system on the transcription of accD TALENs is observed in line WAT-4, MAT-3 and MAT-11. Expression of the LA and RA TALEN arms were verified by RT-PCR. RT-PCR products of mRNA encoding LA and RA using extracted plant RNA were fractionated on agarose gels (Fig 1D). The LA RT-PCR bands were only detected in the plant samples treated with estradiol. This shows tight regulation of expression of the LA- TALEN transgenic lines. Leaky expression of the RA RT-PCR band is visible in the absence of estradiol. However, both LA and RA are required for cleavage of wild-type chloroplast DNA. No bands were visible in the lanes corresponding to WT RNA, which lacks the LA and RA TALENS. Expression of accD TALENs strongly suppress shoots regeneration in tobacco To confirm that induced expression of accD TALEN would provide efficient selection against the wild type cells in WAT-4, MAT-3 and MAT-11 lines, an assay was performed by placing 20 leaf explants from each transgenic line on MS regeneration medium (RM) alone or supplemented with 20 µM 17-β-estradiol (ERM). Leaf explants were passed onto fresh ERM medium every 10-14 days to ensure the concentration of estradiol for the optimal induction of accD TALENs. After four weeks, no regenerated callus or shoots were observed in WAT-4, MAT-3 and MAT-11 on the estradiol regeneration medium (ERM), while numerous shoots were observed in all three lines on the normal regeneration medium. Other transgenic lines were not continued, as their explants were well regenerated and indistinguishable between the RM and ERM plates, similar to the wild type (Fig 2A), consistent with lack of accD TALEN expression. Isolation of homoplasmic accD mutants with inheritable point mutations from transgenic lines expressing both MuPOP and accD TALEN Leaf explants from WAT-4, MAT-3 and MAT-11 were further passed on fresh ERM plates every 10-14 days. After 10 weeks, two green shoots were regenerated from MAT-3 explants, labelled as MAT-3-20E1 and MAT-3-20E2 (20E is the abbreviation for 20 µM estradiol). One more shoot, MAT-3-20E3, was regenerated from the same callus which gave rise to MAT-3- 20E2. Explants not giving rise to callus or shoots became necrotic. As MAT lines have higher base substitution rates in chloroplasts, the inventors predicted these shoots would contain point mutations in the target region of accD gene that were introduced by the error-prone MuPOP These mutations would prevent cleavage by accD TALENs resulting in their positive selection compared to the wild type accD allele, which would be cleaved. The accD point mutations preventing cleavage were located in the accD TALEN binding site (Fig 2B). MAT- 11 explants gave rise to some green-yellowish calli but the inventors were unable to isolate shoots after 12 weeks. The inventors were able to isolate MAT11 shoots when the estradiol concentration was reduced (see below). In contrast to MAT lines, all leaf disks from WAT-4 turned brown and showed no sign of callus or shoot growth by 12 weeks, indicating the induced expression of accD TALENs is lethal to the wild type cells. MAT-3-20E1, MAT-3-20E2 and MAT-3-20E3 were further grown on MS medium without estradiol until they were large enough to be sampled for PCR and Sanger sequencing. From each line, a 300 bp PCR product covering the accD TALENs target region was amplified and sequenced with Sanger technology. Sanger sequencing data revealed that C to T and A to T substitutions were successfully introduced into the DNA binding region of the TALEN left arm. The C to T mutation was discovered in MAT-3-20E1, while the A to T mutation was shared between MAT-3-20E2 and MAT-3-20E3 (Fig 2B). Three point mutations were homoplasmic with no other mutations observed in the rest of the 300 bp amplified sequence. Furthermore, next generation sequencing data confirmed no off-target mutations located elsewhere in the accD gene of MAT-3-20E1 and MAT-3-20E3. At the position revealing point mutations, the percentage of the C to T mutation in MAT3-20E-1 was 97.7% out of the total read coverage of 433, while the percentage of the A to T mutation in MAT3-20E-3 was 98.4% out of the total read coverage of 313 (Fig 7&8). Three T0 accD mutant lines grown to maturity and producing copious seeds exhibited indistinguishable visible phenotypes compared to wild type plants. To investigate if the point mutations were inheritable to the next generation, seeds for the T1 plants were germinated on MS medium supplemented with 20 µM estradiol (Fig 2C). Growth of these seedlings on estradiol medium was consistent with lack of TALEN cleavage of their chloroplast genomes resulting from accD mutations in the TALEN binding site. Sanger sequencing was performed on one seedling randomly selected from each line. All sequenced seedlings showed stable inheritance of the accD point mutations from the parental T0 plants. Estradiol inducible accD-TALEN is an effective and tuneable method for isolating accD mutants The XVE inducible system has been reported to provide tuneable expression regulated by the concentration of estradiol. In separate experiments, use of 2 µM estradiol in Arabidopsis expressing GFP driven by the XVE system showed that expression of GFP decreased after 24 hours. To investigate if changing the estradiol concentration would affect the efficiency of obtaining accD mutant alleles from MAT lines, more leaf explants from MAT-3 and MAT-11 T0 plants were assayed using ERM with 2 µM and 10 µM estradiol, respectively (Table 1). All leaf explants were passaged to fresh ERM every 10-14 days. With lower estradiol concentrations, shoots appeared more rapidly and at a higher frequency using MAT-3 and MAT-11 explants. It took 12 weeks for MAT-3 to give rise to only three shoots when using 20 µM estradiol. When using 2 µM estradiol, the inventors could easily isolate 20 shoots within 8 weeks due to the numerous shoots formed. MAT-11 was not able to give rise to any regenerated shoots within 12 weeks when using 20 µM estradiol. In the assay using 10 µM estradiol, MAT-11 also gave rise to numerous shoots, from which 27 shoots were isolated within 8 weeks. Leaf explants were cultured in the presence of the same concentrations of estradiol for 12 weeks. All isolated shoots were sequenced to identify if any mutations were introduced into the target region of accD. Out of the 20 shoots from MAT-3, point mutations were identified in 16 shoots (80% efficiency) including eight homoplasmic (HM, 35% of total) and eight heteroplasmic (HT, 45% of total) shoots. Four shoots were labelled as “escapes” (20%, ESC) as no mutation was detected in the accD target region. In comparison, the 27 shoots from MAT-11 showed different numbers and ratios of accD mutant and ‘escaped’ shoots lacking mutations in the accD TALEN binding, with lower homoplasmic (5 lines, 19% of total), similar heteroplasmic (11 lines, 42% of total) and higher escaped (10 lines, 39% of total) shoots. Notably, there were two exceptional shoots which were classified as heteroplasmic. One is MAT-3-2E13 which contained a heteroplasmic point mutation in the sequence targeted by RA and a heteroplasmic rearrangement in the upstream to the sequence targeted by the LA. The other is MAT-11- 10E16, which contained a heteroplasmic and a homoplasmic point mutations both located in the sequence targeted by the LA (Fig 5). In summary, with using lower concentrations of estradiol, MAT-3 and MAT-11 lines the inventors were able to rapidly isolate abundant accD mutants accounting for up to 80% of the total number of estradiol-resistant shoots sequenced. These estradiol-resistant shoots contained homoplasmic or heteroplasmic accD mutations in the TALEN binding sites and a small number of shoots only wild type accD alleles that survived the relaxed selection pressure. Additional rounds of selection would be required to purify the heteroplasmic mutants into homoplasmic ones. In summary, a high concentration of estradiol (e.g.20 µM) ensures the isolation of a small number of resistant shoots homoplasmic for accD mutations whilst lowering the estradiol concentration enabled the relatively rapid isolation of numerous resistant shoots but many of these were heteroplasmic or escapes. Table 1 Frequencies of shoots containing homoplasmic (HM), heteroplasmic (HT) and no (ESC) chloroplast accD mutations formed on estradiol regeneration medium. Estradiol Cultured T0 lines No.of conc. time HM HT ESC No. of Explants1shoots (µM) (weeks) MAT-3 20 20 12 3 (100%) 0 0 34 40 2 8 7 (35%) 9 (45%)24 (20%) 205MAT-11 20 20 12 0 0 0 0 40 10 8 5 (19%) 11 (42%)310 (39%) 2651 Explants were passed onto fresh medium every 10-14 weeks. 2 One shoot, MAT-3-2E13, containing a heteroplasmic rearrangement and a heteroplasmic point mutation, was classified as HT mutant. 3 One shoot, MAT-11-10E16, containing two heteroplasmic point mutations, was classified as one HT mutant. 4 Only three shoots were regenerated. 5 Many more shoots were regenerated but not sequenced. Point mutations identified in accD mutants In total forty nine shoots were isolated and sequenced using MAT-3 and MAT-11 T0 plants, of which thirty-five shoots contained accD point mutations representing four types of base substitutions located at different sites in the TALEN binding region of the accD coding sequence. Referring to the adenine in the start codon as the first nucleotide, the point mutations were T734A, A738T, C741T, G742T and G777T, which gave rise to V245I, Q246H, C247C (synonymous codon mutation), E248K and L259F amino acid changes, respectively (Fig 3). The identified base substitutions in accD mutants are consistent with the common types of base substitutions introduced by MuPOP in vitro (26). This strongly suggests that the accD point mutations in the TALEN binding site resulted from the activity of the MuPOP. Analysis of the point mutations in different accD alleles revealed preferences for C741T and G777T mutations. The TALEN LA binds the DNA sequence encoding amino acids encompassing the critical CxxC functional motif (C247 to C250) in the beta-subunit of acetyl Co A carboxylase, which is the product of the accD gene (27). As a result some mutations located in the TALEN LA binding site might be lethal and would not be detectable in the employed assay. C741T is a synonymous mutation for amino acid C247 and would not be expected to have a large influence on accD products activity. G777T is located outside the motif and its recovery in homoplasmic shoots is consistent with a limited impact on accD gene function. The majority of accD mutants harboured a single point mutation that was either homoplasmic or heteroplasmic. As mentioned above, the MAT-3-2E13 and MAT-11-10E16 mutant lines were the two exceptions. MAT-3-2E13 contained a heteroplasmic G777T and a heteroplasmic rearrangement located upstream of the LA binding site in the accD gene. The rearrangement could result from DNA repair involving micro-homology mediated end joining following TALEN cleavage of DNA, which has been reported in plant chloroplasts. MAT-11-10E16 contained a heteroplasmic T734A and a homoplasmic C741T, suggesting coexistence of two mutant genomes with one containing C741T only while the other containing both T734A and C741T. A possible explanation to this result is that the two mutant genomes emerged in the same cell and were simultaneously selected and propagated during organogenesis. From culturing 100 leaf explants (10 petri dishes), the inventors isolated shoots with point mutations located at five different locations in accD, showing an unprecedented high efficiency of isolating chloroplast mutants in an essential tobacco gene, which has not been possible using alternative methods. The data suggests that the inducible TALEN selection method is effective for obtaining single point mutations located at different sites in a small target sequence. Seeds from the transgenic line are effective material for isolating accD mutants Many plant species are recalcitrant to in vitro culture involving the regeneration of plants from explants. Therefore, the inventors investigated if seeds from selfed MAT-3 T0 plants could be used as the starting material for isolating accD mutants. Seeds were sterilised and sown on MS medium containing 100 mg / L kanamycin and 10 mg / l phosphinothricin (PPT). Kanamycin is the selectable marker used to verify the presence of the plastid targeted MuPOP and PPT for the presence of the transgene encoding the inducible TALEN cleaving accD. About seventy-five percent of the seeds germinated into green seedlings the remaining about 25% of seedlings bleached and died on the kanamycin / PPT medium consistent with the absence of the TALEN encoding transgene. This was expected as the T0 plants were not homozygous for the TALEN transgene. The same batch of seeds was also sterilised and sown on MS medium only containing 5 µM estradiol without antibiotics. About half of the seeds failed to germinate into viable seedlings after three weeks, while the remaining seeds germinated and grew into green seedlings (Fig 4). For a single insertion of the TALEN transgene into the nuclear genome, 75% of seedlings will inherit the TALEN transgene, which when expressed would kill them. From the estradiol plate, 12 seedlings with expanded true leaves were sampled and sequenced. Half of the sequenced seedlings were accD mutants, however, they were all heteroplasmic and harboured the G777T substitution only. More seeds were germinated on the MS medium with 20 µM estradiol. At this higher 20 µM estradiol concentration, growth and development of the vast majority of seedlings was arrested showing the efficacy of TALEN-selection. Only one seedling survived, which had no mutations detected in the accD target region (Fig 6) most likely resulting from loss or silencing of the accD TALEN transgene. These data suggests that the selection strength of accD-TALENs is tuneable by different concentrations of estradiol, which is consistent with the results obtained with the regeneration assay using leaf explants (Table 1). The results show that seeds can be used to isolate mutations in the accD gene using the inducible TALEN selection system. This will allow this system to be used in taxa recalcitrant to in vitro tissue culture, such as Arabidopsis, soybean and cereals. Discussion Despite the advances in gene editing methodologies to change genes in the nucleus, the introduction of homoplasmic point mutations into plant chloroplast genes has remained a lengthy and labour intensive process, which is limited to relatively few crops amenable to chloroplast transformation. This has limited the isolation of new mutant alleles of genes encoded by plant organelle genomes (28). Base editor TALE-CD has been shown to be capable of introducing homoplasmic point mutations in both chloroplasts and mitochondria (18, 19, 29), but each TALE-CD can only introduce a single type of base substitution (C:G to A:T) and requires individual transgenic lines of new TALE constructs for each target site. These drawbacks will restrict the usefulness of TALE-CD for forward genetics to isolate multiple alleles of an organelle gene using a single transgenic line. Recently, TALEN has been used to select a wider spectrum of point mutations in the tobacco mitochondrial genome (30). A CaMV35S promoter regulated TALEN was designed to introduce a double strand break (DSB) in the wild type nad9 mitochondrial gene. Mutations in the TALEN binding site located nad9 gene would reduce cleavage by the nad9 TALEN. As a result, the mutant mitochondrial genomes carrying point mutations in the nad9 gene are preferentially amplified following TALEN cleavage of wild type mitochondrial genomes. This results in homoplasmy of mutant nad9 mitochondrial genomes following regeneration of shoots from leaf explants taken from the nad9-TALEN transgenic lines. This method was named TALEN gene-driven mutagenesis (GDM). However, TALEN-GDM has several limitations including 1) only T1 TALEN transgenic line with low cleavage efficiency (up to 20%) could be isolated; 2) When screening for the nad9 mutants, inefficient DSB selection pressure resulted in a high background of wild type shoots (~95% of total sequenced shoots), so a high volume of sequencing for the shoots from each regeneration cycles was necessary until positive mutant identified; 3) up to one year of multiple regeneration cycles in the presence of a chemical mutagen (ethidium bromide, EtBr) was necessary as the spontaneous mutation rate in plant mitochondria is too low to provide selectable point mutations. As a result, these procedures gave rise to a frequency of positive mutant shoots of only 5% of total sequenced shoots. Such a low frequency would also limit the use of TALEN-GDM in chloroplast, which would require higher DSB selection pressure due to the higher genome copy number in chloroplasts (~10,000 per cell) than mitochondria (~150 per cell). In summary, the above method provides an opportunity of selecting more types of point mutations than base editors in plant mitochondria genome, but the process is hazardous and inefficient hindering its widespread use for isolating alleles of organelle genes in crops. CaMV35S is a classic constitutive promoter expected to drive strong transgene expression. The organelle genome targeted by a TALEN driven by CaMV35S will be cut via a double strand break. If the process was efficient and the break not repaired this would lead to cell death as we have observed with inducible expression of the accD TALEN. The transgenic lines constitutively expressing nad9-TALEN were reported to have a wild type phenotype which can be explained by the low ~20% cleavage efficiency of nad9-TALEN. Transgenic cells with mitoTALEN cleavage efficiencies higher than 20% might be lethal and lost during the transformation process before any selectable mutations have emerged in the TALEN binding region. This lethality at high mitoTALEN cleavage rates would bias the selection of suitable transgenic lines expressing mitoTALEN to those with relatively low mitochondrial cleavage rates. Thus these transgenic lines would only confer a weak selection pressure against the wild type mitochondrial genomes. Therefore, the low efficiency of TALEN-GDM in isolating organelle mutants may be attributed to constitutive expression of mitoTALENs, resulting in biased isolation of transgenic lines with weak expression of TALEN. This hypothesis could be even more applicable if the TALEN target region located in essential organelle genes such as accD (chloroplast only), ATP synthase subunit genes and rDNA genes. The present example addresses the above problems by using the XVE inducible system to control the expression of the accD targeted TALEN in the transgenic line. The XVE system provided tight regulation of TALEN expression, which was tuneable depending on the concentration of estradiol used as an inducer. So, the inventors could unbiasedly isolate transgenic lines on the estradiol-free medium. Leaf disks from T0 transgenic seedlings were assayed on the regeneration medium with estradiol to confirm the ideal candidate lines for later regeneration of organelle mutants. The lines were continued for larger scale tissue culture if their regeneration were normal on the estradiol-free medium but strongly inhibited on the estradiol medium, as exhibited by WAT-4, MAT-3, and MAT-11 (Fig 2A). Transgenic lines showing strong estradiol-inducible inhibition in regeneration indicated a strong selection pressure against the wild type chloroplast genomes, resulting in a high efficiency (61-100%) in positive mutants and a low (0-39%) wild type background when screening for the accD mutants (Table 1). Notably, expression of the chloroplast targeted mutator POP provided an advantage of elevated chloroplast mutation rate in MAT-3 and MAT-11, without using any chemical mutagens. Mutator POP has been shown to provide a 51-fold higher mutation rate than the EtBr (WO2023073383) (31). But it was critical to use highly induced accD TALEN for eliminating the wild type background, otherwise the number of wild type shoots would still overwhelm the number of positive mutants, because of the strong purifying selection in the chloroplast. In addition to the inducible expression of accD-TALEN, the inventors consider that the timing for inducing the accD-TALEN expression is also important to ensure the high frequency of accD mutants. It would be more achievable for accD-TALEN to complete the cleavage on all wild type genomes when the plant cell is at the stage containing fewer copies of the organelle genomes. In the present example, the inventors induced accD-TALEN expression at the beginning of the explant regeneration. There were approximately only 9-16 copies of chloroplast genomes for a regenerating plant cell, which is the de-differentiation stage (32, 33). Alternatively, a high frequency of accD mutants should be also achievable if applying estradiol to MAT-3 and MAT-11 lines at the flowering stage, providing a similar effect as AtRPS5A promoter in Arabidopsis. In summary, the system presented here allowed the inventors to isolate the accD mutants from MAT-3 and MAT11 in only 8-12 weeks and a small number of leaf disks in tissue culture. In total five mutant alleles of accD were isolated, which included A:T to T:A, C:G to T:A and G:C to T:A base pair substitutions (Fig 3 and 5 . The inventors also demonstrated the effectiveness of the XVE regulated accD TALEN when using the seeds from T0 MAT-3 and MAT-11 parental lines as the starting material for screening the accD mutants (Fig 4). This feature would facilitate the identification of organelle mutants in many crops recalcitrant to tissue culture. In summary, this example provides an efficient, rapid and economic solution for isolating plant organelle mutants, which is not achievable if the TALEN is regulated by a constitutive promoter, such as the TALEN-GDM system (Table 2). Additionally, this system may provide a novel strategy for transforming plant chloroplasts and mitochondria. As it can confer a robust selection effect against the wild type organelle genome at a high concentration of estradiol. A transgenic organelle genome harbouring a transgene, in place of a wild type organelle genome sequence will not be cleaved, and can be efficiently selected to become homoplasmic in a regenerated shoot or a seedling germinated from seed. This strategy provides an alternative to using antibiotic selection marker genes, such as aadA, in chloroplast transformation. While aadA is only effective in several dicot plant species, the system presented here is applicable to both dicot and monocot species. For mitochondria, where an effective selection marker gene is not available for plants (34), this system provides a new route for mitochondrial transformation in plants, enabling mitochondrial genome modification for valuable traits in crops such as pest resistance, herbicide resistance and cytoplasmic male sterility. The organelle transformants isolated from this system will not contain antibiotic resistance genes in organelle, and the transgenes encoding XVE and TALEN can be removed by backcrossing the organelle transformants with the wild type plant. Table 2 Different plant organelle mutagenesis system based on TALEN. Tools Targeted Mutation Efficiency of Technique for Time gene types isolating isolating homoplasmic organelle mutants mutants cpTALEN rpoB (cp) Random No Tissue culture N / A (35) point homoplasmic mutation or mutants deletion mitoTALEN atp6-1 Large 0-100% Seeds 10-12 weeks (14) and atp6- deletions germination (Arabidopsis 2 (mito) with random only) sizes TALEN- Nad9 Targeted ~5% Tissue culture Up to one GDM (30) (mito) point year mutations ITOMS (this accD (cp) Targeted 19-100% Tissue culture or 8-12 weeks work) point seeds mutations germination and transgenes Sequences SEQ ID NO:1 accD TALEN target sequence in Fig 1A and Fig 3 TGGGTTCAATGCGAAAATTGTTATGGATTAAATTATAAGAAATTTTTGAAATCA Underlines indicate the sequences bound by the left and right TALE domains. SEQ ID NO:2 accD TALEN coding sequence (left arm) ATGGCTTCCTCTGTGATTTCCTCTGCAGCTGTTGCTACTCGCACTAATGTGGCTCAAGC TAGCATGGTTGCACCTTTTAATGGTCTTAAGTCTGCTGTCTCCTTCCCAGTTTCAAGCAA GCAAAACCTTGACATCACTTCCATTGCTAGCAATGGTGGAAGAGTCCAATGCATGGACT ACAAGGATGACGACGATAAAAGTTGGAAGGACGCAAGTGGTTGGTCTAGAATGCATGC GGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGG ATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCG TTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCA CATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAG CACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAAC AGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGA GGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCG TGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCGCTCACGGGAGCACCCCTCA ACCTGACCCCCGACCAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCT GGAAACAGTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGACCACGGCCTGACCCCGGAA CAGGTGGTTGCAATCGCGTCACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAG CGACTCCTGCCCGTCCTGTGCCAGGCCCACGGCCTGACCCCAGACCAGGTTGTGGCC ATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGG TTCTCTGCCAGGCCCACGGCCTGACCCCAGCCCAGGTTGTGGCCATCGCCAGCAACAT AGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGAC CACGGCCTGACCCCCGACCAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAG GCGCTGGAAACAGTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGACCACGGCCTGACCC CAGAACAAGTTGTCGCGATTGCAAGCAACAACGGAGGCAAACAAGCCTTAGAAACAGT CCAGAGATTGTTGCCGGTGCTGTGCCAAGCCCACGGCCTGACCCCGGACCAGGTGGT TGCAATCGCGTCACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTG CCCGTCCTGTGCCAGGCCCACGGCCTGACCCCAGCCCAAGTTGTCGCGATTGCAAGC AACAACGGAGGCAAACAAGCCTTAGAAACAGTCCAGAGATTGTTGCCGGTGCTGTGCC AAGACCACGGCCTGACCCCAGACCAGGTTGTGGCCATCGCCAGCAACATAGGTGGCA AGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGACCACGGCCT GACCCCAGAACAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGA AACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCAGACCAG GTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGA CTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCAGCCCAGGTTGTGGCCATC GCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTC TCTGCCAGGACCACGGCCTGACCCCTGAGCAGGTAGTGGCTATTGCATCCAACGGAG GGGGCAGACCCGCACTGGAGTCAATCGTGGCCCAGCTTTCGAGGCCGGACCCCGCGC TGGCCGCACTCACTAATGATCATCTTGTAGCGCTGGCCTGCCTCGGCGGACGTCCTGC CATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGATCCCAGCTA GTGAAATCTGAATTGGAAGAGAAGAAATCTGAACTTAGACATAAATTGAAATATGTGCCA CATGAATATATTGAATTGATTGAAATCGCAAGAAATTCAACTCAGGATAGAATCCTTGAA ATGAAGGTGATGGAGTTCTTTATGAAGGTTTATGGTTATCGTGGTAAACATTTGGGTGG ATCAAGGAAACCAGACGGAGCAATTTATACTGTCGGATCTCCTATTGATTACGGTGTGA TCGTTGATACTAAGGCATATTCAGGAGGTTATAATCTTCCAATTGGTCAAGCAGATGAAA TGCAAAGATATGTCGAAGAGAATCAAACAAGAAACAAGCATATCAACCCTAATGAATGG TGGAAAGTCTATCCATCTTCAGTAACAGAATTTAAGTTCTTGTTTGTGAGTGGTCATTTC AAAGGAAACTACAAAGCTCAGCTTACAAGATTGAATCATATCACTAATTGTAATGGAGCT GTTCTTAGTGTAGAAGAGCTTTTGATTGGTGGAGAAATGATTAAAGCTGGTACATTGAC ACTTGAGGAAGTGAGAAGGAAATTTAATAACGGCGAGATAAACTTTTAA rbcS chloroplast targeting presequence (DNA) is underlined. SEQ ID NO:3 accD TALEN coding sequence (right arm) ATGGCTTCCTCTGTGATTTCCTCTGCAGCTGTTGCTACTCGCACTAATGTGGCTCAAGC TAGCATGGTTGCACCTTTTAATGGTCTTAAGTCTGCTGTCTCCTTCCCAGTTTCAAGCAA GCAAAACCTTGACATCACTTCCATTGCTAGCAATGGTGGAAGAGTCCAATGCATGGACT ACAAGGATGACGACGATAAAAGTTGGAAGGACGCAAGTGGTTGGTCTAGAATGCATGC GGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGG ATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCG TTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCA CATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAG CACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAAC AGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGA GGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCG TGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCGCTCACGGGAGCACCCCTCA ACCTGACCCCGGACCAGGTGGTTGCAATCGCGTCACACGATGGGGGAAAGCAGGCCC TAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAGGACCACGGCCTGACCCCAGA ACAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCA GAGACTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCAGACCAGGTTGTGGC CATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCG GTTCTCTGCCAGGCCCACGGCCTGACCCCAGCCCAGGTTGTGGCCATCGCCAGCAAC ATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGG ACCACGGCCTGACCCCAGACCAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGC AGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGACCACGGCCTGAC CCCAGAACAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAAC CGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCCGACCAGGTT GTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCCTCT TGCCGGTCTTGTGTCAGGCCCACGGCCTGACCCCCGCCCAGGTTGTCGCTATTGCTAG TAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCCTCTTGCCGGTCTTGTGT CAGGACCACGGCCTGACCCCCGACCAGGTTGTCGCTATTGCTAGTAACGGCGGAGGC AAACAGGCGCTGGAAACAGTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGACCACGGCC TGACCCCGGAACAGGTGGTTGCAATCGCGTCACACGATGGGGGAAAGCAGGCCCTAG AAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAGGCCCACGGCCTGACCCCCGACC AGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCG CCTCTTGCCGGTCTTGTGTCAGGCCCACGGCCTGACCCCCGCCCAGGTTGTCGCTATT GCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCCTCTTGCCGGTC TTGTGTCAGGACCACGGCCTGACCCCAGACCAGGTTGTGGCCATCGCCAGCAACATAG GTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGACCA CGGCCTGACCCCCGAACAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGC GCTGGAAACAGTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGCCCACGGCCTGACCCCT GAGCAGGTAGTGGCTATTGCATCCAACATAGGGGGCAGACCCGCACTGGAGTCAATCG TGGCCCAGCTTTCGAGGCCGGACCCCGCGCTGGCCGCACTCACTAATGATCATCTTGT AGCGCTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCC GCACGCGCCGGAATTGATCAGATCCCAGCTAGTGAAATCTGAATTGGAAGAGAAGAAA TCTGAACTTAGACATAAATTGAAATATGTGCCACATGAATATATTGAATTGATTGAAATC GCAAGAAATTCAACTCAGGATAGAATCCTTGAAATGAAGGTGATGGAGTTCTTTATGAA GGTTTATGGTTATCGTGGTAAACATTTGGGTGGATCAAGGAAACCAGACGGAGCAATTT ATACTGTCGGATCTCCTATTGATTACGGTGTGATCGTTGATACTAAGGCATATTCAGGA GGTTATAATCTTCCAATTGGTCAAGCAGATGAAATGCAAAGATATGTCGAAGAGAATCA AACAAGAAACAAGCATATCAACCCTAATGAATGGTGGAAAGTCTATCCATCTTCAGTAAC AGAATTTAAGTTCTTGTTTGTGAGTGGTCATTTCAAAGGAAACTACAAAGCTCAGCTTAC AAGATTGAATCATATCACTAATTGTAATGGAGCTGTTCTTAGTGTAGAAGAGCTTTTGAT TGGTGGAGAAATGATTAAAGCTGGTACATTGACACTTGAGGAAGTGAGAAGGAAATTTA ATAACGGTGAGATAAACTTTTAA rbcS chloroplast targeting presequence (DNA) is underlined. SEQ ID NO:4 accD TALEN amino acids sequence (left arm) with targeting peptide MASSVISSAAVATRTNVAQASMVAPFNGLKSAVSFPVSSKQNLDITSIASNGGRVQCMDYK DDDDKSWKDASGWSRMHAAPRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKVRSTV AQHHEALVGHGFTHAHIVALSQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSGARAL EALLTDAGELRGPPLQLDTGQLVKIAKRGGVTAMEAVHASRNALTGAPLNLTPDQVVAIASN GGGKQALETVQRLLPVLCQDHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPD QVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQD HGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNNGGKQALETVQRL LPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQA LETVQRLLPVLCQDHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNI GGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQV VAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNGGGRPALESIVAQLSRPDPALA ALTNDHLVALACLGGRPAMDAVKKGLPHAPELIRSQLVKSELEEKKSELRHKLKYVPHEYIE LIEIARNSTQDRILEMKVMEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYS GGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQL TRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF rbcS chloroplast targeting presequence (amino acids) is underlined. Repeat variable diresidues (RVDs) for recognising target accD sequence are in bold. SEQ ID NO:5 accD TALEN amino acid sequence (right arm) with targeting peptide MASSVISSAAVATRTNVAQASMVAPFNGLKSAVSFPVSSKQNLDITSIASNGGRVQCMDYK DDDDKSWKDASGWSRMHAAPRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKVRSTV AQHHEALVGHGFTHAHIVALSQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSGARAL EALLTDAGELRGPPLQLDTGQLVKIAKRGGVTAMEAVHASRNALTGAPLNLTPDQVVAIASH DGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQ VVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDH GLTPDQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNIGGKQALETVQRLLP VLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALE TVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIASHD GGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQ VVAIASNGGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQD HGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGRPALESIVAQL SRPDPALAALTNDHLVALACLGGRPAMDAVKKGLPHAPELIRSQLVKSELEEKKSELRHKLK YVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVI VDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFK GNYKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF rbcS chloroplast targeting presequence (amino acids) is underlined. Repeat variable diresidues (RVDs) for recognising target accD sequence are in bold. SEQ ID NO:6 accD TALEN amino acids sequence (left arm) DYKDDDDKSWKDASGWSRMHAAPRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKV RSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSG ARALEALLTDAGELRGPPLQLDTGQLVKIAKRGGVTAMEAVHASRNALTGAPLNLTPDQVV AIASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHG LTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNIGGKQALETVQRLLPVL CQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNNGGKQALET VQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNG GKQALETVQRLLPVLCQDHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVV AIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGL TPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNGGGRPALESIVAQLSRP DPALAALTNDHLVALACLGGRPAMDAVKKGLPHAPELIRSQLVKSELEEKKSELRHKLKYVP HEYIELIEIARNSTQDRILEMKVMEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDT KAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNY KAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF The +153 N-terminus and +47 C-terminus for TALE are underlined and double underlined, respectively. Repeat variable diresidues (RVDs) for recognising target accD sequence are in bold. SEQ ID NO:7 accD TALEN amino acid sequence (right arm) DYKDDDDKSWKDASGWSRMHAAPRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKV RSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSG ARALEALLTDAGELRGPPLQLDTGQLVKIAKRGGVTAMEAVHASRNALTGAPLNLTPDQVV AIASHDGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGL TPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNIGGKQALETVQRLLPVLC QDHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNIGGKQALETVQR LLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQ ALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIA SHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLT PAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNIGGKQALETVQRLLPVLC QDHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGRPALESIVA QLSRPDPALAALTNDHLVALACLGGRPAMDAVKKGLPHAPELIRSQLVKSELEEKKSELRH KLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDY GVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSG HFKGNYKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF The +153 N-terminus and +47 C-terminus for TALE are underlined and double underlined, respectively. Repeat variable diresidues (RVDs) for recognising target accD sequence are in bold. SEQ ID NO:8 XVE coding sequence ATGAAAGCGTTAACGGCCAGGCAACAAGAGGTGTTTGATCTCATCCGTGATCACATCAG CCAGACAGGTATGCCGCCGACGCGTGCGGAAATCGCGCAGCGTTTGGGGTTCCGTTC CCCAAACGCGGCTGAAGAACATCTGAAGGCGCTGGCACGCAAAGGCGTTATTGAAATT GTTTCCGGCGCATCACGCGGGATTCGTCTGTTGCAGGAAGAGGAAGAAGGGTTGCCG CTGGTAGGTCGTGTGGCTGCCGGTGAACCGTCGAGCGCCCCCCCGACCGATGTCAGC CTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGC GCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTT ACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGC AGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGGATCCGTCTGCTGGAGA CATGAGAGCTGCCAACCTTTGGCCAAGCCCGCTCATGATCAAACGCTCTAAGAAGAAC AGCCTGGCCTTGTCCCTGACGGCCGACCAGATGGTCAGTGCCTTGTTGGATGCTGAGC CCCCCATACTCTATTCCGAGTATGATCCTACCAGACCCTTCAGTGAAGCTTCGATGATG GGCTTACTGACCAACCTGGCAGACAGGGAGCTGGTTCACATGATCAACTGGGCGAAGA GGGTGCCAGGCTTTGTGGATTTGACCCTCCATGATCAGGTCCACCTTCTAGAATGTGC CTGGCTAGAGATCCTGATGATTGGTCTCGTCTGGCGCTCCATGGAGCACCCAGTGAAG CTACTGTTTGCTCCTAACTTGCTCTTGGACAGGAACCAGGGAAAATGTGTAGAGGGCAT GGTGGAGATCTTCGACATGCTGCTGGCTACATCATCTCGGTTCCGCATGATGAATCTGC AGGGAGAGGAGTTTGTGTGCCTCAAATCTATTATTTTGCTTAATTCTGGAGTGTACACAT TTCTGTCCAGCACCCTGAAGTCTCTGGAAGAGAAGGACCATATCCACCGAGTCCTGGA CAAGATCACAGACACTTTGATCCACCTGATGGCCAAGGCAGGCCTGACCCTGCAGCAG CAGCACCAGCGGCTGGCCCAGCTCCTCCTCATCCTCTCCCACATCAGGCACATGAGTA ACAAAGGCATGGAGCATCTGTACAGCATGAAGTGCAAGAACGTGGTGCCCCTCTATGA CCTGCTGCTGGAGATGCTGGACGCCCACCGCCTACATGCGCCCACTAGCCGTGGAGG GGCATCCGTGGAGGAGACGGACCAAAGCCACTTGGCCACTGCGGGCTCTACTTCATC GCATTCCTTGCAAAAGTATTACATCACGGGGGAGGCAGAGGGTTTCCCTGCCACAGTC TGA SEQ ID NO:9 XVE amino acid sequence MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEEHLKALARKGVIEIVSGA SRGIRLLQEEEEGLPLVGRVAAGEPSSAPPTDVSLGDELHLDGEDVAMAHADALDDFDLD MLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGGDPSAGDMRAANLWP SPLMIKRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELV HMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKLLFAPNLLLDRNQGK CVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVL DKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLE MLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATV The amino acids sequence includes LexA binding domain (italic), VP16 transactivation domain (single underlined) and human estradiol receptor regulatory domain (double underlined). SEQ ID NO:10 rbcS pre-sequence for targeting accD TALEN to chloroplast (DNA) ATGGCTTCCTCTGTGATTTCCTCTGCAGCTGTTGCTACTCGCACTAATGTGGCTCAAGC TAGCATGGTTGCACCTTTTAATGGTCTTAAGTCTGCTGTCTCCTTCCCAGTTTCAAGCAA GCAAAACCTTGACATCACTTCCATTGCTAGCAATGGTGGAAGAGTCCAATGCATG SEQ ID NO:11 rbcS pre-sequence for targeting accD TALEN to chloroplast (amino acids) MASSVISSAAVATRTNVAQASMVAPFNGLKSAVSFPVSSKQNLDITSIASNGGRVQCM Sequences in Fig2 Fig 2B SEQ ID NOaccD mutants Sequenced sequence 5’ > 3’ 12 T0 MAT-3-20E1 AATGTGAAA 13 T0 MAT-3-20E2 ATTGCGAAA Point mutations are in bold. Fig 2C SEQ ID NOaccD mutants Sequenced sequence 5’ > 3’ 14 T1 MAT-3-20E1 TTCAATGTG 15 T1 MAT-3-20E2 TTCATTGCG 16 T1 MAT-3-20E3 TTCATTGCG Point mutations are in bold. Primers used in RT-PCR SEQ ID NO Primers Sequences 5’ > 3’ 17 FokI-RT-F GAGGAAGTGAGAAGGAAATTTAATAACG 18 FokI-HSP Term-R GCCACAAATTCATAACACAACAAGC 19 FokI-T3A Term-R TGAACTTGACGAACGTTGTCGA 20 EF1-α-F CACTGGTGGTTTTGAAGCTGGTAT 21 EF1-α-R AAAGGGGATTTTATCAGGGTTGTATCC SEQ ID NO:22 Nicotiana tabacum wild type POP amino acid sequence MAFLGFSVQS SPFKPTSYLW FSPHSFSSSR SFWASSGKAL HRREDCKTQS VENASSSLAV LGDSIKQISS HERKLFSSGL QHKIEEDSTY GWIAETNALK ASKAKSSYNS YKKISAANCN VSASTNRRVK DEFFDVPTEV NTRMMRERIT SSYSATTCIS GGNLSSKSKP PYNPAGGEKK VVGNWREYEN HLPQVSVGLT HSRVNGARSV NKVDGSNVSH YKPLSKGSHL NGQLSSKIME PKLEKVNKLR EGHASDQLRH SVNGTETKVV TVKAKGVIQE RAMNKMEKNV IQAVTADVMN GAEANAKGVI LERATNKMEK NAIESMATDV VNGTKTRIVN DEGTGVSQVS LRERLGAMYD KVHIVDNLSA AKEVVRKLTS QYRHLVHACD TEVAKIDVKQ QTPVDHGEII CFSIYSGPEA DFGDGKSCIW VDVLDGDGKN LLVEFAPFFQ DPSIRKVWHN YSFDNHVIEN YGFKVSGFHA DTMHMARLWD SSRRTSGGYS LEALTGDSTV MRDARPVHAE RLFHGEGLFG KISMKTIFGR KKLKKDGTEG KVTVIPSVEE LQKTERELWI CYSALDSIST LMLYESLKNK LAKRIWTFDG VRKGSMYEFY EKYWRPFGEL LVQMETEGVL VDRAYLAEIE KVAKAEQQVA ANRFRNWAAK YCHDAKYMNV GSDTQLRQLF FGGIQNRKNS DESLPYEKEF KVPNIDKVTE EGKKAPTKFR KIRLHRICDL IDTEMYTASG WPSVSGDALK ALSGKVSADF DILDEADDNA EEDPETSIDE ALATNNEVPS QEPEVSIYGS AYNAFGGGQK GIEACHAIAA LCEMCSIGSL ISNFILPLQG QDVSGENGRI HCSLNINTET GRLSARRPNL QNQPALEKDR YKIRQAFVAA QGNSLIVADY GQLELRILAH LANCKSMLDA FKAGGDFHSR TAMNMYTHIR EAVENGQVLL EWHPQPGEEK PPVPLLKDAF GSERRKAKML NFSIAYGKTT IGLARDWKVS VKEAKETVDR WYRDRKEVSD WQEQRKFEAR EFRRVHTLLG RARWFPSVKN ATGSVKGHIE RAAINTPVQG SAADVAMCAM LEISKNARLE ELGWKLLLQV HDEVILEGPE ESENEAMAIV VDCMSKPFGG KNILRVDLSV DSKCAKNWYS AK Note that positions in bold are those which may be modified as described herein. SEQ ID NO:23 Nicotiana tabacum modified POP amino acid sequence MAFLGFSVQS SPFKPTSYLW FSPHSFSSSR SFWASSGKAL HRREDCKTQS VENASSSLAV LGDSIKQISS HERKLFSSGL QHKIEEDSTY GWIAETNALK ASKAKSSYNS YKKISAANCN VSASTNRRVK DEFFDVPTEV NTRMMRERIT SSYSATTCIS GGNLSSKSKP PYNPAGGEKK VVGNWREYEN HLPQVSVGLT HSRVNGARSV NKVDGSNVSH YKPLSKGSHL NGQLSSKIME PKLEKVNKLR EGHASDQLRH SVNGTETKVV TVKAKGVIQE RAMNKMEKNV IQAVTADVMN GAEANAKGVI LERATNKMEK NAIESMATDV VNGTKTRIVN DEGTGVSQVS LRERLGAMYD KVHIVDNLSA AKEVVRKLTS QYRHLVHACA TAVAKIDVKQ QTPVDHGEII CFSIYSGPEA DFGDGKSCIW VDVLDGDGKN LLVEFAPFFQ DPSIRKVWHN YSFDNHVIEN YGFKVSGFHA DTMHMARLWD SSRRTSGGYS LEALTGDSTV MRDARPVHAE RLFHGEGLFG KISMKTIFGR KKLKKDGTEG KVTVIPSVEE LQKTERELWI CYSALDSIST LMLYESLKNK LAKRIWTFDG VRKGSMYEFY EKYWRPFGEL LVQMETEGVL VDRAYLAEIE KVAKAEQQVA ANRFRNWAAK YCHDAKYMNV GSDTQLRQLF FGGIQNRKNS DESLPYEKEF KVPNIDKVTE EGKKAPTKFR KIRLHRICDL IDTEMYTASG WPSVSGDALK ALSGKVSADF DILDEADDNA EEDPETSIDE ALATNNEVPS QEPEVSIYGS AYNAFGGGQK GIEACHAIAA LCEMCSIGSL ISNFILPLQG QDVSGENGRI HCSLNINTET GRLSARRPNL QNQPALEKDR YKIRQAFVAA QGNSLIVADY GQFELRILAH LANCKSMLDA FKAGGDFHSR TAMNMYTHIR EAVENGQVLL EWHPQPGEEK PPVPLLKDAF GSERRKAKML NFSIAYGKTT IGLARDWKVS VKEAKETVDR WYRDRKEVSD WQEQRKFEAR EFRRVHTLLG RARWFPSVKN ATGSVKGHIE RAAINTPVQG SAADVAMCAM LEISKNARLE ELGWKLLLQV HDEVILEGPE ESENEAMAIV VDCMSKPFGG KNILRVDLSV DSKCAKNWYS AK Note modified positions D390A, E392A and L903F are in bold and underlined. SEQ ID NO:24 +153 TALE N-terminus amino acid sequence APRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVAL SQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSGARALEALLTDAGELRGPPLQLDTG QLVKIAKRGGVTAMEAVHASRNALTGAPLN SEQ ID NO:25 +47 TALE C-terminus amino acid sequence SIVAQLSRPDPALAALTNDHLVALACLGGRPAMDAVKKGLPHAPELIR SEQ ID NO:26 +136 TALE N-terminus amino acid sequence VDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDM IAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAV HAWRNALTGAPLN SEQ ID NO:27 +63 TALE C-terminus amino acid sequence SIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLPHAPALIKRTNRRIPERTSHRVA SEQ ID NO:28 Alternative oxidase 1 (AOX1) presequence amino acid sequence MMMMMSRSGGNRVANTAMFVAKGLSGEVGGLRALYGGGVRSES SEQ ID NO:29 LexA Operator Sequence CGTACTGTACATATAACCACTGGTTTTATATACAGCAGTACTGTACATATAACCACTGGT TTTATATACAGCAGTCGACGTACTGTACATATAACCACTGGTTTTATATACAGCAGTACT GTACATATAACCACTGGTTTTATATACAGCAGTCGA SEQ ID NO:30 minimal CaM35S promoter GACCCTTCCTCTATATAAGGAAGTTCATTTCATTTGGAGAGGACACGCTGAAGCTAGTC SEQ ID NO:31 Pea T3A terminator sequence CAGGCCTCCCAGCTTTCGTCCGTATCATCGGTTTCGACAACGTTCGTCAAGTTCAATGC ATCAGTTTCATTGCCCACACACCAGAATCCTACTAAGTTTGAGTATTATGGCATTGGAAA AGCTGTTTTCTTCTATCATTTGTTCTGCTTGTAATTTACTGTGTTCTTTCAGTTTTTGTTTT CGGACATCAAAATGCAAATGGATGGATAAGAGTTAATAAATGATATGGTCCTTTTGTTCA TTCTCAAATTATTATTATCTGTTGTTTTTACTTTAATGGGTTGAATTTAAGTAAGAAAGGA ACTAACAGTGTGATATTAAGGTGCAATGTTAGACATATAAAACAGTCTTTCACCTCTCTT TGGTTATGTCTTGAATTGGTTTGTTTCTTCACTTATCTGTGTAATCAAGTTTACTATGAGT CTATGATCAAGTAATTATGCAATCAAGTTAAGTACAGTATAGGCTTTTTGTGTCGAG Example 2 In Example 1, out of 10-15 independent transgenic plants, only 1-2 plants could significantly express the accD-TALENs using 17-β-estradiol as inducer (Table 5). Leaf explants from the transgenic plants with good inducibility of accD-TALEN showed strong inhibition in regeneration on the regeneration medium containing 17-β-estradiol (ERM). To improve efficiency, the inventors developed a new TALEN targeting the upstream 5’UTR of the accD gene in tobacco (Figure 9), namely accD-TALEN-UPSTREAM. The construct of accD-TALEN-UPSTREAM resembles the structure of accD-TALEN (as described in Example 1) except that the RVDs recognise the upstream 5’UTR region of accD gene (SEQ ID NO: 52- 55). Together with the XVE system described in Example 1, the two accD-TALEN- UPSTREAM arms were cloned into pCAMBIA3300 vector and then transformed into wild type tobacco plants. Sixteen independent shoots were isolated from the transformation experiment but only five were real transgenic plants as determined by healthy growth in the MS medium containing 10 mg / L phosphinothricin (PPT), which is the selection reagent for the marker gene (Bar) in the pCAMBIA3300 (Figure 10). Other isolated shoots were false positive as they were unable to grow and eventually died in the MS medium containing 10 mg / L PPT. Transgenic plants in this experiment were named as wild-type accD-TALEN-UPSTREAM (WAT-UP). Table 5 Targeting the TALEN to the upstream (5’UTR and start codon region) of accD gene increases the probability of obtaining a transgenic plant showing selection effect. Transformants Total isolated Probability of obtaining a Host plant with estradiol- Construct PPT resistant transgenic plant showing (tobacco) inducible transformants1selection effect phenotype Transgenic XVE-accD tobacco 15 2 (line 3 and 11) 13% TALEN expressing (Example 1) mutator POP wild type 14 1 (line 4) 7% XVE-accD 5 (lines 2, 3, 6, 13, TALEN- wild type 5 2 100% 15) UPSTREAM1The estradiol-inducible phenotype is determined as strong inhibition of regeneration in explants.2All isolated shoots were numbered but only those grew healthy in the 10 mg / L PPT medium were proceeded to estradiol regeneration assay. All five WAT-UP plants (lines 2, 3, 6, 13 and 15) were proceeded to regeneration assay to check the inducibility of accD-TALEN-UPSTREAM on the ERM containing 20 μM 17-β- estradiol (Figure 10). The result showed all five WAT-UP plants could not regenerate in the presence of 20 µM estradiol, indicating successful expression of accD-TALEN-UPSTREAM, and successful cutting of the target sites which leads to diminished plant viability. RT-PCR result also confirmed the induction of accD-TALEN-UPSTREAM in WAT-UP 3, 6 and 13, although unbalanced transcription levels were detected in the left / right arms (LA / RA) (Figure 11). As a result, the chance of obtaining a WAT-UP plant with desirable susceptibility of 17-β- estradiol is 5 / 5 (100%), which is a significant increase compared to the results presented in Example 1 (as shown in Table 5, 2 / 15 and 1 / 14 for MAT and WAT plants, respectively). This result suggests targeting the TALEN to the upstream of the gene and specifically to the 5’UTR could be a better strategy for creating a host plant suitable for selecting organelle mutant. Regardless the unbalanced expression level between LA and RA, accD-TALEN- UPSTREAM can still cleave the target sequence and induce the regeneration inhibition in WAT-UP plants. Others have reported that the TALEN with unbalanced or single expression of LA or RA could still effectively cleave the DNA target (30, 38, 39). Lines such as the five WAT-UP lines created here (lines 2, 3, 6, 13 and 15) will be highly suitable for introducing targeted mutations in the target sites targeted in these lines. For example, these lines may be useful for donor DNA-mediated modification of target sites. By delivering a donor DNA sequence flanked with sequences homologous to the target gene in the chloroplast or mitochondria, a host plant created with the present technology, will allow efficient selection of the homoplasmic mutant plant harbouring the donor sequence. The donor sequence can be designed to contain individual or a combination of point mutation, insertion or deletion in the target gene, for example the donor sequence may contain entire genes destined for knocking into the target site. All methods used are described in Example 1 and the Methods. Sequences (used in Example 2) SEQ ID NO:52 accD-TALEN-UPSTREAM coding sequence (left arm) ATGGCTTCCTCTGTGATTTCCTCTGCAGCTGTTGCTACTCGCACTAATGTGGCTCAAGC TAGCATGGTTGCACCTTTTAATGGTCTTAAGTCTGCTGTCTCCTTCCCAGTTTCAAGCAA GCAAAACCTTGACATCACTTCCATTGCTAGCAATGGTGGAAGAGTCCAATGCATGGACT ACAAGGATGACGACGATAAAAGTTGGAAGGACGCAAGTGGTTGGTCTAGAATGCATGC GGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGG ATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCG TTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCA CATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAG CACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAAC AGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGA GGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCG TGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCGCTCACGGGAGCACCCCTCA ACCTGACCCCAGACCAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCC TCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGACCACGGCCTGACCCCCGA ACAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAG CGCCTCTTGCCGGTCTTGTGTCAGGCCCACGGCCTGACCCCAGACCAGGTTGTGGCC ATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGG TTCTCTGCCAGGCCCACGGCCTGACCCCAGCCCAAGTTGTCGCGATTGCAAGCAACAA CGGAGGCAAACAAGCCTTAGAAACAGTCCAGAGATTGTTGCCGGTGCTGTGCCAAGAC CACGGCCTGACCCCGGACCAGGTGGTTGCAATCGCGTCACACGATGGGGGAAAGCAG GCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAGGACCACGGCCTGACC CCAGAACAAGTTGTCGCGATTGCAAGCAACAACGGAGGCAAACAAGCCTTAGAAACAG TCCAGAGATTGTTGCCGGTGCTGTGCCAAGCCCACGGCCTGACCCCAGACCAGGTTGT GGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTA CCGGTTCTCTGCCAGGCCCACGGCCTGACCCCAGCCCAGGTTGTGGCCATCGCCAGC AACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCC AGGACCACGGCCTGACCCCCGACCAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCA AACAGGCGCTGGAAACAGTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGACCACGGCCT GACCCCAGAACAAGTTGTCGCGATTGCAAGCAACAACGGAGGCAAACAAGCCTTAGAA ACAGTCCAGAGATTGTTGCCGGTGCTGTGCCAAGCCCACGGCCTGACCCCAGACCAG GTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGA CTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCGGCCCAGGTGGTTGCAATC GCGTCACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTC CTGTGCCAGGACCACGGCCTGACCCCCGACCAGGTTGTCGCTATTGCTAGTAACGGC GGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGAC CACGGCCTGACCCCAGAACAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAG GCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCC CCGACCAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGT TCAGCGCCTCTTGCCGGTCTTGTGTCAGGCCCACGGCCTGACCCCTGAGCAGGTAGT GGCTATTGCATCCAACGGAGGGGGCAGACCCGCACTGGAGTCAATCGTGGCCCAGCT TTCGAGGCCGGACCCCGCGCTGGCCGCACTCACTAATGATCATCTTGTAGCGCTGGCC TGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCG GAATTGATCAGATCCCAGCTAGTGAAATCTGAATTGGAAGAGAAGAAATCTGAACTTAG ACATAAATTGAAATATGTGCCACATGAATATATTGAATTGATTGAAATCGCAAGAAATTC AACTCAGGATAGAATCCTTGAAATGAAGGTGATGGAGTTCTTTATGAAGGTTTATGGTTA TCGTGGTAAACATTTGGGTGGATCAAGGAAACCAGACGGAGCAATTTATACTGTCGGAT CTCCTATTGATTACGGTGTGATCGTTGATACTAAGGCATATTCAGGAGGTTATAATCTTC CAATTGGTCAAGCAGATGAAATGCAAAGATATGTCGAAGAGAATCAAACAAGAAACAAG CATATCAACCCTAATGAATGGTGGAAAGTCTATCCATCTTCAGTAACAGAATTTAAGTTC TTGTTTGTGAGTGGTCATTTCAAAGGAAACTACAAAGCTCAGCTTACAAGATTGAATCAT ATCACTAATTGTAATGGAGCTGTTCTTAGTGTAGAAGAGCTTTTGATTGGTGGAGAAATG ATTAAAGCTGGTACATTGACACTTGAGGAAGTGAGAAGGAAATTTAATAACGGCGAGAT AAACTTTTAA rbcS chloroplast targeting presequence (DNA) is underlined. SEQ ID NO:53 accD-TALEN-UPSTREAM coding sequence (right arm) ATGGCTTCCTCTGTGATTTCCTCTGCAGCTGTTGCTACTCGCACTAATGTGGCTCAAGC TAGCATGGTTGCACCTTTTAATGGTCTTAAGTCTGCTGTCTCCTTCCCAGTTTCAAGCAA GCAAAACCTTGACATCACTTCCATTGCTAGCAATGGTGGAAGAGTCCAATGCATGAGCG CATGGAGCCACCCTCAATTCGAGAAAGGGGGTGGCAGCAGAATGCATGCGGCCCCGC GACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCA CGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGT GGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGC GCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATC ACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCC GGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCC GTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCA ATGGAGGCAGTGCATGCATCGCGCAATGCGCTCACGGGAGCACCCCTCAACCTGACC CCAGACCAAGTTGTCGCGATTGCAAGCAACAACGGAGGCAAACAAGCCTTAGAAACAG TCCAGAGATTGTTGCCGGTGCTGTGCCAAGACCACGGCCTGACCCCGGAACAGGTGG TTGCAATCGCGTCACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCT GCCCGTCCTGTGCCAGGCCCACGGCCTGACCCCGGACCAGGTGGTTGCAATCGCGTC ACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTG CCAGGCCCACGGCCTGACCCCGGCCCAGGTGGTTGCAATCGCGTCACACGATGGGGG AAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAGGACCACGG CCTGACCCCGGACCAGGTGGTTGCAATCGCGTCACACGATGGGGGAAAGCAGGCCCT AGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAGGACCACGGCCTGACCCCGGA ACAGGTGGTTGCAATCGCGTCACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCA GCGACTCCTGCCCGTCCTGTGCCAGGCCCACGGCCTGACCCCCGACCAGGTTGTCGC TATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCCTCTTGCCG GTCTTGTGTCAGGCCCACGGCCTGACCCCAGCCCAGGTTGTGGCCATCGCCAGCAAC ATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGG ACCACGGCCTGACCCCCGACCAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACA GGCGCTGGAAACAGTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGACCACGGCCTGAC CCCCGAACAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACA GTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGCCCACGGCCTGACCCCCGACCAGGTT GTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCCTCT TGCCGGTCTTGTGTCAGGCCCACGGCCTGACCCCAGCCCAAGTTGTCGCGATTGCAAG CAACAACGGAGGCAAACAAGCCTTAGAAACAGTCCAGAGATTGTTGCCGGTGCTGTGC CAAGACCACGGCCTGACCCCGGACCAGGTGGTTGCAATCGCGTCACACGATGGGGGA AAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAGGACCACGGC CTGACCCCAGAACAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTC GAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCCGAC CAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGC GCCTCTTGCCGGTCTTGTGTCAGGCCCACGGCCTGACCCCTGAGCAGGTAGTGGCTAT TGCATCCAACAACGGGGGCAGACCCGCACTGGAGTCAATCGTGGCCCAGCTTTCGAG GCCGGACCCCGCGCTGGCCGCACTCACTAATGATCATCTTGTAGCGCTGGCCTGCCTC GGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTG ATCAGATCCCAGCTAGTGAAATCTGAATTGGAAGAGAAGAAATCTGAACTTAGACATAA ATTGAAATATGTGCCACATGAATATATTGAATTGATTGAAATCGCAAGAAATTCAACTCA GGATAGAATCCTTGAAATGAAGGTGATGGAGTTCTTTATGAAGGTTTATGGTTATCGTG GTAAACATTTGGGTGGATCAAGGAAACCAGACGGAGCAATTTATACTGTCGGATCTCCT ATTGATTACGGTGTGATCGTTGATACTAAGGCATATTCAGGAGGTTATAATCTTCCAATT GGTCAAGCAGATGAAATGCAAAGATATGTCGAAGAGAATCAAACAAGAAACAAGCATAT CAACCCTAATGAATGGTGGAAAGTCTATCCATCTTCAGTAACAGAATTTAAGTTCTTGTT TGTGAGTGGTCATTTCAAAGGAAACTACAAAGCTCAGCTTACAAGATTGAATCATATCAC TAATTGTAATGGAGCTGTTCTTAGTGTAGAAGAGCTTTTGATTGGTGGAGAAATGATTAA AGCTGGTACATTGACACTTGAGGAAGTGAGAAGGAAATTTAATAACGGTGAGATAAACT TTTAA rbcS chloroplast targeting presequence (DNA) is underlined. SEQ ID NO:54 accD-TALEN-UPSTREAM amino acids sequence (left arm) with targeting peptide MASSVISSAAVATRTNVAQASMVAPFNGLKSAVSFPVSSKQNLDITSIASNGGRVQCMDYK DDDDKSWKDASGWSRMHAAPRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKVRSTV AQHHEALVGHGFTHAHIVALSQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSGARAL EALLTDAGELRGPPLQLDTGQLVKIAKRGGVTAMEAVHASRNALTGAPLNLTPDQVVAIASN IGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQ VVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDH GLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNNGGKQALETVQRLL PVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNIGGKQALE TVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNN GGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQV VAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDH GLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLP VLCQAHGLTPEQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPAMD AVKKGLPHAPELIRSQLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFF MKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEEN QTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIG GEMIKAGTLTLEEVRRKFNNGEINF rbcS chloroplast targeting presequence (amino acids) is underlined. Repeat variable diresidues (RVDs) for recognising target accD sequence are in bold. SEQ ID NO:55 accD-TALEN-UPSTREAM amino acids sequence (right arm) with targeting peptide MASSVISSAAVATRTNVAQASMVAPFNGLKSAVSFPVSSKQNLDITSIASNGGRVQCMSAW SHPQFEKGGGSRMHAAPRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKVRSTVAQH HEALVGHGFTHAHIVALSQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSGARALEAL LTDAGELRGPPLQLDTGQLVKIAKRGGVTAMEAVHASRNALTGAPLNLTPDQVVAIASNNG GKQALETVQRLLPVLCQDHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVV AIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHG LTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPEQVVAIASHDGGKQALETVQRLLP VLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNIGGKQALE TVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNG GGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQ VVAIASNNGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQD HGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLL PVLCQAHGLTPEQVVAIASNNGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPAM DAVKKGLPHAPELIRSQLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEF FMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEEN QTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIG GEMIKAGTLTLEEVRRKFNNGEINF rbcS chloroplast targeting presequence (amino acids) is underlined. Repeat variable diresidues (RVDs) for recognising target accD sequence are in bold. SEQ ID NO: 64 accD-TALEN-UPSTREAM target site TATAGCGAATGACTATTCATCTATTGTATTTTCATGCAAATAGGGGGCA Example 3 Genetic transformation in plant mitochondrial genome has remained challenging for decades. One major reason is lacking a suitable selectable marker gene allowing efficient positive selection of mitochondrial mutant in plants. A robust selection system with no background of wild type escapes would be favourable for obtaining a plant mutant with a homoplasmic status of the mutagenized mitochondrial genome. Instead of searching for a marker gene specific to plant mitochondria, here the inventors provide another example to illustrate the process of creating two host plants suitable for mitochondrial genetic engineering, by expressing an XVE- TALEN targeting to the atp1 gene (mitochondria located) in transgenic tobacco plants. All methods used in Example 3 are as described in Example 1 and the Methods. To create the construct encoding the XVE-atp1-TALEN, we modified the XVE-accD-TALEN by 1) changing the RVDs coding sequences for recognising the sequences in atp1 gene (Figure 12, SEQ ID NO: 56-59), and 2) changing the rbcS presequences of TALEN left arm (LA) and right arm (RA) to the AOX1 presequence (SEQ ID NO:28) to allow mitochondria- specific targeting of atp1-TALENs. Together with the XVE system described in Example 1, XVE-atp1-TALEN was cloned into pCAMBIA3300 vector and then transformed into wild type tobacco plants. Six independent transgenic plants were isolated from the transformation experiment as determined by healthy growth in the MS medium containing 10 mg / L phosphinothricin (PPT), which is the selection reagent for the marker gene (Bar) in the pCAMBIA3300. Transgenic plants in this experiment were named as wild-type atp1-TALEN (Watp1T). Leaf explants from all Watp1T lines were used in regeneration assay to identify the lines with optimal inducible expression of atp1-TALEN (Figure 13). After two weeks incubation, all six lines could form calli and shoots on RM, but only line 4 and 5 showed strong inhibition in regeneration on the RM with 20 µM estradiol (ERM). To confirm the induced phenotype in lines 4 and 5 correlated with the expression of atp1-TALEN, leaf samples from both RM and ERM were proceeded to RT-PCR analysis (Figure 14). The result suggested clear induction of both LA and RA from atp1-TALEN when the leaf explants were treated with estradiol. Faint bands for LA and RA were visible in samples free from estradiol, indicating an unperfect regulation of atp1-TALEN by the XVE system. However, the level of spontaneous expression did not affect the growth of Watp1T-4 and Watp1T-5 seedlings in the MS medium with 8 mg / L PPT (Figure 15). The result indicates that a high level expression of atp1-TALEN by induction is required to inhibit the cell growth in Watp1T plants. Both Watp1T-4 and Watp1T-5 are promising candidates suitable for mitochondrial transformation. A mitochondrial mutant plant could be efficiently isolated from Watp1T-4 and Watp1T-5 treated with estradiol, if any mutations could be introduced to the region targeted by atp1-TALEN, such as point mutations and / or indels. Such mutations can be created by any methods, such as chemical mutagens, radiation, mutator POP or any DNA delivery methods (e.g. particle bombardment, virus and nanoparticles, etc). Sequences (used in Example 3) SEQ ID NO: 56 atp1-TALEN coding sequence (left arm) ATGATGATGATGATGTCTAGGTCAGGAGGTAACAGGGTAGCTAACACAGCAATGTTTGT GGCAAAGGGTTTGTCAGGTGAAGTTGGTGGTTTGAGAGCATTGTATGGTGGTGGTGTG AGGAGTGAGTCCATGGACTACAAGGATGACGACGATAAAAGTTGGAAGGACGCAAGTG GTTGGTCTAGAATGCATGCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTT CGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGA AGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCC ATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGAC CGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGAT ATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACG GATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAG ATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCG CTCACGGGAGCACCCCTCAACCTGACCCCGGACCAGGTGGTTGCAATCGCGTCACAC GATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAG GACCACGGCCTGACCCCAGAACAAGTTGTCGCGATTGCAAGCAACAACGGAGGCAAAC AAGCCTTAGAAACAGTCCAGAGATTGTTGCCGGTGCTGTGCCAAGCCCACGGCCTGAC CCCAGACCAAGTTGTCGCGATTGCAAGCAACAACGGAGGCAAACAAGCCTTAGAAACA GTCCAGAGATTGTTGCCTGTGCTGTGCCAAGCCCACGGCCTGACCCCCGCCCAGGTT GTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCCTCT TGCCGGTCTTGTGTCAGGACCACGGCCTGACCCCGGACCAGGTGGTTGCAATCGCGT CACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGT GCCAGGACCACGGCCTGACCCCAGAACAAGTTGTCGCGATTGCAAGCAACAACGGAG GCAAACAAGCCTTAGAAACAGTCCAGAGATTGTTGCCGGTGCTGTGCCAAGCCCACGG CCTGACCCCAGACCAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCT CGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCAGC CCAAGTTGTCGCGATTGCAAGCAACAACGGAGGCAAACAAGCCTTAGAAACAGTCCAG AGATTGTTGCCGGTGCTGTGCCAAGACCACGGCCTGACCCCCGACCAGGTTGTCGCTA TTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCCTCTTGCCGGT CTTGTGTCAGGACCACGGCCTGACCCCAGAACAAGTTGTCGCGATTGCAAGCAACAAC GGAGGCAAACAAGCCTTAGAAACAGTCCAGAGATTGTTGCCGGTGCTGTGCCAAGCCC ACGGCCTGACCCCAGACCAAGTTGTCGCGATTGCAAGCAACAACGGAGGCAAACAAGC CTTAGAAACAGTCCAGAGATTGTTGCCTGTGCTGTGCCAAGCCCACGGCCTGACCCCC GCCCAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTC AGCGCCTCTTGCCGGTCTTGTGTCAGGACCACGGCCTGACCCCGGACCAGGTGGTTG CAATCGCGTCACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGC CCGTCCTGTGCCAGGACCACGGCCTGACCCCTGAGCAGGTAGTGGCTATTGCATCCAA CGGAGGGGGCAGACCCGCACTGGAGTCAATCGTGGCCCAGCTTTCGAGGCCGGACCC CGCGCTGGCCGCACTCACTAATGATCATCTTGTAGCGCTGGCCTGCCTCGGCGGACGT CCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGATCCC AGCTAGTGAAATCTGAATTGGAAGAGAAGAAATCTGAACTTAGACATAAATTGAAATATG TGCCACATGAATATATTGAATTGATTGAAATCGCAAGAAATTCAACTCAGGATAGAATCC TTGAAATGAAGGTGATGGAGTTCTTTATGAAGGTTTATGGTTATCGTGGTAAACATTTGG GTGGATCAAGGAAACCAGACGGAGCAATTTATACTGTCGGATCTCCTATTGATTACGGT GTGATCGTTGATACTAAGGCATATTCAGGAGGTTATAATCTTCCAATTGGTCAAGCAGAT GAAATGCAAAGATATGTCGAAGAGAATCAAACAAGAAACAAGCATATCAACCCTAATGA ATGGTGGAAAGTCTATCCATCTTCAGTAACAGAATTTAAGTTCTTGTTTGTGAGTGGTCA TTTCAAAGGAAACTACAAAGCTCAGCTTACAAGATTGAATCATATCACTAATTGTAATGG AGCTGTTCTTAGTGTAGAAGAGCTTTTGATTGGTGGAGAAATGATTAAAGCTGGTACATT GACACTTGAGGAAGTGAGAAGGAAATTTAATAACGGCGAGATAAACTTTTAA AOX1 mitochondria targeting presequence (DNA) is underlined. SEQ ID NO: 57 atp1-TALEN coding sequence (right arm) ATGATGATGATGATGTCTAGGTCAGGAGGTAACAGGGTAGCTAACACAGCAATGTTTGT GGCAAAGGGTTTGTCAGGTGAAGTTGGTGGTTTGAGAGCATTGTATGGTGGTGGTGTG AGGAGTGAGTCCATGGACTACAAGGATGACGACGATAAAAGTTGGAAGGACGCAAGTG GTTGGTCTAGAATGCATGCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTT CGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGA AGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCC ATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGAC CGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGAC ATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACG GATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAG ATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCG CTCACGGGAGCACCCCTCAACCTGACCCCGGACCAGGTGGTTGCAATCGCGTCACAC GATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAG GACCACGGCCTGACCCCGGAACAGGTGGTTGCAATCGCGTCACACGATGGGGGAAAG CAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCCAGGCCCACGGCCTG ACCCCAGACCAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAA ACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCCGCCCAG GTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGAAACAGTTCAGCGCC TCTTGCCGGTCTTGTGTCAGGACCACGGCCTGACCCCAGACCAGGTTGTGGCCATCGC CAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTC TGCCAGGACCACGGCCTGACCCCAGAACAGGTTGTGGCCATCGCCAGCAACATAGGT GGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCAC GGCCTGACCCCAGACCAGGTTGTGGCCATCGCCAGCAACATAGGTGGCAAGCAGGCC CTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCACGGCCTGACCCCG GCCCAGGTGGTTGCAATCGCGTCACACGATGGGGGAAAGCAGGCCCTAGAAACCGTT CAGCGACTCCTGCCCGTCCTGTGCCAGGACCACGGCCTGACCCCAGACCAGGTTGTG GCCATCGCCAGCAACATAGGTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTAC CGGTTCTCTGCCAGGACCACGGCCTGACCCCGGAACAGGTGGTTGCAATCGCGTCAC ACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCCTGTGCC AGGCCCACGGCCTGACCCCAGACCAAGTTGTCGCGATTGCAAGCAACAACGGAGGCA AACAAGCCTTAGAAACAGTCCAGAGATTGTTGCCTGTGCTGTGCCAAGCCCACGGCCT GACCCCCGCCCAGGTTGTCGCTATTGCTAGTAACGGCGGAGGCAAACAGGCGCTGGA AACAGTTCAGCGCCTCTTGCCGGTCTTGTGTCAGGACCACGGCCTGACCCCAGACCAA GTTGTCGCGATTGCAAGCAACAACGGAGGCAAACAAGCCTTAGAAACAGTCCAGAGAT TGTTGCCGGTGCTGTGCCAAGACCACGGCCTGACCCCGGAACAGGTGGTTGCAATCG CGTCACACGATGGGGGAAAGCAGGCCCTAGAAACCGTTCAGCGACTCCTGCCCGTCC TGTGCCAGGCCCACGGCCTGACCCCAGACCAGGTTGTGGCCATCGCCAGCAACATAG GTGGCAAGCAGGCCCTCGAAACCGTCCAGAGACTGTTACCGGTTCTCTGCCAGGCCCA CGGCCTGACCCCTGAGCAGGTAGTGGCTATTGCATCCAACATAGGGGGCAGACCCGC ACTGGAGTCAATCGTGGCCCAGCTTTCGAGGCCGGACCCCGCGCTGGCCGCACTCAC TAATGATCATCTTGTAGCGCTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTG AAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGATCCCAGCTAGTGAAATCTGAAT TGGAAGAGAAGAAATCTGAACTTAGACATAAATTGAAATATGTGCCACATGAATATATTG AATTGATTGAAATCGCAAGAAATTCAACTCAGGATAGAATCCTTGAAATGAAGGTGATG GAGTTCTTTATGAAGGTTTATGGTTATCGTGGTAAACATTTGGGTGGATCAAGGAAACC AGACGGAGCAATTTATACTGTCGGATCTCCTATTGATTACGGTGTGATCGTTGATACTAA GGCATATTCAGGAGGTTATAATCTTCCAATTGGTCAAGCAGATGAAATGCAAAGATATG TCGAAGAGAATCAAACAAGAAACAAGCATATCAACCCTAATGAATGGTGGAAAGTCTAT CCATCTTCAGTAACAGAATTTAAGTTCTTGTTTGTGAGTGGTCATTTCAAAGGAAACTAC AAAGCTCAGCTTACAAGATTGAATCATATCACTAATTGTAATGGAGCTGTTCTTAGTGTA GAAGAGCTTTTGATTGGTGGAGAAATGATTAAAGCTGGTACATTGACACTTGAGGAAGT GAGAAGGAAATTTAATAACGGTGAGATAAACTTTTAA AOX1 mitochondria targeting presequence (DNA) is underlined. SEQ ID NO: 58 atp1-TALEN amino acids sequence (left arm) with targeting peptide MMMMMSRSGGNRVANTAMFVAKGLSGEVGGLRALYGGGVRSESMDYKDDDDKSWKDA SGWSRMHAAPRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGH GFTHAHIVALSQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSGARALEALLTDAGEL RGPPLQLDTGQLVKIAKRGGVTAMEAVHASRNALTGAPLNLTPDQVVAIASHDGGKQALET VQRLLPVLCQDHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNG GKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPDQVV AIASHDGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHG LTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPV LCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNNGGKQALET VQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGG GKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPEQVV AIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPAMDAVKKGLPHAPELIRS QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYGYRGKHLGG SRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWK VYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVR RKFNNGEINF AOX1 mitochondria targeting presequence (amino acids) is underlined. Repeat variable diresidues (RVDs) for recognising target atp1 sequence are in bold. SEQ ID NO: 59 atp1-TALEN amino acids sequence (right arm) with targeting peptide MMMMMSRSGGNRVANTAMFVAKGLSGEVGGLRALYGGGVRSESMDYKDDDDKSWKDA SGWSRMHAAPRRRAAQPSDASPAAQVDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGH GFTHAHIVALSQHPAALGTVAVTYQHIITALPEATHEDIVGVGKQWSGARALEALLTDAGEL RGPPLQLDTGQLVKIAKRGGVTAMEAVHASRNALTGAPLNLTPDQVVAIASHDGGKQALET VQRLLPVLCQDHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIG GKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPDQVV AIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGL TPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVL CQDHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASHDGGKQALETV QRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGG KQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPEQVVA IASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLT PEQVVAIASNIGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPAMDAVKKGLPHAP ELIRSQLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYGYRGK HLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPN EWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLT LEEVRRKFNNGEINF AOX1 mitochondria targeting presequence (amino acids) is underlined. Repeat variable diresidues (RVDs) for recognising target atp1 sequence are in bold. SEQ ID NO: 65 atp1-TALEN target site TATAGCGAATGACTATTCATCTATTGTATTTTCATGCAAATAGGGGGCA Example 4 By delivering a donor DNA construct via particle bombardment, single or multiple mutations can be introduced to the site targeted by TALENs, e.g. accD-TALEN, accD-TALEN- UPSTREAM and atp1-TALEN. In this example, the inventors describe a method of introducing multiple point mutations simultaneously into MAT-3 (accD-TALEN) plants. Plant material A MAT-3 T0 plant was self-pollinated to segregate out the transgene of MuPOP but maintained the accD-TALEN in the T1 generation. A T1 plant harbouring accD-TALEN but no MuPOP was named AT-3 and grown in sterile containers for later use. For WAT-UP and Watp1T lines, the T0 plants grown aseptically can be used. Design of the donor DNA construct A DNA sequence including accD gene (2573 bp, SEQ ID NO: 60) was synthesized, of which nine bases were artificially substituted in the binding region of accD-TALEN (Figure 16). The sequence was cloned into a cloning vector (pUC57-Kan-Simple-Key IIS Free) to generate the donor construct pATR (Figure 17). Both DNA synthesis and cloning into the vector was provided by service from GenScript. The nine substitutions were designed to be synonymous mutations without changing the wild type amino acids in the coding sequence, these mutations should prevent accD-TALEN from binding to the target site. The nine substitutions were flanked by 1.2 kb wild type sequences as the left and right targeting arms to allow homologous recombination between the artificial construct and the tobacco chloroplast genomes. Additionally, different donor DNA constructs can be designed to introduce mutations into the sites targeted by other TALENs, such as accD-TALEN-UPSTREAM and atp1-TALEN from Examples 2 and 3. The procedures to achieve such constructs are similar to the above. Pre-treatment of tobacco leaf material The 2ndto 4thexpanded leaves from the top of AT-3 plants were excised and placed abaxial side upward in the central of 9 cm petri dish with ERM (regeneration medium with 20 µM estradiol, described in Example1). The leaves were bombarded on the same day. Preparation of microprojectiles for bombardment In a 1.5 ml microcentrifuge tube, 50 µl gold particles (60 mg / ml suspension in water) with 1 µm or 0.6 µm diameter were mixed with the following in orders, 5 µl donor construct DNA (1 µg / µl), 50 µl CaCl2and 20 µl of 0.1 M spermidine (free base, tissue culture grade). The tube was continuously vortexed for 3 min. DNA-coated gold particles were pelleted by centrifugation at 10,000 rpm for 5 mins. The supernatant was removed, and the particles were washed by 250 µl of absolute ethanol. Particles were vortexed briefly and spun down for 10 sec at 10,000 rpm. The supernatant was removed, and the gold particles were resuspended in 60 µl of absolute ethanol. Bombardment of plant tissue with microprojectiles The DNA-coated gold particles from the above were dispensed to eight macrocarriers for bombarding eight leaves from AT-3. Bombardments were performed using PDS-1000 / He biolistic device (Bio-Rad) following manufacturer’s manual. For each bombardment, 1100 psi helium pressure and 28 mmhg vacuum were used to shoot each leaf at a distance of 9 cm. Selection of the organelle mutants The leaves were dissected into approximately 3 mm2disks on the next day of bombardment. The leaf explants were incubated on ERM and transferred to fresh ERM medium every two weeks until shoots emerge. The genotype of the shoot can be verified by Sanger sequencing the PCR product from the target region. The homoplasmy status of the organelle mutation can be verified by Next Generation Sequencing with at least a depth of 100 reads. Sequences (used in Example 4) SEQ ID NO: 60 Synthesised DNA sequence including accD coding sequence ATCAAAATCTAAGACTCAAATCTTTCTATTGTTGTCTTGGATCCACAATTAATCCTACGGA TCCTTAGGATTGGTATATTCTTTTCTATCCTGTAGTTTGTAGTTTCCCTGAATCAAGCCAA GTATCACACCTCTTTCTACCCATCCTGTATATTGTCCCCTTTGTTCCGTGTTGAAATAGA ACCTTAATTTATTACTTATTTTTTTATTAAATTTTAGATTTGTTAGTGATTAGATATTAGTAT TAGACGAGATTTTACGAAACAATTATTTTTTTATTTCTTTATAGGAGAGGACAAATCTCTT TTTTCGATGCGAATTTGACACGACATAGGAGAAGCCGCCCTTTATTAAAAATTATATTAT TTTAAATAATATAAAGGGGGTTCCAACATATTAATATATAGTGAAGTGTTCCCCCAGATT CAGAACTTTTTTTCAATACTCACAATCCTTATTAGTTAATAATCCTAGTGATTGGATTTCT ATGCTTAGTCTGATAGGAAATAAGATATTCAAATAAATAATTTTATAGCGAATGACTATTC ATCTATTGTATTTTCATGCAAATAGGGGGCAAGAAAACTCTATGGAAAGATGGTGGTTTA ATTCGATGTTGTTTAAGAAGGAGTTCGAACGCAGGTGTGGGCTAAATAAATCAATGGGC AGTCTTGGTCCTATTGAAAATACCAATGAAGATCCAAATCGAAAAGTGAAAAACATTCAT AGTTGGAGGAATCGTGACAATTCTAGTTGCAGTAATGTTGATTATTTATTCGGCGTTAAA GACATTCGGAATTTCATCTCTGATGACACTTTTTTAGTTAGTGATAGGAATGGAGACAGT TATTCCATCTATTTTGATATTGAAAATCATATTTTTGAGATTGACAACGATCATTCTTTTCT GAGTGAACTAGAAAGTTCTTTTTATAGTTATCGAAACTCGAATTATCGGAATAATGGATT TAGGGGCGAAGATCCCTACTATAATTCTTACATGTATGATACTCAATATAGTTGGAATAA TCACATTAATAGTTGCATTGATAGTTATCTTCAGTCTCAAATCTGTATAGATACTTCCATT ATAAGTGGTAGTGAGAATTACGGTGACAGTTACATTTATAGGGCCGTTTGTGGTGGTGA AAGTCGAAATAGTAGTGAAAACGAGGGTTCCAGTAGACGAACTCGCACGAAGGGCAGT GATTTAACTATAAGAGAAAGTTCTAATGATCTCGAGGTAACTCAAAAATACAGGCATTTG TGGGTACAATGTGAGAACTGTTATGGATTAAATTATAAAAAGTTCCTTAAATCAAAAATG AATATTTGTGAACAATGTGGATATCATTTGAAAATGAGTAGTTCAGATAGAATTGAACTTT TGATCGATCCGGGTACTTGGGATCCTATGGATGAAGACATGGTCTCTCTAGATCCCATT GAATTTCATTCGGAGGAGGAGCCTTATAAAGATCGTATTGATTCTTATCAAAGAAAGACA GGATTAACCGAGGCTGTTCAAACAGGCATAGGCCAACTAAACGGCATTCCCGTAGCAA TTGGGGTTATGGATTTTCAGTTTATGGGGGGTAGTATGGGATCCGTAGTCGGAGAGAA AATCACCCGTTTGATTGAATACGCTGCCAATCAAATTTTACCCCTTATTATAGTGTGTGC TTCTGGGGGGGCGCGCATGCAGGAAGGAAGTTTGAGCTTGATGCAAATGGCTAAAATA TCGTCTGCTTTATATGATTATCAATTAAATAAAAAGTTATTTTATGTATCAATCCTTACATC TCCGACAACTGGTGGAGTGACAGCTAGTTTTGGTATGTTGGGGGATATCATTATTGCCG AACCCAACGCCTACATTGCATTTGCAGGTAAAAGAGTAATTGAACAAACATTGAATAAAA CAGTACCCGAAGGTTCACAAGCAGCTGAATACTTATTCCAGAAGGGTTTATTCGACCTA ATTGTACCACGTAATCTTTTAAAAAGCGTTCTGAGTGAGTTATTTAAGCTCCACGCCTTT TTTCCTTTGAATCAAAAGTCAAGCAAAATCAAGTAGAGCACTAAGTTCAATTATTTTATTT GTGTTTGTAGCAAAAAAGTAGTTAGTTTGTCGGAATCAAAGTAAATAAGATAATAATGGC GCTTTCTTTGGTGATAGAAGATCTAATTGTAGAAAGAATCAAAACTAAAGTTGAGGATAA CTCTTTTTTTGACCTATATTCCTGATTACGAATCAAGAAGCCTTTATCAACAAGAGTGAG TTCTTCCTTTCGTGAAATTAGGAAAATAAAACGAATTTCTTCTTCTTGTCTTAGGTATATA ATTTGAAATTCAAATATAGATAATAGAGTTTTGTATCTTTCTCTATCTCCCGAAAAACCAT TTTAGCTAAAAATTCATGTTGGGTCGGATTCGAACGAATCTTTCGATAATCTGTAAGAAA CTCTTTATCTATTTTTAGAAAATTAGAAGACAAGAACAAAAGACAAAGAAATGAAGAAAA ATAATAAAGTTTATTATGATACATATCTTTCTCATGTAGGGGATGAATA The accD coding sequence is underlined. The substituted nucleotides are in bold. SEQ ID NO: 61 nucleotide sequence of the donor construct in Figure 16 GTACAATGTGAGAACTGTTATGGATTAAATTATAAAAAGTTCCTTAAA SEQ ID NO: 62 animo acid sequence of the wild type and donor constructs in Figure 16 VQCENCYGLNYKKFLK SEQ ID NO: 63 nucleotide sequence of the wild type in Figure 16 GTTCAATGCGAAAATTGTTATGGATTAAATTATAAGAAATTTTTGAAA Methods Construction of TALEN vectors The XVE transcription unit, LexA-mCaM35S promoter, pea T3A terminator were PCR- generated using pER8 derived sequences as the template, Zuo et al. plant journal 2000 (24). The XVE transcription unit, LexA-mCaM35S promoter, pea T3A terminator can be PCR- generated using any plasmid containing XVE inducible system as the template, such as plasmid pER8, which may be obtained from, for example: https: / / lifescience- market.com / plasmid-c-94 / per8-plasmid-p-63727.html. The chloroplast presequence CP-Pre is cloned from petunia rbcS subunit 8 gene. The HSP terminator (HSP Term) is cloned from Arabidopsis heat shock protein 18.2 gene. The left and right arms (LA and RA) of TALEN targeting to accD or atp1 gene were assembled into ptCMV-153-47-VR-NG and ptCMV-153- 47-VR-NG-NI plasmids using Platinum Gate TALEN kit, respectively (37). For accD-TALEN (Example 1), the DNA fragments of LA and RA were PCR-generated using ptCMV-153-47- VR-NG-LA and ptCMV-153-47-VR-NI-RA as templates, respectively. For accD-TALEN- UPSTREAM (Example 2), the DNA fragments of LA and RA were PCR-generated using ptCMV-153-47-VR-NG-LA and ptCMV-153-47-VR-NN-RA as templates, respectively. For atp1-TALEN (Example 3), the DNA fragments of LA and RA were PCR-generated using ptCMV-153-47-VR-NG-LA and ptCMV-153-47-VR-NI-RA as templates, respectively. DNA fragments and their flanking restriction sites are summarized in Table 3 Table 3 Primers for the DNA fragments used for constructing accD-TALEN vector Fragments PCR primers (forward / reverse, 5’>3’) Restriction sites 1. XVE F ACGATCGTAGTTTAAACTGAAGGCGGGAAA (SEQ ID PvuI transcription NO:32) unit R AGAAGACACGTTTGGGATGTTTTACTCCTCATAT (SEQ ID BbsI NO:33) 2. LexA- F ACGTCTCTAAACGCGATAGAAAACAAAATATAGCGCG BsmBI m35S-LA (SEQ ID NO:34) R ACGTCTCACCATTCGAGGCTAGAGTCGACTAG (SEQ ID BsmBI NO:35) 3. CP-Pre-LA F ACGTCTCAATGGCTTCCTCTGTGATTTCCTCTG (SEQ ID BsmBI NO:36) R TCGTCTCTCATGCATTGGACTCTTCCACCATTGCT (SEQ BsmBI ID NO:37) 4. accD- F ACGTCTCACATGGACTACAAGGATGACGACGATAA (SEQ BsmBI TALEN-LA ID NO:38) R ATCGCTCGAGTTAAAAGTTTATCTCGCCGTTATTAAAT XhoI (SEQ ID NO:39) 5. HSP-Term F CCTCGAGATATGAAGATGAAGATG (SEQ ID NO:40) XhoI R GGCGGCCGCTCGACACGATAGGAGGGTCGGT (SEQ ID NotI NO:41) 6. LexA- F CTTTATGCGGCCGCGCGATAGAAAACAAAATATAGCGC NotI m35S-RA (SEQ ID NO:42) R ACGTCTCAATTCGAGGCTAGAGTCGACTAG (SEQ ID BsmBI NO:43) 7. CP-Pre- F ACGTCTCTGAATGGCTTCCTCTGTGATTTCCTCTG (SEQ BsmBI RA ID NO:44) R TCGTCTCTTCCATGCATTGGACTCTTCCACCATTGCT BsmBI (SEQ ID NO:45) 8. accD- F ACGTCTCATGGACTACAAGGATGACGACGATAA (SEQ ID BsmBI TALEN-RA NO:46) R ACGTCTCATTAAAAGTTTATCTCACCGTTATTAAA (SEQ BsmBI ID NO:47) 9. Pea T3A F ACGTCTCATTAACAGGCCTCCCAGCTTTCGT (SEQ ID BsmBI Term NO:48) R AGTACAGTATAGGCTTTTTGTGTCGAGCGATCGA (SEQ PvuI ID NO:49) Fragments 1-5 and 6-9 were cloned into pGEMT-easy vector (Promega, UK) via flanking restriction sites (Table 3) to yield pGEM-XVE-LA and pGEM-RA, respectively. The modules of XVE-LA (assembled fragments 1 to 6) and RA (assemble fragment6 to 9) were cut out using PvuI and NotI restriction enzymes, and then cloned into the binary vector pCAMBIA3300 (www.cambia.org) via PvuI and NotI restriction sites, yielding the final construct pCMABI3300- XVE-accD-TALEN. Plant material and growth conditions Nicotiana tabacum cv. Petit Havana was used for all experiments. For in vitro experiments, plants were grown aseptically on the agar media with modifications depending on the purposes. Different types of media for plant tissue culture are listed in Table 4 Plants grown in petri dishes or Magenta™ GA-7 vessels were incubated in plant growth chambers under standard conditions (24oC with 12-hour day / night cycle, light intensity 50 µE m−2s−1). In the greenhouse, plants were grown in soil under standard conditions (26-28oC, 12-hour day / night cycle and average light intensity 250 µE m−2s−1). Table 4 Media used in plant tissue culture Media type Functions Components MS seeds germination, rooting and 1x or ½x Murashige and Skoog (MS) basal salts asexual propagation and vitamins with 2.5 mM 2-(4-morpholino) ethanesulfonic acid (MES) and 3% w / v sucrose, pH5.8 RM shoots regeneration MS medium supplemented with 4.5 µM benzylaminopurine (BA), 0.5 µM naphthalene acetic acid (NAA) and solidified with 0.7% agar2, pH 5.8 MS20IM Agrobacteria induction MS medium supplemented with 2% w / v sucrose, 100 µM acetosyringone, 1 mM proline and 0.7% agar2, pH 5.25 Plant transformation and isolating transgenic lines Plant explants (leaf disks) were dissected from the wild type or homozygous MuPOP transgenic tobacco plants aseptically grown on MS media. Agrobacteria (GV3101) were transformed with pCMABI3300-XVE-accD-TALEN and grown in a liquid culture to a density of 0.8 at OD600. The Agrobacteria were harvested and resuspended in MS20IM to the same density before incubated with leaf disks at room temperature for 15-20 min. After blotting away excessive Agrobacteria suspension, the leaf disks were transfected by co-cultivating with the remained agrobacteria on the RM media in the dark at 24oC for 3 days. Then the transfected leaf disks were transferred to the RM media supplemented with 10 mg / L phosphinotricin (PPT), 600 mg / L carbenicillin and 200 mg / L cefotaxime, and incubated under standard conditions. Putative transgenic shoots emerged after 3-4 weeks further incubation. Shoots grown to 0.5- 1 cm long were transferred to MS media (supplemented with 10 mg / L PPT, 600 mg / L carbenicillin and 200 mg / L cefotaxime) for rooting. After 1-2 weeks, plantlets with wild-type like phenotype and healthy roots were considered as real transgenic plants, which were selected for later experiments. The transgenic plants were named as wild-type accD-TALEN (WAT) or MuPOP accD-TALEN (MAT) depending on the genotype of the pCMABI3300-XVE- accD-TALEN recipient plant. Identification of the optimal accD-TALEN transgenic lines Transgenic plants resistant and rooting on the MS media containing 10 mg / L PPT were selected for assay to identify the transgenic lines showing optimal expression of accD-TALEN selection system.20 leaf disks from the wild type tobacco plant were used as the negative control, and regenerated on the RM and RM20E (RM supplemented with 20 µM estradiol), respectively.20 leaf disks from each candidate transgenic line were incubated on the RM and RM20E, respectively.4-6 leaf disks were sampled from each media type after 24 hours, which were used for RT-PCR analysis. The rest leaf disks were passed onto fresh media every 10 days. After three weeks, candidate lines were assessed based on the regeneration of their leaf disks. The optimal transgenic line is defined as (36) leaf disks on the RM regenerate normally as the negative control (>10 shoots per leaf disk); (37) leaf disks on the RM20E showed <2 shoot per leaf disk and became pale or necrotic. Usually 10-15 candidate transgenic lines could yield 1-2 optimal lines. RT-PCR Leaf disks sampled from media were disrupted in the TRI reagent (Merck, UK), from which RNA samples were purified using Direct-zol RNA miniprep kit (ZYMO RESEARCH, USA). In a 15 µl reaction, 500 ng RNA from each sample was used for reverse transcription to yield cDNA using GoScript Reverse Transcription kit (Promega, UK). 3 µl from each cDNA synthesis reaction was used in a 20 µl PCR reaction using MyTaq Red Mix (Bioline, UK) with a mixture of primer pairs targeting LA, RA and EF-1α gene (Fig 1D).0.75 µl from each primer stock solution (20 µM) was added to each PCR tube. PCR involved 32 cycles and the products analysed by 2% (W / V) agarose gel electrophoresis. Selection of chloroplast mutants via tissue culture Leaf disks from the optimal MAT lines were cultured on the RM supplemented with estradiol at the desired concentration (2-20 µM). Low concentrations of estradiol (2-5 µM) increased the frequency of regenerated shoots but,decreased the efficiency of achieving homoplasmic organelle mutants. High concentrations of estradiol (10-20 µM) were used to maximise the efficiency of achieving homoplasmic organelle mutants, although the frequencies of regenerated shoots decreased. After 12 weeks, putative organelle mutant shoots were isolated, from which a small leaf piece (0.5 cm2) was sampled for PCR to amplify a 300 bp fragment covering the sequence targeted by accD-TALEN. The PCR product was sent for Sanger sequencing to localise the mutations. Primers for PCR of the accD target region Forward: 5’ ATAGTAGTGAAAACGAGGGTTCC 3’ (SEQ ID NO:50) Reverse: 5’ AATGAAATTCAATGGGATCTAGAGAGA 3’ (SEQ ID NO:51) Selection of chloroplast mutants via seeds germination Optimal MAT lines were grown to mature and setting seeds in soil. Seeds were sterilized using 96% ethanol for 1 min then 10% bleach for 10 min, before plated onto MS containing 10 mg / L PPT and 2-20 µM estradiol. After 2 weeks, the surviving seedlings were isolated, and a small leaf piece was sampled from each seedling for PCR at the accD target region (300 bp). PCR products were sent for Sanger sequencing to confirm the locations of mutations arising from the inducible TALEN selection system. DNA sequencing Sanger sequencing was performed using the Eurofin LightRun Sanger Sequencing service. Next-generation sequencing (NGS) was performed using the Eurofin LightRun NGS service (Illumina MiSeq, 150 bp paired end). Enriched organelle DNA from each plant sample was used as the library for NGS. The plant organelle DNA was enriched using NEBNext Microbiome DNA Enrichment kit (NEB, UK) and then non-exponentially amplified using REPLI-g Mini Kit (Qiagen, UK). The amplified DNA was digested with T7 endonuclease (NEB, UK) for 15 min then cleaned up using JetSeq Clean beads (Bioline, UK).1-2 µg of the cleaned up DNA was sent for NGS. Bioinformatic analysis of NGS data All bioinformatic analysis was performed in Geneious Prime 2020 software. The NGS fastq raw data was filtered and trimmed with criteria minimal length 50 bp and >Q30. The filtered reads mapped to the reference tobacco chloroplast genome (NCBI Z00044.2). An inverted repeat (nucleotide position 130,601 – 155,943) of the reference genome was clipped to allow the mapping quality >90. Typically, 4GB raw data was enough to give a coverage >100. 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Claims

CLAIMS 1. A method of producing a homoplasmic modified plant or part thereof, the method comprising: (a) Modifying plastid or mitochondrial DNA of the plant or part thereof at one or more target sites to produce a modified plant or part thereof having plastid or mitochondrial DNA comprising one or more modified target sites; (b) Introducing into the plant or part thereof, optionally the modified plant or part thereof, an expression construct comprising: one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA operably linked to an inducible system; (c) Inducing expression of the protein from the expression construct; (d) Regenerating the modified plant or part thereof; and (e) Optionally repeating steps c and d.

2. The method of claim 1, wherein the method comprises a further step of selecting the plant or part thereof if it maintains or has improved viability compared to the plant or part thereof of step (a), optionally wherein the further step is performed after inducing expression of the protein step (c) and before regeneration step (d).

3. A method of selecting plants or parts thereof comprising modified plastid or mitochondrial DNA, the method comprising: (a) Providing one or more plants or parts thereof that have undergone a modification process to modify one or more target sites in the plastid or mitochondrial DNA; (b) Introducing into the one or more plants or part thereof an expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non-modified target sites in the plastid or mitochondrial DNA, operably linked to an inducible system; (c) Inducing expression of the protein from the expression construct; (d) Selecting the plants or parts thereof which maintain or have improved viability compared to the plants of step (a); (e) Optionally regenerating the selected plants; and (f) Optionally repeating steps (c) to (e).

4. The method according to any one of the preceding claims, wherein the method produces homoplasmic modified plants or parts thereof.

5. The method according to any one of the preceding claims, wherein the one or more target sites in the plastid or mitochondrial DNA are modified in step (a) of the methods by using an error prone polymerase, optionally by exposing the plastid or mitochondrial DNA to an error prone polymerase, preferably wherein the error prone polymerase is a plant organellar polymerase (POP), more preferably wherein the plant organellar polymerase (POP) comprises an amino acid sequence according to SEQ ID NO:22, or an amino acid sequence having at least 35% identity thereto, or a functional fragment thereof, comprising a modification at position L903, and optionally one or more further modifications at the following positions: D390, E392, R862, E904, and N1065 of SEQ ID NO:22, or positions corresponding thereto.

6. The method according to claim 5, wherein the error prone polymerase is introduced into the plant or part thereof, optionally by transformation, to modify the plastid or mitochondrial DNA at one or more target sites.

7. The method according to any one of the preceding claims, wherein the inducible system is an estradiol inducible system, optionally wherein the estradiol inducible system is an XVE system.

8. The method according to any one of the preceding claims wherein step (b) comprises introducing an expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of non- modified target sites in the plastid or mitochondrial DNA, operably linked to a LexA operator sequence, and an expression construct comprising one or more nucleic acids which encode an XVE system.

9. The method according to any preceding claim, wherein the plastid DNA is proplastid, chloroplast, etioplast, leucoplast, amyloplast, elaioplast, proteinoplast or chromoplast DNA, preferably wherein the plastid DNA is chloroplast DNA 10. The method according to any one of the preceding claims, wherein the plant or part thereof is a dicot or monocot, or a part thereof.

11. The method according to any one of the preceding claims, wherein the plant part is an organ, a tissue, a callus, or a cell of a plant, preferably wherein the plant part is selected from a stem, a leaf, a root, an inflorescence, a flower, a floret, a fruit, a pedicle, a peduncle, a stamen, an anther, a stigma, a style, an ovary, a petal, a sepal, a carpel, a root tip, a root cap, a root hair, a leaf hair, a seed hair, a pollen grain, a microspore, an embryo, a scutellum, an ovule, a cotyledon, a hypocotyl, an epicotyl, xylem, phloem, parenchyma, endosperm, a companion cell, and a guard cell.

12. The method according to any preceding claim, wherein the plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA is selected from: a plastid or mitochondria targeted nuclease, meganuclease, Zinc finger nuclease, CRISPR-Cas system, transcription activator- like effector nuclease (TALEN), restriction enzyme and / or nicking enzyme.

13. The method according to claim 12, wherein the plastid or mitochondria targeted protein capable of cleavage of any non-modified target sites in the plastid or mitochondrial DNA is a plastid or mitochondria targeted TALEN.

14. The method according to any preceding claim, wherein the one or more target sites are in the plastid or mitochondrial genome.

15. The method according to any preceding claim, wherein the one or more target sites are located in a gene, optionally in the coding region of a gene or in a non-coding region of a gene, optionally in a regulatory element.

16. The method according to any preceding claim, wherein the one or more target sites are located in a gene which controls an important agroeconomic trait or within a regulatory element of a gene which controls an important agroeconomic trait, preferably wherein the gene which controls an important agroeconomic trait is selected from: a gene which controls growth, yield, disease resistance, pest resistance and / or sterility.

17. The method according to any preceding claim, wherein the one or more target sites are located in a gene or within a regulatory element of a gene which controls cytoplasmic male sterility when modified.

18. The method according to any preceding claim, wherein step (c) is carried out such that the protein is expressed at an optimal time to produce homoplasmic plants or parts thereof, preferably wherein the optimal time to produce homoplasmic plants is when the copy number of the plastid or mitochondrial genome is low, more preferably wherein step (c) is carried out such that the protein is expressed during germination, leaf formation, flowering, seeding, and / or callus formation.

19. The method according to any preceding claim, wherein step (c) is carried out such that the protein is expressed in an optimal location of the plant or part thereof to produce homoplasmic plants or parts thereof, preferably wherein the optimal location to produce homoplasmic plants or parts thereof is a location in which the copy number of the plastid or mitochondrial genome is low, more preferably wherein step (c) is carried out such that the protein is expressed in a location selected from: the root tip, anther, pollen including pollen mother cells, embryo, callus, and / or shoot meristem.

20. An expression construct comprising one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA, operably linked to an estradiol inducible system.

21. The expression construct according to claim 20, wherein the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA are operably linked to a LexA operator sequence, and optionally wherein the expression construct further comprises one or more nucleic acids which encode components of the estradiol inducible system, preferably wherein the estradiol inducible system is the XVE system.

22. The expression construct according to claims 20 or 21, wherein the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA encode a plastid or mitochondria targeted nuclease, meganuclease, Zinc finger nuclease, CRISPR-Cas system, transcription activator-like effector nuclease (TALEN), restriction enzyme and / or nicking enzyme, preferably wherein the one or more nucleic acids which encode a plastid or mitochondria targeted protein capable of cleavage of a target site in plastid or mitochondrial DNA encode a TALEN.

23. The expression construct according to claim 20, 21, or 22 for use in the method according to any one of claims 1 to 19.

24. A vector comprising the expression construct according to claim 20, 21 or 22.

25. A plastid or mitochondria comprising the expression construct according to claim 20, 21 or 22 or the vector according to claim 24.

26. A plant or part thereof comprising the expression construct according to claim 20, 21 or 22, or the vector according to claim 24.

27. Use of the expression construct according to claim 20, 21 or 22 or the vector according to claim 24 for selecting plants or parts thereof comprising modified plastid or mitochondrial DNA.

28. Use of the expression construct according to claim 20, 21 or 22 or the vector according to claim 24 for producing homoplasmic modified plants or parts thereof, optionally in combination with an error prone polymerase.

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